CN110887905A - Method for detecting key intermediate β -thymidine of anti-HIV drug - Google Patents
Method for detecting key intermediate β -thymidine of anti-HIV drug Download PDFInfo
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Abstract
The invention provides a detection method of β -thymidine, a key intermediate of an anti-HIV drug, which adopts high performance liquid chromatography for analysis and detection, and detects a β -thymidine product finished product after pretreatment according to the following chromatographic conditions, wherein the chromatographic column comprises a C18 column or other equivalent chromatographic columns, a mobile phase comprises methanol aqueous solution, the flow rate is 0.8ml/min-1.2ml/min, the detector comprises an ultraviolet detector, the detection wavelength is 260nm-270nm, and the chromatographic column temperature is 25 ℃ -35 ℃.
Description
Technical Field
The invention relates to the field of β -thymidine detection, in particular to a method for detecting a key intermediate β -thymidine of an anti-HIV drug.
Background
β -thymidine is used as a key intermediate for synthesizing the zidovudine as an anti-AIDS drug, has high technical content and high price, is mainly exported to the United states, India and the like, and brings economic benefit to companies.
Disclosure of Invention
Technical problem to be solved
The invention aims to provide a method for detecting a key intermediate β -thymidine of an anti-HIV drug, which provides a quick, accurate and reliable detection and analysis method for detecting related substances in a β -thymidine product finished product and determining the content of β -thymidine, and reduces blindness.
(II) technical scheme
A method for detecting a key intermediate β -thymidine of an anti-HIV drug adopts high performance liquid chromatography to detect the content of β -thymidine and related substances, and comprises the following steps:
(1) preparing test solution by dissolving β -thymidine product with mobile phase and diluting to obtain solution containing 1mg β -thymidine per 1ml as test solution (requiring fresh preparation);
(2) preparing reference solution by precisely measuring 1ml of β -thymidine reference, placing in a 100ml measuring flask, diluting with mobile phase to 100ml scale, and shaking to obtain reference solution;
(3) β -thymidine was detected by high performance liquid chromatography:
①, detecting conditions that the detector adopted by the high performance liquid chromatography is an ultraviolet detector, the chromatographic conditions are that a chromatographic column is a C18 column or other equivalent chromatographic columns, the flow rate is 0.8ml/min-1.2ml/min, the temperature of the chromatographic column is 25 ℃ to 35 ℃, and the detection wavelength is 260nm-270 nm;
②, the detection method comprises injecting 10 μ l of reference solution into a liquid chromatograph, adjusting detection sensitivity to make the peak height of main component chromatogram peak about 20% of full range, precisely measuring 10 μ l of each of the test solution and the reference solution, injecting into the liquid chromatograph, and recording chromatogram until the retention time of the main component peak is 2.5 times.
Further, in the impurity peaks in the chromatogram of the test solution, the area of a single impurity peak is not more than 0.5 times (0.5%) of the area of the main peak of the control solution, and the sum of the areas of the impurity peaks is not more than 2 times (2.0%) of the area of the main peak of the control solution.
Further, the β -thymidine detection method specifically comprises the following steps:
(1) preparing sample, namely taking 50ml of β -thymidine product finished product, centrifuging for 10 minutes at 18000r/min, taking supernatant, measuring Xml, putting the supernatant into a volumetric flask, adding water to dissolve the supernatant and diluting the supernatant to 100 times of volume, shaking the supernatant evenly, filtering the supernatant by using a 0.45 mu m microporous membrane, taking Yml of filtrate, transferring the Yml of filtrate into the volumetric flask, adding water to dilute the supernatant to 10 times of volume, shaking the filtrate evenly, and filtering the filtrate by using the 0.45 mu m microporous membrane to obtain liquid to be detected;
(2) analyzing and detecting by adopting high performance liquid chromatography, setting six concentration gradients without zero points in the standard working solution, drawing a standard curve by taking β -thymidine as an abscissa and taking a peak area integral value as an ordinate, and calculating the content of β -thymidine in the liquid to be detected according to an external standard method to obtain the content of β -thymidine in a β -thymidine product finished product.
Further, in the step (3), the column was a 4.6mm X250 mm, 5 μm Shim-pack ODS C18 column; the flow rate is 1 ml/min; temperature of the column: 30 ℃; the detection wavelength is 265 nm;
further, the mobile phase is a methanol aqueous solution with the methanol content of 92 percent
(III) advantageous effects
Compared with the prior art, the method has the advantages of being simple in operation, convenient, practical, safe and environment-friendly, having practical guiding significance for β -thymidine production by a fermentation method, being good in precision and high in repeatability, being capable of rapidly and accurately measuring the β -thymidine content in a β -thymidine product finished product, and being beneficial to improving the quality and safety controllability of a key intermediate β -thymidine of an anti-HIV drug.
Detailed Description
The invention is further described below with reference to specific embodiments.
A method for detecting a key intermediate β -thymidine of an anti-HIV drug adopts high performance liquid chromatography to detect the content of β -thymidine and related substances, and comprises the following steps:
(1) preparing test solution by dissolving β -thymidine product with mobile phase and diluting to obtain solution containing 1mg β -thymidine per 1ml as test solution (requiring fresh preparation);
(2) preparing reference solution by precisely measuring 1ml of β -thymidine reference, placing in a 100ml measuring flask, diluting with mobile phase to 100ml scale, and shaking to obtain reference solution;
(3) β -thymidine was detected by high performance liquid chromatography:
①, detecting conditions that the detector adopted by the high performance liquid chromatography is an ultraviolet detector, the chromatographic conditions are that a chromatographic column is a C18 column or other equivalent chromatographic columns, the flow rate is 0.8ml/min-1.2ml/min, the temperature of the chromatographic column is 25 ℃ to 35 ℃, and the detection wavelength is 260nm-270 nm;
②, the detection method comprises injecting 10 μ l of reference solution into a liquid chromatograph, adjusting detection sensitivity to make the peak height of main component chromatogram peak about 20% of full range, precisely measuring 10 μ l of each of the test solution and the reference solution, injecting into the liquid chromatograph, and recording chromatogram until the retention time of the main component peak is 2.5 times.
In order to ensure the product quality, in the impurity peaks in the chromatogram of the test solution, the area of a single impurity peak is not more than 0.5 times (0.5%) of the area of a main peak of a control solution, and the sum of the areas of the impurity peaks is not more than 2 times (2.0%) of the area of the main peak of the control solution.
The detection method of β -thymidine in the β -thymidine product specifically comprises the following steps:
(1) preparing sample, namely taking 50ml of β -thymidine product finished product, centrifuging for 10 minutes at 18000r/min, taking supernatant, measuring Xml, putting the supernatant into a volumetric flask, adding water to dissolve the supernatant and diluting the supernatant to 100 times of volume, shaking the supernatant evenly, filtering the supernatant by using a 0.45 mu m microporous membrane, taking Yml of filtrate, transferring the Yml of filtrate into the volumetric flask, adding water to dilute the supernatant to 10 times of volume, shaking the filtrate evenly, and filtering the filtrate by using the 0.45 mu m microporous membrane to obtain liquid to be detected;
(2) analyzing and detecting by adopting high performance liquid chromatography, setting six concentration gradients without zero points in the standard working solution, drawing a standard curve by taking β -thymidine as an abscissa and taking a peak area integral value as an ordinate, and calculating the content of β -thymidine in the liquid to be detected according to an external standard method to obtain the content of β -thymidine in a β -thymidine product finished product.
EXAMPLE 1 investigation of related substances
Separating target components by high performance liquid chromatography
β -thymidine product is dissolved and diluted with mobile phase to obtain solution containing 1mg of thymidine in 1ml, and the solution is used as sample solution (prepared fresh) and the reference substance solution (1 ml β -thymidine is measured precisely and placed in 100ml measuring flask, diluted to scale with mobile phase and shaken up) to obtain the reference substance solution, 10 microliter of the reference substance solution is injected into liquid chromatograph under the chromatographic condition of content measurement to regulate the detection sensitivity and make the peak height of main component chromatographic peak about 20% of full range, and the sample solution and the reference substance solution are measured precisely and injected into liquid chromatograph separately to record the chromatogram until the retention time of main component peak is 2.5 times.
Second, attribution of chemical components of target components
And (3) analyzing the spectrum impurity, wherein if an impurity peak exists in a chromatogram of the test solution, the area of a single impurity peak is 0.28 times (0.28%) of the area of a main peak of the control solution, and the sum of the areas of the peaks of the impurities is 1.1 times (1.1%) of the area of the main peak of the control solution.
EXAMPLE 2 determination of β -Thymidine content by high Performance liquid chromatography
Instrument, sample and reagent
1. High performance liquid chromatograph: shimadzu high performance liquid chromatograph (LC-20A), ultraviolet detector.
2. An ultrasonic cleaner: KQ-300DE type numerical control ultrasonic cleaner of ultrasonic instrument Limited company in Kunshan city.
β -thymidine standard, β -thymidine sample (sample batch: 20180901), supplied by Jiangxi Chengzhi bioengineering, Inc. water is double distilled water, methanol (chromatographic purity).
System adaptability
The separation degree is more than or equal to 1.5, the relative standard deviation is less than or equal to 2.0 percent, the symmetry factor is less than or equal to 1.3, and the number of theoretical plates is more than or equal to 2000.
Chromatographic conditions
A chromatographic column: a Shim-pack ODS C18 column (4.6 mm. times.250 mm, 5 μm) or other equivalent chromatography column;
mobile phase: methanol is taken as a mobile phase A, water is taken as a mobile phase B, the A phase is 92 percent, and the B phase is 8 percent;
flow rate: 1.0 ml/min;
a detector: an ultraviolet detector;
temperature of the column: 30 ℃;
and (3) detection: 265 nm;
sample introduction amount: 10 μ l.
Preparation of test solution and reference solution
Accurately weighing β -thymidine product 100mg, placing in a 100ml volumetric flask, adding water to dissolve and dilute to scale, shaking up, filtering with 0.45 μm microporous membrane to obtain the sample solution, accurately weighing β -thymidine reference substance 100mg, placing in a 100ml volumetric flask, adding water to dissolve and dilute to scale, shaking up, filtering with 0.45 μm microporous membrane to obtain the reference solution.
The methodology of the detection method provided by the present invention was examined as follows:
1. blank test
When a sample containing no β -thymidine product was prepared and tested according to the method for measuring β -thymidine content, no interference was observed at the β -thymidine peak (β -thymidine peak time of about 17.6 min).
2. Inspection of chromatographic conditions
A chromatographic column: a Shim-pack ODS C18 column (4.6 mm. times.250 mm, 5 μm) or other equivalent chromatography column;
mobile phase: methanol is taken as a mobile phase A, water is taken as a mobile phase B, the A phase is 92 percent, and the B phase is 8 percent;
flow rate: 1.0/min;
a detector: an ultraviolet detector;
temperature of the column: 30 ℃;
detection wavelength: 265 nm;
2.1. investigation of mobile phase
A chromatographic column: a Shim-pack ODS C18 column (4.6 mm. times.250 mm, 5 μm) or other equivalent chromatography column;
flow rate: 1.0/min;
a detector: an ultraviolet detector;
temperature of the column: 30 ℃;
detection wavelength: 265 nm;
the mobile phases were as follows:
| serial number | Mobile phase A% | Mobile phase B% |
| 1 | 88 | 12 |
| 2 | 90 | 10 |
| 3 | 92 | 8 |
| 4 | 94 | 6 |
| 5 | 96 | 4 |
When mobile phase 3 was selected, the separation effect was the best, so mobile phase 3 was selected as the mobile phase for detecting β -thymidine, i.e., methanol was used as mobile phase a, water was used as mobile phase B, 92% of phase a, and 8% of phase B.
2.2. Investigation of detection wavelength
Since β -thymidine has a maximum absorption peak at 265nm, the detection wavelength was chosen to be 265 nm.
