CN116223657A - Method for detecting organic acid in iron sucrose product - Google Patents
Method for detecting organic acid in iron sucrose product Download PDFInfo
- Publication number
- CN116223657A CN116223657A CN202211728370.8A CN202211728370A CN116223657A CN 116223657 A CN116223657 A CN 116223657A CN 202211728370 A CN202211728370 A CN 202211728370A CN 116223657 A CN116223657 A CN 116223657A
- Authority
- CN
- China
- Prior art keywords
- acid
- solution
- mobile phase
- iron sucrose
- eluent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8624—Detection of slopes or peaks; baseline correction
- G01N30/8631—Peaks
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
- G01N2030/3007—Control of physical parameters of the fluid carrier of temperature same temperature for whole column
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
- G01N2030/324—Control of physical parameters of the fluid carrier of pressure or speed speed, flow rate
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a method for detecting organic acid in a ferrosucrose product. The method for detecting the organic acid in the iron sucrose product provided by the invention comprises the following steps: (1) Pretreating the iron sucrose product, which comprises the following steps: mixing the iron sucrose product aqueous solution with the treatment solution, and removing the supernatant to obtain a sample solution; the treatment liquid is an aqueous acid solution; (2) Detecting the sample solution by adopting high performance liquid chromatography, wherein chromatographic conditions comprise: mobile phase a is acetonitrile; mobile phase B is a mixed solution of phosphoric acid aqueous solution and acetonitrile. The detection method can eliminate matrix interference of the sample solution, the separation degree of each obtained organic acid peak meets the detection requirement, the symmetry of chromatographic peak-to-peak type is good, and the detection method has the advantages of high accuracy, good repeatability and good stability in detecting the content of each organic acid.
Description
Technical Field
The invention relates to a method for detecting organic acid in a ferrosucrose product.
Background
Iron deficiency and iron deficiency anemia are one of the major factors responsible for the global disease burden, with more than 20 billions of cases of iron deficiency in 2016. Iron plays an important role in hemoglobin synthesis, oxygen transport, and critical cellular metabolic pathways. In the treatment of iron deficiency or iron deficiency anemia, intravenous iron preparations generally have quick treatment effect, can avoid poor absorption of intestinal medicines, avoid common adverse reactions of digestive tracts when taken orally, and improve medicine tolerance. Intravenous iron preparations have been recommended as a first line for the management of heart failure, chronic kidney disease, inflammatory bowel disease, iron deficiency anemia in the surgical stage and gynaecology and obstetrics.
The iron sucrose injection is one of the intravenous iron preparations which are used in the earliest clinic, and is one of the intravenous iron preparations which are most commonly used in the world. Iron sucrose is a complex of iron hydroxide and sucrose, and is represented by Fe (OH) 3 The compound is composed of a core and sucrose as shells, has a structure similar to ferritin, the sugar shells are dissociated from the iron cores after intravenous injection, the iron cores release iron elements through endocytosis of macrophages to supplement iron required by organisms, and the stability of the sucrose iron injection depends on the stability of the sucrose shells coated outside the iron cores in the medicine (solving Yu, zhang: clinical value comparison of iron isomaltose, iron sucrose and low molecular weight dextran [ J ]]Journal of clinical medicine treatment, 2021,19 (8): 29-33; jin Xuefeng, liu Liucheng, zong et al: iron-sugar complex injection similarity evaluation research progress [ J]Magazine for new medicine, china, 2018,27 (7): 797-802.). The structure of the iron sucrose is as follows:
the ferric saccharate is used as the raw material of the high molecular colloid injection, and the safety and effectiveness are closely related to the structure and composition of the ferric saccharate. Sucrose is reduced into glucose and fructose under alkaline condition, and Maillard reaction is carried out on the glucose and the fructose under alkaline condition to generate 5-hydroxymethylfurfural and furfural, and organic acid (mainly comprising glycolic acid, formic acid, malonic acid, lactic acid and acetic acid) is generated by continuous dehydration and decomposition. The route for degrading the impurity of the sucrose under the alkaline condition is as follows:
in order to ensure the safety and effectiveness of the iron sucrose products, the method for analyzing the content of the organic acid in the iron sucrose products, which is high in accuracy and good in stability, is found to have important significance for quality control, and is a technical problem to be solved in the field of quality research of the iron sucrose products at present.
Currently, chromatography is the most effective method for detecting organic acids. The prior literature does not report a detection method capable of simultaneously detecting and effectively separating five organic acids of glycolic acid, formic acid, malonic acid, lactic acid and acetic acid in iron sucrose products.
Disclosure of Invention
The invention aims to overcome the defect that the existing detection method can not detect and effectively separate five organic acids of glycolic acid, formic acid, malonic acid, lactic acid and acetic acid in the iron sucrose products. Therefore, the invention provides a method for detecting organic acid in the iron sucrose product. The detection method can eliminate matrix interference of the sample solution, the separation degree of the obtained organic acid peaks (glycolic acid, formic acid, malonic acid, lactic acid and acetic acid) meets the detection requirement, the symmetry of chromatographic peak-to-peak type is good, and the detection method has the advantages of high accuracy, good repeatability and good stability for detecting the content of the organic acid.
The invention mainly solves the technical problems through the following technical scheme.
The invention provides a method for detecting organic acid in a ferrosucrose product, which comprises the following steps:
(1) Pretreating the iron sucrose product, which comprises the following steps: mixing the iron sucrose product aqueous solution with the treatment solution, and removing the supernatant to obtain a sample solution;
the treatment liquid is an aqueous acid solution;
(2) Detecting the sample solution by adopting high performance liquid chromatography, wherein chromatographic conditions comprise:
mobile phase a is acetonitrile;
the mobile phase B is a mixed solution of phosphoric acid aqueous solution and acetonitrile;
in the mobile phase B, the volume ratio of the phosphoric acid aqueous solution to the acetonitrile is 2000:20-2000:3, and the mass concentration of the phosphoric acid aqueous solution is 0.05% -0.1%; the mass concentration refers to the percentage of phosphoric acid in the total mass of the phosphoric acid aqueous solution;
the elution conditions of the high performance liquid chromatography are as follows: the eluting time is 13-20min (the eluting time is the difference between the eluting end time and the eluting start time of the eluting stage, the eluting start time is 0 min), the mobile phase A is 0-5v% (0-5 v% refers to the ratio of the volume of the mobile phase A in the eluent to the total volume of the eluent), and the mobile phase B is 95-100v% (95-100 v% refers to the ratio of the volume of the mobile phase B in the eluent to the total volume of the eluent);
the flow rate of the mobile phase A is 0.4-0.6 mL/min;
the flow rate of the mobile phase B is 0.4-0.6 mL/min;
the iron sucrose product contains organic acid, and the organic acid at least contains glycolic acid, formic acid, malonic acid, lactic acid and acetic acid.
