CN112630331A - Method for measuring content of 3-hydroxy-2-butanone by high performance liquid chromatography - Google Patents
Method for measuring content of 3-hydroxy-2-butanone by high performance liquid chromatography Download PDFInfo
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- CN112630331A CN112630331A CN202011462211.9A CN202011462211A CN112630331A CN 112630331 A CN112630331 A CN 112630331A CN 202011462211 A CN202011462211 A CN 202011462211A CN 112630331 A CN112630331 A CN 112630331A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention provides a method for measuring the content of 3-hydroxy-2-butanone by high performance liquid chromatography, which comprises the following steps: (1) preparing a test solution, (2) preparing a reference solution, (3) injecting each test solution and each reference solution into a high performance liquid chromatograph to respectively obtain chromatograms, and (4): the content of 3-hydroxy-2-butanone is calculated by peak area according to an external standard method. Compared with gas chromatography, the method has the characteristics of low cost, short time consumption and simple operation, and is beneficial to popularization and application; proved by methodology, the method provided by the invention is accurate and reliable and has good stability; the method can realize effective separation of the main peak of the 3-hydroxy-2-butanone and the impurity peaks nearby the main peak; the invention can better realize the quality control of the 3-hydroxy-2-butanone.
Description
Technical Field
The invention relates to the field of drug detection, in particular to a method for determining the content of 3-hydroxy-2-butanone by high performance liquid chromatography.
Background
3-hydroxy-2-butanone (C)4H8O2) Also called acetoin and methyl acetyl methanol, the monomer is colorless or light yellow liquid, and the dimer is white crystalline powder, so the cream has pleasant cream fragrance. Acetoin has a low melting point of only 15 ℃, so the dimer becomes liquid when left at room temperature. Its boiling point is 148 deg.C, it can be spontaneously combusted, and can be dissolved in water, ethanol, and propylene glycol. At present, acetoin is widely applied to the fields of medicine, agriculture, chemical industry, food and the like.
Acetoin is an important pharmaceutical intermediate in the pharmaceutical industry. Acetoin and phosgene can be used for synthesizing an important medical intermediate 4-chloro-4, 5-dimethyl-1, 3-dioxolane-2-one (CDMDO), and the CDMDO is used for modifying antibiotic drugs and plays a role in the synthesis of chiral drugs. Acetoin can also be used for synthesizing antibacterial drug of enkexin hydrochloride.
The detection of 3-hydroxy-2-butanone in the food field is reported, for example, the content of 3-hydroxy-2-butanone in wine, fermentation liquor, mature vinegar, bran koji and the like is detected, but these methods cannot effectively separate the 3-hydroxy-2-butanone peak and impurity peaks in the vicinity thereof, and these methods are only suitable for detecting substances with low content of 3-hydroxy-2-butanone. For the detection of high-purity 3-hydroxy-2-butanone, although the China light industry standard QB/T4234-2011 provides a method for detecting the 3-hydroxy-2-butanone by using a gas chromatography, the gas chromatography has the characteristics of high cost, long time consumption and complex operation, and is not beneficial to popularization and application.
Disclosure of Invention
The invention aims to provide a method for measuring the content of 3-hydroxy-2-butanone by using a high performance liquid chromatography, which can effectively separate a main peak of 3-hydroxy-2-butanone and impurity peaks nearby the main peak, is accurate and reliable, has good stability, has the characteristics of low cost, short time consumption and simple operation, and is beneficial to popularization and application.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a method for measuring the content of 3-hydroxy-2-butanone by high performance liquid chromatography comprises the following steps:
(1) preparation of a test solution: taking a proper amount of the product, precisely weighing, adding a mobile phase for dissolving, and quantitatively diluting to prepare a solution containing about 1mg in each 1 ml;
(2) preparation of control solutions: taking a proper amount of 3-hydroxy-2-butanone reference substance, precisely weighing, adding a mobile phase for dissolving, and quantitatively diluting to prepare a solution containing about 1mg in each 1 ml;
(3) respectively injecting the test solution and the reference solution into a high performance liquid chromatograph to respectively obtain chromatograms under the chromatographic conditions that:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filler;
mobile phase: methanol-water-trifluoroacetic acid;
a detector: the detection wavelength is 278 nm;
column temperature: 25 ℃;
sample introduction volume: 20 mu l of the mixture;
flow rate: 1.0 ml/min;
a stripping mode: isocratic elution;
(4) the content of 3-hydroxy-2-butanone is calculated by peak area according to an external standard method.
Further, the volume ratio of methanol, water and trifluoroacetic acid in the mobile phase was 45:55: 0.01.
Further, the column used was a 5 μm, 4.6X 250mm Waters Symmetry C18 column.
Further, the detector for the high school liquid phase analysis was a PDA detector.
Compared with the prior art, the invention has the beneficial effects that:
(1) compared with a gas chromatography, the method adopts a high performance liquid chromatography to detect the 3-hydroxy-2-butanone, has the characteristics of low cost, short time consumption and simple operation, and is beneficial to popularization and application;
(2) the effective separation of the main peak of the 3-hydroxy-2-butanone and the impurity peaks nearby the main peak can be realized;
(3) the methodology proves that the method provided by the invention is accurate and reliable and has good stability;
(4) the quality control of the 3-hydroxy-2-butanone can be better realized.
Drawings
FIG. 1 is a high performance liquid chromatogram of a blank solution of example 1.
FIG. 2 is a high performance liquid chromatogram of the control solution of example 1.
FIG. 3 is a high performance liquid chromatogram of the sample solution of example 1.
Detailed Description
The present invention will be described in further detail with reference to examples. It will also be understood that the following examples are included merely for purposes of further illustrating the invention and are not to be construed as limiting the scope of the invention, as the invention extends to insubstantial modifications and adaptations of the invention following in the light of the principles set forth herein. The specific process parameters and the like of the following examples are also only one example of suitable ranges, and the skilled person can make a selection within the suitable ranges through the description herein, and are not limited to the specific data of the following examples.
Example 1
1. Materials and methods
The instrument comprises the following steps: waters e2695 high performance liquid chromatograph, PDA (photodiode array detector), electronic balance
Reagent: methanol (chromatographic grade), trifluoroacetic acid (chromatographic grade), purified water and a 3-hydroxy-2-butanone reference substance (the purity is more than or equal to 99 percent): supplied by Sigma
Blank solvent: purified water
2. Chromatographic conditions
A chromatographic column: waters Symmetry C184.6X 250mm, 5 μm
Mobile phase: methanol-water-trifluoroacetic acid with volume ratio of 45:55:0.01
A detector: detection wavelength of 278nm
Column temperature: 25 deg.C
Sample introduction volume: 20 μ l
Flow rate: 1.0ml/min
A stripping mode: isocratic elution
3. Test procedure
(1) Preparation of a test solution: taking a proper amount of the product, precisely weighing, adding a mobile phase for dissolving, and quantitatively diluting to prepare a solution containing about 1mg in each 1 ml;
(2) preparation of control solutions: taking a proper amount of 3-hydroxy-2-butanone reference substance, precisely weighing, adding a mobile phase for dissolving, and quantitatively diluting to prepare a solution containing about 1mg in each 1 ml;
(3) and (3) testing the applicability of the system: and (4) injecting 20 mu L of reference substance solution into a liquid chromatograph, and recording the chromatogram. Calculating the applicability parameters of the system, wherein the theoretical plate number of the 3-hydroxy-2-butanone is required to be not less than 2000, the tailing factor is required to be not more than 1.5, the separation degree between a main peak and each impurity is required to be not less than 1.5, the sample introduction is repeated for 6 times, and the RSD value of the peak area of the 3-hydroxy-2-butanone is required to be not more than 2.0 percent;
(4) respectively injecting the test solution and the reference solution into a high performance liquid chromatograph, recording a chromatogram and a peak area, wherein the peak emergence time of the 3-hydroxy-2-butanone reference solution is 3.742min, and the peak emergence time of the test solution is 3.739 mim;
(5) the content of 3-hydroxy-2-butanone is calculated by peak area according to an external standard method.
The blank solution chromatogram under the chromatographic conditions is shown in FIG. 1, the control solution chromatogram is shown in FIG. 2, and the sample solution chromatogram is shown in FIG. 3.
Example 2
The method carries out methodology verification on the content determination method, and verification results show that the method has good system applicability, specificity, linearity, accuracy, precision and durability. Therefore, the method is suitable for detecting the content of the 3-hydroxy-2-butanone. The validation data is summarized as follows:
methodological verification summary sheet
Claims (4)
1. A method for measuring the content of 3-hydroxy-2-butanone by high performance liquid chromatography is characterized by comprising the following steps:
(1) preparation of a test solution: taking a proper amount of the product, precisely weighing, adding a mobile phase for dissolving, and quantitatively diluting to prepare a solution containing about 1mg in each 1 ml;
(2) preparation of control solutions: taking a proper amount of 3-hydroxy-2-butanone reference substance, precisely weighing, adding a mobile phase for dissolving, and quantitatively diluting to prepare a solution containing about 1mg in each 1 ml;
(3) respectively injecting the test solution and the reference solution into a high performance liquid chromatograph to respectively obtain chromatograms under the chromatographic conditions that:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filler;
mobile phase: methanol-water-trifluoroacetic acid;
a detector: the detection wavelength is 278 nm;
column temperature: 25 ℃;
sample introduction volume: 20 mu l of the mixture;
flow rate: 1.0 ml/min;
a stripping mode: isocratic elution;
(4) the content of 3-hydroxy-2-butanone is calculated by peak area according to an external standard method.
2. The method for measuring the content of 3-hydroxy-2-butanone by using the high performance liquid chromatography as claimed in claim 1, wherein the volume ratio of methanol, water and trifluoroacetic acid in the mobile phase is 45:55: 0.01.
3. The method for measuring the content of 3-hydroxy-2-butanone by using the high performance liquid chromatography as claimed in claim 1, wherein the chromatographic column adopts a Waters Symmetry C18 chromatographic column with the specification of 5 μm and 4.6 x 250 mm.
4. The method for measuring the content of 3-hydroxy-2-butanone by high performance liquid chromatography as claimed in claim 1, wherein the detector for high performance liquid chromatography is a PDA detector.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63246659A (en) * | 1986-12-09 | 1988-10-13 | Snow Brand Milk Prod Co Ltd | Determination of carbonyl group-containing organic component by fast liquid chromatograph |
| CN101581707A (en) * | 2009-06-29 | 2009-11-18 | 浙江大学 | Method for simultaneously detecting acetylmethylcar-binol and ligustrazine in table vinegar |
-
2020
- 2020-12-14 CN CN202011462211.9A patent/CN112630331A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63246659A (en) * | 1986-12-09 | 1988-10-13 | Snow Brand Milk Prod Co Ltd | Determination of carbonyl group-containing organic component by fast liquid chromatograph |
| CN101581707A (en) * | 2009-06-29 | 2009-11-18 | 浙江大学 | Method for simultaneously detecting acetylmethylcar-binol and ligustrazine in table vinegar |
Non-Patent Citations (4)
| Title |
|---|
| 刘学彬 等: "高效液相色谱法快速检测麸曲中乙偶姻" * |
| 尹丹丹 等: "基于川芎内生枯草杆菌初步研究川芎中川芎嗪的来源" * |
| 杨宇霞 等: "山西老陈醋中乙偶姻和川芎嗪的高效液相色谱法测定" * |
| 陈继承 等: "食醋中川芎嗪快速检测及其生成机理初探" * |
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