3. Investigation of Linear relationships
An appropriate amount of β -thymidine reference substance is precisely weighed, mobile phases are respectively added to prepare concentrations of 1.003mg/ml, 1.501mg/ml, 2.004mg/ml, 2.531mg/ml, 3.052mg/ml and 4.007mg/ml, a standard curve is drawn by taking β -thymidine concentration as an abscissa and reducing a peak area integral value by 1000 times as an ordinate, and a regression equation is obtained that Y is 6195.9X +102.93, and the result that r is 1 shows that the peak area integral value and the concentration of β -thymidine have a good linear relation in the range of 1.003-4.007 mg/ml (see Table 1).
TABLE 1 β Thymidine Standard Curve measurements
| Numbering | β thymidine (mg/ml) | Integrated value of peak area |
| 1 | 1.003 | 6304892 |
| 2 | 1.501 | 9419097 |
| 3 | 2.004 | 12576716 |
| 4 | 2.531 | 15807584 |
| 5 | 3.052 | 19039081 |
| 6 | 4.007 | 24949802 |
4. Precision test
The same reference solution and the same test solution were sampled for 6 times, respectively, and the peak area values were determined, with RSD of the reference solution and the test solution being 0.19% and 0.48%, respectively (see table 2). The results show that the method is accurate.
TABLE 2 results of precision test
5. Stability test
Taking the test solution, standing for 0, 1, 2, 4, 6, 8, 10 and 12h after preparation, and respectively measuring, wherein the results show that the test solution is stable within 12h, and are shown in Table 3.
TABLE 3 stability test results
6. Repeatability test
The average content of the product is 98.49% and the RSD is 0.10% after 6 times of repeated measurement according to the method for measuring the residual content of β -thymidine in the β -thymidine product finished product by taking the same batch of finished product (20180901) (see Table 4).
TABLE 4 results of the repeatability tests
7. Recovery test
The average recovery rate was 97.92% (n-9) and the RSD was 0.81% (see table 5) when 3 samples with three concentrations were prepared at 80%, 100%, and 120% sample loading ratios, respectively, and the residual content of β -thymidine was measured.
TABLE 5 recovery test
8. Durability examination
The durability of three brands of HPLC chromatographs and three flow rates were examined in this experiment, and the results are shown in Table 6.
TABLE 6 β Thymidine content of β Thymidine in the finished product
The RSD detected by the same chromatographic column of the three instruments is 0.15 percent, and the RSD detected by the same chromatographic column of the same instrument at different flow rates is 0.08 percent. The results show that the method is durable.
The invention provides a reliable analysis method for determination of β -thymidine in an β -thymidine product finished product, has the advantages of simple operation, convenience, practicability, safety, environmental protection and the like, has practical guiding significance for production of β -thymidine by a fermentation method, has good precision and high repeatability, can quickly and accurately determine the content of β -thymidine in a β -thymidine product finished product, and is favorable for improving the quality and safety controllability of a key intermediate β -thymidine of an anti-HIV drug.
Although specific embodiments of the invention have been described above, it will be understood by those skilled in the art that the specific embodiments described are illustrative only and are not limiting upon the scope of the invention, and that equivalent modifications and variations can be made by those skilled in the art without departing from the spirit of the invention, which is to be limited only by the appended claims.
Claims (5)
1. A method for detecting a key intermediate β -thymidine of an anti-HIV drug is characterized in that the content of β -thymidine and related substances are detected by high performance liquid chromatography, and the method comprises the following steps:
(1) preparing test solution by dissolving β -thymidine product with mobile phase and diluting to obtain solution containing 1mg β -thymidine per 1ml as test solution (requiring fresh preparation);
(2) preparing reference solution by precisely measuring 1ml of β -thymidine reference, placing in a 100ml measuring flask, diluting with mobile phase to 100ml scale, and shaking to obtain reference solution;
(3) β -thymidine was detected by high performance liquid chromatography:
①, detecting conditions that the detector adopted by the high performance liquid chromatography is an ultraviolet detector, the chromatographic conditions are that a chromatographic column is a C18 column or other equivalent chromatographic columns, the flow rate is 0.8ml/min-1.2ml/min, the temperature of the chromatographic column is 25 ℃ to 35 ℃, and the detection wavelength is 260nm-270 nm;
②, the detection method comprises injecting 10 μ l of reference solution into a liquid chromatograph, adjusting detection sensitivity to make the peak height of main component chromatogram peak about 20% of full range, precisely measuring 10 μ l of each of the test solution and the reference solution, injecting into the liquid chromatograph, and recording chromatogram until the retention time of the main component peak is 2.5 times.