In the invention, the iron sucrose product can be an iron sucrose solid raw material medicine or an iron sucrose preparation, preferably an iron sucrose solid raw material medicine, and the iron sucrose solid raw material medicine is preferably an iron sucrose solid raw material medicine with the following specification: iron content is 5.3% -5.8% (wherein the content of ferrous iron is less than or equal to 0.4%), sucrose content is 82.0% -88.0%, mass ratio of iron to sucrose is 13.0% -17.0%, the content of glycolic acid is less than or equal to 0.6%, the content of formic acid is less than or equal to 0.5%, the content of malonic acid is less than or equal to 0.1%, the content of lactic acid is less than or equal to 1.0%, the content of acetic acid is less than or equal to 0.2%, and the content of other impurities (one or more of glucose, fructose, furfural and 5-hydroxymethylfurfural) is 3.8-12.7%.
In the present invention, the organic acid is composed of glycolic acid, formic acid, malonic acid, lactic acid and acetic acid.
In the pretreatment, the treatment liquid is preferably one or more of an aqueous hydrochloric acid solution, an aqueous nitric acid solution, an aqueous trifluoroacetic acid solution and an aqueous phosphoric acid solution, preferably an aqueous phosphoric acid solution.
In the pretreatment, the treatment liquid is preferably an aqueous acid solution having a mass concentration (mass concentration means a percentage of phosphoric acid to the total mass of the phosphoric acid aqueous solution) of 0.2% to 5%.
In the pretreatment, the mass to volume ratio of the iron sucrose preparation to water in the aqueous solution of the iron sucrose preparation may be 1:1 to 1000:1g/L, preferably 50:1 to 250:1g/L, more preferably 50:1g/L.
In the pretreatment, the mass-to-volume ratio of the iron sucrose preparation to the treatment liquid may be (1000:1) - (100:1) g/L, preferably (600:1) - (100:1) g/L, more preferably (500:1) g/L.
In the pretreatment, the temperature of the mixing is selected so as to sufficiently mix the aqueous solution of the iron sucrose product with the treatment liquid. The temperature of the mixing may be 10-40 ℃, preferably 20-40 ℃, more preferably 25 ℃.
In the pretreatment, the mixing time is selected so as to sufficiently mix the aqueous solution of the iron sucrose product with the treatment liquid.
In the pretreatment, the mixing may employ means conventional in the art, such as one or more of vortexing, sonication, and standing, preferably vortexing, sonication, and standing.
When vortexing is employed for the mixing, the vortexing time may be from 1 to 5 minutes, preferably from 2 to 5 minutes.
When the mixing is ultrasonic, the time of the ultrasonic may be 10-30 minutes.
When the mixing is carried out by standing, the time for the standing may be 10 to 30 minutes, preferably 15 to 30 minutes.
In the pretreatment, the mixing may be followed by a centrifugation step after which the supernatant is removed.
Preferably, the rotational speed and time of the centrifugation may be those conventional in the art, preferably 3000-6000rpm/min, more preferably 6000rpm/min. The time of the centrifugation is preferably 3 to 15min, more preferably 5 to 10min.
In a preferred embodiment of the present invention, the preprocessing is any of the following cases:
1) At room temperature, dissolving 500mg of iron sucrose product with 10mL of water, adding 1mL of 5% phosphoric acid aqueous solution, swirling for 2min, ultrasonic treating for 10min, standing for 30min, centrifuging for 5min at 6000rpm/min, and removing supernatant;
2) At room temperature, dissolving 500mg of iron sucrose product with 10mL of water, adding 1mL of 5% phosphoric acid aqueous solution, swirling for 2min, ultrasonic treating for 30min, standing for 30min, centrifuging for 5min at 6000rpm/min, and removing supernatant;
3) At room temperature, 500mg of the iron sucrose preparation was dissolved with 10mL of water, 1mL of a 5% aqueous phosphoric acid solution was added, vortexed for 2min, sonicated for 10min, left stand for 30min, centrifuged at 6000rpm/min for 10min, and the supernatant was removed.
In the present invention, the sample solution may be diluted with a diluent after step (1) and before step (2).
In the present invention, the diluent may be various kinds of diluent solvents which can dissolve iron sucrose products and have no influence on the subsequent detection, such as water, isopropanol or isopropanol aqueous solution, preferably, the volume ratio of the isopropanol aqueous solution is 10% (the volume ratio refers to the ratio of isopropanol to the total volume of water and isopropanol).
In the present invention, the mass-to-volume ratio of the iron sucrose preparation to the diluent may be (500:5000) to (500:25) g/L, preferably (500:500) g/L.
In the present invention, the chromatographic conditions further include: the chromatography column may be octadecylsilane chemically bonded silica, preferably YMC-Triart C18, waters xbridge C18 or SHIMADU Shim-pack C18, more preferably YMC-Triart C18. Preferably, the length of the chromatographic column may be 150 to 250mm, such as 250mm. The column may have an inner diameter of 3.5-4.6mm, such as 4.6mm. The particle size of the packing in the column may be 3.5-5 μm, such as 5 μm. More preferably, the chromatographic column has the following specifications: the length is 250mm, the inner diameter is 4.6mm, and the particle size of the filler is 5 mu m.
In the present invention, in the mobile phase B, the volume ratio of the phosphoric acid aqueous solution to the acetonitrile is 2000:7 to 2000:3, for example, 2000:5.
In the present invention, the elution conditions may employ non-uniform flow rates in the art. Preferably, the elution condition is a gradient elution, and the gradient elution is preferable:
0-5v% refers to the ratio of the volume of mobile phase A in the eluent to the total volume of the eluent; 95-100v% refers to the ratio of the volume of mobile phase B in the eluent to the total volume of the eluent.
The gradient elution is further preferably any of the following schemes:
scheme 1
0v% refers to the ratio of the volume of mobile phase a in the eluent to the total volume of the eluent; 100v% refers to the ratio of the volume of mobile phase B in the eluent to the total volume of the eluent;
scheme 2
1v% refers to the ratio of the volume of mobile phase a in the eluent to the total volume of the eluent; 99v% refers to the ratio of the volume of mobile phase B in the eluent to the total volume of the eluent;
scheme 3
5v% refers to the ratio of the volume of mobile phase a in the eluent to the total volume of the eluent; 95v% refers to the ratio of the volume of mobile phase B in the eluent to the total volume of the eluent;
wherein the elution time length is the difference value between the elution end time and the elution start time of the elution stage; for example, the first elution stage is 0min to 15min (including 15 min); the second elution stage is 15min (excluding 15 min) to 20min (including 20 min).
The values at both ends of "→" indicate the start and end points of the change in elution parameters (e.g., elution time, volume ratio of each mobile phase, or flow rate), e.g., "0.4→0.6" indicates that the flow rate is continuously changing from 0.4mL/min to 0.6mL/min at a constant rate during the first elution phase.
In the present invention, the elution conditions may further include further elution, and the elution conditions are:
those skilled in the art will appreciate that the first elution stage is from 0min to 15min (including 15 min); the second elution stage is 15min (excluding 15 min) to 20min (including 20 min); the third elution stage is 20min (excluding 20 min) to 25min (including 25 min); the fourth elution stage is 25min (excluding 25 min) to 30min (including 30 min); the fifth elution stage is 30min (excluding 30 min) to 33min (including 33 min); the sixth elution stage is 33min (excluding 33 min) to 38min (including 38 min);
in the elution condition, 10-90 v% indicates that in the third elution stage, the volume ratio of the mobile phase A in the eluent is continuously changed from 10% to 90% at a constant speed; correspondingly, 90-10v% represents that the mobile phase B continuously and uniformly changes from 90% to 10%; 1.0.fwdarw.1.5 means that the flow rates of mobile phase A and mobile phase B continuously and uniformly vary from 1.0mL/min to 1.5mL/min.
In the detection method of the present invention, the detection wavelength may be 200 to 220nm, preferably 210nm.
In the present invention, the column temperature of the chromatographic column may be 33 to 37 ℃, preferably 35 ℃.
In the detection method of the present invention, the sample injection amount may be selected as usual in the art, and may be 10 to 100. Mu.L, preferably 70. Mu.L.
In a certain preferred scheme, the treatment liquid is phosphoric acid aqueous solution with the mass concentration of 5%;
diluting the sample solution with a diluent after the step (1) and before the step (2), wherein the diluent is an isopropanol water solution, and the volume ratio of the isopropanol water solution is 10%;
the chromatographic conditions are as follows: the chromatographic column is YMC-Triart C18 chromatographic column; the specification of the chromatographic column is as follows: the length is 250mm, the inner diameter is 4.6mm, and the particle size of the filler is 5 mu m; the detection wavelength is 210nm; the column temperature of the chromatographic column is 35 ℃;
the mobile phase A is acetonitrile;
the mobile phase B is a mixed solution of phosphoric acid aqueous solution and acetonitrile; the volume ratio of the phosphoric acid aqueous solution to the acetonitrile is 2000:5; the mass concentration of the phosphoric acid aqueous solution is 0.05% or 0.1%;
the elution conditions are as follows:
in the present invention, the concentration of the test solution (the concentration refers to the ratio of the mass of the iron sucrose preparation to the total volume of the test solution) may be 1 to 20mg/mL, preferably 2.0mg/mL.
In the invention, the solvent in the test solution is various solvents which can dissolve iron sucrose products and have no influence on subsequent detection, such as isopropanol water solution.
In the present invention, the concentration of each organic acid in the sample solution may be a concentration conventional in the art, for example: 2-20 mug/mL.
In the invention, the iron sucrose product is treated according to the method for detecting the organic acid in the iron sucrose product, and the eluent with the retention time of 7-13 min is collected to obtain the glycolic acid, the formic acid, the malonic acid, the lactic acid and the acetic acid.
In the invention, the retention time of the glycolic acid is 6.2-7.9min, preferably 7.5min; the retention time of the formic acid is 6.5-8.2min, preferably 7.8min; the retention time of the malonic acid is 8.3-10.5min, preferably 10.1min; the retention time of the lactic acid is 9.4-11.9min, preferably 11.0min; the retention time of the acetic acid is 10.2 to 12.2min, preferably 11.7min.
In the present invention, the acetonitrile may be acetonitrile solvents commonly used in the art, and generally refers to acetonitrile having a purity of 99.9% or more.
In the present invention, the isopropyl alcohol may be an isopropyl alcohol solvent commonly used in the art, and generally refers to isopropyl alcohol having a purity of 99.9% or more.
In the present invention, the solvent of the phosphoric acid aqueous solution in the mobile phase B is generally referred to as ultrapure water, as known to those skilled in the art.
On the basis of conforming to the common knowledge in the field, the above preferred conditions can be arbitrarily combined to obtain the preferred examples of the invention.
The reagents and materials used in the present invention are commercially available.
The invention has the positive progress effects that: the detection method can eliminate matrix interference of the sample solution, the separation degree of each obtained organic acid peak (glycolic acid, formic acid, malonic acid, lactic acid and acetic acid) meets the detection requirement, the symmetry of chromatographic peak type is good, and the detection method has high accuracy, good repeatability and good stability for detecting the content of each organic acid.
Detailed Description
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
The test object adopted by the invention is solid iron sucrose bulk drug, and the specification is as follows: iron content is 5.3% -5.8% (wherein the content of ferrous iron is less than or equal to 0.4%), sucrose content is 82.0% -88.0%, mass ratio of iron to sucrose is 13.0% -17.0%, the content of glycolic acid is less than or equal to 0.6%, the content of formic acid is less than or equal to 0.5%, the content of malonic acid is less than or equal to 0.1%, the content of lactic acid is less than or equal to 1.0%, the content of acetic acid is less than or equal to 0.2%, and the content of other impurities (one or more of glucose, fructose, furfural and 5-hydroxymethylfurfural) is 3.8-12.7%.
Example 1
Preparing and feeding amount of the mixed organic acid solution: respectively weighing formic acid, acetic acid, glycolic acid, malonic acid and lactic acid at room temperature, placing in the same measuring flask (100 mL), diluting with water to scale, and shaking to obtain reference stock solution 1; precisely measuring 5mL of reference substance stock solution 1, placing in a 50mL measuring flask, diluting with water to scale, and shaking to obtain reference substance stock solution 2; precisely weighing 2mL of reference substance stock solution 2, placing in a 100mL measuring flask, adding 10% isopropanol water solution to dissolve and dilute to scale, and shaking uniformly to obtain reference substance solution of mixed organic acid (glycolic acid concentration is 0.012mg/mL, formic acid concentration is 0.01mg/mL, malonic acid concentration is 0.002mg/mL, lactic acid concentration is 0.02mg/mL, and acetic acid concentration is 0.004 mg/mL). 70. Mu.L of the solution was precisely removed and injected into a liquid chromatograph.
Sample solution preparation and sample injection amount: at room temperature, weighing 500mg of iron sucrose solid bulk drug, adding 10mL of water for dissolution, precisely adding 1mL of 5% phosphoric acid aqueous solution, wherein the mass volume ratio of the iron sucrose solid bulk drug to the 5% phosphoric acid aqueous solution is (500:1), swirling for 2min, fixing the volume to a scale (25 mL) by using water, performing ultrasonic treatment for 10min, standing for 30min, centrifuging for 5min at 600 rpm/min, precisely transferring 1mL to 10mL of measuring flask, adding 10% isopropanol aqueous solution for dilution to form 2.0mg/mL of solution, thus obtaining the sample solution of the iron sucrose solid bulk drug, and precisely transferring 70 mu L of sample solution into a liquid chromatograph.
The control solution mixed with the organic acid and the test solution of the solid iron sucrose bulk drug are eluted by adopting the following chromatographic conditions:
instrument: agilent 1260Infinity II
Chromatographic column: YMC-Triart C18.6X250 mm,5 μm;
mobile phase a is acetonitrile;
mobile phase B is a mixed solution of 0.05% phosphoric acid aqueous solution and acetonitrile; the volume ratio of the 0.05% phosphoric acid aqueous solution to the acetonitrile is 2000:5;
column temperature: 35 ℃;
detection wavelength: 210nm;
the procedure for gradient elution is as follows:
the detection result shows that the retention time of each organic acid (glycolic acid, formic acid, malonic acid, lactic acid and acetic acid) in the reference substance solution of the mixed organic acid under the existing chromatographic condition is 7-12 min. The retention time of glycolic acid was 7.550min; the retention time of formic acid was 7.950min; the retention time of malonic acid is 10.235min; the retention time of lactic acid was 11.184min; the retention time of acetic acid is 11.984min, the separation degree of each organic acid meets the requirement (more than or equal to 1.5), and each peak is good.
In the pretreated sample solution, the sample solution does not have matrix interference on detection of each organic acid, and the retention time of the glycolic acid is 7.577min; the retention time of formic acid was 7.982min; malonic acid was not detected; the retention time of lactic acid was 11.253min; the retention time of acetic acid is 12.039min, the separation degree of organic acid in the sample solution is good, the peak types of the peaks are symmetrical, and the detection requirement is met.
Example 2
Sample preparation and sample introduction amount: at room temperature, weighing 500mg of iron sucrose solid raw material medicine, adding 10mL of water for dissolution, precisely adding 1mL of 5% phosphoric acid aqueous solution, wherein the mass volume ratio of the iron sucrose solid raw material medicine to the 5% phosphoric acid aqueous solution is (500:1), swirling for 2min, fixing the volume of the aqueous solution to a scale (25 mL), performing ultrasonic treatment for 30min, standing for 30min, centrifuging for 5min at 600 rpm/min, precisely transferring into a 1 mL-10 mL measuring flask, and adding 10% isopropanol aqueous solution to dilute to obtain 1.0mg/mL solution. 70. Mu.L of the solution was precisely removed and injected into a liquid chromatograph.
Other detection conditions were the same as in example 1.
The chromatographic conditions were the same as in example 1, except that the ultrasonic time during pretreatment was changed from 10min to 30min. The results show that the substrate of the solution of the test sample does not interfere with the detection of the organic acid, and the retention time of the glycolic acid is 7.576min; the retention time of formic acid was 7.975min; malonic acid was not detected; the retention time of lactic acid was 11.225min; the retention time of acetic acid is 12.006min, the separation degree is good, the peak types of the organic acids are symmetrical, and the measurement requirement is still met.
Example 3
Sample solution preparation and sample injection amount: at room temperature, weighing 500mg of iron sucrose solid raw material medicine, adding 10mL of water for dissolution, precisely adding 1mL of 5% phosphoric acid aqueous solution for dissolution, wherein the mass volume ratio of the iron sucrose solid raw material medicine to the 5% phosphoric acid aqueous solution is (500:1), swirling for 2min, fixing the volume to a scale (25 mL) by using water, performing ultrasonic treatment for 10min, standing for 30min, centrifuging at 3000rpm/min for 10min, precisely transferring 1mL to 10mL measuring flask, and adding 10% isopropanol aqueous solution for dilution to obtain 1.0mg/mL solution. 70. Mu.L of the solution was precisely removed and injected into a liquid chromatograph.
Other detection conditions were the same as in example 1.
The chromatographic conditions were the same as in example 1, except that the centrifugation time in the pretreatment was changed from 5min to 10min. The result shows that the substrate of the solution of the test sample does not interfere with the detection of the organic acid, and the retention time of the glycolic acid is 7.564min; the retention time of formic acid was 7.963min; malonic acid was not detected; the retention time of lactic acid was 11.208min; the retention time of acetic acid is 11.988min, the separation degree of each organic acid is good, the peak type is symmetrical, and the detection requirement can be met.
Example 4
Mobile phase B is a mixed solution of 0.1% phosphoric acid aqueous solution and acetonitrile; the volume ratio of the 0.1% phosphoric acid aqueous solution to the acetonitrile is (2000:5);
other detection conditions were the same as in example 1.
