CN114414680B - Method for measuring related substances in various small molecule anhydrides - Google Patents
Method for measuring related substances in various small molecule anhydrides Download PDFInfo
- Publication number
- CN114414680B CN114414680B CN202111642088.3A CN202111642088A CN114414680B CN 114414680 B CN114414680 B CN 114414680B CN 202111642088 A CN202111642088 A CN 202111642088A CN 114414680 B CN114414680 B CN 114414680B
- Authority
- CN
- China
- Prior art keywords
- anhydride
- solution
- measuring
- diluting
- stock solution
- 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.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 28
- -1 small molecule anhydrides Chemical class 0.000 title claims abstract description 26
- 239000000126 substance Substances 0.000 title claims abstract description 19
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 69
- YNAVUWVOSKDBBP-UHFFFAOYSA-N Morpholine Chemical compound C1COCCN1 YNAVUWVOSKDBBP-UHFFFAOYSA-N 0.000 claims description 66
- 150000008064 anhydrides Chemical class 0.000 claims description 50
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 48
- 239000000243 solution Substances 0.000 claims description 44
- 239000011550 stock solution Substances 0.000 claims description 39
- 238000007865 diluting Methods 0.000 claims description 37
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 32
- 238000005303 weighing Methods 0.000 claims description 26
- LSACYLWPPQLVSM-UHFFFAOYSA-N isobutyric acid anhydride Chemical compound CC(C)C(=O)OC(=O)C(C)C LSACYLWPPQLVSM-UHFFFAOYSA-N 0.000 claims description 22
- 150000008065 acid anhydrides Chemical class 0.000 claims description 20
- 239000012071 phase Substances 0.000 claims description 18
- 239000012535 impurity Substances 0.000 claims description 17
- YHASWHZGWUONAO-UHFFFAOYSA-N butanoyl butanoate Chemical compound CCCC(=O)OC(=O)CCC YHASWHZGWUONAO-UHFFFAOYSA-N 0.000 claims description 16
- 238000001514 detection method Methods 0.000 claims description 16
- WYVAMUWZEOHJOQ-UHFFFAOYSA-N propionic anhydride Chemical compound CCC(=O)OC(=O)CC WYVAMUWZEOHJOQ-UHFFFAOYSA-N 0.000 claims description 16
- FREZLSIGWNCSOQ-UHFFFAOYSA-N 3-methylbutanoyl 3-methylbutanoate Chemical compound CC(C)CC(=O)OC(=O)CC(C)C FREZLSIGWNCSOQ-UHFFFAOYSA-N 0.000 claims description 14
- DUCKXCGALKOSJF-UHFFFAOYSA-N pentanoyl pentanoate Chemical compound CCCCC(=O)OC(=O)CCCC DUCKXCGALKOSJF-UHFFFAOYSA-N 0.000 claims description 14
- PKHMTIRCAFTBDS-UHFFFAOYSA-N hexanoyl hexanoate Chemical compound CCCCCC(=O)OC(=O)CCCCC PKHMTIRCAFTBDS-UHFFFAOYSA-N 0.000 claims description 13
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 11
- 238000000926 separation method Methods 0.000 claims description 11
- 238000010828 elution Methods 0.000 claims description 10
- 239000012488 sample solution Substances 0.000 claims description 10
- 239000000945 filler Substances 0.000 claims description 8
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 8
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 6
- 238000001212 derivatisation Methods 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 239000000741 silica gel Substances 0.000 claims description 5
- 229910002027 silica gel Inorganic materials 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 239000000377 silicon dioxide Substances 0.000 claims description 3
- 239000007791 liquid phase Substances 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 238000004007 reversed phase HPLC Methods 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 239000003814 drug Substances 0.000 abstract description 5
- 238000004458 analytical method Methods 0.000 abstract description 4
- 238000005259 measurement Methods 0.000 abstract description 2
- 238000003908 quality control method Methods 0.000 abstract 1
- 238000011835 investigation Methods 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 7
- 239000012490 blank solution Substances 0.000 description 5
- 239000012085 test solution Substances 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000002253 acid Substances 0.000 description 2
- 239000003470 adrenal cortex hormone Substances 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- KWOLFJPFCHCOCG-UHFFFAOYSA-N Acetophenone Chemical compound CC(=O)C1=CC=CC=C1 KWOLFJPFCHCOCG-UHFFFAOYSA-N 0.000 description 1
- ITPDYQOUSLNIHG-UHFFFAOYSA-N Amiodarone hydrochloride Chemical compound [Cl-].CCCCC=1OC2=CC=CC=C2C=1C(=O)C1=CC(I)=C(OCC[NH+](CC)CC)C(I)=C1 ITPDYQOUSLNIHG-UHFFFAOYSA-N 0.000 description 1
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 1
- PDMMFKSKQVNJMI-BLQWBTBKSA-N Testosterone propionate Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](OC(=O)CC)[C@@]1(C)CC2 PDMMFKSKQVNJMI-BLQWBTBKSA-N 0.000 description 1
- 230000000397 acetylating effect Effects 0.000 description 1
- 229960001138 acetylsalicylic acid Drugs 0.000 description 1
- 229960003234 amiodarone hydrochloride Drugs 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960001102 betamethasone dipropionate Drugs 0.000 description 1
- CIWBQSYVNNPZIQ-XYWKZLDCSA-N betamethasone dipropionate Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@H](C)[C@@](C(=O)COC(=O)CC)(OC(=O)CC)[C@@]1(C)C[C@@H]2O CIWBQSYVNNPZIQ-XYWKZLDCSA-N 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- NNXRFDWZYCXPSP-UHFFFAOYSA-N butyl 3-oxohexanoate Chemical compound CCCCOC(=O)CC(=O)CCC NNXRFDWZYCXPSP-UHFFFAOYSA-N 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 229960001252 methamphetamine Drugs 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- FDDDEECHVMSUSB-UHFFFAOYSA-N sulfanilamide Chemical compound NC1=CC=C(S(N)(=O)=O)C=C1 FDDDEECHVMSUSB-UHFFFAOYSA-N 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229960001712 testosterone propionate Drugs 0.000 description 1
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
- G01N30/26—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
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)
- Spectroscopy & Molecular Physics (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention discloses a method for measuring related substances in various small molecule anhydrides, which belongs to the technical field of medicine analysis. The invention has the advantages of high sensitivity, high precision, accurate content measurement result, good specificity and linear relation and the like, and can be used for quality control of various small molecule anhydrides.
Description
Technical Field
The invention belongs to the technical field of medicine analysis, and particularly relates to a method for measuring related substances in various small molecule anhydrides.
Background
Anhydrides (Anhydrides) are those where an oxyacid removes one or more molecules of water, leaving the remainder. Generally, an inorganic acid is an acid anhydride of an acid formed by directly losing one molecule of water from one molecule of the acid, and the valence of an element determining acidity in the acid anhydride is not changed. Some small molecule anhydrides play an important role in the synthesis and manufacture of the compounds. For example, acetic anhydride is an important acetylating reagent used in the pharmaceutical industry for the manufacture of synthetic milnaconitriles, dibazoles, caffeine and aspirin, sulfanilamide drugs, etc., as well as formulations for the manufacture of hypnone, neohypnone, methamphetamine. Propionic anhydride is used in the pharmaceutical industry as a propionylating agent for the manufacture of medicines, fragrances and special esters for the production of angular sarcin propionate (antibiotic), testosterone propionate (male hormonal deficiency), hydroxymethylandrosterone propionate (anticancer), clobetasol-norubicin-sinate (adrenocortical hormone), betamethasone dipropionate (adrenocortical hormone), etc. Butyric anhydride is a main raw material for producing butyl butyrylacetate and perfume, and also for producing amiodarone hydrochloride. The isobutyric anhydride can be used for producing Monumpiravir (antiviral drug), and has wide application in essence and spice, organic solvent, medical intermediate and pesticide.
Anhydride has wide application in the pharmaceutical industry and is a starting material or a key reagent in the production of most raw materials. However, anhydride has the characteristics of high activity, easy hydrolysis and the like, and the development of an analysis method is greatly limited. At present, the main analysis method for anhydride is gas chromatography, but under the condition of high temperature, cross reaction occurs among anhydrides, and the accuracy of a detection result is affected.
Disclosure of Invention
The invention aims to: in order to solve the problems, ensure the quality of anhydride and avoid introducing new impurities in the production process of medicines, the invention can effectively analyze and detect related substances in target anhydride after a derivatization reaction by searching a proper derivatization agent.
The technical scheme is as follows: the invention adopts the following technical scheme:
the invention provides a method for measuring related substances in a plurality of small molecule anhydrides, which comprises the following steps: acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride, n-caproic anhydride; preparing a sample solution by taking morpholine as a derivatization reagent when measuring related substances in small molecule anhydride, and then detecting by using a high performance liquid chromatography; the preparation method of the test sample solution comprises the following steps:
weighing anhydride to be measured, placing the anhydride to be measured into a measuring flask, and adding morpholine, wherein the molar ratio of the anhydride to be measured to the morpholine is 1:1.5 to 1:10, the molar ratio of anhydride to morpholine to be measured is preferably 1:2, diluting with acetonitrile until the content of anhydride to be detected is 20 mg/ml-1 mg/ml, uniformly mixing, standing at room temperature for 3-7 min, preferably standing at room temperature for 5min, and obtaining anhydride stock solution to be detected; during detection, the anhydride stock solution to be detected is measured, placed in a measuring flask, diluted to 500 mug/ml-0.1 mug/ml by water and evenly mixed, thus obtaining the acid anhydride.
The preparation method of the preferred test solution comprises the following steps:
weighing 200mg of anhydride to be detected, placing the anhydride to be detected into a 20ml measuring flask, adding 0.2ml of morpholine, diluting to a scale with acetonitrile, shaking uniformly, and standing at room temperature for 5 minutes to obtain anhydride stock solution to be detected; precisely measuring 1ml of anhydride stock solution to be measured, placing into a 10ml measuring flask, diluting with water to scale, and shaking.
The detection method of the high performance liquid chromatography comprises the following steps: the chromatographic column with octadecylsilane chemically bonded silica gel as filler has ultraviolet absorption detector and detection wavelength of 205-215 nm and column temperature of 20-30 deg.c, and the process includes gradient elution with water as mobile phase A and acetonitrile as mobile phase B; the flow rate of the mobile phase is 0.6 ml/min-1.0 ml/min.
Wherein, the chromatographic column taking octadecylsilane chemically bonded silica as a filler is ACE Excel 5super C18, the specification is 4.6mm multiplied by 250mm, and the particle size of the filler is 5 mu m.
Wherein, the gradient elution procedure is as follows,
wherein the flow rate of the mobile phase is preferably 0.8ml/min.
Wherein the detection wavelength of the ultraviolet detector of the liquid chromatograph is preferably 210nm.
Wherein the column temperature of the chromatographic column is 25 ℃.
The invention provides a method for measuring related substances in various small molecule anhydrides, which comprises the following specific operation processes:
(1) Preparing an anhydride separation degree solution, a sample solution and a system applicability solution;
(2) Setting high performance liquid phase detection conditions: the chromatographic column uses octadecylsilane chemically bonded silica gel as a filler, water as a mobile phase A, acetonitrile as a mobile phase B, the flow rate is 0.8ml/min, gradient elution is carried out, the detection wavelength of a detector is 210nm, and the column temperature of the chromatographic column is 25 ℃;
(3) And respectively sucking the acid anhydride separation degree solution, the sample solution and the system applicability solution, and injecting into a high performance liquid chromatograph for HPLC analysis.
The preparation method of the acid anhydride separation degree solution comprises the following steps:
weighing acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride and n-caproic anhydride separately, placing in different measuring flasks, and adding morpholine respectively, wherein the ratio of acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride, n-caproic anhydride to morpholine is 1:1.5 to 1:10, diluting with acetonitrile to a scale, shaking uniformly, standing, respectively weighing the above solutions, placing in a same measuring flask, diluting with water to the scale, shaking uniformly, and diluting with water twice when in use to obtain an anhydride resolution solution;
the preparation method of the preferable acid anhydride separation degree solution comprises the following steps:
respectively weighing 20mg of acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride and n-caproic anhydride, placing into different 20ml measuring flasks, respectively adding 0.2ml of morpholine, diluting to scale with acetonitrile, shaking, standing at room temperature for 5 minutes, respectively weighing 1ml, placing into the same 100ml measuring flask, diluting to scale with water, shaking; precisely weighing 5ml, placing in a 10ml measuring flask, diluting with water to scale, and shaking;
the preparation method of the system applicability solution comprises the following steps:
(S1) respectively weighing acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride and n-caproic anhydride, placing the mixture in different measuring bottles, and respectively adding morpholine; the ratio of acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride, n-caproic anhydride and morpholine is 1:1.5 to 1:10, diluting with acetonitrile to a scale, shaking uniformly, standing, respectively weighing the solutions, placing the solutions in the same measuring flask, diluting with water to the scale, shaking uniformly, and taking the solutions as impurity stock solution;
weighing anhydride to be measured, placing the anhydride into a measuring bottle, adding morpholine, diluting to a scale with acetonitrile, shaking uniformly, and standing at room temperature for 3-7 minutes to serve as anhydride stock solution to be measured;
and (S2) respectively measuring the impurity stock solution and the anhydride stock solution to be measured, placing the impurity stock solution and the anhydride stock solution to be measured in a measuring flask, diluting the solution by 10 times with water, and shaking the solution uniformly to obtain the acid anhydride stock solution.
The preferred system applicability solution is formulated as follows:
(S1) respectively weighing 20mg of acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride and n-caproic anhydride, placing into different 20ml measuring bottles, respectively adding 0.2ml of morpholine, diluting to a scale with acetonitrile, shaking, standing at room temperature for 5 minutes, respectively weighing 1ml, placing into the same 100ml measuring bottle, diluting to the scale with water, shaking to be uniform and taking as an impurity stock solution;
weighing 200mg of anhydride to be detected, placing the anhydride to be detected into a 20ml measuring flask, adding 0.2ml of morpholine, diluting to a scale with acetonitrile, shaking uniformly, and standing at room temperature for 5 minutes to obtain anhydride stock solution to be detected;
and (S2) respectively measuring 1ml of the impurity stock solution and 1ml of the anhydride stock solution to be measured, placing the mixture into a 10ml measuring flask, diluting the mixture to a scale with water, and shaking the mixture uniformly to obtain the acid anhydride stock solution.
The beneficial effects are that:
the invention discloses a method for measuring related substances in various small-molecule anhydrides, which has the advantages of high sensitivity, wide linear range and strong specificity, and can be used for controlling the quality of the related substances in various small-molecule anhydrides. The invention adopts the derivatization reaction to greatly improve the defects of poor specificity and accuracy caused by easy hydrolysis of anhydride and easy cross reaction at high temperature, and lays a foundation for the quality evaluation of various small molecule anhydrides. The invention selects morpholine as the derivative agent, which can react rapidly and completely at normal temperature; and morpholine can react with various anhydrides and can be effectively separated. The invention can effectively separate various isomer anhydride compounds.
Drawings
FIG. 1 is a chromatogram of a hollow white solution of example 1;
FIG. 2 is a chromatogram of the acid anhydride resolution solution of example 1;
FIG. 3 is a chromatogram of the system applicability solution in example 2.
FIG. 4 is a chromatogram of the sample solution in example 2.
Detailed Description
The following is a further detailed description of the method of the present invention for detecting a substance of interest in a wide variety of small molecule anhydrides, in two examples.
Example 1:
(1) High performance liquid chromatography conditions
The column was octadecylsilane chemically bonded silica gel column, model ACE Excel 5super C18 (4.6 mm. Times.250 mm,5 μm), water as mobile phase A, acetonitrile as mobile phase B, gradient elution was carried out according to the following Table 1 at a flow rate of 0.8ml per minute; the detection wavelength is 210nm; the column temperature was 25 ℃.
TABLE 1 gradient elution
(2) Solution preparation
Each anhydride stock solution: and (3) respectively weighing about 20mg of acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride and n-caproic anhydride, placing into different 20ml measuring bottles, respectively adding 0.2ml of morpholine, diluting to a scale with acetonitrile, shaking uniformly, and standing at room temperature for 5 minutes to obtain the finished product.
Acid anhydride resolution solution: weighing 1ml respectively, placing into the same 100ml measuring flask, diluting with water to scale, and shaking; precisely weighing 5ml, placing in a 10ml measuring flask, diluting with water to scale, and shaking.
Blank solution: taking 0.2ml of morpholine, placing the morpholine into a 20ml measuring flask, diluting the morpholine to a scale with acetonitrile, shaking the flask uniformly, standing the flask at room temperature for 5 minutes, measuring 1ml of morpholine, placing the flask into a 100ml measuring flask, diluting the flask to the scale with water, and shaking the flask uniformly; precisely weighing 5ml, placing in a 10ml measuring flask, diluting with water to scale, and shaking.
And precisely measuring 5 mu l of each of the anhydride separation degree solution and the blank solution, injecting into a liquid chromatograph, and recording a chromatogram.
(3) Separation degree investigation
A blank solution and an acid anhydride isolation solution were prepared and measured by the method described in the examples. The patterns are shown in fig. 1 and 2. The result shows that the chromatographic conditions of the invention are adopted, the blank solution is free from interference, and chromatographic peaks and adjacent peaks can be effectively separated after the acid anhydride is derived, so that the method has good specificity.
(4) Linear investigation
The respective acid anhydride stock solutions were prepared by the method described in the examples, and the respective acid anhydride stock solutions were precisely measured in appropriate amounts, and diluted with water to prepare a series of concentrations as linear solutions of the respective components. The results of the measurements are shown in Table 2 below. The result shows that the linear relation of each component to be detected is good by adopting the chromatographic condition of the invention.
Table 2 results of linear investigation
(5) Solution stability investigation
After the acid anhydride isolation solution was left at room temperature for 24 hours, 5. Mu.l of the solution was precisely sucked up and injected into a liquid chromatograph, and a chromatogram was recorded. The results are shown in Table 3. The result shows that the acid anhydride separation degree solution is placed for 24 hours at room temperature, the peak area of each component to be detected is compared with that of the component to be detected for 0 hour, the ratio is between 90.0% and 110.0%, the solution stability is good in the method, and the test solution is stable in the room temperature for at least 24 hours.
TABLE 3 results of solution stability investigation
| Map name | Peak area of 0h | Area of peak at 24h | Ratio to 0h peak area |
| Acetic anhydride | 316.964 | 317.546 | 100.2％ |
| Propionic anhydride | 262.182 | 259.614 | 99.0％ |
| N-butyric anhydride | 215.640 | 215.594 | 100.0％ |
| Isobutyric anhydride | 185.204 | 186.564 | 100.7％ |
| N-valeric anhydride | 190.290 | 194.647 | 102.3％ |
| Isopentanic anhydride | 196.514 | 198.431 | 101.0％ |
| N-caproic acid anhydride | 143.754 | 144.842 | 100.8％ |
Example 2: the method of the invention is used for analyzing the isobutyric anhydride.
(1) High performance liquid chromatography conditions
The chromatographic column is octadecylsilane chemically bonded silica gel column, the model is ACE Excel 5super C18 (4.6 mm. Times.250 mm,5 μm), water is used as mobile phase A, acetonitrile is used as mobile phase B, gradient elution is carried out according to the above table 1, and the flow rate is 0.8ml per minute; the detection wavelength is 210nm; the column temperature was 25 ℃.
(2) Solution preparation
Test solution: weighing about 200mg of isobutyric anhydride, placing into a 20ml measuring flask, adding 0.2ml of morpholine, diluting to a scale with acetonitrile, shaking uniformly, and standing at room temperature for 5 minutes to obtain isobutyric anhydride stock solution; precisely measuring 1ml, placing in a 10ml measuring flask, diluting with water to scale, and shaking.
Impurity stock solution: respectively weighing about 20mg of acetic anhydride, propionic anhydride and n-butyric anhydride, placing into different 20ml measuring flasks, respectively adding 0.2ml of morpholine, diluting to scale with acetonitrile, shaking, standing at room temperature for 5min, respectively weighing 1ml, placing into the same 100ml measuring flask, diluting to scale with water, shaking to obtain impurity stock solution;
system applicability solution: measuring 1ml of each of the impurity stock solution and the isobutyric anhydride stock solution, placing into a 10ml measuring flask, diluting with water to scale, and shaking.
(3) Molar ratio of morpholine
Referring to the preparation method of the sample solution in the example 2, about 200mg of isobutyric anhydride is respectively weighed and put into different 20ml measuring flasks, 0.15ml, 0.2ml and 0.3ml of morpholine are respectively added, diluted to the scale by acetonitrile, shaken uniformly and placed at room temperature for 5 minutes to be used as a stock solution of isobutyric anhydride; precisely measuring 1ml respectively, placing into a 10ml measuring flask, diluting with water to scale, and shaking. The test results show that the mole ratio of the added morpholine is 1:1.5 to 1: in the range of 3, the recovery rate of the main peak is 100.9% -101.2%, and the absolute difference of the impurity contents is less than 0.02%, namely the reaction is complete.
(4) Separation degree investigation
Test solutions were prepared and measured by the method described in example 2. The map is shown in figure 3. The results show that the chromatographic conditions of the invention are adopted, the blank solution is free from interference, and the isobutyric anhydride peak can be effectively separated from the adjacent peaks and from the impurity peaks, thus the method has good specificity.
(5) Sensitivity investigation
The impurity stock solutions were prepared by the method of example 2, and the respective impurity control stock solutions were precisely measured in appropriate amounts, and the respective impurity control stock solutions were gradually diluted with a diluent and measured, and the detection sensitivity was examined using a signal-to-noise ratio of about 10:1 as a quantitative limit. The results are shown in Table 4, and the results show that the method has good sensitivity.
TABLE 4 sensitivity investigation results
In conclusion, the method provided by the invention is used for measuring related substances in various small-molecule anhydrides, and has the advantages of high sensitivity, wide linear range and strong specificity.
It should be understood that the above examples of the present invention are provided for clarity of illustration only and are not intended to limit the embodiments of the present invention. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. Any modification, equivalent replacement, improvement, etc. which come within the spirit and principles of the invention are desired to be protected by the following claims.
Claims (6)
1. A method for determining related substances in a plurality of small molecule anhydrides, which is characterized in that the small molecule anhydride comprises: acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride, n-caproic anhydride; preparing a sample solution by taking morpholine as a derivatization reagent when related substances in the small molecule anhydride are measured, and then detecting by using a reversed-phase high-performance liquid chromatography;
the preparation method of the sample solution comprises the following steps:
weighing anhydride to be measured, placing the anhydride to be measured into a measuring flask, and adding morpholine, wherein the molar ratio of the anhydride to be measured to the morpholine is 1:1.5 to 1:10, diluting with acetonitrile until the content of anhydride to be detected is 20 mg/ml-1 mg/ml, uniformly mixing, and standing at room temperature to obtain anhydride stock solution to be detected; during detection, the anhydride stock solution to be detected is measured, the anhydride stock solution to be detected is placed in a measuring flask, diluted with water until the content of the anhydride to be detected is 500 mug/ml to 0.1 mug/ml, and the anhydride stock solution to be detected is evenly mixed to obtain the acid anhydride;
the detection method of the high performance liquid chromatography comprises the following steps:
adopting octadecylsilane chemically bonded silica as a chromatographic column of a filler, wherein a detector is an ultraviolet absorption detector, the detection wavelength is 205-215 nm, the column temperature is 20-30 ℃, a mobile phase A is water, a mobile phase B is acetonitrile, the flow rate of the mobile phase is 0.6-1.0 ml/min, and gradient elution is carried out;
the chromatographic column taking octadecylsilane chemically bonded silica as a filler is ACE Excel 5super C18, the specification is 4.6mm multiplied by 250mm, and the particle size of the filler is 5 mu m;
the gradient elution is carried out according to the following elution program:
。
2. The method for measuring a substance of interest in a plurality of small molecule anhydrides according to claim 1, wherein: the flow rate of the mobile phase is 0.8ml/min.
3. The method for measuring a substance of interest in a plurality of small molecule anhydrides according to claim 1, wherein: the detection wavelength of the ultraviolet absorption detector is 210nm.
4. The method for measuring a substance of interest in a plurality of small molecule anhydrides according to claim 1, wherein: the column temperature of the chromatographic column is 25 ℃.
5. The method for measuring a substance of interest in a plurality of small molecule anhydrides according to claim 1, wherein: the operation process is as follows,
(1) Preparing an anhydride separation degree solution, a sample solution and a system applicability solution;
(2) Setting high performance liquid phase detection conditions: the chromatographic column uses octadecylsilane chemically bonded silica gel as a filler, water as a mobile phase A, acetonitrile as a mobile phase B, the flow rate is 0.8ml/min for gradient elution, the detection wavelength is 210nm, and the column temperature of the chromatographic column is 25 ℃;
(3) And respectively sucking the acid anhydride separation degree solution, the sample solution and the system applicability solution, and injecting into a high performance liquid chromatograph for HPLC analysis.
6. The method for measuring a substance of interest in a plurality of small molecule anhydrides according to claim 5,
the preparation method of the acid anhydride separation degree solution comprises the following steps: respectively weighing acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride and n-caproic anhydride, placing into different measuring flasks, and respectively adding morpholine, wherein the molar ratio of the acetic anhydride, the propionic anhydride, the n-butyric anhydride, the isobutyric anhydride, the n-valeric anhydride, the isovaleric anhydride, the n-caproic anhydride and the morpholine is 1:1.5 to 1:10, diluting with acetonitrile to a scale, shaking uniformly, standing, respectively weighing the above solutions, placing in a same measuring flask, diluting with water to the scale, shaking uniformly, and diluting with water twice when in use to obtain an anhydride resolution solution;
the preparation method of the system applicability solution comprises the following steps:
(S1) respectively weighing acetic anhydride, propionic anhydride, n-butyric anhydride, isobutyric anhydride, n-valeric anhydride, isovaleric anhydride and n-caproic anhydride, placing the mixture in different measuring bottles, and respectively adding morpholine; the molar ratio of the acetic anhydride, the propionic anhydride, the n-butyric anhydride, the isobutyric anhydride, the n-valeric anhydride, the isovaleric anhydride, the n-caproic anhydride and the morpholine is 1:1.5 to 1:10, diluting with acetonitrile to a scale, shaking uniformly, standing, respectively weighing the solutions, placing the solutions in the same measuring flask, diluting with water to the scale, shaking uniformly, and taking the solutions as impurity stock solution;
weighing anhydride to be measured, placing the anhydride into a measuring flask, adding morpholine, diluting to a scale with acetonitrile, shaking uniformly, and standing at room temperature to serve as anhydride stock solution to be measured;
and (S2) respectively measuring the impurity stock solution and the anhydride stock solution to be measured, placing the impurity stock solution and the anhydride stock solution to be measured in a measuring flask, diluting the solution by 10 times with water, and shaking the solution uniformly to obtain the acid anhydride stock solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111642088.3A CN114414680B (en) | 2021-12-29 | 2021-12-29 | Method for measuring related substances in various small molecule anhydrides |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111642088.3A CN114414680B (en) | 2021-12-29 | 2021-12-29 | Method for measuring related substances in various small molecule anhydrides |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114414680A CN114414680A (en) | 2022-04-29 |
| CN114414680B true CN114414680B (en) | 2023-10-13 |
Family
ID=81269901
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111642088.3A Active CN114414680B (en) | 2021-12-29 | 2021-12-29 | Method for measuring related substances in various small molecule anhydrides |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114414680B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115684394B (en) * | 2022-10-17 | 2024-06-21 | 浙江圣兆药物科技股份有限公司 | Method for detecting trace acetic anhydride in glacial acetic acid |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09286816A (en) * | 1996-02-20 | 1997-11-04 | Nitto Boseki Co Ltd | Acylated polyallylamine and its production |
| RU2124201C1 (en) * | 1996-10-23 | 1998-12-27 | Закрытое акционерное общество "Электроизолит" | Method of determining anhydride groups in organic acid anhydrides |
| JP2003149221A (en) * | 2001-11-07 | 2003-05-21 | Kanegafuchi Chem Ind Co Ltd | Quantitative determination method for acetic anhydride |
| CN113049699A (en) * | 2021-03-17 | 2021-06-29 | 北京鑫开元医药科技有限公司 | Method for detecting biphenyl anhydride and related substances thereof and application |
-
2021
- 2021-12-29 CN CN202111642088.3A patent/CN114414680B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09286816A (en) * | 1996-02-20 | 1997-11-04 | Nitto Boseki Co Ltd | Acylated polyallylamine and its production |
| RU2124201C1 (en) * | 1996-10-23 | 1998-12-27 | Закрытое акционерное общество "Электроизолит" | Method of determining anhydride groups in organic acid anhydrides |
| JP2003149221A (en) * | 2001-11-07 | 2003-05-21 | Kanegafuchi Chem Ind Co Ltd | Quantitative determination method for acetic anhydride |
| CN113049699A (en) * | 2021-03-17 | 2021-06-29 | 北京鑫开元医药科技有限公司 | Method for detecting biphenyl anhydride and related substances thereof and application |
Non-Patent Citations (1)
| Title |
|---|
| Product class 1: pyrylium salts;Balaban, T. S. 等;Science of Synthesis;第14卷;11-200 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114414680A (en) | 2022-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111983074B (en) | Method for determining peramivir intermediate isomer by using high performance liquid chromatography | |
| CN114414680B (en) | Method for measuring related substances in various small molecule anhydrides | |
| CN104749269A (en) | Method for determining enantiomer impurity in alogliptin crude drug and preparation by virtue of HPLC | |
| CN108152418B (en) | HPLC detection method for ketorolac tromethamine or/and impurities in preparation of ketorolac tromethamine | |
| CN114113405B (en) | High performance liquid chromatography analysis method for glycerophosphorylcholine and isomer thereof | |
| CN113049687B (en) | Method for detecting ambroxol hydrochloride raw material and injection related substances | |
| CN114624339A (en) | Analysis method for determining indobufen impurities | |
| CN110646520A (en) | Method for separating and/or detecting impurities in atorvastatin calcium | |
| CN109142585B (en) | Method for detecting isomer of sodium pantothenate | |
| CN110221009B (en) | Method for measuring butylphthalide related substances by HPLC | |
| CN114354789B (en) | Method for simultaneously measuring cabozantinib analogue and related substances thereof | |
| CN116265937A (en) | Detection method and application of oseltamivir phosphate related impurities | |
| CN1967253B (en) | UV spectrophotometry and HPLC combined method for determining content of phenylpropanol | |
| CN109001342B (en) | High performance liquid chromatography method for detecting N-2, 3-dimethoxybenzyl piperonylethylamine and salt content thereof | |
| CN111679004A (en) | Quality control method of probucol | |
| CN114594168A (en) | Method for detecting indobufen impurity | |
| CN116735774B (en) | Method for detecting content of genotoxic impurities in propionofovir fumarate bulk drug | |
| CN114354788B (en) | Method for measuring related substances in Monnpiravir raw material and preparation thereof | |
| CN116754705B (en) | Method for detecting acetic acid and acetate content | |
| CN113686977B (en) | Method for measuring related substances in compound fenbendazole preparation | |
| CN116539765B (en) | Method for detecting related substances of teprenone capsules | |
| CN112578045B (en) | Detection method for simultaneously determining content of 8 compounds in glipizide drugs | |
| CN114280169B (en) | Method for determining isomer in monabivir | |
| CN114397386B (en) | Method for determining 3R-3-tert-butyldimethylsiloxy-5-oxo-6-triphenylphosphine alkene caproic acid methyl ester isomer | |
| CN115469039B (en) | Butylphthalide and detection method of related substances thereof |
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 | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20220429 Assignee: Suzhou Zhengji Pharmaceutical Co.,Ltd. Assignor: JIANGSU ZENJI PHARMACEUTICALS LTD. Contract record no.: X2024980015147 Denomination of invention: A method for determining related substances in various small molecule anhydrides Granted publication date: 20231013 License type: Common License Record date: 20240913 Application publication date: 20220429 Assignee: Rizhao Zhengji Pharmaceutical Co.,Ltd. Assignor: JIANGSU ZENJI PHARMACEUTICALS LTD. Contract record no.: X2024980015140 Denomination of invention: A method for determining related substances in various small molecule anhydrides Granted publication date: 20231013 License type: Common License Record date: 20240913 |
|
| EE01 | Entry into force of recordation of patent licensing contract |