CN114047262B - A method for separating and determining easily hydrolyzed large polar compounds - Google Patents
A method for separating and determining easily hydrolyzed large polar compounds Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 29
- 150000001875 compounds Chemical class 0.000 title claims abstract description 28
- 239000000243 solution Substances 0.000 claims abstract description 50
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 45
- 239000012535 impurity Substances 0.000 claims abstract description 40
- 229960004150 aciclovir Drugs 0.000 claims abstract description 34
- MKUXAQIIEYXACX-UHFFFAOYSA-N aciclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCO)C=N2 MKUXAQIIEYXACX-UHFFFAOYSA-N 0.000 claims abstract description 34
- 239000000523 sample Substances 0.000 claims abstract description 23
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 11
- 239000012488 sample solution Substances 0.000 claims abstract description 11
- 238000001514 detection method Methods 0.000 claims abstract description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 9
- 238000010829 isocratic elution Methods 0.000 claims abstract description 9
- 239000000741 silica gel Substances 0.000 claims abstract description 9
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 9
- 238000004811 liquid chromatography Methods 0.000 claims abstract description 7
- 239000003085 diluting agent Substances 0.000 claims abstract description 5
- 238000002347 injection Methods 0.000 claims abstract description 5
- 239000007924 injection Substances 0.000 claims abstract description 5
- QNXFUWFRTWSSOK-UHFFFAOYSA-N n-acetyl-n-(6-oxo-3,7-dihydropurin-2-yl)acetamide Chemical compound O=C1NC(N(C(C)=O)C(=O)C)=NC2=C1NC=N2 QNXFUWFRTWSSOK-UHFFFAOYSA-N 0.000 claims description 28
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 16
- WBDLSXCAFVEULB-UHFFFAOYSA-N acetonitrile;methylsulfinylmethane Chemical compound CC#N.CS(C)=O WBDLSXCAFVEULB-UHFFFAOYSA-N 0.000 claims description 15
- YXSLALYOAHCVAN-UHFFFAOYSA-N 8-acetyl-2-amino-3,7-dihydropurin-6-one Chemical compound N1C(N)=NC(=O)C2=C1N=C(C(=O)C)N2 YXSLALYOAHCVAN-UHFFFAOYSA-N 0.000 claims description 9
- 239000012086 standard solution Substances 0.000 claims description 6
- -1 dihydroxypropyl Chemical group 0.000 claims description 2
- 230000008569 process Effects 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 6
- 230000007062 hydrolysis Effects 0.000 abstract description 4
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 4
- 239000007853 buffer solution Substances 0.000 abstract description 3
- 238000005070 sampling Methods 0.000 abstract description 2
- 239000011550 stock solution Substances 0.000 description 18
- 238000007865 diluting Methods 0.000 description 17
- XUKUURHRXDUEBC-SXOMAYOGSA-N (3s,5r)-7-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyheptanoic acid Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-SXOMAYOGSA-N 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 11
- 239000007788 liquid Substances 0.000 description 11
- 239000012071 phase Substances 0.000 description 9
- 230000006872 improvement Effects 0.000 description 8
- 239000012490 blank solution Substances 0.000 description 5
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 4
- 238000002013 hydrophilic interaction chromatography Methods 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
- 238000005303 weighing Methods 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 206010061598 Immunodeficiency Diseases 0.000 description 2
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- 239000002994 raw material Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- QGQXAMBOYWULFX-LZWSPWQCSA-N 2-morpholin-4-ylethyl (e)-6-(4,6-dihydroxy-7-methyl-3-oxo-1h-2-benzofuran-5-yl)-4-methylhex-4-enoate Chemical compound OC=1C=2C(=O)OCC=2C(C)=C(O)C=1C\C=C(/C)CCC(=O)OCCN1CCOCC1 QGQXAMBOYWULFX-LZWSPWQCSA-N 0.000 description 1
- 206010048461 Genital infection Diseases 0.000 description 1
- 208000009889 Herpes Simplex Diseases 0.000 description 1
- 208000000903 Herpes simplex encephalitis Diseases 0.000 description 1
- 208000037018 Herpes simplex virus encephalitis Diseases 0.000 description 1
- 208000007514 Herpes zoster Diseases 0.000 description 1
- 241000700584 Simplexvirus Species 0.000 description 1
- HDOVUKNUBWVHOX-QMMMGPOBSA-N Valacyclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCOC(=O)[C@@H](N)C(C)C)C=N2 HDOVUKNUBWVHOX-QMMMGPOBSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 150000008209 arabinosides Chemical class 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- GILZZWCROUGLIS-UHFFFAOYSA-N n-(9-acetyl-6-oxo-3h-purin-2-yl)acetamide Chemical compound N1C(NC(=O)C)=NC(=O)C2=C1N(C(C)=O)C=N2 GILZZWCROUGLIS-UHFFFAOYSA-N 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 238000005220 pharmaceutical analysis Methods 0.000 description 1
- 150000003834 purine nucleoside derivatives Chemical class 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000037384 skin absorption Effects 0.000 description 1
- 231100000274 skin absorption Toxicity 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 229940093257 valacyclovir Drugs 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
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- 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
- G01N30/60—Construction of the 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
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- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
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Abstract
The invention discloses a method for separating and measuring easily-hydrolyzed large polar compounds, which comprises the following steps of dissolving acyclovir in a diluent to form a sample solution, detecting a standard substance solution and the sample solution by adopting high performance liquid chromatography, wherein the liquid chromatography comprises a chromatographic column, a hybrid silica gel column, a mobile phase A, a buffer solution, a mobile phase B, acetonitrile, isocratic elution, a mobile phase flow rate of 0.4-0.6 ml/min, a column temperature of 25-35 ℃, a detection wavelength of UV254nm, a sample injection amount of 20 μl and a running time of 50min. The method can inhibit hydrolysis in the process of sampling the easily-hydrolyzed large polar compound impurity, truly reflects the detection amount of the impurity, has high precision, operates at room temperature, has good reproducibility, and can effectively separate and determine the content of the easily-hydrolyzed large polar compound.
Description
Technical Field
The invention relates to the technical field of pharmaceutical analysis, in particular to a method for separating and measuring easily-hydrolyzed large polar compounds, and specifically relates to a method for separating and measuring the impurity content of easily-hydrolyzed large polar compounds in acyclovir.
Background
Acyclovir (Acyclovir) is an antiviral drug.
The chemical structural formula:
Acyclovir is a synthetic purine nucleoside analogue. Is mainly used for various infections caused by herpes simplex virus, and can be used for treating HSV infections caused by primary or recurrent skin, mucous membrane, external genital infections and immunodeficiency. Is the first choice for treating HSV encephalitis, and has better effect on reducing morbidity and mortality than the arabinoside. It can also be used for treating herpes zoster, EB virus, and immunodeficiency complicated with varicella. Topical application to skin only, acyclovir has less skin absorption.
In the quality control of acyclovir, partial impurities are extremely unstable, and can be rapidly hydrolyzed in a system such as water, weak acid or weak base, for example, diacetylguanine (impurity A) in the acyclovir can be rapidly hydrolyzed and deacylated in the water to be changed into monoacetylguanine (impurity B) and guanine, so that the contents of diacetylguanine, monoacetylguanine and guanine in acyclovir cannot be truly reflected in the detection process.
Chinese patent CN202010731803.X reports that the comparison of qualified finished products by ultraviolet dual-wavelength absorbance ratio detection method can rapidly and accurately control the reaction end point and the quality of the finished products, and the ultraviolet spectrophotometry has poor specificity and cannot accurately quantify the content of monoacetylguanine in the reaction liquid and diacetylguanine.
The ultra high pressure liquid chromatography (UPLC) method for measuring the purity of diacetylguanine (the research of the detection method of the beta-furfront, liu Chunyan, mo Wenjuan, N-2, 9-diacetylguanine, international medical and health guide, volume 19 in 2013, 1675-1677) is reported by the Yuanfeng et al, and the better separation can be realized due to the high column efficiency of the UPLC, the short chromatographic column and the short analysis time, wherein the retention time of the monoacetylguanine and the diacetylguanine is only 1.1 minutes and 2.0 minutes. The ultrahigh pressure liquid chromatography analysis speed is high, so that the hydrolysis can be inhibited. At the same time we repeated the paper using HPLC detection, but the diacetylguanine retention time was 2 minutes, the monoacetylguanine and diacetylguanine co-flowed, the diacetylguanine and diacetylguanine chromatographic peaks were fully coincident, only one chromatographic peak was shown in the liquid chromatogram, indicating that under the reference conditions, diacetylguanine was completely free from monoacetylguanine separation.
Therefore, there is a need for a high performance liquid chromatography method for separating and determining impurities in acyclovir, which can solve the problems, inhibit degradation of the easily hydrolyzed large polar compound impurities in acyclovir, completely separate related impurities, and has the advantages of simple operation and good reproducibility.
Disclosure of Invention
Because the ultra-high pressure liquid chromatograph is high in price and the equipment is not equipped in a general medicine enterprise, the method for analyzing the easily hydrolyzed large polar compound by using the common HPLC has certain practicability, meanwhile, the HILIC chromatographic column is skillfully utilized to be different from the common C-18 chromatographic column, the hydrolysis of the diacetyl guanine in the chromatographic analysis process is reduced when the HILIC chromatographic column is used for a large proportion of organic phases, meanwhile, the large polar compound is kept strongly, the problem that the diacetyl guanine is unstable and cannot be quantified in the analysis process is solved, and the method has innovativeness.
The method is also suitable for detecting the EP impurity Q in the acyclovir Wei Chengpin, and the European pharmacopoeia 10.0 (EP 10.0) analysis method is adopted to detect the impurity Q in the acyclovir Wei Chengpin, so that the impurity Q is hydrolyzed under the chromatographic condition to generate acyclovir, the accuracy in the verification of the analysis method cannot meet the requirement of the regulation, and the fact that the content of the impurity Q in the acyclovir Wei Chengpin cannot be accurately quantified by the EP10.0 analysis method is explained. By adopting the analysis method for detecting the large polar compounds which are easy to hydrolyze, the hydrolysis problem of the impurity Q in liquid chromatography analysis can be well inhibited.
In order to solve the technical problems, the invention provides a method for separating and measuring easily hydrolyzed large polar compounds, which comprises the following steps:
Dissolving a sample containing a large polar compound which is easy to hydrolyze in a diluent to form a sample solution, and detecting the standard substance solution and the sample solution by adopting high performance liquid chromatography;
Wherein the diluent is 10% dimethyl sulfoxide acetonitrile solution;
liquid chromatography conditions:
The chromatographic column is a hybrid silica gel column, the mobile phase A is trifluoroacetic acid solution, the mobile phase B is acetonitrile, isocratic elution is adopted, the flow rate of the mobile phase is 0.4-0.6 ml/min, the column temperature is 25-35 ℃, the detection wavelength is UV254nm, the sample injection amount is 20 mu l, and the running time is 50min.
As a preferred aspect of the above technical solution, the method for separating and determining a readily hydrolyzable large polar compound provided by the present invention further includes some or all of the following technical features:
As an improvement of the above technical scheme, the sample containing the easily hydrolyzable large polar compound is acyclovir or diacetylguanine.
As an improvement of the technical scheme, the sample containing the easily-hydrolyzed large polar compound is acyclovir, wherein the sample contains an impurity A, an impurity B and an impurity Q, and the specific names and chemical structures of the compounds are as follows:
Impurity A is diacetylguanine with structural formula
Impurity B is monoacetylguanine with structural formula
The impurity Q is acyclovir EP impurity Q, and the structural formula is
Acyclovir has the structural formula:
As an improvement of the technical scheme, in the process of the method for separating and measuring the easily hydrolyzed large polar compound, the following solution needs to be prepared:
Dissolving 10mg of impurity B in a 100ml volumetric flask, diluting to scale with 10% dimethyl sulfoxide acetonitrile solution, and shaking;
Mixing standard solution, namely placing 10mg of diacetylguanine sample into a 100ml measuring flask, dissolving 0.5ml of impurity B stock solution with 10% dimethyl sulfoxide acetonitrile solution, diluting to scale, and shaking uniformly;
Taking 10mg of diacetylguanine sample, dissolving the diacetylguanine sample in a 100ml volumetric flask by using a 10% dimethyl sulfoxide acetonitrile solution, diluting the solution to a scale, and shaking the solution uniformly;
blank solution 10% dimethyl sulfoxide acetonitrile solution.
As an improvement of the technical scheme, the isocratic elution is specifically performed according to the following volume ratio of 96-94% of A+4-6% of B.
As an improvement of the technical scheme, the isocratic elution is specifically performed according to the following volume ratio of 95% A+5% B.
As an improvement of the technical scheme, the hybrid silica gel column is a bonded dihydroxypropyl hybrid silica gel column.
As an improvement of the technical scheme, the pH value of the buffer solution is 3.0-4.0.
As an improvement of the technical scheme, the pH value of the buffer solution is regulated by trifluoroacetic acid.
Compared with the prior art, the method has the advantages that the problems can be solved, the degradation of the easily-hydrolyzed large polar compound impurities can be restrained, the content of the easily-hydrolyzed large polar compound is truly embodied, the method is simple, acyclovir and the impurities can be completely separated, the precision is high, the room-temperature operation is performed, the solution reproducibility is good, and the reagent is cheap.
The foregoing description is only an overview of the present invention, and is intended to be implemented in accordance with the teachings of the present invention in order that the same may be more clearly understood and appreciated, as well as the other objects, features and advantages of the present invention, as described in detail below in connection with the preferred embodiments.
Drawings
In order to more clearly illustrate the technical solution of the embodiments of the present invention, the drawings of the embodiments will be briefly described below.
FIG. 1 is a liquid chromatogram of a blank solution prepared in example 1;
FIG. 2 is a liquid chromatographic analysis chart of the mixed standard solution of example 1;
FIG. 3 is a linear plot of impurity A in the linear solution prepared in example 1;
FIG. 4 is a linear plot of impurity B in the linear solution prepared in example 1;
FIG. 5 is a graph of the mixed solution prepared in example 2;
FIG. 6 is a graph of a sample solution prepared in example 2.
Detailed Description
The following detailed description of the invention, which is a part of this specification, illustrates the principles of the invention by way of example, and other aspects, features, and advantages of the invention will become apparent from the detailed description.
Example 1
1) Preparing a solution:
Blank solution 10 dimethyl sulfoxide acetonitrile solution;
and (3) weighing 20mg of impurity A reference substance in a 100ml measuring flask, precisely weighing, dissolving in DMSO, diluting to a scale, and uniformly mixing.
Dissolving 10mg of impurity B in a 100ml volumetric flask, diluting to scale with 10% dimethyl sulfoxide acetonitrile solution, and shaking;
Mixing standard solution, namely placing 10mg of diacetylguanine sample into a 100ml measuring flask, dissolving 0.5ml of impurity B stock solution with 10% dimethyl sulfoxide acetonitrile solution, diluting to scale, and shaking uniformly;
Taking 10mg of diacetylguanine sample, dissolving the diacetylguanine sample in a 100ml volumetric flask by using a 10% dimethyl sulfoxide acetonitrile solution, diluting the solution to a scale, and shaking the solution uniformly;
and (3) precisely removing 14.0ml to 100ml of the impurity B stock solution from the linear stock solution A, adding acetonitrile to dilute to a scale, and shaking uniformly to obtain the product.
And (3) accurately transferring the impurity linear 10% solution to a 10ml measuring flask by taking 0.5ml of the linear stock solution A, diluting to a scale by adding acetonitrile, and uniformly mixing.
And (3) accurately transferring the impurity linear 20% solution to a 10ml measuring flask by taking 1.0ml of the linear stock solution A, diluting to a scale by adding acetonitrile, and uniformly mixing.
And (3) accurately transferring the impurity linear 40% solution to a 10ml measuring flask by taking 2.0ml of the linear stock solution A, diluting to a scale by adding acetonitrile, and uniformly mixing.
And (3) accurately transferring the linear stock solution A4.0 ml into a 10ml measuring flask, diluting with acetonitrile to a scale, and mixing uniformly.
And (3) accurately transferring the impurity linear 100% solution into a 10ml measuring flask by taking 5.0ml of the linear stock solution A, adding acetonitrile to dilute to a scale, and uniformly mixing.
Impurity linear 200% solution, co-linear stock solution A
And (3) precisely transferring 1.0ml to 100ml of the impurity A stock solution to the linear stock solution B, adding acetonitrile to dilute to a scale, and shaking uniformly to obtain the product.
Diacetylguanine linear LOQ solution, taking quantitative limiting solution.
The diacetyl guanine linear solution-1 is prepared by precisely transferring 1.0ml of the linear stock solution B into a 10ml measuring flask, adding acetonitrile to dilute to a scale, and uniformly mixing.
Diacetylguanine linear solution-2, precisely transferring linear stock solution B4.0 ml into a 10ml measuring flask, diluting with acetonitrile to scale, and mixing.
Diacetylguanine linear solution-3, precisely transferring linear stock solution B5.0 ml into a 10ml measuring flask, diluting with acetonitrile to scale, and mixing.
Diacetylguanine linear solution-4. Co-linear stock solution B
2) And detecting the standard substance solution and the sample solution by adopting high performance liquid chromatography, wherein the conditions of the high performance liquid chromatography are as follows:
High performance liquid chromatograph, agilent 1260VWD;
Chromatographic column YMC HILIC hybrid silica gel column (250 mm. Times.4.6 mm. Times.5 μm);
mobile phase a trifluoroacetic acid solution (ph=3.0);
Mobile phase B, acetonitrile;
isocratic elution: a: b=5:95
The flow rate of the mobile phase is 0.5ml/min;
Column temperature is 30 ℃;
the detection wavelength is UV254nm;
the sample injection amount is 20 μl.
FIG. 1 is a liquid chromatogram of a blank solution prepared in example 1, and FIG. 2 is a liquid chromatogram of a mixed standard solution of example 1.
A linear test:
and (5) sequentially sampling according to the sequence of the blank sample and the linear solution.
Drawing a standard curve:
Analysis was performed under the set chromatographic conditions. Calibration curves were drawn from standard series mass concentrations and peak areas. And (3) carrying out linear regression by taking the proportion of the linear solution as an abscissa (%) and the peak area as an ordinate (Y) to obtain a regression equation and a correlation coefficient. In the measurement range, each impurity and valacyclovir have good linear relation, and the linear correlation coefficient R is not less than 0.9990. The data are shown in tables 1-2. FIG. 3 is a linear plot of the impurity A in the linear solution prepared in example 1, and FIG. 4 is a linear plot of the impurity B in the linear solution prepared in example 1;
TABLE 1 liquid chromatography impurity A chromatographic peak analysis results for example 1 Linear solution
TABLE 2 liquid chromatography impurity B chromatography peak analysis results for example 1 Linear solution
Example 2
1) Preparing a solution:
Blank solution, 10% dimethyl sulfoxide acetonitrile solution;
Taking 10mg of impurity Q, dissolving the impurity Q in a 10ml volumetric flask by using a 10% dimethyl sulfoxide acetonitrile solution, diluting to a scale, and shaking uniformly;
Acyclovir Wei Zhubei liquid, namely weighing a 10mg acyclovir sample in a 10ml measuring flask, precisely weighing, dissolving with a 10% dimethyl sulfoxide acetonitrile solution, diluting to a scale, and uniformly mixing.
Mixing standard solution, namely taking 1ml of impurity Q stock solution, 0.1ml of acyclovir stock solution into a 10ml measuring flask, dissolving with 10% dimethyl sulfoxide acetonitrile solution, diluting to scale, and shaking uniformly;
taking a sample solution, namely taking 10mg of acyclovir sample, dissolving the acyclovir sample in a 10ml volumetric flask by using a 10% dimethyl sulfoxide acetonitrile solution, diluting the acyclovir sample to a scale, and shaking the acyclovir sample uniformly;
2) And detecting the standard substance solution and the sample solution by adopting high performance liquid chromatography, wherein the conditions of the high performance liquid chromatography are as follows:
High performance liquid chromatograph, agilent 1260VWD;
Chromatographic column YMQ HILIC hybrid silica gel column (250 mm. Times.4.6 mm. Times.5 μm);
mobile phase a trifluoroacetic acid solution (ph=3.0);
Mobile phase B, acetonitrile;
isocratic elution: a: b=5:95
The flow rate of the mobile phase is 0.5ml/min;
Column temperature is 30 ℃;
the detection wavelength is UV254nm;
the sample injection amount is 20 μl.
FIG. 5 is a liquid chromatogram of the mixed solution prepared in example 2, and FIG. 6 is a liquid chromatogram of the sample solution of example 2.
The mixed solution is continuously injected for 6 needles, the areas of the impurity Q and the acyclovir Wei Feng are not obviously changed, which indicates that the impurity Q is not degraded into acyclovir under the chromatographic condition, and the result is as follows:
TABLE 3 stability investigation results of example 2 mixed solution
| Component (A) | 1 | 2 | 3 | 4 | 5 | 6 | Average value of | RSD% |
| Acyclovir | 37.962 | 38.256 | 38.115 | 38.194 | 37.998 | 38.537 | 38.177 | 0.55 |
| Impurity Q | 35.404 | 35.144 | 34.716 | 34.397 | 34.249 | 34.585 | 34.749 | 1.27 |
。
The present invention can be realized by the respective raw materials listed in the present invention, and the upper and lower limits and interval values of the respective raw materials, and the upper and lower limits and interval values of the process parameters (such as temperature, time, etc.), and examples are not listed here.
While the invention has been described with respect to the preferred embodiments, it will be understood that the invention is not limited thereto, but is capable of modification and variation without departing from the spirit of the invention, as will be apparent to those skilled in the art.
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| CN104359992A (en) * | 2014-12-02 | 2015-02-18 | 湖北华世通潜龙药业有限公司 | Method for analyzing acyclovir for injection by adopting high performance liquid chromatography (HPLC) |
| CN110940754A (en) * | 2019-12-18 | 2020-03-31 | 湖北省宏源药业科技股份有限公司 | High performance liquid chromatography method for separating and measuring impurities in valacyclovir hydrochloride |
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| JP3160439B2 (en) * | 1993-09-03 | 2001-04-25 | 住化ファインケム株式会社 | Method for producing acetylguanine |
| US5565565A (en) * | 1994-08-04 | 1996-10-15 | Syntex (U.S.A.) Inc. | Preparation of N-9 substituted guanine compounds |
| CN111879713B (en) * | 2020-07-27 | 2022-05-31 | 湖北省宏源药业科技股份有限公司 | Method for judging reaction end point of diacetylguanine and controlling quality of finished product |
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| CN104359992A (en) * | 2014-12-02 | 2015-02-18 | 湖北华世通潜龙药业有限公司 | Method for analyzing acyclovir for injection by adopting high performance liquid chromatography (HPLC) |
| CN110940754A (en) * | 2019-12-18 | 2020-03-31 | 湖北省宏源药业科技股份有限公司 | High performance liquid chromatography method for separating and measuring impurities in valacyclovir hydrochloride |
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