The results showed that the retention time of each organic acid (glycolic acid, formic acid, malonic acid, lactic acid, acetic acid) in the mixed organic acid control solution was 7 to 12min. The retention time of glycolic acid was 7.610min; the retention time of formic acid was 7.983min; the retention time of malonic acid is 10.237min; the retention time of lactic acid was 10.880min; the retention time of acetic acid was 11.607min. The organic acids have good separation degree (the minimum separation degree is 1.57), the symmetry factor is 1.07-1.17, and the peak-to-peak type is good.
The substrate of the solution of the test sample does not interfere with the detection of each organic acid, wherein the retention time of the glycolic acid is 7.620min; the retention time of formic acid was 7.967min; malonic acid was not detected; the retention time of lactic acid was 10.943min; the retention time of acetic acid is 11.627min, the peak types of the organic acids are symmetrical, and the separation degree is good.
Therefore, the mass concentration of the phosphoric acid aqueous solution in the mobile phase B was changed from 0.05% to 0.1%, and the results showed that the conditions still satisfied the measurement requirements.
EXAMPLE 5 screening of treatment fluid in pretreatment step
The procedure for preparing the sample solution was the same as in example 1, and the type or amount of the treatment solution in the pretreatment step was adjusted only, and the detection was carried out using the chromatographic conditions of example 1.
The detection result shows that the organic acid in 500mg of iron sucrose solid bulk drug is fully dissolved, and the use amount of various acid aqueous solutions with the mass concentration of 5% is not less than 1mL.
Comparative example 1
Gas chromatography:
chromatographic column: agilent 123-7033 DB-WAX 30m,0.32 μm,0.5 μm
Carrier gas: nitrogen gas
Flow rate: 1.5mL/min
Sample inlet temperature: 200 DEG C
Detector temperature: 220 DEG C
Split ratio: 10:1
Sample injection amount: 1 mu L
Heating program
| Rate of temperature rise | Temperature (. Degree. C.) | Retention time (min) |
| 0 | 40 | 2 |
| 5 | 150 | 5 |
| 10 | 200 | 0 |
Preparing a reference substance solution of mixed organic acid: respectively weighing formic acid, acetic acid, glycolic acid, malonic acid and lactic acid, respectively placing into measuring bottles, adding 10% methanol for dissolution, diluting to scale, and shaking to obtain each organic acid stock solution. And respectively transferring 1mL of each stock solution into the same measuring flask, and uniformly mixing to obtain the reference substance solution of the mixed organic acid.
In the existing GC conditions, the control solution only detects that the acetic acid retention time is 4.856min; the retention time of the glycolic acid is 5.953min; malonic acid retention time was 18.626min; the retention time of lactic acid was 20.407min for four organic acids, but formic acid could not be detected, and thus the requirements for five target organic acid determinations could not be met.
Comparative example 2
Liquid chromatography
Chromatographic column: a chromatographic column using octadecylsilane chemically bonded silica as a filler; SHIMADZU Shim-pack gist C18-AQ 4.6 x 250mm,5 μm
Mobile phase A is acetonitrile (the purity of acetonitrile is above 99.9 percent);
mobile phase B was a 0.05% aqueous phosphoric acid-acetonitrile mixture (0.05% aqueous phosphoric acid: acetonitrile=1000:5).
The gradient elution procedure was as follows:
column temperature: 35 ℃;
flow rate: 0.5mL/min;
detection wavelength: 210nm.
Sample solution preparation and sample injection amount: weighing a test sample, adding 3mL of isopropanol water solution with the volume ratio of 10% for vortex dissolution, adding 0.3mL of orthophosphoric acid (namely undiluted phosphoric acid solution) for vortex for 1.5min, carrying out ultrasonic treatment for 45min, and adding the isopropanol water solution with the volume ratio of 10% for dilution to obtain 1.0mg/mL solution. Precisely removing 20 mu L of the solution and injecting the solution into a liquid chromatograph;
preparing a control solution of mixed organic acid and feeding amount: respectively weighing formic acid, acetic acid, glycolic acid, malonic acid and lactic acid, respectively placing into a measuring flask, adding water for dissolution, diluting to scale, and shaking to obtain each organic acid stock solution. And respectively transferring 1mL of each stock solution into the same measuring flask, and uniformly mixing to obtain the reference substance solution of the mixed organic acid.
As can be seen from the chromatographic results of the control solution of mixed organic acids obtained by the detection method of the comparative example, in the existing conditions, the collection retention time of each organic acid is 6-11 min, wherein the retention time of glycolic acid is 6.168min; the retention time of formic acid was 6.585min; the retention time of malonic acid is 8.352min; the retention time of lactic acid was 9.500min; the retention time of acetic acid is 10.216min, the separation degree between organic acids meets the requirement, and the peak type of each organic acid is good;
after pretreatment of the sample solution, the substrate of the sample solution interferes with the determination of the organic acid, and the five organic acids cannot be effectively separated, so that the method does not meet the requirements of the determination of the five target organic acids.
Comparative example 3
Sample solution preparation and sample injection amount: at room temperature, weighing 500mg of solid iron sucrose raw material, adding 10mL of water for dissolution, metering the volume to a scale (the scale is 25 mL) by using water, precisely transferring the solution into a 1 mL-10 mL measuring flask, adding 10% by volume of isopropanol aqueous solution for dilution to form 1.0mg/mL solution, obtaining a sample solution of the solid iron sucrose raw material, precisely transferring 70 mu L of the sample solution, and injecting the sample solution into a liquid chromatograph.
The chromatographic conditions of example 1 were used for the test.
The chromatographic result shows that the aqueous solution of the solid iron sucrose bulk drug is not pretreated, the obtained sample solution matrix interferes with the detection of organic acid, and the peak types of the organic acid are asymmetric. Therefore, the sample solution which is not pretreated cannot meet the requirements of five target organic acid determination.
Comparative example 4
Liquid chromatography
Instrument: thermo Ultimate3000
Chromatographic column: YMC-Triart C18.6X250 mm,5 μm;
mobile phase a is acetonitrile;
mobile phase B is 0.1% phosphoric acid aqueous solution;
column temperature: 35 ℃;
detection wavelength: 210nm;
the procedure for gradient elution is as follows:
column temperature: 35 ℃;
flow rate: 1.0mL/min;
detection wavelength: 210nm.
The test solution and the control solution were prepared in the same manner as in example 1
Under the existing chromatographic conditions, the retention time of glycolic acid in the reference solution is 3.090min, the retention time of formic acid is 3.370min, the retention time of malonic acid is 3.763min, the retention time of lactic acid is 3.970min, and the retention time of acetic acid is 4.277min. Wherein the separation degree of malonic acid and lactic acid is 1.42, and the peak type of malonic acid is asymmetric.
Under the existing chromatographic condition, the retention time of the glycolic acid in the sample solution is 3.090min; the retention time of formic acid was 3.373min; malonic acid was not detected, the retention time of lactic acid was 4.027min; the retention time of acetic acid was 4.290min and the matrix interfered with the detection of each organic acid.
Therefore, when the mobile phase B is 0.1% phosphoric acid aqueous solution and the initial ratio of the mobile phase B to the mobile phase A is (95:5) phosphoric acid aqueous solution, the detection requirements of the product on five target organic acids are not met.
Comparative example 5
Instrument: thermo Ultimate3000
Chromatographic column: YMC-Triart C18.6X250 mm,5 μm;
mobile phase a is acetonitrile;
mobile phase B is 0.1% phosphoric acid aqueous solution
Column temperature: 35 ℃;
detection wavelength: 210nm;
the procedure for gradient elution is as follows:
column temperature: 35 ℃;
detection wavelength: 210nm.
The test solution and the control solution were prepared in the same manner as in example 1.
Under the existing chromatographic conditions, the retention time of glycolic acid in the reference substance solution is 6.883min, the retention time of formic acid is 7.453min, the retention time of malonic acid is 8.183min, the retention time of lactic acid is 8.667min, the retention time of acetic acid is 9.233min, the separation degree between organic acids meets the requirement (more than or equal to 1.5), and the peak type of each organic acid is good.
The retention time of the glycolic acid in the sample solution under the existing chromatographic condition is 6.890min; the retention time of formic acid was 7.457min; malonic acid was not detected, the retention time of lactic acid was 8.773min; the retention time of acetic acid was 9.257min and the substrate interfered with the detection of organic acids.
Therefore, when the mobile phase B is 0.1% phosphoric acid aqueous solution and the initial ratio of the mobile phase B to the mobile phase A is (95:5), the detection requirements of the product on five target organic acids are still not met under the condition of changing the flow velocity gradient.
Comparative example 6
Instrument: thermo Ultimate3000
Chromatographic column: YMC-Triart C18.6X250 mm,5 μm;
mobile phase a is acetonitrile;
mobile phase B is 0.1% phosphoric acid aqueous solution
Column temperature: 35 ℃;
detection wavelength: 210nm;
the procedure for gradient elution is as follows:
column temperature: 35 ℃;
detection wavelength: 210nm.
The test solution and the control solution were prepared in the same manner as in example 1
Under the existing chromatographic conditions, the retention time of each organic acid (glycolic acid, formic acid, malonic acid, lactic acid and acetic acid) in the reference substance solution is 6.8-9.4 min. The retention time of glycolic acid is 6.883min, the retention time of formic acid is 7.453min, the retention time of malonic acid is 8.203min, the retention time of lactic acid is 8.693min, the retention time of acetic acid is 9.417min, the separation degree among organic acids meets the requirement (more than or equal to 1.5), and the peak-to-peak type is good.
Under the existing chromatographic conditions, the retention time of each organic acid (glycolic acid, formic acid, malonic acid, lactic acid and acetic acid) in the sample solution is 6.8-9.4 min. The retention time of glycolic acid was 6.890min; the retention time of formic acid was 7.453min; malonic acid was not detected, the retention time of lactic acid was 8.780min; the retention time of acetic acid was 9.440min, each peak-to-peak type was good, but the matrix still interfered with the detection of organic acids.
Therefore, the mobile phase B is 0.1% phosphoric acid aqueous solution, the ratio of the mobile phase B to the mobile phase A is (99:1), and the detection requirements of the product on five target organic acids cannot be met.
Claims (12)
1. The method for detecting the organic acid in the iron sucrose product comprises the following steps:
(1) Pretreating the iron sucrose product, which comprises the following steps: mixing the iron sucrose product aqueous solution with the treatment solution, and removing the supernatant to obtain a sample solution;
the treatment liquid is an aqueous acid solution;
(2) Detecting the sample solution by adopting high performance liquid chromatography, wherein chromatographic conditions comprise:
mobile phase a is acetonitrile;
the mobile phase B is a mixed solution of phosphoric acid aqueous solution and acetonitrile;
in the mobile phase B, the volume ratio of the phosphoric acid aqueous solution to the acetonitrile is 2000:20-2000:3, and the mass concentration of the phosphoric acid aqueous solution is 0.05% -0.1%; the mass concentration refers to the percentage of phosphoric acid in the total mass of the phosphoric acid aqueous solution;
the elution conditions of the high performance liquid chromatography are as follows: the eluting time is 13-20min, the mobile phase A is 0-5v%, and 0-5v% refers to the ratio of the volume of the mobile phase A in the eluent to the total volume of the eluent; the mobile phase B is 95-100v%, and 95-100v% refers to the ratio of the volume of the mobile phase B in the eluent to the total volume of the eluent;
the flow rate of the mobile phase A is 0.4-0.6 mL/min;
the flow rate of the mobile phase B is 0.4-0.6 mL/min;
the iron sucrose product contains organic acid, and the organic acid at least contains glycolic acid, formic acid, malonic acid, lactic acid and acetic acid.
2. The method for detecting organic acid in iron sucrose products according to claim 1, wherein one or more of the following conditions are satisfied:
(1) The iron sucrose product is an iron sucrose bulk drug or an iron sucrose preparation; the iron sucrose solid raw material medicine is preferably selected from the following specifications: iron content is 5.3-5.8%, sucrose content is 82.0-88.0%, mass ratio of iron to sucrose is 13.0-17.0%, hydroxyacetic acid content is less than or equal to 0.6%, formic acid content is less than or equal to 0.5%, malonic acid content is less than or equal to 0.1%, lactic acid content is less than or equal to 1.0%, acetic acid content is less than or equal to 0.2%,
the content of other impurities is 3.8-12.7%;
(2) The organic acid consists of glycolic acid, formic acid, malonic acid, lactic acid and acetic acid.
