CN111650290A - Analysis method of tenofovir alafenamide hemifumarate related substances - Google Patents
Analysis method of tenofovir alafenamide hemifumarate related substances Download PDFInfo
- Publication number
- CN111650290A CN111650290A CN202010303085.6A CN202010303085A CN111650290A CN 111650290 A CN111650290 A CN 111650290A CN 202010303085 A CN202010303085 A CN 202010303085A CN 111650290 A CN111650290 A CN 111650290A
- Authority
- CN
- China
- Prior art keywords
- tenofovir alafenamide
- solution
- alafenamide hemifumarate
- methanol
- substances
- 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.)
- Pending
Links
Images
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
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention relates to the technical field of pharmaceutical analysis, and particularly relates to a high-performance liquid phase detection method for related substances of tenofovir alafenamide hemifumarate. The invention discloses a method for simultaneously separating and analyzing 10 related impurities of tenofovir alafenamide. The method can effectively and accurately determine various related substances and contents in tenofovir alafenamide, and ensure the product quality.
Description
Technical Field
The invention relates to the technical field of chemical drug analysis methods, in particular to an analysis method for determining related substances of tenofovir alafenamide hemifumarate.
Background
Tenofovir Alafenamide hemifumarate (Tenofovir Alafenamide Fumarate), chemical name 9- [ (R) -2- [ ((S) - [ [ (S) -1- (isopropoxycarbonyl) ethyl]Amino group]Phenoxy phosphinyl) methoxy group]Propyl radical]Adenine hemifumarate of formula C21H29O5N6P·1/2(C4H4O4) Molecular weight of 534.51, and the structural formula is as follows:
hepatitis b virus infection is a worldwide infectious disease, and it is reported by the world health organization that about 20 million people worldwide have been infected with hepatitis b virus, and about one million people die each year from cirrhosis or liver failure due to hepatitis b infection. At present, the nucleotide drugs applied to anti-hepatitis B virus in China only comprise lamivudine, adefovir dipivoxil, telbivudine, entecavir, tenofovir disoproxil and other drugs, and in the process of treating hepatitis B by anti-nucleotide drugs, drug resistance is a problem which is relatively troublesome, and the development of tenofovir alafenamide provides a new choice for clinical treatment of hepatitis B infection. Related substances such as initial raw materials, intermediates, byproducts, chiral isomers and the like can be introduced in the production process of the tenofovir alafenamide raw material drug, and byproducts or other degradation products can be generated in the production, storage and transportation processes of the raw material drug and the preparation, so that the determination of the related substances is very important to the control of the drug quality. At present, no literature is available for specifically measuring related substances of tenofovir alafenamide hemifumarate raw materials and preparations, so that appropriate chromatographic conditions are searched, corresponding chromatographic parameters are adjusted, and corresponding impurities are necessarily controlled.
Disclosure of Invention
The invention provides an analysis method for determining related substances of tenofovir alafenamide hemifumarate, and solves the problems of more related substances of tenofovir alafenamide and difficult separation.
The technical scheme of the invention comprises the following steps:
the high performance liquid chromatography method for determining related substances of tenofovir alafenamide hemifumarate comprises the following steps:
(1) preparing a tenofovir alafenamide hemifumarate analysis solution;
(2) adopting high performance liquid chromatography, wherein the chromatographic conditions comprise: the chromatographic column is an octadecyl bonded silica gel chromatographic column, a solution with a volume ratio of phosphate buffer solution to methanol of 92:8 is used as a mobile phase A, and a solution with a volume ratio of phosphate buffer solution to methanol of 30:70 is used as a mobile phase B. The gradient elution process is as follows: the proportion of the A phase and the B phase in the mobile phase is 0, 15-20, 35-38, 40-45, 50 and 60 minutes, the proportion of the A phase is 90-95%, 40-45%, 30-35%, 0%, 90-95% and 90-95%, and the detection wavelength is 260 nm.
(3) And (3) sample determination: injecting the analysis solution prepared in the step (1) into a liquid chromatograph, recording a chromatogram, and separating the following related substances:
in a preferred scheme, the analysis method of the tenofovir alafenamide hemifumarate related substances mainly comprises the following steps:
(1) preparing an analytical solution
Dissolving a sample by using a single solvent of methanol and acetonitrile or a mixed solution of methanol-water, acetonitrile-water, a methanol-phosphate aqueous solution and an acetonitrile-phosphate aqueous solution to prepare an analysis solution.
(2) Chromatographic conditions
Adopting high performance liquid chromatography, wherein the chromatographic conditions comprise: the chromatographic column is an octadecyl bonded silica gel chromatographic column, a solution with a volume ratio of phosphate buffer solution to methanol of 92:8 is used as a mobile phase A, a solution with a volume ratio of phosphate buffer solution to methanol of 30:70 is used as a mobile phase B, the detection wavelength is 260nm, and gradient elution is carried out.
According to the invention, the length of the column is 150mm, the diameter is 4.6mm and the packing particle size is 5 μm.
According to the invention, the packing material of the chromatographic column is octadecyl bonded silica gel, preferably the chromatographic column is ACE ExcelC 18.
According to the invention, in a phosphate buffer solution, the concentration of sodium dihydrogen phosphate is 15-25 mmol/L; preferably, the concentration of sodium dihydrogen phosphate is 20 mmol/L.
According to the invention, the concentration of the sodium hydroxide solution for adjusting the pH value is 0.1-1.0 mol/L; preferably, the concentration of the sodium hydroxide solution is 0.5 mol/L.
According to the invention, in a phosphate buffer solution, a 0.5mol/L sodium hydroxide solution is used for adjusting the pH value to be 4-7, and preferably, the pH value is 6.0.
According to the invention, the flow rate is 0.8-1.5 ml/min; preferably, the flow rate is 1.0 ml/min.
According to the invention, the column temperature is 10-30 ℃; preferably, the column temperature is 20 ℃.
According to the invention, the gradient elution process is: the proportion of the A phase and the B phase in the mobile phase is 0, 15-20, 35-38, 40-45, 50 and 60 minutes, and the proportion of the A phase is 90-95%, 40-45%, 30-35%, 0%, 90-95% and 90-95%
The sample is tenofovir alafenamide hemifumarate bulk drug or a preparation thereof, and the preparation is tablets, capsules, granules, suppositories, pills, ointments, creams, pastes, inhalants, sprays, aerosols, gels, powders, syrups, liniments, paints, plastics, tinctures, patches, oral solutions, implants, films, lotions, rinses, decocted ointments, pastes, lotions or tea.
Has the advantages that:
the inventor refers to and has effectively developed the method for determining related substances of tenofovir alafenamide hemifumarate, can realize the simultaneous separation of 10 impurities in one system, and provides an effective means for the quality control of bulk drugs and preparations.
Drawings
FIG. 1 is a mixed solution chromatogram of example 1;
FIG. 2 is a mixed solution chromatogram of example 2;
FIG. 3 is a mixed solution chromatogram of example 3;
FIG. 4 is a mixed solution chromatogram of example 4.
Detailed Description
Hereinafter, preferred embodiments of the present invention will be described in detail. The experimental methods of the preferred embodiments, which do not indicate specific conditions, are generally performed according to conventional conditions, and the examples are given for better illustration of the present invention, but the present invention is not limited to the examples. Those skilled in the art can make insubstantial modifications and adaptations to the embodiments described above while remaining within the scope of the invention.
Example 1
(1) Instrumentation and high performance liquid chromatography conditions
High performance liquid chromatograph: a Thermo U3000 high performance liquid chromatography system and workstation;
a chromatographic column: column of Welch XB-phenyl (4.6X 150mm, 5 μm) on phenylsilane-bonded silica
20mmo/L sodium dihydrogen phosphate buffer solution (pH6.0) is used as a mobile phase A, 20mmo/L sodium dihydrogen phosphate buffer solution (pH4.0) -methanol (35:65) is used as a mobile phase B, the proportions of the two phases A and B in the mobile phase are 0, 20, 38, 45, 50 and 60 minutes, the proportion of the phase A is 90%, 40%, 30%, 0%, 90% and 90%, the detection wavelength is 260nm, the flow rate is 1.0ml/min, and the column temperature is 30 ℃.
(2) Experimental procedure
Respectively taking an impurity A-an impurity J reference substance and a proper amount of tenofovir alafenamide hemifumarate, and preparing a mixed solution containing 1mg of tenofovir alafenamide and 10 micrograms of the other impurity reference substances in each 1ml by using 80% methanol solution;
and (5) taking 5 mu l of the analysis solution, injecting the analysis solution into a liquid chromatograph, and recording a chromatogram. The results are shown in FIG. 1. The impurity H and the impurity I are completely coincided with the API, and the peak shape of the impurity B is poorer under the condition, so the chromatographic condition still needs to be adjusted and optimized.
Example 2
(1) Instrumentation and high performance liquid chromatography conditions
High performance liquid chromatograph: a Thermo U3000 high performance liquid chromatography system and workstation;
a chromatographic column: intersil ODS-3 (4.6X 150mm, 5 μm) octadecylsilane chemically bonded silica column
20mmo/L sodium dihydrogen phosphate buffer solution (pH6.0) -methanol (92:8) is used as a mobile phase A, 20mmo/L sodium dihydrogen phosphate buffer solution (pH5.0) -methanol (35:65) is used as a mobile phase B, the proportions of the A phase and the B phase in the mobile phase are respectively set according to the time points of 0, 15, 35, 40, 50 and 60 minutes, the proportion of the A phase is 92%, 45%, 35%, 0%, 92% and 92%, the detection wavelength is 260nm, the flow rate is 1.0ml/min, and the column temperature is 15 ℃.
(3) Experimental procedure
Respectively taking an impurity A-an impurity J reference substance and a proper amount of tenofovir alafenamide hemifumarate, and preparing a mixed solution containing 1mg of tenofovir alafenamide and 10 micrograms of the other impurity reference substances in each 1ml by using 80% methanol solution;
and (5) taking 5 mu l of the analysis solution, injecting the analysis solution into a liquid chromatograph, and recording a chromatogram. The results are shown in FIG. 2. The tenofovir alafenamide and each impurity can be completely separated.
Example 3
(1) Instrumentation and high performance liquid chromatography conditions
High performance liquid chromatograph: a Thermo U3000 high performance liquid chromatography system and workstation;
a chromatographic column: ACE Excel C18 (4.6X 150mm, 5 μm) octadecylsilane chemically bonded silica chromatographic column
20mmo/L sodium dihydrogen phosphate buffer solution (pH6.0) -methanol (92:8) is used as a mobile phase A, 20mmo/L sodium dihydrogen phosphate buffer solution (pH5.0) -methanol (35:65) is used as a mobile phase B, the proportions of the A phase and the B phase in the mobile phase are 0, 15, 35, 40, 50 and 60 minutes, the proportion of the A phase is 90%, 40%, 30%, 0%, 90% and 90%, the detection wavelength is 260nm, the flow rate is 1.5ml/min, and the column temperature is 30 ℃.
(2) Experimental procedure
Respectively taking an impurity A-an impurity J reference substance and a proper amount of tenofovir alafenamide hemifumarate, and preparing a mixed solution containing 1mg of tenofovir alafenamide and 10 micrograms of the rest impurity reference substances in each 1ml by using methanol;
and (5) taking 5 mu l of the analysis solution, injecting the analysis solution into a liquid chromatograph, and recording a chromatogram. The results are shown in FIG. 3. The tenofovir alafenamide and each impurity can be completely separated.
Example 4
(1) Instrumentation and high performance liquid chromatography conditions
High performance liquid chromatograph: a Thermo U3000 high performance liquid chromatography system and workstation;
a chromatographic column: ACE Excel C18 (4.6X 150mm, 5 μm) octadecylsilane chemically bonded silica chromatographic column
20mmo/L sodium dihydrogen phosphate buffer solution (pH6.0) -methanol (92:8) is used as a mobile phase A, 20mmo/L sodium dihydrogen phosphate buffer solution (pH6.0) -methanol (35:65) is used as a mobile phase B, the proportions of the A phase and the B phase in the mobile phase are 0, 15, 35, 40, 50 and 60 minutes, the proportion of the A phase is 95%, 45%, 35%, 0%, 95% and 95%, the detection wavelength is 260nm, the flow rate is 1.0ml/min, and the column temperature is 20 ℃.
(2) Experimental procedure
Respectively taking an impurity A-an impurity J reference substance and a proper amount of tenofovir alafenamide hemifumarate, and preparing a mixed solution containing 1mg of tenofovir alafenamide and 10 micrograms of the rest impurity reference substances in each 1ml by using methanol;
and (5) taking 5 mu l of the analysis solution, injecting the analysis solution into a liquid chromatograph, and recording a chromatogram. The results are shown in FIG. 4. The tenofovir alafenamide and each impurity can be completely separated.
As shown in the following table, the separation degree of the method in the embodiment 1 is poor, the impurities H and I are wrapped by API, the separation in the embodiments 2-4 is good, and the complete separation of 10 impurities can be realized in the same system.
Table 1 comparison table of impurity separation ratios in examples 1 to 4
Note: impurity G appears as two chromatographic peaks in the chromatogram due to its isomer.
Claims (10)
1. An analysis method for determining related substances of tenofovir alafenamide hemifumarate is characterized in that:
(1) preparing a tenofovir alafenamide hemifumarate analysis solution;
(2) adopting high performance liquid chromatography, wherein the chromatographic conditions comprise: the chromatographic column is an octadecyl bonded silica gel chromatographic column, a solution with a volume ratio of phosphate buffer solution to methanol of 92:8 is used as a mobile phase A, and a solution with a volume ratio of phosphate buffer solution to methanol of 30:70 is used as a mobile phase B. The gradient elution process is as follows: the proportion of the A phase and the B phase in the mobile phase is 0, 15-20, 35-38, 40-45, 50 and 60 minutes, the proportion of the A phase is 90-95%, 40-45%, 30-35%, 0%, 90-95% and 90-95%, and the detection wavelength is 260 nm;
(3) and (3) sample determination: injecting the analysis solution prepared in the step (1) into a liquid chromatograph, recording a chromatogram, and separating the following related substances:
impurity I
2. The assay method for assaying tenofovir alafenamide hemifumarate related substances according to claim 1, wherein the step (1) of preparing the assay solution comprises the following specific steps: dissolving a sample by using a single solvent of methanol and acetonitrile or a mixed solution of methanol-water, acetonitrile-water, a methanol-phosphate aqueous solution and an acetonitrile-phosphate aqueous solution to prepare an analysis solution.
3. The assay for the determination of tenofovir alafenamide hemifumarate related substance of claim 1, wherein the length of the column is 150mm, the diameter is 4.6mm, and the particle size of the packing is 5 μm.
4. The assay for the determination of a substance related to tenofovir alafenamide hemifumarate of claim 1, wherein the chromatography column is an ACE Excel C18 chromatography column.
5. The analysis method for determining substances related to tenofovir alafenamide hemifumarate according to claim 1, wherein the concentration of sodium dihydrogen phosphate in a phosphate buffer solution is 15 to 25 mmol/L.
6. The analytical method for determining substances related to tenofovir alafenamide hemifumarate according to claim 1, wherein the concentration of the sodium hydroxide solution for adjusting the pH value is 0.1 to 1.0 mol/L.
7. The assay method for assaying tenofovir alafenamide hemifumarate-related substances according to claim 1, wherein the pH is adjusted to 4 to 7 with 0.5mol/L sodium hydroxide solution in phosphate buffer.
8. The analytical method for determining substances related to tenofovir alafenamide hemifumarate according to claim 1, wherein the flow rate is 1.2 to 1.8 ml/min.
9. The analytical method for determining substances related to tenofovir alafenamide hemifumarate according to claim 1, wherein the column temperature is 15 to 40 ℃.
10. The assay method for determining tenofovir alafenamide hemifumarate related substances according to claim 1, wherein the sample is a raw material drug of tenofovir alafenamide hemifumarate or a preparation thereof, and the preparation is tablet, capsule, granule, suppository, pill, ointment, cream, paste, inhalation, spray, aerosol, gel, powder, syrup, liniment, film coating, tincture, patch, oral solution, implant, film, lotion, rinse, decoction, ointment, lotion, tea.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010303085.6A CN111650290A (en) | 2020-04-16 | 2020-04-16 | Analysis method of tenofovir alafenamide hemifumarate related substances |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010303085.6A CN111650290A (en) | 2020-04-16 | 2020-04-16 | Analysis method of tenofovir alafenamide hemifumarate related substances |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111650290A true CN111650290A (en) | 2020-09-11 |
Family
ID=72352145
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010303085.6A Pending CN111650290A (en) | 2020-04-16 | 2020-04-16 | Analysis method of tenofovir alafenamide hemifumarate related substances |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111650290A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113777186A (en) * | 2021-08-12 | 2021-12-10 | 北京鑫开元医药科技有限公司 | Method for detecting impurities in propane fumarate tenofovir |
-
2020
- 2020-04-16 CN CN202010303085.6A patent/CN111650290A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113777186A (en) * | 2021-08-12 | 2021-12-10 | 北京鑫开元医药科技有限公司 | Method for detecting impurities in propane fumarate tenofovir |
| CN113777186B (en) * | 2021-08-12 | 2023-06-13 | 北京鑫开元医药科技有限公司 | Method for detecting impurities in propionofovir fumarate |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Recent applications of hydrophilic interaction liquid chromatography in pharmaceutical analysis | |
| CN107064350B (en) | Method for detecting suspected genotoxic impurity of tofacitinib citrate | |
| CN107655987B (en) | HPLC detection method for tenofovir alafenamide and isomer thereof | |
| CN108802255B (en) | Method for determining content of ganoderma lucidum triterpene component in compound preparation | |
| CN106822203B (en) | Radix angelicae pubescentis granules and preparation method and quality control method thereof | |
| Lopes et al. | Quantification of carbamazepine and its active metabolite by direct injection of human milk serum using liquid chromatography tandem ion trap mass spectrometry | |
| CN103323540A (en) | Content determination method for amino acids in Jinshuibao preparation | |
| CN110261531B (en) | Method for detecting related substances in loxoprofen or sodium salt thereof | |
| CN111650290A (en) | Analysis method of tenofovir alafenamide hemifumarate related substances | |
| CN114252532B (en) | Method for establishing fingerprint of Fangfengsheng particles and fingerprint thereof | |
| CN108490083B (en) | Quality detection method of Suhuang cough-relieving capsule | |
| CN109239253B (en) | High performance liquid detection method for impurities of abacavir | |
| CN101816753B (en) | Method for detecting quality of compound preparation for treating cold | |
| CN110907583B (en) | Method for separating related substances in loxoprofen or sodium salt thereof | |
| CN112147239B (en) | Method for detecting nucleoside content in polyporus umbellatus | |
| KR102110966B1 (en) | Process for Producing Deoxypodophilotoxin in Quantity | |
| CN109342579B (en) | HPLC fingerprint detection method for traditional Chinese medicine for relaxing bowel | |
| CN113655135A (en) | Quantitative detection method and limiting method of aristolochic acid I | |
| CN107655986B (en) | Detection method of related substances of vipatavir | |
| CN103800397B (en) | The technique of macroporous resin purification Herba Ephedrae total alkaloids | |
| CN106290596A (en) | The method that separation analysis succinum love song Ge Lieting and preparation thereof have related substance | |
| CN101377475A (en) | Quality control method suitable for solancarpine, solamargine and preparation thereof | |
| CN112209882A (en) | Levoornidazole impurity C and preparation method and application thereof | |
| CN114034805B (en) | Metabolic kinetic analysis method of pentafluorouracil drugs | |
| Kouno et al. | Direct sample injection into the high-performance liquid chromatographic column in theophylline monitoring |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 211100 building 7, life science and Technology Innovation Park, 568 longmian Avenue, Jiangning District, Nanjing City, Jiangsu Province Applicant after: Jiangsu ailikang Pharmaceutical Technology Co.,Ltd. Address before: 211100 building 7, life science and Technology Innovation Park, 568 longmian Avenue, Jiangning District, Nanjing City, Jiangsu Province Applicant before: JIANGSU ALICORN PHARMACEUTICAL Co.,Ltd. |
|
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |