CN109758423A

CN109758423A - Use the method for vitamin K1 fat emulsion injection treatment coagulation disorders

Info

Publication number: CN109758423A
Application number: CN201910200165.6A
Authority: CN
Inventors: 傅卫国; 梁庆; 王伟
Original assignee: XI'AN ANJIAN PHARMACEUTICAL CO Ltd
Current assignee: Xi'an Anjian Pharmaceutical Co ltd
Priority date: 2019-03-16
Filing date: 2019-03-16
Publication date: 2019-05-17
Anticipated expiration: 2039-03-16
Also published as: CN109758423B

Abstract

The present invention relates to the methods for using vitamin K1 fat emulsion injection treatment coagulation disorders.Specifically, including vitamin K1, soybean oil, phosphatide, glycerol, water the present invention relates to emulsion composition.The emulsion composition is prepared by the techniques such as preparation water phase, the oily phase of preparation, preparation thick cream, high-pressure emulsification, pressure sterilizing, further includes carrying out the operation in relation to substance especially impurity B detection to experience pressure sterilizing gained emulsion composition.The invention further relates to the pharmaceutical applications of emulsion composition, the disease for coagulation disorders caused by prothrombin, VII, IX and X dyssynthesis caused by the pharmaceutical intervention of vitamin K deficiency or interference vitamin k activity.The invention further relates to use the method in relation to substance especially impurity B in HPLC method measurement vitamin K1 composition.One or more technical effects as used in the description are presented in emulsion composition of the present invention.

Description

Use the method for vitamin K1 fat emulsion injection treatment coagulation disorders

Technical field

The invention belongs to pharmaceutical technology fields, are related to a kind of fat emulsion injection, contain vitamin K1 more particularly to one kind Fat emulsion injection.The invention further relates to the methods for preparing fat emulsion injection of the present invention.The present invention is further It is related to the purposes of prepared vitamin K1 fat emulsion injection, in particular for vitamin K deficiency or interference vitamin k activity Pharmaceutical intervention caused by coagulation disorders disease caused by prothrombin, VII, IX and X dyssynthesis.Further, The present invention relates to the methods for carrying out quality testing to the vitamin K1 fat emulsion injection.

Background technique

Vitamin K1 (Vitamin K1, Phytonadione) is a kind of clear thick liquid of Yellow-to-orange, odorless Or it is almost odorless, it meets light and easily decomposes.Vitamin K1 is readily soluble in chloroform, ether or vegetable oil, slightly molten in ethanol, in water In it is insoluble.The chemical name of vitamin K1 are as follows: 2- methyl -3- (3,7,11,15- tetramethyl -2- hexadecene base) -1,4- naphthalene two The mixture of the trans- and cis-isomer of ketone, molecular formula: C31H46O2, molecular weight: 450.71, structural formula are as follows:

Currently, version Chinese Pharmacopoeia in 2015 has recorded the bulk pharmaceutical chemicals and injection of vitamin K1 (Vitamin K1), the U.S. Pharmacopeia USP40 has recorded the bulk pharmaceutical chemicals, emulsion for injection and tablet of vitamin K1 (Phytonadione).

Vitamin K1 is a kind of liposoluble vitamin, is widely present in green plants, with phylloquinone (phylloquinone) form exists, can also be artificial synthesized.Vitamin K1 is the yellow for purifying out from alfalfa earliest Oily liquids, and it is named as vitamin K1, people just do hemostatic using vitamin K1 very early.The medical value that it stops blooding is 20 The discovery thirties in century.For the conspicuous Garrick of the biochemist of Denmark up to the illness of nurse research chicken internal haemorrhage, he has found chicken in injury Haemophilia can not condense, and until certain feed is added in food, find to stop blooding immediately in this feed rich in one kind later Substance, the substance of this hemostasis is exactly vitamin K1.Hereafter vitamin K1 is gradually applied to various hemorrhagic diseases.Through more The basic research and clinical use in year, the pharmacological activity other than vitamin K1 hemostasis are gradually found: treatment respiratory disease, solution Convulsion analgesic, prevention and treatment osteoporosis, treatment hepatitis, beauty, antitumor etc..Its pharmacological activity is definite, and toxic side effect is light, therefore answers extensively For clinic, existing vitamin K1 is clinical commonly used drug.

Vitamin K1 is vitamin K class medicine, and vitamin K is substance necessary to liver composition-factor II, VII, Ⅸ, Ⅹ, dimension Raw element K shortage can cause these coagulation factor dyssynthesis or exception, clinical visible hemorrhagic tendency and prolonged prothrombin. Clinically vitamin K1 can be used for bleeding caused by vitamin K deficiency, as caused by obstructive jaundice, leak, chronic diarrhea etc. Bleeding, Hypoprothrombinemia caused by Coumarins, sodium salicylate etc., prevents hemorrhage of newborn and prolonged application wide spectrum is anti- Internal vitamin K deficiency caused by raw element.Basic research and clinical use through many years, it was found that vitamin K1 can prevent and treat breathing Tract disease, relieving spasm and pain and treatment hepatitis.In addition also related with osteoporosis and beauty.Vitamin K1 pharmacological activity is definite, poison It is secondary light, therefore it is widely used in clinic, it is now clinical commonly used drug.

Human body cannot synthesize or secrete vitamin K1, and required vitamin K1 is mainly derived from food, big in enteron aisle Enterobacteria can also synthesize vitamin K1 as human body utilization.Vitamin K1 is the main component for forming prothrombing in blood of human body, It can promote liver manufacture factor, when tissue bleeding, blood platelet is destroyed, and thrombokinase and calcium ion, blood coagulation are released Proenzyme can be changed into fibrin ferment with thrombokinase and calcium binding, and fibrin ferment can be catalyzed plasma soluble fibrinogen It is changed into insoluble fibrin, reaches haemostatic effect.

Vitamin K1 can release smooth muscle spasmodic contraction, scavenging activated oxygen, and have parahormone sample anti-inflammatory effect, resistance Hinder the release of the chemical mediators such as histamine, mitigate respiratory mucosa inflammation, reduce exudation, expand blood vessel, improves blood fortune, mitigate the heart Dirty front and back load corrects heart failure；Excited respiratory center inhibits cerebral cortex, and improving brain microcirculation prevents brain edema, is conducive to exhale It inhales failure to correct, therefore can be relieved and improve the clinical symptoms of infantile pneumonia.In addition vitamin K1 can enhance certain antibiotic Effect, to inhibit growth of pathogenic bacteria, also has a better effect pertussis.

Vitamin K1 can directly act on a variety of visceral smooth muscles, terminate a variety of illness by the smooth muscle of relaxation spasm Caused internal organs pain, such as cholecystalgia, renal colic, ureter and cystolith colic pain etc., effective percentage can reach 80~90%, analgesic Effect is fast, persistent, safety, curative effect are high.

The shortage of vitamin K1 can cause blood not solidify, because the patient that vitamin K1 lacks bleeding is mostly new life in clinic Youngster.Newborn is born, and 1~2 day diet is limited, and the intake of vitamin K is few, and vitamin content is low in breast milk, in addition nothing in enteron aisle Bacterium influences the synthesis of vitamin K, leads to vitamin K deficiency, and liver is not mature enough, and liver synthesizes prothrombin, VII, IX, X Process be obstructed, cruor time extending leads to bleeding, and this disease is hemorrhagic disease of the newborn.Just early in nineteen fifty-two Dam etc. Describe the generation and avitaminous relationship of hemorrhagic disease of the newborn (HDN).Three kinds can be divided into according to disease time: early Hair style haemorrhage, typical haemorrhage, delayed.Be deficient in vitamin in body K1 when newborn is just born, the Pure breast feeding time compared with Long newborn's meeting is fast-developing to be lacked to serious vitamin K1: children can occur typically or 2 for first week after birth Occurs delayed haemorrhage in a month, both of these case morbidity is anxious, Yi Yinqi intracranials hemorrhage damage, disability rate height, the death rate are also It is high.The disease incidence in China is up to 0.5~2%, and the disease incidence of Area in Yantai Region is up to 9.03%, and China is vitamin K deficiency The hotspot of hemorrhagic disease.Medical research show newborn birth give vitamin K1 can prevent vitamin K1 lack property go out Hemorrhagic disease.The administration route of vitamin K1 also influences drug effect.Oral vitamin K1 is only capable of preventing typical haemorrhage, to delayed Haemorrhage is invalid, and Longhan has found that vitamin K1 with 2mg intramuscular injection drug effect sustainable 2 months or more, takes orally relatively poor.2000 Year, Hengxian County, China promotes newborn's birth, and interior routine is primary using vitamin K1 intramuscular injection for 24 hours, and result of study shows: baby Vitamin K deficiency number of hospitalized significantly reduces, vitamin K deficiency patient decline.In addition, because human liver can synthesize fibrin ferment Original, hepatic disfunction person or surgery large operation person, haemorrhage can be prevented by being both needed to give a certain amount of vitamin K1.

Infantile respiratory disease occupies pediatric disease disease incidence 70%, and winter-spring season disease incidence is higher, be mainly shown as pneumonia, Asthma, bronchitis, pertussis.

Infantile pneumonia refers to the lung inflammation as caused by various different pathogens and other factors, and lung microcirculation is caused to hinder Hinder, the reasons such as anoxic, infection, fever make blood that highly enriched, high aggregation, highly viscous state be presented, and can cause and aggravate microcirculation Obstacle.Changeable intracellular (cAMP/cGMP) ratio of vitamin K1, releases smooth muscle spasmodic and shrinks, scavenging activated oxygen, and With parahormone sample anti-inflammatory effect, the release of the chemical mediators such as histamine is hindered, mitigates respiratory mucosa inflammation, reduces exudation, Have the function that cough-relieving is coughed to relieving asthma；The also expansible blood vessel of vitamin K1 improves blood fortune, mitigates load before and after heart, corrects heart failure； Excited respiratory center inhibits cerebral cortex, and improving brain microcirculation prevents brain edema, can reduce airway resistance, increases alveolar ventilation, The ventilation ventilation failure for alleviating crisis life, is conducive to exhale the correction that declines, therefore vitamin K1 can be relieved and improve infantile pneumonia Clinical symptoms, while can also substantially reduce the case fatality rate of severe pneumonia shorten the hospital stays, to aminophylline, fenazil, fill in rice The measures nonresponder such as pine, oxygen uptake reapplies, and can still receive promising result, and someone assists severe pneumonia of infants 100 with dimension life Plain K1 treats 50, obtains preferable effect.

Vitamin K1 treats bronchiolitis and bronchial asthma report is also more, and the country, which has, to be studied from October, 1998 extremely October in 2001,41 children asthmatic brouchitis case intravenous applications vitamin K1s of institute treated asthmatic bronchitis, Satisfactory effect.After child vein instillation vitamin K1, infant is quiet rapidly, falls asleep quickly, and cough and asthma significantly reduces, and breathing is smooth.

Vitamin K1 also has a better effect pertussis.Pertussis is that the acute respiratory as caused by Bordetella pertussis passes It catches an illness.Infectiousness is strong, is more common in infant with liver source property blood coagulation disorders when pertussis is broken out and causes the point of respiratory mucosa Shape bleeding, this generates paroxysmal spasmodic cough to the cause of disease, and respiratory tract petechial hemorrhage is related with venous stasis or blood vessel toxic lesion.Though vitamin K1 Pathogen cannot be directly acted on, but it can enhance the effect of certain antibiotic, to inhibit growth of pathogenic bacteria.

Animal experiment study proves within 1974, vitamin K1 excised tea fresh leaves are shunk automatically and acetylcholine caused by intestines receive Contracting has apparent inhibiting effect, and clinical test shows in recent years: vitamin K1 can directly act on a variety of visceral smooth muscles, pass through pine The smooth muscle of relaxation spasm, and internal organs pain caused by a variety of illness is terminated, such as cholecystalgia, renal colic, ureter and cystolith Colic pain etc., effective percentage can reach 80~90%, and clinical vitamin K1 treatment children's gastrointestinal convulsion abdominal pain also has good Effect.

Vitamin K1 can be used to treat or prevent disease related with osteoporosis or illness.Nineteen sixty, Bouckaert etc. People reports that vitamin K can promote the union of rat and rabbit.Pettifor in 1975 etc. has found pregnant woman oral anti-coagulants institute Baby nose dysosteogenesis is produced, reports influence of the vitamin K deficiency to human body bone development for the first time, vitamin K participates in people's bone generation The hypothesis thanked；Pettifor thinks that replenishing vitamins K can promote bon e formation, reduces bone catabolism, has actively to osteoporosis Preventive and therapeutic action, mechanism of action is that glutaminic acid residue that the confactor as enzyme is catalyzed in certain protein turns It turns to γ-Hydroxy GIutaminic Acid residue (Gla), this albuminoid is also known as vitamin K dependent protein matter, γ-hydroxyl after activation Glutaminic acid residue has in conjunction with Ca⁺⁺Characteristic, so that vitamin k-dependent protein be made to play its bioactivity, when vitamin K lacks Weary, which is hydroxylated obstacle, therefore bioactivity also reduces.More bases and clinical tests illustrate vitamin K and The relationship of osteoporosis.Separately there is what anti-coagulants can block internal vitamin K to recycle the clear proof for causing vitamin K deficiency. There is within 1975 side effect of the clinical report anti-coagulants to bone metabolism, hereafter crowd and animal experiment are continuously available confirmation, long diaphysis After bone, the end growth too early calcification of bone plate.The above test prompt vitamin K deficiency can interfere bone tissue eubolism.

Vitamin K1 facilitates the treatment or prevention of liver inflammatory disease.It is thin that research confirms that vitamin K1 can enhance lesion liver The effect of born of the same parents' vigor and absorbability.Also it has been reported that and " swashs and raise therapy " treatment hepatopathy, vitamin K energy using megavitamin K Promote the Activity of hepatocytes of impending death, enhances the absorbability to nutriment, accelerate histiocytic recovery.

Version Chinese Pharmacopoeia in 2010 recorded vitamin K 1 injection (referring to 2010 version Chinese Pharmacopoeia two the 910th Page), this is a kind of sterilizing aqueous dispersions in yellow liquid that main ingredient concentration is 10mg/ml, allows micro- aobvious muddiness, needs to prevent To freeze and save, if there is oil droplet is precipitated or is layered, 70-80 DEG C can be heated under dark conditions, shaking makes its natural cooling, if Clarity normally may continue to show using, these information that the injection that the pharmacopeia is recorded is a kind of to keep drug straight using solubilizer Connect the injection being dissolved in the water, rather than the normally understood emulsion of art of pharmacy because emulsion occur oil droplet be precipitated or point It can not be by the way that clinically-acceptable clarity can be reached after being heated to 70-80 DEG C and shaking after layer.

WO2011/153513A2 discloses the micro emulsion composition of liposoluble vitamin K, it is stated that the micro emulsion composition is alternative (Phytonadione Injectable Emulsion, USP, may be simply referred to as the vitamin K1 emulsion for injection of United States Pharmacopeia PIE-USP, former name are vitamin K 1 injection) it is used for clinical treatment.PIE-USP is a kind of aqueous dispersions, wherein containing Polyoxyethylated fatty acid (also known as Cremophor), this is a kind of potent surfactant.Although nomenclature of drug is recorded as infusing It penetrates and uses cream, but it is Merck & Co., Inc. that the original of vitamin K1 emulsion for injection (USP), which grinds product,Its thing Any oil is not contained in reality, therefore is different from the vitamin K1 fat emulsion of meaning of the invention.Other manufacturers are also using being similar to The surfactant of Cremophor comes solubilized vitamin K1, such as the HCO-60 using high hlb.In WO2011/153513A2 The shortcomings that describing using such surfactant, and by not using such reagent, but a microemulsion is prepared to substitute Existing PIE-USP, the micro emulsion is for example containing chain three in 0.2~1% vitamin K1,0.5~2% soybean oil, 0.5~2% Acid glyceride, 4~25% phosphatide and 10% sucrose, phosphatide are 13-25 times of vitamin K1 amount.However it is this with larger amount of Phosphatide and it is less amount of oil constitute micro emulsion be a kind of critical state between solution and emulsion metastable state, be easy because Outside cause such as pressure sterilizing and be detached from this metastable state, such as in the preparation embodiment of WO2011/153513A2, need With 0.22 μm of membrane filtration degerming.In addition, being advantageous containing a certain amount of oily such as soybean oil, such as the soybean of 10% amount Oil, can be provided for human body can not or inconvenient intake energy.Compared to other types oil for injection (such as safflower oil, Olive oil etc.) for soybean oil safety and validity sufficiently demonstrate, in the form of fat emulsion injection provide Large dosage of soybean oil clinically used many decades.The present inventor team obtains the Chinese patent of power in the early time ZL201210030051.X (CN102552134B) discloses a kind of based on using vitamin made of soybean oil, phosphatide, glycerol K1 Fat Emulsion shows that excellent results are presented in one or more aspects in these Fat Emulsions, and the full content of the patent passes through reference It is incorporated herein.

People, which still expect to have, assigns more excellent performance vitamin K 1 injection such as vitamin K1 fat emulsion injection use In clinic, and expect that this vitamin K 1 injection such as vitamin K1 fat emulsion injection has good quality testing side Method.

Summary of the invention

The purpose of the present invention is provide a kind of new vitamin K 1 injection such as vitamin K1 Fat Emulsion injection for clinic Liquid, this injection have beneficial effect documented by least one such as present invention.The inventors discovered that by including about The vitamin K 1 injection that the special formulation of 10% soybean oil is prepared is that a kind of performance is fully able to meet clinical application need The Fat Emulsion asked.It is accomplished the present invention is based on this discovery.

For this purpose, first aspect present invention provides emulsion composition, it includes: vitamin K1, soybean oil, phosphatide, glycerol, Water.

Emulsion composition according to a first aspect of the present invention, it includes the vitamin K1s of 0.45-2%, preferably comprise 0.7- 1.5% vitamin K1 preferably comprises the vitamin K1 of 0.9-1.1%, preferably comprises about 1.0% vitamin K1.According to this The emulsion composition of invention first aspect, it includes the vitamin K1s of 0.5-2%, preferably comprise the vitamin of 0.75-1.5% K1 preferably comprises the vitamin K1 of 0.9-1.1%, preferably comprises about 1.0% vitamin K1.

Emulsion composition according to a first aspect of the present invention, it includes 7.5~12.5% soybean oil, preferably comprise 8~ 12% soybean oil preferably comprises 9~11% soybean oil, preferably comprises about 10% soybean oil.

Emulsion composition according to a first aspect of the present invention, it includes 0.8~2% phosphatide, preferably comprise 1.0~ 1.5% phosphatide preferably comprises about 1.2% phosphatide.

Emulsion composition according to a first aspect of the present invention, it includes 1.5~3% glycerol, preferably comprise 1.75~ 2.5% glycerol preferably comprises 1.95~2.45% glycerol, preferably comprises about 2.2% glycerol.

Emulsion composition according to a first aspect of the present invention, it includes:

Emulsion composition according to a first aspect of the present invention, pH value are 7.0~9.0.In one embodiment, pH value It is 7.0~8.5.In one embodiment, pH value is 7.5~9.0.In one embodiment, pH value is 7.5~8.5.

Emulsion composition according to a first aspect of the present invention, when it is by under the Chinese Pharmacopoeia annex VI H of version two in 2010 Method measurement when, pH value be 7.0~9.0.In one embodiment, pH value is 7.0~8.5.In one embodiment, PH value is 7.5~9.0.In one embodiment, pH value is 7.5~8.5.

Emulsion composition according to a first aspect of the present invention, wherein also including pH adjusting agent, dosage is to adjust the emulsion The amount of the pH value of composition to 7.0~9.0.In one embodiment, the dosage of the pH adjusting agent is to adjust the emulsion group Close the amount of the pH value to 7.0~8.5 of object.In one embodiment, the dosage of the pH adjusting agent is to adjust emulsion combination The amount of the pH value of object to 7.5~9.0.In one embodiment, the dosage of the pH adjusting agent is to adjust the emulsion composition PH value to 7.5~8.5 amount.In one embodiment, the pH adjusting agent is selected from sodium hydroxide, hydrochloric acid and its water-soluble Liquid.

Emulsion composition according to a first aspect of the present invention, when it is attached according to Pharmacopoeia of People's Republic of China 2010 version two When recording the wet process measuring method in Ⅸ E granularity and determination of particle size distribution third method (light scattering method), the grain of 70% or more milk particle Diameter is at 1.0 μm or less.For example, in one embodiment, the partial size of 80% or more milk particle is at 1.0 μm or less；In a reality It applies in scheme, the partial size of 90% or more milk particle is at 1.0 μm or less.For example, in one embodiment, 70% or more milk particle Partial size between 0.01~1.0 μm；In one embodiment, the partial size of 80% or more milk particle 0.01~1.0 μm it Between；In one embodiment, the partial size of 90% or more milk particle is between 0.01~1.0 μm.For example, in an embodiment In, the partial size of 70% or more milk particle is between 0.05~1.0 μm；In one embodiment, the grain of 80% or more milk particle Diameter is between 0.05~1.0 μm；In one embodiment, the partial size of 90% or more milk particle is between 0.05~1.0 μm.Example Such as, in one embodiment, the partial size of 70% or more milk particle is between 0.1~1.0 μm；In one embodiment, The partial size of 80% or more milk particle is between 0.1~1.0 μm；In one embodiment, the partial size of 90% or more milk particle exists Between 0.1~1.0 μm.For example, in one embodiment, be greater than in partial size and the milk particle sum equal to 0.5 μm in, partial size is big In 1 μm of milk particle less than 20%, preferably less than 15%, less than 10%, less than 5%.

Emulsion composition according to a first aspect of the present invention, when it is attached according to Pharmacopoeia of People's Republic of China 2010 version two When recording the wet process measuring method in Ⅸ E granularity and determination of particle size distribution third method (light scattering method), the average grain diameter of milk particle is little In 0.4 μm, such as average grain diameter is 0.05~0.4 μm, 0.1~0.4 μm, 0.15~0.4 μm, 0.2~0.4 μm, 0.25~0.4 μm, 0.3~0.4 μm, 0.05~0.35 μm, 0.05~0.3 μm, 0.05~0.25 μm, 0.05~0.2 μm, 0.05~0.15 μm, 0.05~0.1 μm, 0.1~0.35 μm, 0.15~0.3 μm or 0.2~0.3 μm.

Emulsion composition according to a first aspect of the present invention, when it is attached according to Pharmacopoeia of People's Republic of China 2010 version two When recording the wet process measuring method in Ⅸ E granularity and determination of particle size distribution third method (light scattering method), D90 value (the i.e. particle of milk particle Cumulative distribution be 90% partial size) be not more than 0.6 μm, such as D90 value be 0.1~0.6 μm, 0.1~0.5 μm, 0.1~0.4 μm, 0.1~0.3 μm, 0.1~0.2 μm, 0.2~0.6 μm, 0.3~0.6 μm, 0.4~0.6 μm, 0.3~0.6 μm, 0.2~0.5 μm, Or 0.3~0.4 μm.

Emulsion composition according to a first aspect of the present invention, when it is attached according to Pharmacopoeia of People's Republic of China 2010 version two When recording the wet process measuring method in Ⅸ E granularity and determination of particle size distribution third method (light scattering method), it cannot detect greater than 5 μm Milk particle.

Emulsion composition according to a first aspect of the present invention is oil-in-water type emulsion for injection.

Emulsion composition according to a first aspect of the present invention is the fat emulsion injection made of pressure sterilizing.

Emulsion composition according to a first aspect of the present invention, the soybean oil are the other soybean oils of injection stage.

Emulsion composition according to a first aspect of the present invention, the phosphatide are the other phosphatide of injection stage.In an embodiment party In case, the phosphatide is soybean lecithin or egg yolk lecithin.

Emulsion composition according to a first aspect of the present invention, the glycerol are the other glycerol of injection stage.

Emulsion composition according to a first aspect of the present invention, the water are water for injection.