3. The method for detecting organic acid in iron sucrose products according to claim 1, wherein one or more of the following conditions are satisfied:
(1) In the pretreatment, the treatment liquid is hydrochloric acid aqueous solution, nitric acid aqueous solution,
One or more of an aqueous trifluoroacetic acid solution and an aqueous phosphoric acid solution, preferably an aqueous phosphoric acid solution;
(2) In the pretreatment, the treatment liquid is an aqueous solution of an acid with a mass concentration of 0.2% -5%;
(3) In the pretreatment, the mass-to-volume ratio of the iron sucrose product to water in the aqueous solution of the iron sucrose product is 1:1-1000:1g/L, preferably 50:1-250:1g/L, more preferably 50:1g/L;
(4) In the pretreatment, the mass-to-volume ratio of the iron sucrose product to the treatment liquid is (1000:1) - (100:1) g/L, preferably (600:1) - (100:1) g/L, more preferably (500:1) g/L;
(5) In the pretreatment, the temperature of the mixing is 10-40 ℃, preferably 20-40 ℃,
more preferably 25 ℃;
(6) In the pretreatment, the mixing adopts one or more of vortex, ultrasonic and standing;
(7) In the pretreatment, the mixing is followed by a centrifugation step after which the supernatant is removed.
4. A method for detecting organic acids in iron sucrose products according to claim 3, characterized in that it fulfils one or more of the following conditions:
(1) When the mixing is by vortexing, the vortexing time is 1-5min, preferably 2-5min;
(2) When the mixing adopts ultrasonic, the ultrasonic time is 10-30min;
(3) When the mixing is carried out by standing, the standing time is 10-30min, preferably 15-30min;
(4) The rotational speed of the centrifugation is 3000-6000rpm/min;
(5) The centrifugation time is 3-15min, preferably 5-10min.
5. The method for detecting organic acid in iron sucrose products according to claim 1, wherein the pretreatment is any of the following conditions:
(1) At room temperature, dissolving 500mg of iron sucrose product with 10mL of water, adding 1mL of 5% phosphoric acid aqueous solution, swirling for 2min, ultrasonic treating for 10min, standing for 30min, centrifuging for 5min at 6000rpm/min, and removing supernatant;
(2) At room temperature, dissolving 500mg of iron sucrose product with 10mL of water, adding 1mL of 5% phosphoric acid aqueous solution, swirling for 2min, ultrasonic treating for 30min, standing for 30min, centrifuging for 5min at 6000rpm/min, and removing supernatant;
(3) At room temperature, 500mg of the iron sucrose preparation was dissolved with 10mL of water, 1mL of a 5% aqueous phosphoric acid solution was added, vortexed for 2min, sonicated for 10min, left stand for 30min, centrifuged at 6000rpm/min for 10min, and the supernatant was removed.
6. The method for detecting organic acids in iron sucrose products according to claim 1, wherein the sample solution is diluted with a diluent after step (1) and before step (2), said dilution satisfying one or more of the following conditions:
(1) The diluent is water, isopropanol or isopropanol water solution, preferably the volume ratio of the isopropanol water solution is 10%;
(2) The mass volume ratio of the iron sucrose product to the diluent is (500:5000) - (500:25) g/L, preferably (500:500) g/L.
7. The method for detecting organic acids in iron sucrose products as set forth in claim 1 wherein said chromatographic conditions further include: the chromatographic column is octadecylsilane chemically bonded silica chromatographic column, preferably YMC-Triart C18 chromatographic column, waters xbridge C18 chromatographic column or SHIMDU Shim-pack C18 chromatographic column, more preferably YMC-Triart C18 chromatographic column;
preferably, the chromatographic column also satisfies one or more of the following conditions:
(1) The length of the chromatographic column is 150-250 mm, such as 250mm;
(2) The inner diameter of the chromatographic column is 3.5-4.6mm, such as 4.6mm;
(3) The particle size of the filler in the chromatographic column is 3.5-5 μm, such as 5 μm;
more preferably, the chromatographic column has the following specifications: the length is 250mm, the inner diameter is 4.6mm, and the particle size of the filler is 5 mu m.
8. The method for detecting organic acids in iron sucrose products according to claim 1, wherein the volume ratio of phosphoric acid aqueous solution to acetonitrile in mobile phase B is 2000:7-2000:3, such as 2000:5.
9. The method for detecting organic acids in iron sucrose products according to claim 1, wherein the elution conditions are non-uniform elution, preferably gradient elution, preferably:
0-5v% refers to the ratio of the volume of mobile phase A in the eluent to the total volume of the eluent; 95-100v% refers to the ratio of the volume of mobile phase B in the eluent to the total volume of the eluent;
the gradient elution is further preferably any of the following schemes:
scheme 1
0v% refers to the ratio of the volume of mobile phase a in the eluent to the total volume of the eluent; 100v% refers to the ratio of the volume of mobile phase B in the eluent to the total volume of the eluent;
scheme 2
1v% refers to the ratio of the volume of mobile phase a in the eluent to the total volume of the eluent; 99v% refers to the ratio of the volume of mobile phase B in the eluent to the total volume of the eluent;
scheme 3
5v% refers to the ratio of the volume of mobile phase a in the eluent to the total volume of the eluent; 95v% refers to the ratio of the volume of mobile phase B in the eluent to the total volume of the eluent;
"0.4.fwdarw.0.6" means that the flow rate was continuously constant from 0.4mL/min to 0.6mL/min during the first elution phase.
10. The method for detecting organic acids in iron sucrose products according to claim 1, wherein the detection method satisfies one or more of the following conditions:
(1) The detection wavelength is 200-220nm, preferably 210nm;
(2) The column temperature of the chromatographic column is 33-37 ℃, preferably 35 ℃;
(3) The sample amount is 10-100. Mu.L, preferably 70. Mu.L.
11. The method for detecting organic acid in iron sucrose products according to claim 1, wherein in the detection method, the treatment liquid is a phosphoric acid aqueous solution with a mass concentration of 5%;
diluting the sample solution with a diluent after the step (1) and before the step (2), wherein the diluent is an isopropanol water solution, and the volume ratio of the isopropanol water solution is 10%;
the chromatographic conditions are as follows: the chromatographic column is YMC-Triart C18 chromatographic column; the specification of the chromatographic column is as follows: the length is 250mm, the inner diameter is 4.6mm, and the particle size of the filler is 5 mu m; the detection wavelength is 210nm; the column temperature of the chromatographic column is 35 ℃;
the mobile phase A is acetonitrile;
the mobile phase B is a mixed solution of phosphoric acid aqueous solution and acetonitrile; the volume ratio of the phosphoric acid aqueous solution to the acetonitrile is 2000:5; the mass concentration of the phosphoric acid aqueous solution is 0.05% or 0.1%;
the elution conditions are as follows:
12. the method for detecting organic acids in iron sucrose products according to claim 1, wherein the sample solution satisfies one or more of the following conditions:
(1) The concentration of the sample solution is 1-20mg/mL, preferably 2.0mg/mL;
(2) The solvent in the sample solution is isopropanol water solution;
(3) The concentration of each organic acid in the test solution is 2-20 mug/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211728370.8A CN116223657B (en) | 2022-12-30 | 2022-12-30 | Method for detecting organic acid in iron sucrose product |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211728370.8A CN116223657B (en) | 2022-12-30 | 2022-12-30 | Method for detecting organic acid in iron sucrose product |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116223657A true CN116223657A (en) | 2023-06-06 |
| CN116223657B CN116223657B (en) | 2023-09-15 |
Family
ID=86575933
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211728370.8A Active CN116223657B (en) | 2022-12-30 | 2022-12-30 | Method for detecting organic acid in iron sucrose product |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116223657B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018043240A1 (en) * | 2016-09-02 | 2018-03-08 | Ｓｂｉファーマ株式会社 | Aqueous formulation containing 5-aminolevulinic acid or the like |
| WO2019189717A1 (en) * | 2018-03-30 | 2019-10-03 | 持田製薬株式会社 | Method for producing tetrahydronaphthylurea derivative |
| WO2021060395A1 (en) * | 2019-09-25 | 2021-04-01 | 持田製薬株式会社 | Sustained-release pharmaceutical composition |
| WO2022140654A1 (en) * | 2020-12-23 | 2022-06-30 | The Broad Institute, Inc. | Compositions and methods related to bicyclo[2.2.1] heptanamine-containing compounds |
-
2022
- 2022-12-30 CN CN202211728370.8A patent/CN116223657B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018043240A1 (en) * | 2016-09-02 | 2018-03-08 | Ｓｂｉファーマ株式会社 | Aqueous formulation containing 5-aminolevulinic acid or the like |
| WO2019189717A1 (en) * | 2018-03-30 | 2019-10-03 | 持田製薬株式会社 | Method for producing tetrahydronaphthylurea derivative |
| WO2021060395A1 (en) * | 2019-09-25 | 2021-04-01 | 持田製薬株式会社 | Sustained-release pharmaceutical composition |
| WO2022140654A1 (en) * | 2020-12-23 | 2022-06-30 | The Broad Institute, Inc. | Compositions and methods related to bicyclo[2.2.1] heptanamine-containing compounds |
Non-Patent Citations (2)
| Title |
|---|
| KEIJI GAMOH等: "Improved liquid chromatography/mass spectrometric analysis of low molecular weight carboxylic acids by ion exclusion separation with electrospray ionization", RAPID COMMUNICATIONS IN MASS, pages 685 - 689 * |
| 袁启凤等: "百香果品种‘紫香1号’果实糖、酸和维生素成分分析", 中国果树, vol. 4, pages 43 - 47 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116223657B (en) | 2023-09-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113049699B (en) | Method for detecting biphenyl anhydride and related substances thereof and application | |
| CN102293772A (en) | Compound injection, and preparation method and quality control method thereof | |
| CN110715995A (en) | Method for detecting impurities of multi-vitamin injection | |
| CN116223657B (en) | Method for detecting organic acid in iron sucrose product | |
| CN103833773A (en) | Cefathiamidine compound | |
| CN111239265B (en) | Liquid chromatography method for determining clavulanic acid related substances in amoxicillin and potassium clavulanate pharmaceutical composition | |
| CN102070540B (en) | 1-oxygen-2-(1-ethoxy)-3-methyl-quinoxaline, and preparation method and application thereof | |
| CN113820417A (en) | Method for separating and measuring piroxicam and impurities thereof | |
| CN109946398B (en) | Method for detecting dalbavancin and impurities thereof | |
| CN109917045B (en) | HPLC method for simultaneously measuring contents of 5 components in prepared rhizoma cibotii decoction pieces | |
| CN111077235B (en) | Method for determining 2- [ (2-methyl-5-bromophenyl) methyl ] -5- (4-fluorophenyl) thiophene impurity | |
| CN111855841B (en) | Method for measuring related substances in voglibose raw material and preparation | |
| CN111521714B (en) | Method for separating and measuring capecitabine and impurities thereof | |
| CN104130270B (en) | Celbenin crystal formulations and preparation method thereof | |
| CN103724359B (en) | A kind of amorphous cefotetan acid and prepared the method for Cefotetan Disodium and containing the pharmaceutical composition of this Cefotetan Disodium by it | |
| CN108152432A (en) | A kind of Luo Tafuxin pieces efficient liquid phase detection method | |
| CN110530997B (en) | Method for detecting 9- (2-hydroxypropyl) adenine enantiomer | |
| CN113125608B (en) | Impurity detection method for edaravone sodium chloride injection | |
| CN110627771B (en) | Preparation method of esomeprazole thioether | |
| CN116840378B (en) | Method for detecting content of monohydrate of ropivacaine/meloxicam salt | |
| CN116973473A (en) | Related substance detection method of felodipine | |
| CN111018804A (en) | Preparation method of amoxicillin impurity E | |
| CN108658905B (en) | Method for reducing related substances in potassium sodium dehydroandroan drographolide succinate raw material medicine finished product | |
| CN110850012B (en) | Detection method of 1- (2, 3-dichlorophenyl) piperazine hydrochloride and related substances thereof | |
| CN105906559B (en) | A kind of one-step synthesis of pharmaceutical grade metadoxine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |