CN109123006A - A kind of day lily health protection tea and preparation method thereof and detection method - Google Patents

Abstract

The present invention relates to the technical fields of health food, the invention discloses a kind of day lily health protection teas for preventing alcoholic fatty liver, entitled a kind of day lily health protection tea and preparation method thereof and detection method, the health protection tea are made of raw material from the following weight: 500 parts of day lily；It is losing weight, on the evaluation of result of three aspect of pathology of liver function blood lipid and liver, health protection tea of the present invention has protective effect to the hepatic tissue cell of alcoholic fatty liver.The invention also provides the preparation methods of the health protection tea, and carry out the detection method of assay to health protection tea using high performance liquid chromatography and gas chromatography.

Description

A kind of day lily health protection tea and preparation method thereof and detection method

Technical field

The present invention relates to the technical fields of health food, and in particular to a kind of day lily health care for preventing alcoholic fatty liver Tea and preparation method.

Background technique

Day lily (Hemerocallis fulva) also known as day lily, popular name datlily are planted for Liliaceae perennial herb Object, day lily is fresh and sweet delicious, meat and vegetables and excellent, and in Chinese medicine more than 3000 years dietotherapy history, day lily is listed in common food Treat one of food.Chinese medicine think day lily have calming the liver blood-nourishing, swelling diuretic, antibacterial anti-inflammatory, hemostasis, ease pain, promote lactation, stomach invigorating and The function of tranquilizing the mind can treat hepatitis, yellow subcutaneous ulcer, defecate lower blood, flu, dysentery, urinary tract infections, dizziness, tinnitus, palpitaition, pain in the loins, water The various diseases such as swollen, hypogalactia, painful swelling of joints.

Alcoholic fatty liver disease (alcohol fatty liver disease, AFLD) refers to since heavy drinking is led The liver cell inner lipid accumulation of cause is more than the 5% of liver weight in wet base, or histologically per unit area has 1/3 or more liver thin Born of the same parents occur steatosis, that is, show as the fat deposition in liver cell, if further development, it will cause alcoholic hepatitis, Alcoholic fibrosis and alcoholic cirrhosis, these belong to alcoholic liver disease, can induce liver under conditions of serious excessive drinking Meronecrosis or even liver failure.Alcoholic liver disease is on the rise in the disease incidence of China in recent years, is only second to hepatitis Poison, alcohol have become the second largest factor for leading to hepar damnification, but for AFLD, there has been no treatment methods very outstanding at present. And relative to drug therapy, the prevention of health care tea more has its special advantage, such as toxic side effect is small, convenient to take, in advance It is anti-first, and as tea-drinking, the psychological repellence of drinking person is small.

In the prior art, application No. is 2014102074300, a kind of entitled dendrobium nobile Dealcoholic liver-protecting tea and its system The Chinese invention patent of Preparation Method discloses a kind of dendrobium nobile Dealcoholic liver-protecting tea and preparation method thereof, by the raw material of following parts by weight Be made: dendrobium nobile 20~30, white tea 80~100, penthorum chinense pursh 16~22, coralhead plant stem with leaf 10~15, wolfberry leaf 6~12, Common Leafflower Herb 3~6, Pomelo peel 4~8, pueraria lobata 6~9, Milk Thistle grass 1~3, pale reddish brown Chinese Ixeris 2~5, chamomile 1.5~2.5, water chestnut leaf 1~2, gardenia 2 ~3, balm 0.5~1.5, spearmint 1~2, appropriate amount of water.Dendrobium tea of the present invention has clearing heat and detoxicating, Dealcoholic sobering-up, removing jaundice The effect of wet, liver protection, can speed up alcohol metabolism, reduce ethanol in blood concentration, and can have protective effect to liver, The damage reduced to liver can be lowered, long-term drinking, which helps to drink, causes alcoholic fatty liver, alcoholic hepatitis, alcoholic liver Fibrosis and alcoholic cirrhosis, it is without side-effects.Application No. is 2012100426503, a kind of entitled blood-snow tea drop The extract of rouge liver-protectivity and its Chinese invention patent of application, which disclose, belongs to natural drug preparation technical field A kind of extract of lethariella cladonioides active site for reducing fat and protecting liver and its application in the drug of preparation prevention and treatment nonalcoholic fatty liver. The extract is extracted using the dry coarse powder of blood-snow tea as raw material by water, alcohol precipitating, trypsase and papain hydrolysis, thoroughly Analysis, drying and other steps are made.

Have effects that certain health protection tea to alcoholic fatty liver in the prior art, is usually made of plurality of raw materials, at This height, and effect is general.And the product made of a kind of raw material, majority are existed with medicament forms, for treating Alcoholic rouge The shortcomings that fat liver, this mode, also refers in discussion above, and if toxic side effect is larger, the most key is the heart of patient It is very poor to manage compliance.So it is few to be currently badly in need of a kind of raw material for preparing, significant effect, and highly-safe health protection tea.The present invention mentions The health protection tea of confession can solve current this problem.

Applicant Gansu Lv Yuan agriculture and forestry science and technology Co., Ltd is found in June, 2005, is located at Gansu Province Qingyang City Zhenyuan County Upper Xiao South Street 10,11,600,000 yuan of registered capital.It is one and is absorbed in agricultural product research, exploitation, production and sale and agricultural production Product New technical use is promoted, the agricultural product intensive processing enterprise based on e-commerce, foreign trade.Company have the present age compared with Four production lines of advanced activated carbon production apparatus and debittering almond, more than 8700 tons of almond of year purchase processing, produce various active carbons 3265 tons, 2000 tons of debittering almond, major product has southern beam board debittering almond, rye benevolence, shell gold carbon, air cleaning charcoal etc. A kind of 3 major class more than 20, wherein " Nan Liang " board debittering almond, day lily, that purple perilla has obtained China Green Food Development Center is " green Color food certificate ".It is cited as within 2009 " Qingyang City agriculture industrialization emphasis leading enterprise "；It is cited as " Gansu Province's agriculture within 2013 Industry industrialization emphasis leading enterprise "；" first batch of national level forestry industrialization emphasis faucet enterprise was regarded as by the State Administration of Forestry in 2014 Industry ", and win Chinese Enterprise investigation credit assessment centers " AAA grades of credit business " honorary title.Applicant studies by many years, The technical solution of day lily health protection tea of the present invention is obtained.

Summary of the invention

Technical problems based on background technology, the invention proposes a kind of day lily guarantors for preventing alcoholic fatty liver Strong tea and preparation method.

The object of the present invention is to provide a kind of health protection teas for preventing alcoholic fatty liver disease.

It is a further object of the present invention to provide the preparation methods of the health protection tea.

It is a still further object of the present invention to provide the detection methods of the health protection tea.

The present invention also provides the health purposes of the health protection tea composition.

The purpose of foregoing invention, is realized in the following way:

A kind of day lily health protection tea preventing alcoholic fatty liver, the health protection tea are made of day lily extract.

The day lily health protection tea of the described prevention alcoholic fatty liver, the health protection tea the preparation method comprises the following steps:

S1: taking day lily, the day lily after after 35~40 DEG C of 4~6h of drying, taking drying in baking oven is set, using air-flow crushing Machine is pulverized, and the stream pressure of airslide disintegrating mill is 3000~3400kPa, and grade frequency is 70~90Hz, when crushing Between be 45~55min, obtain day lily Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 10~14 times of amounts is added, and carries out steam distillation extraction 8~12h, extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；After steam distillation Residue T4 retains, spare；

S3: taking the resulting residue T4 of S2 step, is added the water of 6~8 times of amounts, and heating and refluxing extraction 1~3 time, every time 3 ~5h, obtained extracting solution merge with the resulting Aqueous extracts T3 of S2 step, and concentration obtains medicinal extract T5；Residue T6 after water extraction Retain, it is spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 1~3 time with 6~10 times of 70~80% alcohol refluxs of amount, is mentioned every time 2~4h of time is taken, combined extract recycles ethyl alcohol, and concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves The l-Alanine of 0.05~0.15% parts by weight is added in hair oil T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder.

The health protection tea it is preferred the preparation method comprises the following steps:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, is carried out using airslide disintegrating mill It pulverizes, the stream pressure of airslide disintegrating mill is 3200kPa, and grade frequency 80Hz, grinding time 50min obtain chrysanthemum Dish Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves The l-Alanine of 0.1% parts by weight is added in hair oil T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder.

A kind of detection method for the day lily health protection tea preventing alcoholic fatty liver, which is by day lily extract It is made, which prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, is carried out using airslide disintegrating mill It pulverizes, the stream pressure of airslide disintegrating mill is 3200kPa, and grade frequency 80Hz, grinding time 50min obtain chrysanthemum Dish Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves The l-Alanine of 0.1% parts by weight is added in hair oil T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

Using the content of rp-hplc determination obtusifoline, methyl Rhein, steps are as follows:

(1) chromatographic condition: chromatographic column: C₁₈, mobile phase: volume ratio is the methanol -0.04moL/L biphosphate of 15:80:5 The miscible fluid of -0.1% trifluoroacetic acid of potassium, column temperature be 40~50 DEG C, Detection wavelength be 280~290nm, flow velocity be 1.0~ 2.0mL·min^-1；

(2) prepared by reference substance solution: precision weighs 8.00~12.00mg of obtusifoline reference substance, methyl Rhein pair It according to 5.00~7.00mg of product, sets in 100mL measuring bottle, 0.1% trifluoroacetic acid aqueous solution is added to dissolve constant volume, precision measures 4~6mL, It sets in 25mL measuring bottle, is diluted to scale with 0.1% trifluoroacetic acid aqueous solution and shakes up to get reference substance solution；

(3) preparation of test solution: precision weighs 0.4~0.6g of sample, sets in 100mL measuring bottle, adds 0.1% trifluoro second Aqueous acid dissolution, 25~35min of ultrasound are settled to scale with 0.1% trifluoroacetic acid aqueous solution after cooling, shake up, 0.45 μm Miillpore filter filtration, takes subsequent filtrate to get test solution；

(4) accurate to draw reference substance solution and each 5~20 μ L of test solution, high performance liquid chromatograph is injected, is surveyed It is fixed.

The detection method of the day lily health protection tea of the prevention alcoholic fatty liver, which extracted by day lily Object is made, which prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, is carried out using airslide disintegrating mill It pulverizes, the stream pressure of airslide disintegrating mill is 3200kPa, and grade frequency 80Hz, grinding time 50min obtain chrysanthemum Dish Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves The l-Alanine of 0.1% parts by weight is added in hair oil T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

Using the content of rp-hplc determination obtusifoline, methyl Rhein, preferred steps are as follows:

(1) chromatographic condition: chromatographic column: C₁₈, mobile phase: volume ratio is the methanol -0.04moL/L biphosphate of 15:80:5 The miscible fluid of -0.1% trifluoroacetic acid of potassium, column temperature are 45 DEG C, Detection wavelength 286nm, flow velocity 1.5mLmin^-1；

(2) prepared by reference substance solution: precision weighs obtusifoline reference substance 10.00mg, methyl Rhein reference substance 6.00mg is set in 100mL measuring bottle, and 0.1% trifluoroacetic acid aqueous solution is added to dissolve constant volume, and precision measures 5mL, sets in 25mL measuring bottle, Scale is diluted to 0.1% trifluoroacetic acid aqueous solution to shake up to get reference substance solution；

(3) preparation of test solution: precision weighs sample 0.5g, sets in 100mL measuring bottle, adds 0.1% trifluoroacetic acid water Solution dissolution, ultrasonic 30min are settled to scale with 0.1% trifluoroacetic acid aqueous solution after cooling, shake up, 0.45 μm of miillpore filter Filtration, takes subsequent filtrate to get test solution；

(4) accurate to draw reference substance solution and each 10 μ L of test solution, high performance liquid chromatograph is injected, is measured.

A kind of detection method for the day lily health protection tea preventing alcoholic fatty liver, which is by day lily extract It is made, which prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, is carried out using airslide disintegrating mill It pulverizes, the stream pressure of airslide disintegrating mill is 3200kPa, and grade frequency 80Hz, grinding time 50min obtain chrysanthemum Dish Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves The l-Alanine of 0.1% parts by weight is added in hair oil T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

5 hydroxymethyl furfural content is measured using gas chromatography, steps are as follows:

(1) chromatographic condition and system suitability: chromatographic column: HP-5 column is to fix to be crosslinked 5% methyl-polysiloxane Phase；Injector temperature: 210~230 DEG C；Detector temperature: 210~230 DEG C；Split ratio: 14~16:2；Temperature programming: initial 65~75 DEG C, with 4~6 DEG C of min^-1Speed, rise to 135~145 DEG C；Carrier gas is nitrogen, 10~15mLmin of flow^-1, flow velocity is 1.0~3.0mLmin^-1；5~20 μ L of sample volume；

(2) preparation of test solution: taking 1.00~3.00g of health protection tea, accurately weighed, is placed in 100mL stuffed conical flask, 30~50mL normal hexane, 15~25min of ultrasound, filtration is added, filtrate adds normal hexane constant volume to get test sample into 50mL measuring bottle Solution；

(3) preparation of reference substance solution: taking 5 hydroxymethyl furfural reference substance, accurately weighed, adds normal hexane, and concentration, which is made, is 0.50~1.50mgmL^-15 hydroxymethyl furfural reference substance solution；

(4) measure: precision draws test solution, each 5~15 μ L of reference substance solution, injects gas chromatograph, is surveyed It is fixed.

The detection method of the day lily health protection tea of the prevention alcoholic fatty liver, which extracted by day lily Object is made, which prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, is carried out using airslide disintegrating mill It pulverizes, the stream pressure of airslide disintegrating mill is 3200kPa, and grade frequency 80Hz, grinding time 50min obtain chrysanthemum Dish Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves The l-Alanine of 0.1% parts by weight is added in hair oil T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

5 hydroxymethyl furfural content is measured using gas chromatography, preferred steps are as follows:

(1) chromatographic condition and system suitability: chromatographic column: HP-5 column is to fix to be crosslinked 5% methyl-polysiloxane Phase；Injector temperature: 220 DEG C；Detector temperature: 220 DEG C；Split ratio: 15:2；Temperature programming: initial 70 DEG C, with 5 DEG C min^-1Speed, rise to 140 DEG C；Carrier gas is nitrogen, flow 12mLmin^-1, flow velocity 2.0mLmin^-1；10 μ of sample volume L；

(2) preparation of test solution: taking health protection tea 2.00g, accurately weighed, is placed in 100mL stuffed conical flask, is added 40mL normal hexane ultrasound 20min, filtration, filtrate add normal hexane constant volume to get test solution into 50mL measuring bottle；

(3) preparation of reference substance solution: taking 5 hydroxymethyl furfural reference substance, accurately weighed, adds normal hexane, and concentration, which is made, is 1.00mg·mL^-15 hydroxymethyl furfural reference substance solution；

(4) measure: precision draws test solution, each 10 μ L of reference substance solution, injects gas chromatograph, is measured.

A kind of application of the health protection tea made of day lily in the day lily health protection tea of preparation prevention alcoholic fatty liver, The health protection tea is made of day lily extract；The health protection tea prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, is carried out using airslide disintegrating mill It pulverizes, the stream pressure of airslide disintegrating mill is 3200kPa, and grade frequency 80Hz, grinding time 50min obtain chrysanthemum Dish Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves The l-Alanine of 0.1% parts by weight is added in hair oil T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

Using the content of rp-hplc determination obtusifoline, methyl Rhein, steps are as follows:

(1) chromatographic condition: chromatographic column: C₁₈, mobile phase: volume ratio is the methanol -0.04moL/L biphosphate of 15:80:5 The miscible fluid of -0.1% trifluoroacetic acid of potassium, column temperature are 45 DEG C, Detection wavelength 286nm, flow velocity 1.5mLmin^-1；

(2) prepared by reference substance solution: precision weighs obtusifoline reference substance 10.00mg, methyl Rhein reference substance 6.00mg is set in 100mL measuring bottle, and 0.1% trifluoroacetic acid aqueous solution is added to dissolve constant volume, and precision measures 5mL, sets in 25mL measuring bottle, Scale is diluted to 0.1% trifluoroacetic acid aqueous solution to shake up to get reference substance solution；

(3) preparation of test solution: precision weighs sample 0.5g, sets in 100mL measuring bottle, adds 0.1% trifluoroacetic acid water Solution dissolution, ultrasonic 30min are settled to scale with 0.1% trifluoroacetic acid aqueous solution after cooling, shake up, 0.45 μm of miillpore filter Filtration, takes subsequent filtrate to get test solution；

(4) accurate to draw reference substance solution and each 10 μ L of test solution, high performance liquid chromatograph is injected, is measured；

And/or 5 hydroxymethyl furfural content is measured using gas chromatography, steps are as follows:

(1) chromatographic condition and system suitability: chromatographic column: HP-5 column is to fix to be crosslinked 5% methyl-polysiloxane Phase；Injector temperature: 220 DEG C；Detector temperature: 220 DEG C；Split ratio: 15:2；Temperature programming: initial 70 DEG C, with 5 DEG C min^-1Speed, rise to 140 DEG C；Carrier gas is nitrogen, flow 12mLmin^-1, flow velocity 2.0mLmin^-1；10 μ of sample volume L；

(2) preparation of test solution: taking health protection tea 2.00g, accurately weighed, is placed in 100mL stuffed conical flask, is added 40mL normal hexane ultrasound 20min, filtration, filtrate add normal hexane constant volume to get test solution into 50mL measuring bottle；

(3) preparation of reference substance solution: taking 5 hydroxymethyl furfural reference substance, accurately weighed, adds normal hexane, and concentration, which is made, is 1.00mg·mL^-15 hydroxymethyl furfural reference substance solution；

(4) measure: precision draws test solution, each 10 μ L of reference substance solution, injects gas chromatograph, is measured.

It is specifically described by following experimental study and analyzes technical solution of the present invention and technical effect:

Experiment one: the health protection tea screening experiment of the prevention effect of mouse alcoholic fatty liver is studied

1 material and method

1.1 experimental animal

SPF grades of cleaning grade mouse 70,20~22g of weight are provided, animal by Gansu university of TCM Medical experimental center Quality certification number: SCXK (sweet) 2011-0001-0001011；Experimental Establishment quality certification number: SYXK (sweet) 2011-0001- 0000314；It raises in Gansu university of TCM Medical experimental center, animal feeding is under standard conditions, 12h light and shade alternating, and (22 ± 2) DEG C, drinking-water of freely ingesting.Animal adaptive feeding in above-mentioned environment starts to test after 1 week.

1.2 experiment reagent

White wine: 55 degree of strong, colourless liquor distilled from sorghum white wine of Red Star board, Hongxing Co., Ltd. Beijing's production, production licence QS110015010349, specification: 500mL/ bottles；

Tested health protection tea LA: prescription: day lily 1000g；

Preparation method: taking dry day lily, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h, extracting solution closes And be concentrated, it is freeze-dried, crushes, obtain tested health protection tea LA.

Tested health protection tea LB: prescription: day lily 1000g；

Preparation method: taking dry day lily, and 8 times of 75% alcohol refluxs of amount are added and extract 2 times, each extraction time 3h, Combined extract recycles ethyl alcohol, and concentration obtains medicinal extract, is freeze-dried, and crushes, obtains tested health protection tea LB.

Tested health protection tea LC: prescription: day lily 1000g；

Preparation method: S1: taking day lily, the day lily after after 37 DEG C of drying 5h, taking drying in baking oven is set, using air-flow powder Broken machine is pulverized, and the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, and grinding time is 50min obtains day lily Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves Hair oil T2 is mixed, and is freeze-dried, and is crushed, is obtained tested health protection tea LC.

Tested health protection tea LD: prescription: day lily 1000g；

Preparation method: taking day lily, the day lily after after 37 DEG C of drying 5h, taking drying in baking oven is set, using air-flow crushing Machine is pulverized, and the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, grinding time 50min, Day lily Ultramicro-powder is obtained, 8 times of 35% alcohol refluxs of amount are added and extract 2 times, each extraction time 3h, combined extract recycles second Alcohol, concentration, obtains medicinal extract, is freeze-dried, and crushes, obtains tested health protection tea LD.

1.3 experimental groups and given low

Mouse is randomly divided into following 6 groups, be respectively as follows: blank control group, model control group, tested health protection tea LA group, by Examination health protection tea LB group, tested health protection tea LC group, tested health protection tea LD group, every group 10；

The tested tested health protection tea LA of health protection tea LA group stomach-filling, dosage 6.5g/10mL/Kg；Tested health protection tea LB group stomach-filling Tested health protection tea LB, dosage 6.5g/10mL/Kg；The tested tested health protection tea LC of health protection tea LC group stomach-filling, dosage 6.5g/ 10mL/Kg；The tested tested health protection tea LD of health protection tea LD group stomach-filling, dosage 6.5g/10mL/Kg；Blank control group and model pair The physiological saline of Isodose (10mL/Kg) is gavaged according to group.In addition to blank control group, other groups gavage white wine, and dosage is 10mL/kg is continuously gavaged 6 weeks.

1.4 Testing index and method

It before blood sampling, to mouse fasting 12h, then takes a blood sample through mouse vena orbitalis posterior clump, centrifugal blood 20min separates blood Clearly, alanine aminotransferase (ALT), aspartate transaminase (AST) and the triglycerides (TG) of each group mouse are detected.

The liver organization of each group mouse is won, is weighed, and calculate liver index.

Liver index=liver quality (g)/weight (g) × 100%；

The hepatic tissue and one section of colonic tissue for acquiring the same position of all mouse, are fixed with 4% formaldehyde, after HE dyeing, Analyze the pathology testing result of each group mouse liver and colonic tissue.

1.5 statistical method

Statistical analysis is carried out to the Testing index of each group mouse using the variance analysis method of SPSS12.0 software.

2 results

The analysis of 2.1 each group mouse weight increments, liver index and biochemical indicator

It is shown by the comparative analysis to each group mouse weight increment, liver index and biochemical indicator: with blank control group It compares, the body weight increase amount of model group mouse significantly reduces (P < 0.05), and liver index dramatically increases (P < 0.05), Serum ALT, AST and TG also significantly increases (P < 0.05)；And compared with model control group, the body weight increase amount of each dosage group mouse of HLJDT is equal It dramatically increases (P < 0.05), liver index is substantially reduced (P < 0.05), and Serum ALT, AST and TG content are substantially reduced (P < 0.05), wherein influence of the HLJDT to ALT and TG content shows dose dependent (table 1).

1 each group mouse liver index of table and Analysis of Biochemical

Note: compared with blank control group^*P < 0.05；Compared with model control group^△P < 0.05.

2.2 liver pathology

(1) lobuli hepatis structure of blank control group mouse is clear, and liver rope queueing discipline has no inflammatory cell infiltration；See Bright book attached drawing 1.

(2) model control group mouse lobuli hepatis boundary is fuzzy, has fat drop in heaps, has the liver of a large amount of inflammatory infiltrations thin Born of the same parents；See Figure of description 2.

(3) tested health protection tea LA group mouse lobuli hepatis boundary is fuzzy, has the fat drop of aggregation, there is the liver of more inflammatory infiltration Cell；See Figure of description 3.

(4) tested health protection tea LB group mouse lobuli hepatis boundary is fuzzy, has the fat drop of aggregation, there is the liver of more inflammatory infiltration Cell；See Figure of description 4.

(5) fat of tested health protection tea LC group mouse becomes and swelling of liver cell degree is more substantially reduced than model control group, Each dosage group mouse liver cell, which does not observe fat drop in heaps, to be occurred, and lobuli hepatis and portal area have no inflammatory cell infiltration； See Figure of description 5.

(6) tested health protection tea LD group mouse lobuli hepatis boundary is fuzzy, has the fat drop of aggregation, there is the liver of more inflammatory infiltration Cell；See Figure of description 6.

3 conclusions

Experimental studies results are shown: compared with model control group, mouse liver index significantly drops tested health protection tea LC group result Low (P < 0.05), Serum ALT, AST and TG content significantly reduce (P < 0.05), illustrate that tested health protection tea LC can significantly drop The concentration of low serum transaminase, the effect with stronger reparation alcoholic liver damage；Tested health protection tea LC group can simultaneously The content of serum TG is significantly reduced, liver pathology is the results show that tested health protection tea LC all has significantly alcoholic liver injury Ground improves repair.

Experiment two: obtusifoline in rp-hplc determination health protection tea of the present invention, methyl Rhein contain Amount

1 instrument, reagent and material

1.1 instrument

DIONEX high performance liquid chromatograph is equipped with P680 efficient liquid phase and pumps, ASI-100 autosampler, and UVD170U is ultraviolet Detector, Chromeleon work station.

1.2 reagents and material

Obtusifoline reference substance (Shaanxi Biao Pu Pharmaceutical Technology Co., Ltd provides, lot number 170123-201603), methyl Rhein reference substance (Shaanxi Biao Pu Pharmaceutical Technology Co., Ltd provides, lot number 170205-201605).

Health protection tea (sample) of the present invention: prescription: day lily 1000g；

Preparation method: S1: taking day lily, the day lily after after 37 DEG C of drying 5h, taking drying in baking oven is set, using air-flow powder Broken machine is pulverized, and the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, and grinding time is 50min obtains day lily Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves Hair oil T2 is mixed, and is freeze-dried, and is crushed, is obtained health protection tea of the present invention (sample).

2 methods and result

2.1 chromatographic condition

Chromatographic column: C₁₈, specification: 4.6mm × 200mm, 5 μm, mobile phase: volume ratio is the methanol -0.04moL/ of 15:80:5 The miscible fluid of -0.1% trifluoroacetic acid of L potassium dihydrogen phosphate, column temperature are 45 DEG C, Detection wavelength 286nm, flow velocity 1.5mL min^-1。

The preparation of 2.2 solution

2.2.1 prepared by reference substance solution

Precision weighs obtusifoline reference substance 10.00mg and methyl Rhein reference substance 6.00mg, sets 100mL measuring bottle In, add 0.1% trifluoroacetic acid aqueous solution to dissolve constant volume, precision measures 5mL, sets in 25mL measuring bottle, water-soluble with 0.1% trifluoroacetic acid Liquid is diluted to scale and shakes up to get reference substance solution.

2.2.2 the preparation of test solution

Precision weighs sample 0.5g, sets in 100mL measuring bottle, and 0.1% trifluoroacetic acid aqueous solution is added to dissolve, and ultrasonic 30min is cold But it is settled to scale with 0.1% trifluoroacetic acid aqueous solution afterwards, shaken up, 0.45 μm of miillpore filter filtration takes subsequent filtrate to get for examination Product solution.

2.3 standard curve

2 μ L of reference substance solution, 4 μ L, 8 μ L, 16 μ L, 32 μ L, 64 μ L are drawn, high performance liquid chromatograph is injected, with sample volume (X) be abscissa, chromatographic peak area (Y) be ordinate, following regression equation:

Obtusifoline: Y=3.1587X-5.2685, r=0.9998；

Methyl Rhein: Y=2.1652X-6.2542, r=0.9999.

Show that obtusifoline sample volume linear relationship in 0.02015~0.5635 μ g range is good, methyl Rhein Sample volume linear relationship in 0.01525~0.4585 μ g range is good.

Reference substance solution and 10 μ L of test solution is taken to inject high performance liquid chromatograph, high-efficient liquid phase chromatogram is shown in specification Attached drawing 7 and attached drawing 8.

2.4 precision test

Test solution is taken, is repeated sample introduction 5 times, the peak area RSD of obtusifoline is 1.22%, the peak of methyl Rhein Area RSD is 0.95%, shows that precision is good.

2.5 stability test

Test solution is taken, 1h, 2h, 4h, 8h, 16h sample introduction after preparation, measure obtusifoline and methyl Rhein contains Amount, the RSD of obtusifoline are 1.15%, and the RSD of methyl Rhein is 1.28%, are shown in 16h, test solution stability Well.

2.6 reappearance test

Precision weighs 5 parts of same batch sample, test solution is prepared according to the method provided by the invention, according to the present invention The content of chromatographic condition the measurement obtusifoline and methyl Rhein of offer, obtusifoline RSD is 2.11%, methyl rheum officinale Sour RSD is 1.85%, shows that reproducibility is good.

2.7 sample-adding recovery tests

Accurately weighed health protection tea powder of the present invention, is separately added into obtusifoline and methyl Rhein reference substance is appropriate, presses Method operation provided by the invention, measurement, average recovery rate is respectively 100.67% and 100.01%, and RSD is respectively 1.71% With 2.03% (n=6), it the results are shown in Table 2 and table 3.

2 obtusifoline of table is loaded recovery test result (n=6)

3 methyl Rhein of table is loaded recovery test result (n=6)

2.8 sample size

Measurement weighs the health protection tea of the present invention of different lot numbers respectively, accurately weighed, prepares according to the method provided by the invention Test solution measures according to chromatographic condition provided by the invention, the results are shown in Table 4.

The content (n=3) of obtusifoline, methyl Rhein in 4 three batches of samples of table

3 conclusions

The result shows that the HPLC method is simple, and it is easily operated, it can be used as obtusifoline, methyl in health protection tea of the present invention The quantitative detecting method of Rhein.

Experiment three: gas chromatography measures 5 hydroxymethyl furfural in health protection tea of the present invention and contains quantifier elimination

1 instrument and reagent

Gas chromatograph；Flame ionization ditector (FID)；HP Innowax (30m × 0.25mm, 0.25 μm) elasticity Quartz capillary chromatographic column.

5 hydroxymethyl furfural reference substance is won Science and Technology Ltd. by hundred Aurion of Beijing and is provided, lot number: 0625-201603.

Health protection tea (sample) of the present invention: prescription: day lily 1000g；

Preparation method: S1: taking day lily, the day lily after after 37 DEG C of drying 5h, taking drying in baking oven is set, using air-flow powder Broken machine is pulverized, and the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, and grinding time is 50min obtains day lily Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves Hair oil T2 is mixed, and is freeze-dried, and is crushed, is obtained health protection tea of the present invention (sample).

Acetonitrile, chromatographically pure, redistilled water, self-control, other reagents are that analysis is pure.

2 experimental methods and result

2.1 chromatographic conditions and system suitability

Chromatographic column: HP-5 column, model: 30m × 0.32mm × 0.25 μm is solid to be crosslinked 5% methyl-polysiloxane Determine phase；Injector temperature: 220 DEG C；Detector temperature: 220 DEG C；Split ratio: 15:2；Temperature programming: initial 70 DEG C, with 5 DEG C min^-1Speed, rise to 140 DEG C；Carrier gas is nitrogen, flow 12mLmin^-1, flow velocity 2.0mLmin^-1；10 μ of sample volume L；Under this condition, 5 hydroxymethyl furfural can reach baseline separation with other components, be greater than 1.8 with adjacent chromatographic peak separating degree； Number of theoretical plate is calculated by 5 hydroxymethyl furfural peak should be not less than 15000；As a result see Figure of description 9, attached drawing 10.

The preparation of 2.2 test solutions

Health protection tea 2.00g is taken, it is accurately weighed, it is placed in 100mL stuffed conical flask, 40mL normal hexane ultrasound 20min, filter is added It crosses, filtrate adds normal hexane constant volume to get test solution into 50mL measuring bottle.

The preparation of 2.3 reference substance solutions

5 hydroxymethyl furfural reference substance is taken, it is accurately weighed, add normal hexane, it is 1.00mgmL that concentration, which is made,^-15- hydroxyl first The reference substance solution of base furfural.

2.4 linear relationships are investigated

It is accurate respectively to draw a certain amount of mass concentration as 1.00mgmL^-15 hydroxymethyl furfural reference substance stock solution, add Normal hexane is diluted to a series of reference substance solution of mass concentrations.Each 10 μ L of above-mentioned reference substance solution is drawn respectively, by this hair The chromatographic condition of bright offer is measured.

Using integrating peak areas value as ordinate (Y), it is abscissa (X, μ g) that 5 hydroxymethyl furfural, which compares quality, draws mark Directrix curve, as a result as follows:

Y=856.245X-6.238, r=0.9998, linear relationship is good in 0.0528~5.368 μ g.

2.5 precision test

It is 0.5mgmL that precision, which draws mass concentration,^-1Reference substance solution repeat sample introduction 5 times, measure 5 hydroxymethyl furfural Peak area RSD be 0.85%.

2.6 stability test

Health protection tea 2.00g is taken, it is accurately weighed, according to method provided by the invention processing, surveyed respectively at 0,2,4,8,16,32h Determine the peak area of 5 hydroxymethyl furfural in test solution.The result shows that test solution is basicly stable in 32h, RSD is 1.05%.

2.7 repeated experiment

Health protection tea is taken, parallel sampling 6 times is accurately weighed, according to method provided by the invention processing, in color provided by the invention It is measured under spectral condition, as a result the RSD of 5 hydroxymethyl furfural content is 1.03%.

The experiment of 2.8 sample recovery rates

It is appropriate to weigh 5 hydroxymethyl furfural reference substance for precision respectively, is added in health protection tea, by preparation provided by the invention, Chromatographic condition provided by the invention is measured, and calculates the rate of recovery.It the results are shown in Table 5.

55 hydroxymethyl furfural sample recovery rate experimental result of table

2.9 assay

4 batches of health protection teas of the present invention are taken respectively, by legal system available test sample solution below " 2.1.2 " item, in given chromatostrip It is measured under part, the content of 5 hydroxymethyl furfural (C10H20O) is calculated with external standard method, the results are shown in Table 6.

The assay result of 5 hydroxymethyl furfural in 6 sample of table

3 conclusions

This experiment establishes the assay of volatile component 5 hydroxymethyl furfural in health protection tea of the present invention with gas-chromatography Method can effectively detect the important activity ingredient in health protection tea of the present invention.Detection method precision of the invention is high, repeatability Good, stability is good, can be used for the quality testing of this product.

Figure of description:

Fig. 1 is blank control group mouse liver histopathology inspection result (200 ×).

Fig. 2 is model control group murine liver tissue pathological examination result (200 ×).

Fig. 3 is tested health protection tea LA group murine liver tissue pathological examination result (200 ×).

Fig. 4 is tested health protection tea LB group murine liver tissue pathological examination result (200 ×).

Fig. 5 is tested health protection tea LC group murine liver tissue pathological examination result (200 ×).

Fig. 6 is tested health protection tea LD group murine liver tissue pathological examination result (200 ×).

Fig. 7 is reference substance solution HPLC chromatogram, in which: No. 1 peak is obtusifoline chromatographic peak, and No. 2 peaks are that methyl is big Yellow acid chromatographic peak.

Fig. 8 is HPLC chromatogram of sample solution, in which: No. 1 peak is obtusifoline chromatographic peak, and No. 2 peaks are that methyl is big Yellow acid chromatographic peak.

Fig. 9 is reference substance gas chromatogram, wherein No. 1 peak is 5 hydroxymethyl furfural chromatographic peak.

Figure 10 is test sample gas chromatogram, wherein No. 1 peak is 5 hydroxymethyl furfural chromatographic peak.

Specific embodiment

Combined with specific embodiments below the present invention is made further to explain.

Embodiment 1:

Prescription: day lily 1000g；

Preparation method: S1: taking day lily, the day lily after after 37 DEG C of drying 5h, taking drying in baking oven is set, using air-flow powder Broken machine is pulverized, and the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, and grinding time is 50min obtains day lily Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, Extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation is protected It stays, it is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h is obtained Extracting solution merge with the resulting Aqueous extracts T3 of S2 step, be concentrated, obtain medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times, each extraction time 3h, is closed with 8 times of 75% alcohol refluxs of amount And extracting solution, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and what addition S2 step obtained waves Hair oil T2 is mixed, and is freeze-dried, and is crushed, is obtained health protection tea of the present invention.

Healthcare function: nourishing and protecting liver.

Suitable population: the crowd of advanced low-grade alcoholic fatty liver.

Eating method and amount: 2 times a day, each 5g is brewed with warm water, slow to drink.

The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto, Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.

Claims (8)

1. a kind of day lily health protection tea for preventing alcoholic fatty liver, which is characterized in that the health protection tea is by day lily extract It is made.

2. the day lily health protection tea of prevention alcoholic fatty liver as described in claim 1, which is characterized in that the system of the health protection tea Preparation Method are as follows:

S1: taking day lily, set after 35~40 DEG C of 4~6h of drying, taken in baking oven it is dry after day lily, using airslide disintegrating mill into Row pulverizes, and the stream pressure of airslide disintegrating mill is 3000~3400kPa, and grade frequency is 70~90Hz, and grinding time is 45~55min obtains day lily Ultramicro-powder T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 10~14 times of amounts are added, carry out steam distillation extract 8~ 12h, extraction obtains volatile oil T2, spare；Aqueous extracts T3 after steam distillation retains, spare；Residue after steam distillation T4 retains, spare；

S3: taking the resulting residue T4 of S2 step, is added the water of 6~8 times of amounts, and heating and refluxing extraction 1~3 time, every time 3~ 5h, obtained extracting solution merge with the resulting Aqueous extracts T3 of S2 step, and concentration obtains medicinal extract T5；Residue T6 after water extracts is protected It stays, it is spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 1~3 time with 6~10 times of 70~80% alcohol refluxs of amount, when extracting every time Between 2~4h, combined extract, recycle ethyl alcohol, concentration, obtain medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and the volatile oil that S2 step obtains is added The l-Alanine of 0.05~0.15% parts by weight is added in T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder.

3. the day lily health protection tea of prevention alcoholic fatty liver as claimed in claim 2, which is characterized in that the system of the health protection tea Preparation Method are as follows:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, carries out ultra micro using airslide disintegrating mill It crushes, the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, grinding time 50min, and it is super to obtain day lily Micro mist T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, extracts Volatile oil T2 is obtained, it is spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation retains, It is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h, what is obtained mentions It takes liquid to merge with the resulting Aqueous extracts T3 of S2 step, is concentrated, obtains medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times with 8 times of 75% alcohol refluxs of amount, each extraction time 3h, merging mentions Liquid is taken, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and the volatile oil that S2 step obtains is added The l-Alanine of 0.1% parts by weight is added in T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder.

4. a kind of detection method for the day lily health protection tea for preventing alcoholic fatty liver, which is by day lily extract system At the health protection tea prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, carries out ultra micro using airslide disintegrating mill It crushes, the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, grinding time 50min, and it is super to obtain day lily Micro mist T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, extracts Volatile oil T2 is obtained, it is spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation retains, It is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h, what is obtained mentions It takes liquid to merge with the resulting Aqueous extracts T3 of S2 step, is concentrated, obtains medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times with 8 times of 75% alcohol refluxs of amount, each extraction time 3h, merging mentions Liquid is taken, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and the volatile oil that S2 step obtains is added The l-Alanine of 0.1% parts by weight is added in T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

It is characterized in that, step is such as using the content of rp-hplc determination obtusifoline, methyl Rhein Under:

(1) chromatographic condition: chromatographic column: C₁₈, mobile phase: volume ratio is the methanol -0.04moL/L potassium dihydrogen phosphate-of 15:80:5 The miscible fluid of 0.1% trifluoroacetic acid, column temperature are 40~50 DEG C, and Detection wavelength is 280~290nm, and flow velocity is 1.0~2.0mL min^-1；

(2) prepared by reference substance solution: precision weighs 8.00~12.00mg of obtusifoline reference substance, methyl Rhein reference substance 5.00~7.00mg is set in 100mL measuring bottle, and 0.1% trifluoroacetic acid aqueous solution is added to dissolve constant volume, and precision measures 4~6mL, sets In 25mL measuring bottle, scale is diluted to 0.1% trifluoroacetic acid aqueous solution and is shaken up to get reference substance solution；

(3) preparation of test solution: precision weighs 0.4~0.6g of sample, sets in 100mL measuring bottle, adds 0.1% trifluoroacetic acid water Solution dissolution, 25~35min of ultrasound are settled to scale with 0.1% trifluoroacetic acid aqueous solution after cooling, shake up, 0.45 μm of micropore Filter membrane filtration, takes subsequent filtrate to get test solution；

(4) accurate to draw reference substance solution and each 5~20 μ L of test solution, high performance liquid chromatograph is injected, is measured.

5. as claimed in claim 4 prevention alcoholic fatty liver day lily health protection tea detection method, the health protection tea be by Day lily extract is made, which prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, carries out ultra micro using airslide disintegrating mill It crushes, the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, grinding time 50min, and it is super to obtain day lily Micro mist T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, extracts Volatile oil T2 is obtained, it is spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation retains, It is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h, what is obtained mentions It takes liquid to merge with the resulting Aqueous extracts T3 of S2 step, is concentrated, obtains medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times with 8 times of 75% alcohol refluxs of amount, each extraction time 3h, merging mentions Liquid is taken, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and the volatile oil that S2 step obtains is added The l-Alanine of 0.1% parts by weight is added in T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

It is characterized in that, step is such as using the content of rp-hplc determination obtusifoline, methyl Rhein Under:

(1) chromatographic condition: chromatographic column: C₁₈, mobile phase: volume ratio is the methanol -0.04moL/L potassium dihydrogen phosphate-of 15:80:5 The miscible fluid of 0.1% trifluoroacetic acid, column temperature are 45 DEG C, Detection wavelength 286nm, flow velocity 1.5mLmin^-1；

(2) prepared by reference substance solution: precision weighs obtusifoline reference substance 10.00mg, methyl Rhein reference substance 6.00mg, It sets in 100mL measuring bottle, 0.1% trifluoroacetic acid aqueous solution is added to dissolve constant volume, precision measures 5mL, sets in 25mL measuring bottle, with 0.1% Trifluoroacetic acid aqueous solution is diluted to scale and shakes up to get reference substance solution；

(3) preparation of test solution: precision weighs sample 0.5g, sets in 100mL measuring bottle, adds 0.1% trifluoroacetic acid aqueous solution Dissolution, ultrasonic 30min are settled to scale with 0.1% trifluoroacetic acid aqueous solution after cooling, shake up, 0.45 μm of miillpore filter filtration, Take subsequent filtrate to get test solution；

(4) accurate to draw reference substance solution and each 10 μ L of test solution, high performance liquid chromatograph is injected, is measured.

6. a kind of detection method for the day lily health protection tea for preventing alcoholic fatty liver, which is by day lily extract system At the health protection tea prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, carries out ultra micro using airslide disintegrating mill It crushes, the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, grinding time 50min, and it is super to obtain day lily Micro mist T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, extracts Volatile oil T2 is obtained, it is spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation retains, It is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h, what is obtained mentions It takes liquid to merge with the resulting Aqueous extracts T3 of S2 step, is concentrated, obtains medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times with 8 times of 75% alcohol refluxs of amount, each extraction time 3h, merging mentions Liquid is taken, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and the volatile oil that S2 step obtains is added The l-Alanine of 0.1% parts by weight is added in T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

It is characterized in that, measuring 5 hydroxymethyl furfural content using gas chromatography, steps are as follows:

(1) chromatographic condition and system suitability: chromatographic column: HP-5 column, to be crosslinked 5% methyl-polysiloxane as stationary phase； Injector temperature: 210~230 DEG C；Detector temperature: 210~230 DEG C；Split ratio: 14~16:2；Temperature programming: initial 65~ 75 DEG C, with 4~6 DEG C of min^-1Speed, rise to 135~145 DEG C；Carrier gas is nitrogen, 10~15mLmin of flow^-1, stream Speed is 1.0~3.0mLmin^-1；5~20 μ L of sample volume；

(2) preparation of test solution: taking 1.00~3.00g of health protection tea, accurately weighed, is placed in 100mL stuffed conical flask, is added 30~50mL normal hexane 15~25min of ultrasound, filtration, filtrate add normal hexane constant volume molten to get test sample into 50mL measuring bottle Liquid；

(3) preparation of reference substance solution: taking 5 hydroxymethyl furfural reference substance, accurately weighed, adds normal hexane, and it is 0.50 that concentration, which is made, ~1.50mgmL^-15 hydroxymethyl furfural reference substance solution；

(4) measure: precision draws test solution, each 5~15 μ L of reference substance solution, injects gas chromatograph, is measured.

7. as claimed in claim 6 prevention alcoholic fatty liver day lily health protection tea detection method, the health protection tea be by Day lily extract is made, which prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, carries out ultra micro using airslide disintegrating mill It crushes, the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, grinding time 50min, and it is super to obtain day lily Micro mist T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, extracts Volatile oil T2 is obtained, it is spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation retains, It is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h, what is obtained mentions It takes liquid to merge with the resulting Aqueous extracts T3 of S2 step, is concentrated, obtains medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times with 8 times of 75% alcohol refluxs of amount, each extraction time 3h, merging mentions Liquid is taken, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and the volatile oil that S2 step obtains is added The l-Alanine of 0.1% parts by weight is added in T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

It is characterized in that, measuring 5 hydroxymethyl furfural content using gas chromatography, steps are as follows:

(1) chromatographic condition and system suitability: chromatographic column: HP-5 column, to be crosslinked 5% methyl-polysiloxane as stationary phase； Injector temperature: 220 DEG C；Detector temperature: 220 DEG C；Split ratio: 15:2；Temperature programming: initial 70 DEG C, with 5 DEG C of min^-1's Speed rises to 140 DEG C；Carrier gas is nitrogen, flow 12mLmin^-1, flow velocity 2.0mLmin^-1；10 μ L of sample volume；

(2) preparation of test solution: taking health protection tea 2.00g, accurately weighed, is placed in 100mL stuffed conical flask, and 40mL is being added just Hexane ultrasound 20min, filtration, filtrate add normal hexane constant volume to get test solution into 50mL measuring bottle；

(3) preparation of reference substance solution: taking 5 hydroxymethyl furfural reference substance, accurately weighed, adds normal hexane, and concentration, which is made, is 1.00mg·mL^-15 hydroxymethyl furfural reference substance solution；

(4) measure: precision draws test solution, each 10 μ L of reference substance solution, injects gas chromatograph, is measured.

8. a kind of application of health protection tea made of day lily in the day lily health protection tea of preparation prevention alcoholic fatty liver, It is characterized in that, which is made of day lily extract；The health protection tea prepares with the following method:

S1: taking day lily, sets the day lily after after 37 DEG C of drying 5h, taking drying in baking oven, carries out ultra micro using airslide disintegrating mill It crushes, the stream pressure of airslide disintegrating mill is 3200kPa, grade frequency 80Hz, grinding time 50min, and it is super to obtain day lily Micro mist T1；

S2: taking the resulting day lily Ultramicro-powder T1 of S1 step, and the water of 12 times of amounts is added, and carries out steam distillation and extracts 10h, extracts Volatile oil T2 is obtained, it is spare；Aqueous extracts T3 after steam distillation retains, spare；Residue T4 after steam distillation retains, It is spare；

S3: taking the resulting residue T4 of S2 step, the water of 7 times of amounts is added, heating and refluxing extraction 2 times, each 4h, what is obtained mentions It takes liquid to merge with the resulting Aqueous extracts T3 of S2 step, is concentrated, obtains medicinal extract T5；Residue T6 after water extracts retains, spare；

S4: the residue T6 for taking S3 step to obtain, it is extracted 2 times with 8 times of 75% alcohol refluxs of amount, each extraction time 3h, merging mentions Liquid is taken, ethyl alcohol is recycled, concentration obtains medicinal extract T7；

S5: the medicinal extract T7 that medicinal extract T5, S4 step that S3 step obtains obtains is uniformly mixed, and the volatile oil that S2 step obtains is added The l-Alanine of 0.1% parts by weight is added in T2, mixes, and is freeze-dried, and crushes, obtains health-care tea powder；

Using the content of rp-hplc determination obtusifoline, methyl Rhein, steps are as follows:

(1) chromatographic condition: chromatographic column: C₁₈, mobile phase: volume ratio is the methanol -0.04moL/L potassium dihydrogen phosphate-of 15:80:5 The miscible fluid of 0.1% trifluoroacetic acid, column temperature are 45 DEG C, Detection wavelength 286nm, flow velocity 1.5mLmin^-1；

(2) prepared by reference substance solution: precision weighs obtusifoline reference substance 10.00mg, methyl Rhein reference substance 6.00mg, It sets in 100mL measuring bottle, 0.1% trifluoroacetic acid aqueous solution is added to dissolve constant volume, precision measures 5mL, sets in 25mL measuring bottle, with 0.1% Trifluoroacetic acid aqueous solution is diluted to scale and shakes up to get reference substance solution；

(3) preparation of test solution: precision weighs sample 0.5g, sets in 100mL measuring bottle, adds 0.1% trifluoroacetic acid aqueous solution Dissolution, ultrasonic 30min are settled to scale with 0.1% trifluoroacetic acid aqueous solution after cooling, shake up, 0.45 μm of miillpore filter filtration, Take subsequent filtrate to get test solution；

(4) accurate to draw reference substance solution and each 10 μ L of test solution, high performance liquid chromatograph is injected, is measured；

And/or 5 hydroxymethyl furfural content is measured using gas chromatography, steps are as follows:

(1) chromatographic condition and system suitability: chromatographic column: HP-5 column, to be crosslinked 5% methyl-polysiloxane as stationary phase； Injector temperature: 220 DEG C；Detector temperature: 220 DEG C；Split ratio: 15:2；Temperature programming: initial 70 DEG C, with 5 DEG C of min^-1's Speed rises to 140 DEG C；Carrier gas is nitrogen, flow 12mLmin^-1, flow velocity 2.0mLmin^-1；10 μ L of sample volume；

(2) preparation of test solution: taking health protection tea 2.00g, accurately weighed, is placed in 100mL stuffed conical flask, and 40mL is being added just Hexane ultrasound 20min, filtration, filtrate add normal hexane constant volume to get test solution into 50mL measuring bottle；

(3) preparation of reference substance solution: taking 5 hydroxymethyl furfural reference substance, accurately weighed, adds normal hexane, and concentration, which is made, is 1.00mg·mL^-15 hydroxymethyl furfural reference substance solution；

(4) measure: precision draws test solution, each 10 μ L of reference substance solution, injects gas chromatograph, is measured.