Emulsion composition according to a first aspect of the present invention is substantially prepared by a method comprising the following steps It arrives:

(a) water soluble adjuvant (especially glycerol) and all or part of water are mixed and made into water phase；

(b) oil-soluble auxiliary material (especially soybean oil and phosphatide) and vitamin K1 are mixed and made into oily phase；

(c) oily be mutually stirred with water phase is made to prepare thick cream, optional adds water to full dose, and optional is adjusted with pH adjusting agent Medical fluid pH value；

(d) by thick lotion carry out high-pressure emulsification, filter, packing, pressure sterilizing to get,

Optional, the operation in relation to substance especially impurity B detection is carried out to emulsion composition obtained by experience pressure sterilizing, Include the following steps:

(1) it is measured according to " Chinese Pharmacopoeia " 2015 editions four 0512 sections of general rule about the specification of high performance liquid chromatography, It is protected from light when operation；

(2) octadecylsilane chemically bonded silica chromatographic column (such as 5 μm of filler particle size, column internal diameter chromatographic condition: are used 4.6mm, 200~300mm of column length), 25~35 DEG C of column temperature (such as 28~32 DEG C), mobile phase A is the anhydrous second of volume ratio 90:10 Alcohol-water mixtures, Mobile phase B are dehydrated alcohol, and flow velocity is 0.8~1.2ml/min (such as 1.0ml/min)；Detection wavelength is 265~275nm (such as 270nm)；

(3) eluted using mobile phase A and Mobile phase B by such as Gradient: first time period (such as initial 20~ In 40 minutes sections such as 25~35 minutes sections, such as during 0 minute to 30 minutes of entire elution process) flowing Phase are as follows: mobile phase A is 100% and Mobile phase B is 0, in second time period (such as next 5~20 minutes section such as 8 In~15-min period, such as during 30 minutes to 40 minutes of entire elution process) by mobile phase A be 100% and stream Dynamic phase B was 0 to be changed into that mobile phase A is 0 and Mobile phase B is 100%, at the third period (such as next 10~30 minutes In period such as 15~25 minutes sections, such as during 40 minutes to 60 minutes of entire elution process) by mobile phase A It is 100% to be changed into that mobile phase A is 100% and Mobile phase B is 0 for 0 and Mobile phase B；

(4) measuring method: precision amount emulsion composition 1ml sets in 10ml measuring bottle, with isopropanol to scale, shakes up, As test solution；Precision measures above-mentioned test solution 1ml, sets in 100ml measuring bottle, with isopropanol to scale, shakes It is even, as contrast solution；

(5) accurate respectively to measure above-mentioned test solution and each 20 μ l of contrast solution, it is injected separately into chromatography, records color Spectrogram is to 60min；

(6) response of each impurity peaks in test solution chromatogram is read, and calculates separately each impurity peak response value phase For the percentage of contrast solution vitamin K1 main peak response；It is 1 calculating with the relative retention time at vitamin K1 peak, for Relative retention time is that the impurity peaks at 0.52 ± 0.03 are named as impurity B, if impurity B exists, calculates its response multiplied by school Percentage of 0.7 resulting value of positive divisor relative to contrast solution main peak response, thus calculates impurity B relative to vitamin K1 Percentage.

(1) vitamin K1, soybean oil, phosphatide, glycerol are weighed by recipe quantity, it is spare；

(2) stainless cylinder of steel is added in the water for injection that will prepare total amount about 60-90% (for example, about 80%), and recipe quantity is added Glycerol stirs evenly, and water phase is made；

(3) soybean oil of recipe quantity is added in another stainless cylinder of steel, is passed through nitrogen stream, the vitamin K1 of recipe quantity is added And phosphatide, stirring to each material dissolution, oil phase is made；

(4) it is passed through nitrogen stream, oil is mutually slowly added in the water phase of high-speed stirred, continuation high-speed stirred is in due course, obtains colostrum；

(5) adjust the pH value of the colostric fluid to prescribed limit with pH adjusting agent, then plus water for injection be settled to theoretical prescription Amount, stirs evenly, and adjusts solution ph to prescribed limit with pH adjusting agent when necessary；

(6) under nitrogen flowing, make colostrum high pressure homogenization under 40-70MPa (such as 55-60MPa) pressure, then in 5- Low pressure is homogenized under 25MPa (such as 12-18MPa) pressure, cooling, then with 10 μm of membrane filtration；

(7) filtrate is filling in ampoule bottle, nitrogen charging sealing, pressure sterilizing to get,

Emulsion composition according to a first aspect of the present invention, wherein also including alkanoic acid or its alkali metal salt such as sodium salt.

Emulsion composition according to a first aspect of the present invention, the alkanoic acid or its alkali metal salt such as sodium salt be oleic acid or its Alkali metal salt such as sodium salt.In one embodiment, 0.01~0.05% oleic acid or its alkali gold are included in emulsion composition Belong to salt such as sodium salt, such as includes 0.01~0.045%, 0.01~0.04%, 0.01~0.035% or 0.01~0.03% Oleic acid or its alkali metal salt such as sodium salt, such as comprising 0.015~0.05%, 0.02~0.05%, 0.025~0.05%, Or 0.03~0.05% oleic acid or its alkali metal salt such as sodium salt, such as comprising 0.015~0.045%, 0.02~0.04%, 0.025~0.035% oleic acid or its alkali metal salt such as sodium salt.It has been unexpectedly discovered that the oil comprising Optimum Acid or its alkali metal salt such as sodium salt can assign emulsion composition some or certain excellent characteristics.It is oiliness object in view of oleic acid Matter is first added in oily phase with oiliness material of course in emulsion preparation process, for example, with soybean oil or vitamin K1 or Phosphatide adds together.Certainly, with its alkali metal salt such as sodium salt in use, with aqueous materials be added in water phase for example directly It is added in water.

Emulsion composition according to a first aspect of the present invention, the alkanoic acid or its alkali metal salt such as sodium salt be fatty acid or Its alkali metal salt such as sodium salt.In one embodiment, in emulsion composition comprising 0.01~0.05% fatty acid or its Alkali metal salt such as sodium salt, such as comprising 0.01~0.045%, 0.01~0.04%, 0.01~0.035% or 0.01~ 0.03% fatty acid or its alkali metal salt such as sodium salt, such as comprising 0.015~0.05%, 0.02~0.05%, 0.025~ 0.05% or 0.03~0.05% fatty acid or its alkali metal salt such as sodium salt, such as include 0.015~0.045%, 0.02 ~0.04%, 0.025~0.035% fatty acid or its alkali metal salt such as sodium salt.According to the present invention, the fatty acid or its Alkali metal salt such as sodium salt be include oleic acid, linoleic acid, the combination of palmitinic acid three or its alkali metal salt such as sodium salt.According to this Invention, the fatty acid or its alkali metal salt such as sodium salt be include that oleic acid, linoleic acid, the combination of palmitinic acid three or its alkali are golden Belonging to salt such as sodium salt and three's weight ratio is 70~88:5~15:2~10.According to the present invention, the fatty acid or its alkali gold Belonging to salt such as sodium salt to be includes oleic acid, linoleic acid, the combination of palmitinic acid three or its alkali metal salt such as sodium salt and three's weight Amount is than being 72~85:8~13:2~6.It has been unexpectedly discovered that the fatty acid of the said combination comprising Optimum or its Alkali metal salt such as sodium salt can assign emulsion composition some or certain excellent characteristics.It is oiliness object in view of these fatty acid Matter is first added in oily phase with oiliness material of course in emulsion preparation process, for example, with soybean oil or vitamin K1 or Phosphatide adds together.Certainly, with its alkali metal salt such as sodium salt in use, with aqueous materials be added in water phase for example directly It is added in water.

Emulsion composition involved in emulsion composition according to a first aspect of the present invention or either side of the present invention or Vitamin K1 composition, wherein the alkanoic acid is added together with oiliness material.For example, the alkanoic acid be with soybean oil and/ Or vitamin K1 and/or phosphatide add together.Or wherein the alkali metal salt of the alkanoic acid such as sodium salt is and aqueous materials It adds together.For example, the alkali metal salt of the alkanoic acid such as sodium salt is added together with water.

Emulsion composition according to a first aspect of the present invention obtains finished product in experience pressure sterilizing during the preparation process Afterwards, further include being carried out to the emulsion composition in relation to the operation of substance especially impurity B detection, include the following steps:

Further, second aspect of the present invention provides the method for emulsion composition described in preparation first aspect present invention, Itself the following steps are included:

Method according to a second aspect of the present invention comprising following steps:

Further, third aspect present invention provides the pharmaceutical applications of emulsion composition described in first aspect present invention, For prothrombin, VII, IX and X dyssynthesis caused by the pharmaceutical intervention of vitamin K deficiency or interference vitamin k activity The disease of caused such as, but not limited to following coagulation disorders: it is coagulated caused by the anti-coagulants such as cumarin or indene-dione derivative Hemase is former insufficient；Hypoprothrombinemia caused by antibiotic treatment；It is low that disease causes absorption or synthesis vitamin K obstacle to cause Factor mass formed by blood stasis, such as obstructive jaundice, leak, sprue, ulcerative colitis, celiaca, Bowel resection, pancreas capsule Property fibrosis and regional enteritis；Other drugs interfere vitamin K metabolism, caused Hypoprothrombinemia, such as salicylate. Vitamin K1 fat emulsion injection produced by the present invention when in use can each 10mg, 1-2 times daily, total amount is not in 24 hours More than 40mg, dosage and frequency are adjusted according to prothrombin time response or clinical symptoms；Usage mode is intravenously administrable, Infusion can not have to dilution, direct intravenous injection, or with 5% glucose injection dilute after intravenous drip.

Further, fourth aspect present invention provides outstanding using related substance in HPLC method measurement vitamin K1 composition It is the method for impurity B, and this method includes following operation:

Method according to a fourth aspect of the present invention, the vitamin K1 composition are the emulsion groups comprising soybean oil and phosphatide Close object.

Method according to a fourth aspect of the present invention, the vitamin K1 composition includes: vitamin K1, soybean oil, phosphatide, Glycerol, water.

Method according to a fourth aspect of the present invention, the vitamin K1 composition includes the vitamin K1 of 0.45-2%, excellent Choosing includes the vitamin K1 of 0.7-1.5%, preferably comprises the vitamin K1 of 0.9-1.1%, preferably comprises about 1.0% vitamin K1。

Method according to a fourth aspect of the present invention, the vitamin K1 composition includes 7.5~12.5% soybean oil, excellent Choosing includes 8~12% soybean oil, preferably comprises 9~11% soybean oil, preferably comprises about 10% soybean oil.

Method according to a fourth aspect of the present invention, the vitamin K1 composition include 0.8~2% phosphatide, are preferably wrapped Containing 1.0~1.5% phosphatide, about 1.2% phosphatide is preferably comprised.

Method according to a fourth aspect of the present invention, the vitamin K1 composition include 1.5~3% glycerol, are preferably wrapped Containing 1.75~2.5% glycerol, 1.95~2.45% glycerol is preferably comprised, preferably comprises about 2.2% glycerol.

Method according to a fourth aspect of the present invention, the vitamin K1 composition includes:

Method according to a fourth aspect of the present invention, described its pH value of vitamin K1 composition are 7.0~9.0.In a reality It applies in scheme, pH value is 7.0~8.5.In one embodiment, pH value is 7.5~9.0.In one embodiment, pH value It is 7.5~8.5.

Method according to a fourth aspect of the present invention, the vitamin K1 composition is when it is by Chinese Pharmacopoeia 2010 version two When method under annex VI H measures, pH value is 7.0~9.0.In one embodiment, pH value is 7.0~8.5.At one In embodiment, pH value is 7.5~9.0.In one embodiment, pH value is 7.5~8.5.

Method according to a fourth aspect of the present invention, also includes pH adjusting agent in the vitamin K1 composition, and dosage is Adjust the amount of the pH value to 7.0~9.0 of the emulsion composition.In one embodiment, the dosage of the pH adjusting agent is to adjust Save the amount of the pH value to 7.0~8.5 of the emulsion composition.In one embodiment, the dosage of the pH adjusting agent is to adjust The amount of the pH value of the emulsion composition to 7.5~9.0.In one embodiment, the dosage of the pH adjusting agent is to adjust to be somebody's turn to do The amount of the pH value of emulsion composition to 7.5~8.5.In one embodiment, the pH adjusting agent is selected from sodium hydroxide, salt Acid and its aqueous solution.

Method according to a fourth aspect of the present invention, the vitamin K1 composition shine Pharmacopoeia of People's Republic of China when it When wet process measuring method in two annex of version in 2010, Ⅸ E granularity and determination of particle size distribution third method (light scattering method), 70% The partial size of above milk particle is at 1.0 μm or less.For example, in one embodiment, the partial size of 80% or more milk particle is at 1.0 μm Below；In one embodiment, the partial size of 90% or more milk particle is at 1.0 μm or less.For example, in one embodiment, The partial size of 70% or more milk particle is between 0.01~1.0 μm；In one embodiment, the partial size of 80% or more milk particle exists Between 0.01~1.0 μm；In one embodiment, the partial size of 90% or more milk particle is between 0.01~1.0 μm.For example, In one embodiment, the partial size of 70% or more milk particle is between 0.05~1.0 μm；In one embodiment, 80% The partial size of above milk particle is between 0.05~1.0 μm；In one embodiment, the partial size of 90% or more milk particle is 0.05 Between~1.0 μm.For example, in one embodiment, the partial size of 70% or more milk particle is between 0.1~1.0 μm；At one In embodiment, the partial size of 80% or more milk particle is between 0.1~1.0 μm；In one embodiment, 90% or more cream The partial size of grain is between 0.1~1.0 μm.For example, in one embodiment, it is greater than in partial size and milk particle equal to 0.5 μm is total In number, milk particle of the partial size greater than 1 μm is less than 20%, preferably less than 15%, less than 10%, be less than 5%.

Method according to a fourth aspect of the present invention, the vitamin K1 composition shine Pharmacopoeia of People's Republic of China when it When wet process measuring method in two annex of version in 2010, Ⅸ E granularity and determination of particle size distribution third method (light scattering method), milk particle Average grain diameter be not more than 0.4 μm, such as average grain diameter be 0.05~0.4 μm, 0.1~0.4 μm, 0.15~0.4 μm, 0.2~ 0.4 μm, 0.25~0.4 μm, 0.3~0.4 μm, 0.05~0.35 μm, 0.05~0.3 μm, 0.05~0.25 μm, 0.05~0.2 μ M, 0.05~0.15 μm, 0.05~0.1 μm, 0.1~0.35 μm, 0.15~0.3 μm or 0.2~0.3 μm.

Method according to a fourth aspect of the present invention, the vitamin K1 composition shine Pharmacopoeia of People's Republic of China when it When wet process measuring method in two annex of version in 2010, Ⅸ E granularity and determination of particle size distribution third method (light scattering method), milk particle D90 value (i.e. particulate accumulation be distributed as 90% partial size) no more than 0.6 μm, such as D90 value is 0.1~0.6 μm, 0.1~0.5 μm, 0.1~0.4 μm, 0.1~0.3 μm, 0.1~0.2 μm, 0.2~0.6 μm, 0.3~0.6 μm, 0.4~0.6 μm, 0.3~0.6 μm, 0.2~0.5 μm or 0.3~0.4 μm.

Method according to a fourth aspect of the present invention, the vitamin K1 composition shine Pharmacopoeia of People's Republic of China when it It, cannot when wet process measuring method in two annex of version in 2010, Ⅸ E granularity and determination of particle size distribution third method (light scattering method) Detect the milk particle greater than 5 μm.

Method according to a fourth aspect of the present invention, the vitamin K1 composition are oil-in-water type emulsion for injection.

Method according to a fourth aspect of the present invention, the vitamin K1 composition is the Fat Emulsion made of pressure sterilizing Injection.

Method according to a fourth aspect of the present invention, the soybean oil of the vitamin K1 composition are that injection stage is other big Soya-bean oil.

Method according to a fourth aspect of the present invention, the phosphatide of the vitamin K1 composition are the other phosphorus of injection stage Rouge.In one embodiment, the phosphatide is soybean lecithin or egg yolk lecithin.

Method according to a fourth aspect of the present invention, the glycerol of the vitamin K1 composition are that injection stage is other sweet Oil.

The water of method according to a fourth aspect of the present invention, the vitamin K1 composition is water for injection.

Method according to a fourth aspect of the present invention, the vitamin K1 composition are substantially to pass through to include the following steps What method was prepared:

(d) by thick lotion carry out high-pressure emulsification, filter, packing, pressure sterilizing to get.

Method according to a fourth aspect of the present invention, the vitamin K1 composition are such as any implementation of first aspect present invention Emulsion composition described in scheme.

The documents and materials that the present invention quotes from, entire contents are incorporated herein by reference.In either present invention face, Any one or more technical characteristics of any embodiment can be combined in any other embodiments of this aspect, can also To be combined in any embodiment of another aspect, as long as this combination is not in contradiction.

The general sense that there are the various terms that the present invention uses those skilled in the art routinely to understand, is generally containing with this When justice is inconsistent, it is subject to the present invention.

During the present invention prepares emulsion composition, the group is measured and/or adjusted before composition encapsulates and sterilizes It is very easy for closing the pH value of object；And lead to (after becoming seal-packed emulsion finished product) after composition encapsulates and sterilizes Chang Buyi is again adjusted the pH value of the composition.It will be appreciated by those skilled in the art that the pH value of composition can after before sterilization When can be varied, either slightly increase or slightly reduce, therefore pH value being adjusted during matching liquid, Ke Yishi When deviateing target ph, so that the pH value after composition sterilizing falls into as far as possible or close to purpose pH value or range.From this Said in a meaning, in the present invention, when referring to the pH value of emulsion composition, if be not indicated otherwise, refer to sterilization treatment it PH value afterwards；Such as below in preparation example 1, when describing composite formula, refer to that " appropriate pH adjusting agent adjusts pH value to 8.0 " Refer in formula or appropriate with suitable pH adjusting agent is added during liquid, medical fluid is made to handle and pass through through corresponding preparation method After pressure sterilizing obtains final emulsion composition, the pH value of the emulsion composition is 8.0 or so, such as pH value is 8.0 ± 0.2 In the range of, such as pH value is in the range of 8.0 ± 0.1.

In the present invention, convenient to write, vitamin K1 can simply be written as VK1.

Present invention generally provides be a kind of liquid composition, main carriers therein are water, and solute includes liquid Substance (such as glycerol) and/or solid matter (such as egg yolk lecithin in semi-solid).Therefore, for convenience, exist In the present invention, when in the case where indicating percent concentration using " % " is arrived, if not otherwise indicated, refer to weight/volume percentage Number；Such as.For containing " vitamin K1 1%, soybean oil 10%, egg yolk lecithin 1.2%, glycerol 2.2%, appropriate amount of water add To 100% " formula, refer in every 100ml formula containing vitamin K1 to be 1g, soybean oil 10g, egg yolk lecithin 1.2g, Glycerol 2.2g and the water for adding to 100ml volume.

It will be appreciated by those skilled in the art that milk particle partial size is usually that statistical way is calculated or provided.For example, phrase " partial size of 70% or more milk particle is at 1.0 μm or less " can refer to according to some way (such as microscopic method, light scattering method Deng) partial size of the milk particle particle of measurement certain amount, such as about 1000 milk particle particles are measured, partial size is then fallen in 1.0 μm About 1000 populations that population in following range is calculated divided by statistics, obtained percentage are greater than 70%.Other classes Also there is similar meaning like term.

In the present invention, phrase " milk particle partial size is at 0.5 μm or so " refers to the partial size of milk particle 0.2 if not otherwise indicated μm~1.0 μm between.

In the present invention, term " 10% fat emulsion injection ", if not otherwise indicated, refer to according to method of preparing emulsion, The emulsion being injected intravenously substantially by being prepared including following formula: soybean oil 10%, egg yolk lecithin 1.2%, Glycerol 2.2%, appropriate amount of water adds to 100%.

The present invention is the emulsion composition of injection, and one of the various materials are preferably matched with injection rank material Material.In emulsion composition of the invention, vitamin K1 exists as main ingredient or active constituent.It is combined in emulsion of the invention In object, soybean oil is used usually as oily phase solvent, it also has the function of providing energy certainly.In emulsion composition of the present invention Mesolecithal lecithin usually plays the effect of emulsifier, and glycerol is usually the effect for playing osmotic pressure regulator；Further, Water for injection is used as water phase solvent.

Unquestionably, the auxiliary material used in the present invention, including soybean oil, be need to meet it is necessarily required, for example, preparing When fat emulsion for injection, these auxiliary materials can should be used for injection.This is well known to those skilled in the art.

Phosphatide of the present invention is the phosphatide selected from following purification: egg yolk lecithin, soybean lecithin, cephalin, Hydrogenated yolk lecithin, hydrogenated soy phosphatidyl choline etc..They can be commercially, it is also possible to usual method, it is such as organic molten It is prepared by agent partition method.Namely raw phospholipid is dissolved in cold normal hexane-acetone, acetone, mistake are slowly added under stirring Filter recycling insoluble matter, after repeating this operation, steaming solvent can be obtained by refined phosphatide.Its main ingredient includes phosphatidyl gallbladder Alkali, phosphatidyl-ethanolamine etc..It can also illustrate out phosphatidylinositols, phosphatidylserine, sphingomyelins etc. as other phosphatide.This The signified phosphatide of invention is preferably egg yolk lecithin, soybean lecithin, more preferably egg yolk lecithin.

In the present invention, it refers to " Chinese Pharmacopoeia ", refers to Pharmacopoeia of People's Republic of China, can refer to various versions Chinese Pharmacopoeia；Particularly, as do not indicated especially, refer to the version of version in 2010；As not especially indicate, refer to 2010 version two Related content.

The pH adjusting agent that the present invention uses includes but is not limited to sodium hydroxide solution, hydrochloric acid solution, such as 1M sodium hydroxide Solution, 1M hydrochloric acid solution.In the present invention, refer to emulsion composition of the present invention or for the composition of control pH value when, The pH value is the final ph of composition, rather than the pH value of material among it.Such as emulsion of the present invention, dispensing The pH value measured after the pH value and sterilizing that are measured before into ampoule bottle is possible to have slight variations, mentions in the present invention And pH value refer to sterilizing after emulsion pH value.Therefore in production practice, when carrying out ingredient before sterilization, can suitably examine Consider this sterilizing front and back possible variation of pH value；For example, if pH value can reduce by 0.2 unit after sterilization, and target ph is 8.0, then the pH value of prepared emulsion composition, which is adjusted to 8.2, before sterilizing is advisable.

Fat emulsion provided by the invention can be prepared as follows:

Vitamin K1, soybean oil, phosphatide, glycerol are weighed by recipe quantity, it is spare；The water for injection of total amount about 80% will be prepared Stainless cylinder of steel is added, the glycerol of recipe quantity is added while stirring；The soybean oil of recipe quantity is added in another stainless cylinder of steel, is passed through Nitrogen stream, is added the vitamin K1 and phosphatide of recipe quantity, and stirring to phosphatide is dissolved；It is passed through nitrogen stream, oil is mutually slowly added to height In the water phase of speed stirring, it is in due course to continue high-speed stirred, adjusts solution ph to prescribed limit with sodium hydroxide solution, then this is first Cream is settled to theoretical recipe quantity, stirs evenly；Under nitrogen flowing, make colostrum high under 40-70MPa (such as 55-60MPa) pressure Pressure homogenizing, then low pressure is homogenized under 5-25MPa (such as 12-18MPa) pressure, it is cooling, then with 10 μm of membrane filtration；Filtrate It is filling in ampoule bottle, nitrogen charging sealing, pressure sterilizing to get.

Fat emulsion containing vitamin K1 of the invention is suitable for medicinal, for example oral and injection uses.Particularly, this hair Bright emulsion composition can suitable injection administration use, milk particle diameter therein is small, and average grain diameter has between 0.1~1 μm Meet the general quality requirement of drug.

Fat emulsion containing vitamin K1 of the invention can be infused by the approach and mode medication of injection, including muscle It penetrates, intravenous injection, approach and the mode such as intravenous drip.For example, when giving people's used time, the fat of the invention containing vitamin K1 Emulsion, dosage for each person every day can be 0.1mg~50mg, preferably 1mg~40mg.It can once a day or repeatedly Administration, can also be administered once for more days.It, can the direct vein of fat emulsion by the present invention containing vitamin K1 in intravenously administrable Injection, disposably intravenous injection or intravenous drip after it can also being diluted with infusion diluent appropriate.It is described Infusion diluent include electrolyte solution such as physiological saline, glucose solution, xylitol solution, fructose soln, woods grignard T₃ With glycerine etc..Particularly, the infusion diluent is glucose solution such as 5% glucose injection and, for example, commercially available 10% fat emulsion injection for nutritional supplement.

The fat emulsion that the present invention contains vitamin K1 has good pharmacy performance, meets medicinal requirements.For example, this The vitamin K1 emulsion composition that inventive embodiments 1~15 are prepared according to state food and drug administration about The rule of the anaphylaxis of injection systemic administration emulsion, hemolytic and vascular stimulation test, muscle irritation, subcutaneous irritation test Model is tested, as the result is shown: the vitamin K1 fat emulsion injection of 4mg/kg and 1mg/kg is to cavy without allergenic effect；Dimension Raw element K1 fat emulsion injection 0.2mg/kg~1mg/kg is to family's rabbit erythrocyte without obvious hemolysis in vitro and agglutination；1mg/ml Vitamin K1 fat emulsion injection 1mg/min intravenously administrable, 1 time a day, continuous 7 days, to rabbit auricular vein without obvious stimulation Effect；1ml is subcutaneously injected in the vitamin K1 fat emulsion injection of 10mg/ml, 1 time a day, continuous 7 days, no obvious stimulation effect.

Although excellent traditional performance as described herein is presented in some emulsion compositions prepared by the present invention, these creams Agent composition encounters some transnormal situations sometimes.For example, although fat emulsion injection prepared by the present invention combines Object is used directly for being injected intravenously, however clinically also often expects with injection solvent such as 5% glucose injection dilution Intravenous drip is afterwards to reduce injection speed, however this dilution may be especially such as its milk particle grain to the immanent structure of emulsion Diameter has an impact, therefore is necessary to the consideration of being diluted property of emulsion；In another example although emulsion composition of the present invention can be with It is saved in the shady place lower than 20 DEG C, however there may come a time when lower than 20 DEG C of shady place for this legal provisions can be special close to 0 DEG C Be not there are many fat emulsion injection be usually require 2~8 DEG C at a temperature of save, storage and transportational process in this hair Bright fat emulsion injection is easy to coexist in same environment with other Fat Emulsions, and this 2~8 DEG C of common environment are very easy to Drug is set unintentionally to be exposed to freezing point temperature hereinafter, may result in emulsion creaming in turn, therefore to emulsion composition of the present invention Injection carries out freezing-thawing test and is exposed to influence of the freezing point temperature or less to emulsion to investigate emulsion being necessary.Below to this The emulsion composition of invention preparation carries out the dilution of 5% glucose injection and freezing and thawing test is investigated.5% glucose injection Dilution test is investigated: the various emulsion composition injections for taking specific embodiment of the invention part to prepare add to 50 times of volumes It in 5% glucose injection, is uniformly mixed, stands 5 hours, measure milk particle average grain diameter and be denoted asIn addition it measures without dilute The milk particle average grain diameter for the emulsion composition released simultaneously is denoted asAverage grain diameter percent change is calculated as follows:The average grain diameter percent change is got over closer to 0 change of size Small, possibly even since source of error causes the percentage for negative value in substantially unconverted situation, and the average grain diameter becomes Change percentage to be positive value and show that average grain diameter is significantly increased when larger；The results show that the whole emulsion combinations of preparation example 1~9 Object, Ex121a~Ex123a of preparation example 12 and Ex121b~Ex123b emulsion composition, the whole emulsion compositions of preparation example 13, The oleic acid or fatty acid dosage of these emulsion compositions, which are respectively less than, is equal to 0.01% or unused oleic acid or fatty acid, they Average grain diameter percent change is in 125~203% ranges and oleic acid or the fewer average grain diameter variation of fatty acid dosage is presented The bigger trend of percentage, for example, the average grain diameter percent change of Ex121, Ex122 and Ex123 three be respectively 128.4%, 167.2%, 202.7%；And the whole emulsion compositions of preparation example 10, the whole emulsion compositions of preparation example 11, preparation example 12 it Ex124a~Ex128a and the whole emulsion compositions of Ex124b~Ex128b emulsion composition, preparation example 14, preparation example 15 are all Emulsion composition, the oleic acid or fatty acid dosage of these emulsion compositions are more than or equal to 0.02% in 0.02~0.5% range, Their average grain diameter percent change can be sayed in -9.7~14.1% a small range and irregularities, such as preparation example 11 The average grain diameter percent change of Ex111a~Ex114a be respectively 4.3%, 2.7%, -6.3%, 8.2%；According to this A little results are it is found that in emulsion composition of the present invention, and addition is more than or equal to 0.02% oleic acid or fatty acid for dimension in emulsion It is beneficial that emulsion composition, which is held, in terms of the milk particle grain size stability after being diluted with glucose injection.Freezing and thawing test is examined Examine: 1 day at a temperature of emulsion composition is set -10 DEG C, then 1 day at a temperature of setting 25 DEG C, so circulation three times, freeze through this three by measurement Melt the milk particle average grain diameter after recycling and is denoted asAnd see whether that there are demulsifying phenomenons, it in addition measures without at Frozen-thawed cycled The milk particle average grain diameter of the emulsion composition of reason is simultaneously denoted asAverage grain diameter percent change is calculated as follows:The average grain diameter percent change is got over closer to 0 change of size Small, possibly even since source of error causes the percentage for negative value in substantially unconverted situation, and the average grain diameter becomes Change percentage to be positive value and show that average grain diameter is significantly increased when larger；The results show that the whole emulsion compositions of preparation example 10, The whole emulsion compositions of Ex124a~Ex128a and Ex124b~Ex128b emulsion composition of preparation example 12, preparation example 14, this The oleic acid or fatty acid dosage of a little emulsion compositions are more than or equal to 0.06% in 0.06~0.5% range, their average grain Diameter percent change is in 443~818% ranges and oleic acid or the bigger average grain diameter percent change of fatty acid dosage is presented Bigger trend, for example, the average grain diameter percent change of Ex124a~Ex128a is respectively 452.2%, 524.1%, 585.6%, 647.3%, 802.7%；And the whole emulsion compositions of preparation example 1~9, the whole emulsion compositions of preparation example 11, system The whole emulsion compositions of Ex121a~Ex123a and Ex121b~Ex123b emulsion composition of standby example 12, preparation example 13, preparation The whole emulsion compositions of example 15, the oleic acid or fatty acid dosage of these emulsion compositions are respectively less than 0.06% or unused oleic acid Or fatty acid, their average grain diameter percent change can be sayed in -14.5~33.4% a small range and irregularities, example As the average grain diameter percent change of Ex111a~Ex114a of preparation example 11 is respectively 8.6%, 18.3%, -9.5%, 22.9%；In addition, it is observed that the whole emulsion compositions of oleic acid or fatty acid dosage less than or equal to 0.05% do not have demulsification existing As occurring, and the whole emulsion compositions of oleic acid or fatty acid dosage more than or equal to 0.06% have different degrees of demulsifying phenomenon And it shows that oleic acid or the bigger demulsification of fatty acid dosage are more serious, has different degrees of small oil droplet that display is precipitated on emulsion liquid level Demulsification；According to these results it is found that in emulsion composition of the present invention, addition is less than or equal to 0.05% oleic acid or rouge in emulsion Fat acid is beneficial in terms of undergoing the milk particle grain size stability of freezing-thawing test and avoiding demulsification for maintaining emulsion composition.

In addition, can be used for detecting the related of emulsion composition of the present invention the present invention provides a kind of unique HPLC method The content of substance impurity B especially therein, which, which is unique in that, can be used for Accurate Determining and quantitating vitamin K1 The content of impurity B in the composition of emulsion composition and other formulas.Specific method and result are recorded in hereafter.Use the present invention The HPLC method of test example 5 measures the impurity B content in preparation example 1~15 in vitamin K1 bulk pharmaceutical chemicals used, as the result is shown dimension life Impurity B content in plain K1 bulk pharmaceutical chemicals is 0.013%；Using the HPLC method of test example 5 of the present invention, 1~15 institute of preparation example is measured Whole emulsion compositions are obtained, impurity B content is in 0.01~0.1% range；The whole emulsion combinations of 1~15 gained of preparation example Object undergo 15 ± 1 DEG C shading, after closed, shady place saves 24 months, measure its impurity B content 0.3~0.6% In range, each emulsion composition impurity B content therein compared with it is 0 month rises appreciably；Use the HPLC of test example 5 of the present invention Method, the commercially available vitamin K 1 injection of five kinds of measurement (are the sterilizing aqueous dispersions of 1ml:10mg specification rather than emulsion of the invention Composition, respectively by Jiangsu, Shandong, Zhejiang enterprise produce) in impurity B content, as the result is shown commercially available nearly production period and The content of impurity B is significantly larger than the content of impurity B in each emulsion composition in the injection of nearly validity period, nearly production period and closely has The content of impurity B is respectively 0.2%~0.6% or so and 0.8%~1.5% or so in the injection of effect phase, or even part is looked forward to The content of impurity B has been more than 1.5% in the commercially available injection of industry, according to two vitamins recorded of " Chinese Pharmacopoeia " version in 2015 K1 injection quality standard, it is known that the commercially available injection of nearly effect phase does not meet its quality standard.According to the above results as it can be seen that this hair Bright emulsion composition quality standard is better than commercially available injection.Impurity B content in emulsion composition and commercially available injection is slow It is slow to increase, but impurity B is increased speed much smaller than in commercially available injection in emulsion composition, within validity period, impurity B Content within the acceptable range.In summary, impurity B content in different dosage forms product increases, and only increases speed Difference, in order to assess the quality of product, it is very useful for being monitored to the impurity B content in emulsion composition.

Certain or certain excellent effects is presented in the record of context according to the present invention, emulsion composition prepared by the present invention.

Specific embodiment

Following embodiment further illustrates the present invention, rather than limits the present invention.In following example, when using pH It unless otherwise noted, is adjusted with 1M sodium hydroxide solution or 1M hydrochloric acid solution when material is adjusted in regulator, Its dosage is that the pH value of material especially solution is made to be adjusted to specified value (such as in ± 0.1 range of indicated value) or range, Such as be 6.5 in the pH value of current material solution and when target ph is 8.0, be first adjusted to 8.0 with 1M sodium hydroxide solution ± 0.1, if adjusted excessively, it can be adjusted back with 1M hydrochloric acid solution, these operations are the general technical ability that those skilled in the art have.Under Literary preparation step in order to citing purpose, and the comparability based on each citing and make some specific description, art technology Personnel can therefrom summarize the method for obtaining present invention preparation pharmaceutical composition according to existing knowledge completely.In the context of the invention It prepares in the specific example of emulsion composition, when needing to add alkanoic acid similar such as oleic acid in formula, such as not especially Illustrate, such alkanoic acid in emulsion preparation process is added with soybean oil；Emulsion combination is prepared in the context of the invention In the specific example of object, when the alkali gold for needing to add similar alkanoic acid such as the alkali metal salt such as sodium salt of oleic acid in formula Belong to salt such as sodium salt when, if not otherwise specified, the alkali metal salt of such alkanoic acid such as sodium salt be in emulsion preparation process with Water adds together.

A, composition preparation example part: emulsion composition is prepared

Preparation example 1: emulsion composition is prepared

Composite formula:

Preparation method:

(1) vitamin K1, soybean oil (injection), egg yolk lecithin, glycerol (injection) are weighed by recipe quantity, it is standby With.

(2) preparation of water phase: stainless cylinder of steel is added in the water for injection for preparing total amount about 80%, prescription is added while stirring The glycerol (injection) of amount.

(3) preparation of oily phase: the soybean oil (injection) of recipe quantity is added in another stainless cylinder of steel, nitrogen is passed through Stream, is added the vitamin K1 and egg yolk lecithin of recipe quantity, and stirring to egg yolk lecithin is dissolved.

(4) preparation of colostrum: being passed through nitrogen stream, and oil is mutually slowly added in the water phase of high-speed stirred, continues high-speed stirred At least after five minutes, solution ph is adjusted with 1M sodium hydroxide solution (suitably increase by 0.1 on the basis of in specified value in specified value PH unit, so that the pH value after emulsion sterilizing reaches defined value), then colostrum is settled to theoretical amount, it stirs evenly.

(5) it is passed through nitrogen stream, colostrum is under 55-60MPa pressure after high-pressure homogeneous 4 times, 12-18MPa low pressure homogeneous 1 time (so that most (micro- sem observation has 95% or more) milk particle granularities are between 0.2~0.8um) is cooled down afterwards, then with 10 μm Membrane filtration.

(6) filtrate is filling seals after 1ml brown glass ampoules bottle, inflated with nitrogen, 118 DEG C of moist heat sterilization 25min to get.

As a kind of drug, hereafter can further by the sterilising prods after lamp inspection labeling, packaging, after the full review of sampling is qualified To obtain the final product.Additionally as drug, the capacity of ampoule bottle can be replaced, in order to provide the single-dose preparations of various dose.

Preparation example 2: emulsion composition is prepared

Composite formula:

Preparation method: it is carried out with reference to the method recorded in preparation example 1.

Preparation example 3: emulsion composition is prepared

Composite formula:

Preparation example 4: emulsion composition is prepared

Composite formula:

Preparation method: vitamin K1, soybean oil, egg yolk lecithin and the glycerol in prescription are heated at 70~80 DEG C and abundant It is uniformly mixed, 20ml water for injection is added into obtained solution, stirs, water for injection is added to be allowed into 100ml, use This solution of homogenizer preliminary homogenisation adjusts pH value to specified value when necessary.Then, under the pressure of 1100Bar, NS1001L is used Type high pressure homogenizer (Italian GEA Niro Soavi company) be homogenized 7 times (so that most milk particle granularity 0.2~ Between 0.8um), the fat emulsion for being 10mg containing vitamin K1 in every 1ml is made.This fat emulsion can be potted in small ampoule In bottle, every can fill 1ml, 2ml, 5ml, 10ml, 20ml etc., then sterilize 30 minutes through 115 DEG C to get the rouge containing vitamin K1 Fat emulsion injection.This injection can directly intramuscular injection or intravenous injection, can also be with after the dilution of 5% glucose injection Intravenous drip, intravenous drip after can also being diluted with 10% fat emulsion injection.

Preparation example 5: emulsion composition is prepared

Composite formula:

Preparation method: it is carried out with reference to the method recorded in preparation example 1.In addition, referring to above-mentioned preparation example 5 it is different be only that will tie up life Plain K1 inventory is changed to 0.45g, and the composition of preparation is tested through the various detection/evaluation methods of the present invention, as the result is shown this Emulsion and the basic indifference of 5 emulsion of preparation example.

Preparation example 6: emulsion composition is prepared

Composite formula:

Preparation example 7: emulsion composition is prepared

Composite formula:

Preparation example 8: emulsion composition is prepared

Composite formula:

Preparation method: it is carried out with reference to the method recorded in preparation example 1.In addition, referring to above-mentioned preparation example 8 it is different be only that will tie up life Plain K1 inventory is changed to 0.7g, and the composition of preparation is tested through the various detection/evaluation methods of the present invention, as the result is shown this cream Agent and the basic indifference of 8 emulsion of preparation example.

Preparation example 9: emulsion composition is prepared

Composite formula:

Preparation example 10: emulsion composition is prepared

Composite formula a:

Preparation method: it is carried out with reference to the method recorded in preparation example 1.Gained emulsion composition can be labeled as Ex10a.

Composite formula b:

Preparation method: it is carried out with reference to the method recorded in preparation example 1.* fatty acid be include oleic acid: linoleic acid: palmitinic acid three With the combination of weight ratio 79:11:4, be related to herein fatty acid such ratio statement when such as it is not specified have and this Identical meanings.Gained emulsion composition can be labeled as Ex10b.

Preparation example 11: emulsion composition is prepared

Different amounts of oleic acid is added in a composition: respectively referring to the formula and preparation method of preparation example 1~9, the difference is that with dimension Raw element K1, soybean oil, egg yolk lecithin these oily matters also added (in every 100ml emulsion composition) together 0.02g, 0.03g, 0.04g, 0.05g, 0.025g, 0.035g, 0.045g, 0.03g, 0.04g oleic acid (that is, make emulsion composition respectively Middle oleic acid concentration is in 0.02~0.05% range), 9 kinds of emulsion compositions are obtained, they can be respectively labeled as Ex111a (reference The gained of preparation example 1), Ex112a (referring to 2 gained of preparation example) ... Ex119a (referring to 9 gained of preparation example).Add in b composition Add different amounts of fatty acid: respectively referring to the formula and preparation method of preparation example 1~9, the difference is that with vitamin K1, soybean oil, egg Yellow lecithin these oily matters together also (in every 100ml emulsion composition) addition 0.02g, 0.03g, 0.04g, 0.05g, 0.025g, 0.035g, 0.045g, 0.03g, 0.04g fatty acid (that is, make fatty acid concentration in emulsion composition exist respectively In 0.02~0.05% range, three kinds of oily ratios of fatty acid are 79:11:4), 9 kinds of emulsion compositions are obtained, they can distinguish Labeled as Ex111b (referring to the gained of preparation example 1), Ex112b (referring to 2 gained of preparation example) ... Ex119b is (referring to preparation example 9 Gained).

Preparation example 12: emulsion composition is prepared

Different amounts of oleic acid is added in a composition: respectively referring to the formula and preparation method of preparation example 1, the difference is that giving birth to dimension Plain K1, soybean oil, egg yolk lecithin these oily matters together also (in every 100ml emulsion composition) addition 0.0025g, 0.005g, 0.01g, 0.06g, 0.09g, 0.15g, 0.2g, 0.5g oleic acid (that is, make oleic acid concentration in emulsion composition respectively In 0.06~0.5% range in 0.0025~0.01% range less than 0.02% and greater than 0.05%), obtain 8 kinds of emulsions Composition, they can be respectively labeled as Ex121a (obtained by addition 0.0025g oleic acid), Ex122a (addition 0.005g oleic acid institute ) ... Ex128a (obtained by addition 0.5g oleic acid).Different amounts of fatty acid is added in b composition: respectively referring to preparation example 1 Formula and preparation method, unlike with vitamin K1, soybean oil, these oily matters of egg yolk lecithin also (in every 100ml In emulsion composition) addition 0.0025g, 0.005g, 0.01g, 0.06g, 0.09g, 0.15g, 0.2g, 0.5g fatty acid (that is, point Do not make in emulsion composition in 0.0025~0.01% range of the fatty acid concentration less than 0.02% and greater than 0.05% In 0.06~0.5% range, three kinds of oily ratios of fatty acid are 79:11:4), 8 kinds of emulsion compositions are obtained, they can be marked respectively Be denoted as Ex121b (addition 0.0025g fatty acid obtained by), Ex122b (obtained by addition 0.005g fatty acid) ... Ex128b (adds Add obtained by 0.5g fatty acid).

Preparation example 13: emulsion composition is prepared

Different amounts of oleic acid is added in a composition: respectively referring to the formula and preparation method of preparation example 2~9, the difference is that with dimension Raw element K1, soybean oil, egg yolk lecithin these oily matters also add 0.0075g (in every 100ml emulsion composition) together Oleic acid (that is, oleic acid concentration in emulsion composition is made to be 0.0075% respectively), obtains 8 kinds of emulsion compositions, they can distinguish Labeled as Ex131a (referring to the gained of preparation example 2), Ex132a (referring to 3 gained of preparation example) ... Ex138a is (referring to preparation example 9 Gained).Different amounts of fatty acid is added in b composition: respectively referring to the formula and preparation method of preparation example 2~9, the difference is that with dimension Raw element K1, soybean oil, egg yolk lecithin these oily matters also add 0.0075g (in every 100ml emulsion composition) together Fatty acid (that is, fatty acid concentration in emulsion composition is made to be 0.0075% respectively, three kinds of oily ratios of fatty acid are 79:11: 4) 8 kinds of emulsion compositions, are obtained, they can be respectively labeled as Ex131b (referring to 2 gained of preparation example), Ex132b (referring to preparation 3 gained of example) ... Ex138b (referring to 9 gained of preparation example).

Preparation example 14: emulsion composition is prepared

Different amounts of oleic acid is added in a composition: respectively referring to the formula and preparation method of preparation example 2~9, the difference is that with dimension Also addition 0.125g is oily (in every 100ml emulsion composition) together for raw element K1, soybean oil, egg yolk lecithin these oily matters Acid (that is, oleic acid concentration in emulsion composition is made to be 0.125% respectively), obtains 8 kinds of emulsion compositions, they can be marked respectively For Ex141a (referring to the gained of preparation example 2), Ex142a (referring to 3 gained of preparation example) ... Ex148a is (referring to 9 institute of preparation example ).Different amounts of fatty acid is added in b composition: respectively referring to the formula and preparation method of preparation example 2~9, the difference is that giving birth to dimension These oily matters of plain K1, soybean oil, egg yolk lecithin also add 0.125g fat (in every 100ml emulsion composition) together Acid (that is, fatty acid concentration in emulsion composition is made to be 0.125% respectively, three kinds of oily ratios of fatty acid are 79:11:4), obtains To 8 kinds of emulsion compositions, they can be respectively labeled as Ex141b (referring to 2 gained of preparation example), Ex142b (referring to 3 institute of preparation example ) ... Ex148b (referring to 9 gained of preparation example).

Preparation example 15: emulsion composition is prepared

The formula and preparation method of Ex111b are respectively referred to, the difference is that three kinds of oily ratios of fatty acid used are 70:5:2,88: 15:10,70:15:10 and 88:5:2；Respectively refer to the formula and preparation method of Ex112b, unlike used three kinds of fatty acid it is oily Ratio is 70:5:2,88:15:10,70:15:10 and 88:5:2；The formula and preparation method of Ex113b are respectively referred to, the difference is that institute It is 70:5:2,88:15:10,70:15:10 and 88:5:2 with three kinds of oily ratios of fatty acid；Respectively refer to the formula of Ex112b And preparation method, the difference is that three kinds of oily ratios of fatty acid used are 72:8:2,85:13:6,72:13:6,85:8:2,80:10:4.

B, detection method part: the method for detecting emulsion composition

Following detection method example provides the method that can be used for detecting emulsion composition, these methods can be also used for measuring Emulsion composition can also compare other the one of the method for the present invention preparation through handling the internal performance for example after high-temperature process A little emulsion compositions for being used for comparative test.

Detection method example 1: the pH value of emulsion composition is detected

Method: according to method documented by two annex VIH of Chinese Pharmacopoeia version in 2010, directly measurement obtained cream of the invention Agent composition or various other samples.It is the mean value of 3 parallel determinations when providing pH value in present invention test.This hair The pH value of some emulsion compositions of bright preparation respectively value described in the formula referring to various preparating examples (with formula sign value Between poor less than 0.1 pH unit).

Detection method example 2: the granularity and distribution of emulsion composition are detected

Method: two annex of Pharmacopoeia of People's Republic of China version in 2010, Ⅸ E granularity and determination of particle size distribution third are shone Wet process measurement in method (light scattering method), is carried out using laser diffraction particle size distribution instrument, records and count no less than 500 milk particles The partial size of particle.Calculate partial size the percentage that 1.0 μm of milk particle numbers below account for the total statistical number of milk particle (herein can be with " percentile 1 " indicates).Calculate simultaneously be greater than in partial size and the milk particle sum equal to 0.5 μm in, partial size is greater than 1 μm of cream The percentage (can be indicated herein with " percentile 2 ") of grain.In addition it can calculate each emulsion composition sample Average grain diameter, pass through this detection method example 2 calculate D10, D50 and D90 and accordingly calculating milk particle across footpath be also easy.

Generally, as emulsion that can be used for intravenous injection, partial size accounts for the total statistical number of milk particle in 1.0 μm of milk particle numbers below Percentage, i.e. percentile 1 should be 70% or more, preferably 80% or more, preferably 90% or more.In addition, it is general and Speech, as emulsion that can be used for intravenous injection, be greater than in partial size and the milk particle sum equal to 0.5 μm in, partial size is greater than 1 μm of milk particle Percentage, i.e. percentile 2 should be less than 20%, preferably less than 15%, less than 10%, be less than 5%.

Detection method example 3: the method for vitamin K1 content in detection emulsion composition

(1) it is measured according to high performance liquid chromatography (2010 editions two annex V D of Chinese Pharmacopoeia)；

(2) chromatographic condition and system suitability test: being filler with octadecylsilane chemically bonded silica, with alcohol-water (95:5) is mobile phase, and column temperature is 35 DEG C, Detection wavelength 254nm, and theoretical cam curve should be not less than with vitamin K1 calculating 1500；

(3) preparation of test solution: precision measurement emulsion composition is appropriate, adds isopropanol to dissolve and is quantitatively made every The solution for being 0.05mg containing vitamin K1 in 1ml, as test solution；

(4) preparation of reference substance solution: it is appropriate to weigh vitamin K1 reference substance, accurately weighed, and 10% Fat Emulsion is added to inject 1ml (is prepared, be a difference in that and do not add vitamin K1) in liquid with reference to preparation example 1 of the present invention, adds isopropanol to dissolve and quantifies system At the solution for being 0.05mg containing vitamin K1 in every 1ml, as reference substance solution；

(5) measuring method: taking each 20 μ l of test solution and control solution respectively, injects liquid chromatograph, records chromatography Figure, by external standard method with the content of vitamin K1 in calculated by peak area test sample.

The content of vitamin K1 can with % (w/v) perhaps mg/ml indicate or be equivalent to be formulated labelled amount percentage Number indicates.

Detection method example 4: the content of glycerol in detection emulsion composition

Method: precision measures emulsion composition 2.5ml, sets in conical flask, adds water 100ml, adds bromocresol purple indicator solution 5 Drop, shakes up, if aobvious acidity, is added dropwise 0.1mol/L sodium hydroxide solution, makes solution in bluish violet；If aobvious alkalinity, should first be added dropwise It is just in yellow that 0.5mol/L sulfuric acid solution, which is adjusted to solution, then 0.1mol/L sodium hydroxide solution, which is added dropwise, makes solution be in bluish violet, Add 0.7% potassium metaperiodate solution (face with newly match) 100ml, sets in 37~40 DEG C of water-bath and keep the temperature 15 minutes, and constantly shake, add 1,2-PD 3ml is placed 5 minutes, be titrated to solution just with 0.1mol/L sodium hydroxide titration liquid and be in bluish violet to get.Often The 0.1mol/L sodium hydroxide titration liquid of 1ml is equivalent to the C3H8O3 of 9.210mg.

The content of glycerol can with % (w/v) perhaps mg/ml indicate or be equivalent to be formulated labelled amount percentage table Show.

Detection method example 5: the method for triglyceride content in detection emulsion composition

(1) it is measured according to high performance liquid chromatography (two annex VD of Chinese Pharmacopoeia version in 2010)；

(2) chromatographic condition and system suitability: being filler with silica gel, n-hexane-isopropanol-acetic acid (98.9:1: It 0.1) is mobile phase, evaporative light scattering detector (atomization gas: N2, atomization gas pressure: 240Pa, evaporator temperature: 60 DEG C) inspection It surveys, the separating degree of triglyceride and oleic acid should be greater than 2, and the relative standard deviation of the peak area of triglyceride should be not more than 3.0%；

(3) preparation of system suitability solution: taking triglyceride and each 10mg of oleic acid, until using in 25ml measuring bottle Flowing phased soln simultaneously be diluted to scale, shake up to get；

(4) preparation of reference substance solution: taking soybean oil reference substance about 0.35g, accurately weighed, sets in 50ml measuring bottle, with just Hexane and isopropanol etc. hold mixed solution and dissolve and be diluted to scale, as stock solution；Precision measures stock solution 3ml and 4ml, point It does not set in 25ml measuring bottle, is diluted to scale with mobile phase, shakes up, as reference substance solution 1 and reference substance solution 2；

(5) preparation of test solution: precision measures emulsion composition 4ml, sets in 50ml measuring bottle, with n-hexane and isopropyl Alcohol etc. holds mixed solution and dissolves and be diluted to scale, shakes up；Precision measures 3ml, sets in 25ml measuring bottle, is diluted to quarter with mobile phase Degree, shake up to get；

(6) measuring method: accurate respectively to measure reference substance solution 1, reference substance solution 2, each 10 μ l of test solution is alternately infused Enter in liquid chromatograph, through chromatographic isolation obtain calculated by peak area to get.

The content of triglyceride can perhaps mg/ml be indicated or is formulated the hundred of labelled amount with being equivalent to % (w/v) Fraction representation.

Detection method example 6: the content of fatty acid measuring method in emulsion composition

It takes emulsion composition 10ml to set in 50ml conical flask, the methanol solution 2ml of 10% sodium hydroxide is added, in a water bath Reflux 30 minutes, is added the methanol solution 2ml of boron trifluoride, then flow back 30 minutes, and normal heptane 40ml is added, and continues 5 points of reflux Clock is let cool, and saturated sodium chloride solution 10ml is added, and is vibrated 15 seconds, is placed, and is drawn upper liquid, is washed with water 3 times, each 2ml has Machine layer is filtered by anhydrous sodium sulfate, and subsequent filtrate is as test solution.The oleic acid through esterification treatment, sub- oil in advance are taken respectively Acid, palmitinic acid reference substance solution and each 1ul of test solution inject gas chromatograph, and nitrogen buffer gas is into sample temperature 250 DEG C, detector temperature is 280 DEG C.Temperature programming, keeps 5min to rise to 240 DEG C with 6 DEG C per minute by 60 DEG C of initial temperature, 25min measurement is kept, the content of various fatty acid is calculated by the areas of peak normalization method that correction factor is not added.

The various emulsion compositions for being added to oleic acid or fatty acid prepared by the present invention are measured, have been measured wherein oily The amount of one of acid, linoleic acid, palmitinic acid or three are coincide with its theory addition respectively.

C, test example part

Test example 1: influence of the soybean oil mass to formula in vitamin K1 formula is investigated

The emulsion composition containing vitamin K1 is prepared with 6 different formulations, formula and preparation method are referring to preparation example 1, no With place be using soybean oil amount, the amount of the soybean oil used in 6 formulas is respectively 0%, 2.5%, 5%, 7.5%, colostrum and eventually cream are respectively obtained in 10%, 15% different samples, the step described in preparation example 1 (4) and step (6). The colostrum of different prescriptions is set and observes delamination after standing 6 hours in colorimetric cylinder.Whole cream set in centrifuge tube with 10000rpm × Delamination is observed after 10min centrifugal treating, while according to the percentile 1 of the whole cream of the method for " detection method example 2 " measurement (" percentage 1 ") and percentile 2 (" percentage 2 ").As a result it see the table below:

As it can be seen that the amount for the about 7-15% soybean oil advocated using the present invention is beneficial for forming more stable emulsion.

Test example 2: the content of the content of vitamin K1 after measurement emulsion composition is placed at high temperature

The emulsion composition of preparation is divided in brown ampoule, is placed in 50 DEG C of baking ovens and places 2 months, then with above The content (mg/ml) of the method measurement vitamin K1 of the detection method example 3, in addition measures the respective sample without high-temperature treatment In vitamin K1 content, thus calculate the remaining percentage (%) of vitamin K1 in reagent after high temperature is placed 2 months.Part It is that 94.7, preparation example 2 is 93.9, makes that the result of vitamin K1 remnants percentage (%), which is respectively as follows: preparation example 1, in emulsion composition Standby example 3 is 96.1, preparation example 4 is 95.2, preparation example 5 is 96.7, preparation example 6 is 94.3, preparation example 7 is 97.1, preparation example 8 is 95.8, it is 93.8 that preparation example 9, which is 94.8, preparation example 10a,.

In addition, the method measurement through being same as above table result, VK1 remnants' percentage of the whole emulsion compositions of 11 gained of preparation example In 95.2~98.7% ranges；VK1 remnants' percentage of the whole emulsion compositions of the gained of preparation example 12 94.3~ In 97.8% range；VK1 remnants' percentage of the whole emulsion compositions of 13 gained of preparation example is in 94.8~97.8% ranges； VK1 remnants' percentage of the whole emulsion compositions of 14 gained of preparation example is in 95.3~98.5% ranges；15 gained of preparation example VK1 remnants' percentage of whole emulsion compositions is in 95.36~98.4% ranges；These are the result shows that in emulsion composition It is middle add different amounts of oleic acid or mixed fatty acid for VK1 chemical stability substantially without influence.

Test example 3: the pharmaceutical property of emulsion composition is investigated

1、The content of vitamin K1:It is measured according to method illustrated above, the percentage of formula mark content is accounted for the content of test Number meter.It is that 99.7, preparation example 2 is that the result of vitamin K1 content (%), which is respectively as follows: preparation example 1, in the emulsion composition of part 98.9, preparation example 3 be 100.1, preparation example 4 be 99.2, preparation example 5 is 99.7, preparation example 6 is 100.3, preparation example 7 is 99.1, Preparation example 8 is 99.8, preparation example 9 is 99.6, preparation example 10a is 101.8.In addition, the method measurement through being same as above table result, preparation The VK1 content of example 10b and the whole emulsion compositions of 11~15 gained of preparation example is in 99.1~102.8% ranges, with reason It is identical by feeding intake.

2, glycerol content:It is measured according to method illustrated above.In terms of the percentage that the content of test accounts for formula mark content.Portion Dividing the result of glycerol content (%) in emulsion composition to be respectively as follows: preparation example 1 is that 98.9%, preparation example 2 is 102.3%, prepares Example 3 is 100.7%, preparation example 4 is 99.8%, preparation example 5 is 99.3%, preparation example 6 is 101.2%, preparation example 7 is 99.9%, preparation example 8 is 97.8%, preparation example 9 is 101.3%, preparation example 10a is 100.2%.In addition, through being same as above table result Method measurement, the glycerol content of preparation example 10b and the whole emulsion compositions of the gained of preparation example 11~15 99.0~ In 102.4% range, feed intake with theory identical.

3, triglyceride content:It is measured according to method illustrated above.Soybean oil mark in formula is accounted for the content of test to contain The percentage meter of amount.It is 97.7%, preparation example 2 that the result of glycerol content (%), which is respectively as follows: preparation example 1, in the emulsion composition of part For 98.6%, preparation example 3 be 104.7%, preparation example 4 is 97.8%, preparation example 5 is 99.8%, preparation example 6 is 104.5%, system Standby example 7 is 101.9%, preparation example 8 is 99.8%, preparation example 9 is 97.3%, preparation example 10a is 104.5%.In addition, through being same as above The method of table result measures, and the soybean oil content of preparation example 10b and the whole emulsion compositions of 11~15 gained of preparation example exists In 98.1~102.8% ranges, feed intake with theory identical.

Test example 4: the standard stability of emulsion composition is investigated

Preparation of the vitamin K1 emulsion composition prepared by the present invention as injection, using " shading, it is closed, in cool place It is beneficial for saving under conditions of place's (being no more than 20 DEG C) ", this regulation is also that pharmacopeia infuses injection especially emulsion-type Penetrate the most basic requirement of liquid.

Three parameters (i.e. VK1 content, glycerol content, triglyceride content) and pH described in the above test example Value, milk particle granularity be index, investigate preparation example 1 to preparation example 15 whole emulsion compositions 15 ± 1 DEG C shading, it is closed, Shady place saves 24 months.The situation of change between value and 0 month initial value when comparing 24 months, as the result is shown: (1) whole samples PH value respectively compared with its initial value, variation is respectively less than 0.15 pH value unit, and can connect in the human body of pH value 7~9 By in the range of.(2) the milk particle granularity of whole samples: at 0 month, 85% or more milk particle partial size is at 0.5 μm or so；Be greater than and In milk particle sum equal to 0.5 μm, the milk particle greater than 1 μm is less than 6%, and the milk particle greater than 5 μm is not detected；At 24 months, 85% with On milk particle partial size at 0.5 μm or so；Be greater than and milk particle sum equal to 0.5 μm in, milk particle greater than 1 μm is less than 6%, not Detection is greater than 5 μm of milk particle.

(3) the vitamin K1 content of whole samples is respectively compared with its initial value, initial value 95.7%~ Between 102.3%, such as preparation example 1 is 96.9%, Ex111a 98.4%, Ex111b 98.1%.

(4) glycerol content of whole samples is respectively compared with its initial value, initial value 95.2%~104.3% it Between, such as preparation example 4 is 99.7%, Ex111a 97.1%, Ex111b 97.4%.

(5) triglyceride content of whole samples is respectively compared with its initial value, initial value 94.8%~ Between 106.3%, such as preparation example 6 is 98.4%, Ex111a 99.3%, Ex111b 99.0%.

(6) in addition, in various specific examples of the invention, oleic acid used can be replaced with the enuatrol of equivalent, used Three kinds of sour fatty acid mixeds can be replaced with three kinds of sodium soaps of equivalent, and it has been found that after such substitution gained Composition is compared with original use oleic acid or fatty acid mixed resulting composition, in terms of whole pharmaceutical properties of the present invention Two class compositions have essentially identical result；For example, after above-mentioned (5) middle enuatrol or three kinds of sodium soap substitutions all The triglyceride content of sample is respectively compared with its initial value, between the 94.7%~106.5% of initial value, such as Ex111a uses that enuatrol result is 99.4%, to use corresponding amount fatty acid mixed sodium result instead be 99.2% to Ex111b instead.

From the above results, it can be seen that vitamin K1 fat emulsion made from this paper has good pharmaceutical property.

Test example 5: the related substance detection of emulsion composition

Using the related substance especially impurity B in HPLC method measurement emulsion composition, such as above obtained by each preparation example Emulsion composition, the specific method is as follows:

(2) octadecylsilane chemically bonded silica chromatographic column (i.e. usually said C18 column, such as filler chromatographic condition: are used The pillar of 5 μm of granularity, column internal diameter 4.6mm, 200~300mm of column length such as 250mm, what this example actually used is that specification is 5 μ M-4.6mm × 250mm, brand are the C18 column of Agilent ZORBAX SB-C18), 25~35 DEG C of column temperature (such as 28~32 DEG C, The column temperature of this example actual use is 30 DEG C), mobile phase A is dehydrated alcohol-water mixed liquid of volume ratio 90:10, and Mobile phase B is Dehydrated alcohol, flow velocity are 0.8~1.2ml/min (flow velocity of this example actual use is 1.0ml/min)；Detection wavelength is 265 ~275nm (wavelength of this example actual use is 270nm)；

(3) it is eluted using mobile phase A and Mobile phase B by such as Gradient: the mobile phase during 0 minute to 30 minutes Are as follows: mobile phase A is 100% and Mobile phase B is 0, is 100% by mobile phase A during 30 minutes to 40 minutes and Mobile phase B is 0 is changed into that mobile phase A is 0 and Mobile phase B is 100%, is 0 by mobile phase A during 40 minutes to 60 minutes and Mobile phase B is 100% is changed into that mobile phase A is 100% and Mobile phase B is 0；

The present invention attempts to purify impurity B in test, it has been found that obtained impurity B is unstable, it is unsuitable It is used as detection working reference substance, the present inventor has carried out correction factor measuring and calculating to the impurity in HPLC test thus, calculates Obtaining its correction factor is 0.7, it is possible thereby in the related substance detection of emulsion composition of the present invention, using the correction up factor Self-control method investigates the content in relation to substance.Although the present inventor not yet knows the chemical structure of the impurity B at present, it is believed that with The continuous development of science and technology, understand that its chemical structure and other performance in the future, however the present invention passes through in regulation liquid Two parameters of relative retention time and correction factor determined under phase chromatographic condition, completely can be qualitative to impurity B progress, simultaneously Using its response under the conditions of HPLC of the present invention, the impurity B can be quantified completely.After measured, above-mentioned HPLC method Whether it measures vitamin K1 content or measures the content of impurity B, all have excellent methodology performance, be fully able to meet Measurement demand, such as the rate of recovery of vitamin K1 and impurity B reach 99% or more.

Claims

1. emulsion composition, it includes:

The emulsion composition is prepared by a method comprising the following steps to obtain:

(c) oily be mutually stirred with water phase is made to prepare thick cream, optional adds water to full dose, optional use pH adjusting agent regulating liquid medicine PH value；

Optional, the operation in relation to substance especially impurity B detection is carried out to emulsion composition obtained by experience pressure sterilizing, including Following steps:

(1) it is measured, operates about the specification of high performance liquid chromatography according to " Chinese Pharmacopoeia " 2015 editions four 0512 sections of general rule When be protected from light；

(2) octadecylsilane chemically bonded silica chromatographic column (such as 5 μm of filler particle size, column internal diameter 4.6mm, column chromatographic condition: are used Long 200~300mm), 25~35 DEG C of column temperature (such as 28~32 DEG C), dehydrated alcohol-water that mobile phase A is volume ratio 90:10 mixes Liquid is closed, Mobile phase B is dehydrated alcohol, and flow velocity is 0.8~1.2ml/min (such as 1.0ml/min)；Detection wavelength be 265~ 275nm (such as 270nm)；

(3) it is eluted using mobile phase A and Mobile phase B by such as Gradient: at (such as initial 20~40 points of first time period In clock time section such as 25~35 minutes sections, such as during 0 minute to 30 minutes of entire elution process) mobile phase Are as follows: mobile phase A is 100% and Mobile phase B is 0, second time period (such as next 5~20 minutes section such as 8~ In 15-min period, such as during 30 minutes to 40 minutes of entire elution process) by mobile phase A be 100% and flowing Phase B is 0 to be changed into that mobile phase A is 0 and Mobile phase B is 100%, in the third period (such as when 10~30 minutes next Between in section such as 15~25 minutes sections, such as during 40 minutes to 60 minutes of entire elution process) by mobile phase A be 0 and Mobile phase B be 100% to be changed into that mobile phase A is 100% and Mobile phase B is 0；

(4) measuring method: precision amount emulsion composition 1ml sets in 10ml measuring bottle, with isopropanol to scale, shakes up, as Test solution；Precision measures above-mentioned test solution 1ml, sets in 100ml measuring bottle, with isopropanol to scale, shakes up, and makees For contrast solution；

(5) accurate respectively to measure above-mentioned test solution and each 20 μ l of contrast solution, it is injected separately into chromatography, records chromatogram To 60min；

(6) read test solution chromatogram in each impurity peaks response, and calculate separately each impurity peak response value relative to The percentage of contrast solution vitamin K1 main peak response；It is 1 calculating with the relative retention time at vitamin K1 peak, for opposite Retention time be 0.52 ± 0.03 at impurity peaks be named as impurity B, if impurity B exist, calculate its response multiplied by correction because Percentage of sub 0.7 resulting value relative to contrast solution main peak response, thus calculates hundred of impurity B relative to vitamin K1 Score.

2. emulsion composition according to claim 1, it is characterised in that:

It includes 0.45-2%, 0.7-1.5%, 0.9-1.1% or 1.0% vitamin K1s；

It includes 7.5~12.5% soybean oils, preferably comprise 8~12% soybean oil, preferably comprise 9~11% soybean Oil preferably comprises about 10% soybean oil；

It includes 0.8~2% phosphatide, preferably comprise 1.0~1.5% phosphatide, preferably comprise about 1.2% phosphatide；

It includes 1.5~3% glycerol, preferably comprise 1.75~2.5% glycerol, preferably comprise 1.95~2.45% it is sweet Oil preferably comprises about 2.2% glycerol；

Its pH value is 7.0~9.0,7.0~8.5,7.5~9.0 or 7.5~8.5；

When it is measured by the lower method of the Chinese Pharmacopoeia annex VI H of version two in 2010, pH value for 7.0~9.0,7.0~ 8.5,7.5~9.0 or 7.5~8.5；

It wherein also include pH adjusting agent, dosage is to adjust the pH value of the emulsion composition to 7.0~9.0,7.0~8.5,7.5 ~9.0 or 7.5~8.5 amount.

3. emulsion composition according to claim 1, it is characterised in that:

It includes:

4. emulsion composition according to claim 1, it is characterised in that:

When it shines two annex of Pharmacopoeia of People's Republic of China version in 2010, Ⅸ E granularity and determination of particle size distribution third method (light Scattering method) in wet process measuring method when, the partial size of 70%, 80% or 90% or more milk particle at 1.0 μm hereinafter, 70%, The partial size of 80% or 90% or more milk particle is between 0.01~1.0 μm, the partial size of 70%, 80% or 90% or more milk particle Between 0.05~1.0 μm, the partial size of 70%, 80% or 90% or more milk particle is between 0.1~1.0 μm；

Be greater than in partial size and equal to 0.5 μm milk particle sum in, partial size greater than 1 μm milk particle be less than 20%, preferably less than 15%, Less than 10%, less than 5%；

When it shines two annex of Pharmacopoeia of People's Republic of China version in 2010, Ⅸ E granularity and determination of particle size distribution third method (light Scattering method) in wet process measuring method when, the average grain diameter of milk particle is not more than 0.4 μm, for example, average grain diameter be 0.05~0.4 μm, 0.1~0.4 μm, 0.15~0.4 μm, 0.2~0.4 μm, 0.25~0.4 μm, 0.3~0.4 μm, 0.05~0.35 μm, 0.05~ 0.3 μm, 0.05~0.25 μm, 0.05~0.2 μm, 0.05~0.15 μm, 0.05~0.1 μm, 0.1~0.35 μm, 0.15~0.3 μm or 0.2~0.3 μm；

When it shines two annex of Pharmacopoeia of People's Republic of China version in 2010, Ⅸ E granularity and determination of particle size distribution third method (light Scattering method) in wet process measuring method when, milk particle D90 value (i.e. particulate accumulation be distributed as 90% partial size) no more than 0.6 μm, example As D90 value be 0.1~0.6 μm, 0.1~0.5 μm, 0.1~0.4 μm, 0.1~0.3 μm, 0.1~0.2 μm, 0.2~0.6 μm, 0.3~0.6 μm, 0.4~0.6 μm, 0.3~0.6 μm, 0.2~0.5 μm or 0.3~0.4 μm；

When it shines two annex of Pharmacopoeia of People's Republic of China version in 2010, Ⅸ E granularity and determination of particle size distribution third method (light Scattering method) in wet process measuring method when, cannot detect the milk particle greater than 5 μm.

5. emulsion composition according to claim 1, it is characterised in that:

It is oil-in-water type emulsion for injection；

It is the fat emulsion injection made of pressure sterilizing；

The soybean oil is the other soybean oil of injection stage；

The phosphatide is the other phosphatide of injection stage；

The phosphatide is soybean lecithin or egg yolk lecithin；

The glycerol is the other glycerol of injection stage；

The water is water for injection；

It is prepared by a method comprising the following steps to obtain:

(2) stainless cylinder of steel is added in the water for injection that will prepare total amount about 60-90% (for example, about 80%), and the sweet of recipe quantity is added Oil stirs evenly, and water phase is made；

(3) soybean oil of recipe quantity is added in another stainless cylinder of steel, is passed through nitrogen stream, the vitamin K1 and phosphorus of recipe quantity is added Oil phase is made in rouge, stirring to each material dissolution；

(5) adjust the pH value of the colostric fluid to prescribed limit with pH adjusting agent, then plus water for injection be settled to theoretical recipe quantity, stir It mixes uniformly, adjusts solution ph to prescribed limit with pH adjusting agent when necessary；

(6) under nitrogen flowing, make colostrum high pressure homogenization under 40-70MPa (such as 55-60MPa) pressure, then in 5-25MPa (example Such as 12-18MPa) low pressure is homogenized under pressure, and it is cooling, then with 10 μm of membrane filtration；

6. emulsion composition according to claim 1, it is characterised in that:

It wherein also include alkanoic acid or its alkali metal salt such as sodium salt；

The alkanoic acid is oleic acid or its alkali metal salt such as sodium salt；

It include 0.01~0.05% oleic acid or its alkali metal salt such as sodium salt in emulsion composition, such as comprising 0.01~ 0.045%, 0.01~0.04%, 0.01~0.035% or 0.01~0.03% oleic acid or its alkali metal salt such as sodium salt, Such as oleic acid or its alkali gold comprising 0.015~0.05%, 0.02~0.05%, 0.025~0.05% or 0.03~0.05% Belong to salt such as sodium salt, such as the oleic acid comprising 0.015~0.045%, 0.02~0.04%, 0.025~0.035% or its alkali gold Belong to salt such as sodium salt；

The alkanoic acid is fatty acid or its alkali metal salt such as sodium salt；

It include 0.01~0.05% fatty acid or its alkali metal salt such as sodium salt in emulsion composition, such as comprising 0.01~ 0.045%, 0.01~0.04%, 0.01~0.035% or 0.01~0.03% fatty acid or its alkali metal salt such as sodium Salt, for example, comprising 0.015~0.05%, 0.02~0.05%, 0.025~0.05% or 0.03~0.05% fatty acid or Its alkali metal salt such as sodium salt, such as include 0.015~0.045%, 0.02~0.04%, 0.025~0.035% fatty acid Or its alkali metal salt such as sodium salt；

The fatty acid is the combination for including oleic acid, linoleic acid, palmitinic acid three；The alkali metal salt of the fatty acid such as sodium salt It is the combination for including oleic acid, linoleic acid, palmitinic acid three alkali metal salt such as sodium salt；

The fatty acid is the combination for including oleic acid, linoleic acid, palmitinic acid three, the alkali metal salt of the fatty acid such as sodium salt Being includes the combination of oleic acid, linoleic acid, palmitinic acid three alkali metal salt such as sodium salt, and three's weight ratio be 70~88:5~ 15:2~10；

The fatty acid is the combination for including oleic acid, linoleic acid, palmitinic acid three, the alkali metal salt of the fatty acid such as sodium salt Being includes the combination of oleic acid, linoleic acid, palmitinic acid three alkali metal salt such as sodium salt, and three's weight ratio be 72~85:8~ 13:2~6；

The alkanoic acid is to be added in oily phase in emulsion preparation process with oiliness material；Alternatively, the alkali metal salt of alkanoic acid is for example Sodium salt, which is added in water phase in emulsion preparation process with aqueous materials, to be for example directly appended in water.

7. the method for preparing emulsion composition described in claim 1-6 comprising following steps:

Alternatively, itself the following steps are included:

8. the pharmaceutical applications of emulsion composition described in claim 1-6 for vitamin K deficiency or interfere vitamin k activity It is such as, but not limited to following coagulation disorders caused by prothrombin caused by pharmaceutical intervention, VII, IX and X dyssynthesis Disease: factor caused by the anti-coagulants such as cumarin or indene-dione derivative is insufficient；Low blood coagulation caused by antibiotic treatment Proenzyme mass formed by blood stasis；Disease causes to absorb or synthesis vitamin K obstacle leads to Hypoprothrombinemia, such as obstructive jaundice, leak, mouth Inflammatory diarrhea, ulcerative colitis, celiaca, Bowel resection, cystic fibrosis of the pancreas and regional enteritis；Other drugs interference Vitamin K metabolism, caused Hypoprothrombinemia, such as salicylate；For example, the fat emulsion injection when in use can be with Each 10mg, 1-2 times daily, total amount is no more than 40mg in 24 hours, and dosage and frequency are responded according to prothrombin time Or clinical symptoms adjustment；Usage mode is intravenously administrable, and infusion can not have to dilution, direct intravenous injection, or with 5% grape Intravenous drip after sugared injection dilution.

9. this method includes as follows using the method in relation to substance especially impurity B in HPLC method measurement vitamin K1 composition Operation:

10. method according to claim 9, the vitamin K1 composition is the emulsion composition comprising soybean oil and phosphatide； For example, the vitamin K1 composition is any one of claim 1-6 emulsion composition.