2. The method for detecting the key intermediate β -thymidine in anti-HIV medicine according to claim 1, wherein the area of the single impurity peak in the chromatogram of the test solution is not more than 0.5 times (0.5%) of the area of the main peak in the control solution, and the sum of the areas of the impurity peaks is not more than 2 times (2.0%) of the area of the main peak in the control solution.
3. The method for detecting the key intermediate β -thymidine of the anti-HIV drug according to claim 1, wherein the method for detecting β -thymidine comprises the following steps:
(1) preparing sample, namely taking 50ml of β -thymidine product finished product, centrifuging for 10 minutes at 18000r/min, taking supernatant, measuring Xml, putting the supernatant into a volumetric flask, adding water to dissolve the supernatant and diluting the supernatant to 100 times of volume, shaking the supernatant evenly, filtering the supernatant by using a 0.45 mu m microporous membrane, taking Yml of filtrate, transferring the Yml of filtrate into the volumetric flask, adding water to dilute the supernatant to 10 times of volume, shaking the filtrate evenly, and filtering the filtrate by using the 0.45 mu m microporous membrane to obtain liquid to be detected;
(2) analyzing and detecting by adopting high performance liquid chromatography, setting six concentration gradients without zero points in the standard working solution, drawing a standard curve by taking β -thymidine as an abscissa and taking a peak area integral value as an ordinate, and calculating the content of β -thymidine in the liquid to be detected according to an external standard method to obtain the content of β -thymidine in a β -thymidine product finished product.
4. The method for detecting β -thymidine as a key intermediate of anti-HIV drug in step (3), wherein the chromatographic column in step (3) is a Shim-pack ODS C18 column of 4.6mm x 250mm and 5 μm, the flow rate is 1ml/min, the temperature of the chromatographic column is 30 ℃, and the detection wavelength is 265 nm.
5. The method for detecting the key intermediate β -thymidine of anti-HIV drugs in claim 1, wherein the mobile phase is an aqueous solution of methanol with a methanol content of 92%.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103149301A (en) * | 2013-03-07 | 2013-06-12 | 通化华夏药业有限责任公司 | Method for simultaneously determining contents of 10 nucleoside components in sowthistle-leaf ixeris seedling injection by utilizing HPLC-DAD (High Performance Liquid Chromatography-Diode Array detector) method |
| CN109932441A (en) * | 2019-03-01 | 2019-06-25 | 贵州瑞和制药有限公司 | A kind of method for building up of easypro liver injection for curing HPLC finger-print |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103149301A (en) * | 2013-03-07 | 2013-06-12 | 通化华夏药业有限责任公司 | Method for simultaneously determining contents of 10 nucleoside components in sowthistle-leaf ixeris seedling injection by utilizing HPLC-DAD (High Performance Liquid Chromatography-Diode Array detector) method |
| CN109932441A (en) * | 2019-03-01 | 2019-06-25 | 贵州瑞和制药有限公司 | A kind of method for building up of easypro liver injection for curing HPLC finger-print |
Non-Patent Citations (2)
| Title |
|---|
| 吕慧慧等: "HPLC-DAD同时测定金蝉花中8种核苷类成分的含量", 《实用药物与临床》 * |
| 孔凤利等: "HPLC指纹图谱结合主成分分析研究僵蚕不同溶剂提取物的化学组分", 《中国中医药科技》 * |
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