CN111537593B - Method for measuring content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis extracting solution - Google Patents
Method for measuring content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis extracting solution Download PDFInfo
- Publication number
- CN111537593B CN111537593B CN202010495542.6A CN202010495542A CN111537593B CN 111537593 B CN111537593 B CN 111537593B CN 202010495542 A CN202010495542 A CN 202010495542A CN 111537593 B CN111537593 B CN 111537593B
- Authority
- CN
- China
- Prior art keywords
- tetrahydropalmatine
- tetrahydroberberine
- solution
- reference substance
- chiral
- 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
- AEQDJSLRWYMAQI-KRWDZBQOSA-N tetrahydropalmatine Chemical compound C1CN2CC(C(=C(OC)C=C3)OC)=C3C[C@H]2C2=C1C=C(OC)C(OC)=C2 AEQDJSLRWYMAQI-KRWDZBQOSA-N 0.000 title claims abstract description 65
- 238000000034 method Methods 0.000 title claims abstract description 48
- AEQDJSLRWYMAQI-UHFFFAOYSA-N Tetrahydropalmatine Natural products C1CN2CC(C(=C(OC)C=C3)OC)=C3CC2C2=C1C=C(OC)C(OC)=C2 AEQDJSLRWYMAQI-UHFFFAOYSA-N 0.000 title claims abstract description 47
- 241000218176 Corydalis Species 0.000 title claims abstract description 43
- LYFPBWOBIJJASK-INIZCTEOSA-N (S)-(-)-Canadine Natural products O(C)c1c(OC)ccc2c1C[N+]1[C@H](c3c(cc4OCOc4c3)CC1)C2 LYFPBWOBIJJASK-INIZCTEOSA-N 0.000 title claims abstract description 38
- PQECCKIOFCWGRJ-UHFFFAOYSA-N Tetrahydro-berberine Natural products C1=C2C3CC4=CC=C(OC)C(O)=C4CN3CCC2=CC2=C1OCO2 PQECCKIOFCWGRJ-UHFFFAOYSA-N 0.000 title claims abstract description 38
- VZTUIEROBZXUFA-UHFFFAOYSA-N canadine Chemical compound C1=C2C3CC4=CC=C(OC)C(OC)=C4CN3CCC2=CC2=C1OCO2 VZTUIEROBZXUFA-UHFFFAOYSA-N 0.000 title claims abstract description 38
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 title claims abstract description 32
- 238000004458 analytical method Methods 0.000 claims abstract description 23
- 238000001514 detection method Methods 0.000 claims abstract description 14
- 238000001819 mass spectrum Methods 0.000 claims abstract description 13
- 238000000738 capillary electrophoresis-mass spectrometry Methods 0.000 claims abstract description 12
- 238000011049 filling Methods 0.000 claims abstract description 10
- ODLHGICHYURWBS-LKONHMLTSA-N trappsol cyclo Chemical compound CC(O)COC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](COCC(C)O)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)COCC(O)C)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1COCC(C)O ODLHGICHYURWBS-LKONHMLTSA-N 0.000 claims abstract description 8
- 239000000243 solution Substances 0.000 claims description 72
- 239000013558 reference substance Substances 0.000 claims description 50
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 38
- 239000000523 sample Substances 0.000 claims description 23
- UOTMYNBWXDUBNX-UHFFFAOYSA-N 1-[(3,4-dimethoxyphenyl)methyl]-6,7-dimethoxyisoquinolin-2-ium;chloride Chemical compound Cl.C1=C(OC)C(OC)=CC=C1CC1=NC=CC2=CC(OC)=C(OC)C=C12 UOTMYNBWXDUBNX-UHFFFAOYSA-N 0.000 claims description 22
- 229960003207 papaverine hydrochloride Drugs 0.000 claims description 22
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 19
- 235000019253 formic acid Nutrition 0.000 claims description 19
- AEQDJSLRWYMAQI-QGZVFWFLSA-N (13ar)-2,3,9,10-tetramethoxy-6,8,13,13a-tetrahydro-5h-isoquinolino[2,1-b]isoquinoline Chemical compound C1CN2CC(C(=C(OC)C=C3)OC)=C3C[C@@H]2C2=C1C=C(OC)C(OC)=C2 AEQDJSLRWYMAQI-QGZVFWFLSA-N 0.000 claims description 18
- 239000007864 aqueous solution Substances 0.000 claims description 17
- 239000012488 sample solution Substances 0.000 claims description 14
- 239000008151 electrolyte solution Substances 0.000 claims description 13
- 238000002360 preparation method Methods 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 239000000126 substance Substances 0.000 claims description 12
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical compound [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 claims description 10
- 239000012085 test solution Substances 0.000 claims description 9
- VZTUIEROBZXUFA-INIZCTEOSA-N (S)-canadine Chemical compound C1=C2[C@@H]3CC4=CC=C(OC)C(OC)=C4CN3CCC2=CC2=C1OCO2 VZTUIEROBZXUFA-INIZCTEOSA-N 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 8
- 229910021641 deionized water Inorganic materials 0.000 claims description 8
- 239000007789 gas Substances 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- 238000005303 weighing Methods 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 7
- HQVFCQRVQFYGRJ-UHFFFAOYSA-N formic acid;hydrate Chemical compound O.OC=O HQVFCQRVQFYGRJ-UHFFFAOYSA-N 0.000 claims description 6
- 150000002500 ions Chemical class 0.000 claims description 6
- 239000003607 modifier Substances 0.000 claims description 6
- 239000012088 reference solution Substances 0.000 claims description 5
- 238000000926 separation method Methods 0.000 claims description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 230000001419 dependent effect Effects 0.000 claims description 4
- 238000010586 diagram Methods 0.000 claims description 4
- 238000000132 electrospray ionisation Methods 0.000 claims description 4
- 239000005350 fused silica glass Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 238000012544 monitoring process Methods 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 3
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 2
- XQYZDYMELSJDRZ-UHFFFAOYSA-N papaverine Chemical compound C1=C(OC)C(OC)=CC=C1CC1=NC=CC2=CC(OC)=C(OC)C=C12 XQYZDYMELSJDRZ-UHFFFAOYSA-N 0.000 claims 2
- 229930008281 A03AD01 - Papaverine Natural products 0.000 claims 1
- 229960001789 papaverine Drugs 0.000 claims 1
- 238000005406 washing Methods 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 11
- 230000008901 benefit Effects 0.000 abstract description 6
- 239000003814 drug Substances 0.000 abstract description 6
- 238000005516 engineering process Methods 0.000 abstract description 3
- 230000005764 inhibitory process Effects 0.000 abstract description 3
- 238000005259 measurement Methods 0.000 abstract description 3
- 239000003960 organic solvent Substances 0.000 abstract description 3
- 230000009467 reduction Effects 0.000 abstract description 3
- 238000005251 capillar electrophoresis Methods 0.000 description 11
- 241000620709 Corydalis ambigua Species 0.000 description 4
- 229910001873 dinitrogen Inorganic materials 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 238000004811 liquid chromatography Methods 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 208000002193 Pain Diseases 0.000 description 2
- 239000008186 active pharmaceutical agent Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000003821 enantio-separation Methods 0.000 description 2
- 238000000227 grinding Methods 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 230000036407 pain Effects 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 235000021419 vinegar Nutrition 0.000 description 2
- 239000000052 vinegar Substances 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 201000000736 Amenorrhea Diseases 0.000 description 1
- 206010001928 Amenorrhoea Diseases 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- 208000005171 Dysmenorrhea Diseases 0.000 description 1
- 206010013935 Dysmenorrhoea Diseases 0.000 description 1
- 206010061876 Obstruction Diseases 0.000 description 1
- 206010042674 Swelling Diseases 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 231100000540 amenorrhea Toxicity 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000002306 biochemical method Methods 0.000 description 1
- 238000005460 biophysical method Methods 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000000038 chest Anatomy 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 229940097362 cyclodextrins Drugs 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 235000017807 phytochemicals Nutrition 0.000 description 1
- 229930000223 plant secondary metabolite Natural products 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000013074 reference sample Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 230000008736 traumatic injury Effects 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44756—Apparatus specially adapted therefor
- G01N27/44791—Microapparatus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44717—Arrangements for investigating the separated zones, e.g. localising zones
- G01N27/4473—Arrangements for investigating the separated zones, e.g. localising zones by electric means
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44743—Introducing samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2866—Grinding or homogeneising
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Electrochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dispersion Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
A method for measuring the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis tuber extracting solution belongs to the technical field of measurement of traditional Chinese medicine components. The method adopts capillary electrophoresis-mass spectrometry combined analysis, and realizes the direct determination of the contents of two pairs of chiral enantiomers of tetrahydropalmatine and tetrahydroberberine in a complicated sample system of corydalis tuber extract by using hydroxypropyl-beta-cyclodextrin as a chiral selector by means of a partial filling technology. The method has the advantages of simple operation, high sensitivity, low detection cost (few samples, chiral selectors and organic solvents are consumed), and high practicability, can directly determine the contents of two pairs of chiral enantiomers of tetrahydropalmatine and tetrahydroberberine in a complicated sample system of the corydalis tuber extracting solution, simultaneously ensures that the two pairs of chiral enantiomers are well separated, avoids ionization inhibition and sensitivity reduction caused by the chiral selectors reaching a mass spectrum detector, and can objectively, comprehensively and accurately evaluate the quality of the corydalis tuber extracting solution.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine component content determination, and particularly relates to a method for determining the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in a corydalis extracting solution.
Background
Rhizoma corydalis is corydalis tuber (corydalis tuber ) of PapaveraceaeCorydalis yanhusuoW.T. Wang), is a clinically widely used traditional Chinese medicine, has the effects of activating blood circulation, promoting qi circulation and relieving pain, and can be used for treating chest and hypochondrium, abdominal pain, thoracic obstruction, cardialgia, amenorrhea, dysmenorrhea, postpartum stasis, traumatic injury, swelling and pain and the like. Corydalis tuber is also a common medicinal flavor in a traditional Chinese medicine prescription, and corydalis tuber needs to be extracted in the production process of a plurality of Chinese patent medicines, so that the establishment of the quality analysis method of the corydalis tuber extracting solution is helpful for the quality evaluation and control of corydalis tuber and related traditional Chinese medicine preparations. Tetrahydropalmatine and tetrahydroberberine are important active ingredients in rhizoma corydalis in 2015 edition of Chinese pharmacopoeiaRhizoma corydalis content determination with tetrahydropalmatine as index component. Tetrahydropalmatine and tetrahydroberberine have a chiral carbon in each structure, and each has a pair of chiral enantiomers, i.e. l-tetrahydropalmatine and d-tetrahydropalmatine, and (a), (b)R) -tetrahydroberberine with (S) -tetrahydroberberine. Multiple studies show that the two pairs of chiral enantiomers have different pharmacodynamic and pharmacokinetic properties, so that the overall efficacy of corydalis tuber is affected by the high or low proportion of each chiral enantiomer. Therefore, it is necessary to establish a suitable analytical method for separately determining the content of each chiral enantiomer.
Document (A)Journal of Chromatography B, 2005, 826, 108-113; Journal of Chromatography A, 2006, 1117, 163-169; Analytical and Bioanalytical Chemistry2006, 384, 939-945) discloses a method for determining l-tetrahydropalmatine and d-tetrahydropalmatine by liquid chromatography using a chiral chromatography column. Document (A)Chirality, 2012, 24, 239-244; Analytical and Bioanalytical Chemistry, 2006, 384, 939-945; Journal of Liquid Chromatography & Related Technologies2014, 37, 26-38) disclose determining by liquid chromatography using a chiral chromatography column (c: (c)R) -tetrahydroberberine with (S) -a process for tetrahydroberberine. However, these methods are mainly used for analysis of racemate (i.e. mixture of chiral enantiomers), and are not suitable for corydalis ambigua extract because the components in corydalis ambigua extract are complex and many of them interfere with detection. In addition, the methods cannot simultaneously determine two pairs of chiral enantiomers, and have the problems that chiral chromatographic columns are difficult to prepare and are expensive. The capillary electrophoresis method for chiral analysis has the advantages of multiple chiral selectors (various cyclodextrins, chiral surfactants and the like) and low analysis cost, and the addition amount of the chiral selectors can be flexibly adjusted according to the properties of the components to be detected, so that simultaneous analysis of multiple pairs of chiral enantiomers can be realized. Document (A)Phytochemical Analysis, 1999, 10, 175-180; Journal of Biochemical and Biophysical Methods, 2007, 70, 71-76; Electrophoresis. 2010, 31, 2049-2054; University of North China newspaper, 2014, 15, 35-38) discloses a method for determining chiral enantiomers of tetrahydropalmatine or tetrahydroberberine by capillary electrophoresis-ultraviolet detection. However, because many components in the corydalis ambigua extractive solution interfere with the ultraviolet measurement, none of the above methods can measure the content of chiral enantiomers in the corydalis ambigua extractive solution, which is a complex sample system.
The capillary electrophoresis-mass spectrometry method can utilize the advantage of strong specificity of a mass spectrometer detector, avoid the interference of other components on the components to be detected, and simultaneously utilize the advantage of high sensitivity of the mass spectrometer detector, and avoid sample pretreatment operations such as concentration and enrichment. However, the mass spectrometer requires that the background electrolyte solution used in capillary electrophoresis has good volatility, and the capillary electrophoresis methods for measuring chiral enantiomers of tetrahydropalmatine and tetrahydroberberine disclosed in the existing documents do not meet the requirement. Therefore, it is necessary to search a suitable volatile background electrolyte solution based on the prior art, and study and design a capillary electrophoresis-mass spectrometry combined method capable of simultaneously determining two pairs of chiral enantiomers of tetrahydropalmatine and tetrahydroberberine.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to design and provide a method for measuring the content of chiral enantiomers of tetrahydropalmatine and tetrahydroberberine in a corydalis extracting solution. The method adopts a partial filling technology (partial filling technique) to avoid the problems of ionization inhibition and sensitivity reduction caused by the fact that a chiral selective agent (hydroxypropyl-beta-cyclodextrin) reaches a mass spectrum detector. The method has the advantages of simple operation, high sensitivity, low detection cost (few samples, chiral selectors and organic solvents are consumed), and high practicability, and can be used for directly determining the contents of two pairs of chiral enantiomers of tetrahydropalmatine and tetrahydroberberine in a complicated sample system of the corydalis tuber extracting solution so as to objectively, comprehensively and accurately evaluate the quality of the corydalis tuber extracting solution.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for measuring the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in a corydalis extracting solution is characterized by comprising the following steps:
(1) Preparation of control solutions: respectively preparing l-tetrahydropalmatine, d-tetrahydropalmatine and (B)R) -tetrahydroberberine, (ii) tetrahydroberberine: (iii)S) -four reference substance solutions of tetrahydroberberine and an internal standard reference substance solution of papaverine hydrochloride with known concentration, precisely sucking the four reference substance solutions, mixing, adding formic acid water solution with volume fraction of 0.01 to 10% for dilution by different times to obtain mixed solutions with different concentrations, and respectively adding the internal standard reference substance solution of papaverine hydrochloride to ensure that the concentrations of papaverine hydrochloride in the mixed solutions are the same and are within 10 to 100 mu g/mL to obtain reference substance solutions with different concentrations for later use;
(2) Preparation of a test solution: adding the above papaverine hydrochloride internal standard reference solution into rhizoma corydalis extract to make the concentration of papaverine hydrochloride be the same as that of papaverine hydrochloride in the reference solution obtained in step (1), and making into test solution;
(3) Determination of the standard curve: respectively taking the control solution with different concentrations prepared in the step (1), analyzing by using a capillary electrophoresis-mass spectrometer to obtain a capillary electrophoresis-mass spectrogram, obtaining peak areas Ax of four control substances in the control solution and peak areas As of an internal standard control substance from the diagram, respectively calculating by using a formula fx/s = Ax/As to obtain relative peak areas, and respectively calculating by using a least square regression method by using the relative peak areas As independent variables and the concentrations of the four control substances As dependent variables to obtain a standard curve;
(4) And (3) sample analysis: analyzing the sample solution obtained in the step (2) by using a capillary electrophoresis-mass spectrometer to obtain a capillary electrophoresis-mass spectrogram, obtaining peak areas Ax ' of four components to be detected in the sample solution and a peak area As ' of an internal standard reference substance from the diagram, and (3) respectively calculating relative peak areas by using a formula fx/S ' = Ax '/As ', and respectively calculating the concentrations of four components to be measured, namely l-tetrahydropalmatine, d-tetrahydropalmatine, (R) -tetrahydroberberine and (S) -tetrahydroberberine according to the standard curve obtained in the step (3).
Rhizoma corydalis in the rhizoma corydalis extracting solutionThe method for measuring the content of chiral enantiomers of the Sofosetyl and tetrahydroberberine is characterized in that l-tetrahydropalmatine, d-tetrahydropalmatine and (I), (II) and (III) in the step (1)R) -tetrahydroberberine and (b)S) The preparation method of four reference substance solutions of the tetrahydroberberine comprises the following steps: precisely weighing l-tetrahydropalmatine, d-tetrahydropalmatine, and (A) and (B)R) -tetrahydroberberine and (a)S) -4-component tetrahydroberberine reference substances are respectively dissolved by formic acid water solution with volume fraction of 0.01 to 10 percent and volume is fixed to prepare each reference substance solution with known concentration; the preparation method of the papaverine hydrochloride internal standard reference substance solution comprises the following steps: precisely weighing papaverine hydrochloride reference substance, dissolving with water, and diluting to desired volume to obtain internal standard reference substance solution with known concentration.
The method for determining the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in the corydalis tuber extracting solution is characterized in that the conditions of capillary electrophoresis-mass spectrometry combined determination in the step (4) are as follows: the capillary is a fused quartz capillary, the length is 75 to 120cm, the inner diameter is 50 to 75cm, and the outer diameter is 360 mu m; the background electrolyte solution is formic acid aqueous solution containing hydroxypropyl-beta-cyclodextrin and ammonium formate; the capillary flushing and filling procedure before analysis comprises the steps of sequentially keeping the pressure of deionized water at 0.5min 40psi, the pressure of 0.1 mol/L sodium hydroxide aqueous solution at 4 min 40psi, the pressure of deionized water at 0.5min 40psi, the pressure of formic acid aqueous solution containing ammonium formate at 4 min 40psi and the filling length of a background electrolyte solution at 40-70 cm; the sample injection condition is that the pressure sample injection is 0.5 psi, the sample injection time is 3 to 20 s, the separation voltage is plus 15 to 30 kV, and the column temperature is 20 to 30 ℃; the capillary electrophoresis-mass spectrometry interface chemical modifier solution is a methanol water solution containing formic acid, and the flow rate is 0.5 to 3 mu L/min; the mass spectrum detection parameters are a positive ion multi-reaction monitoring mode, the voltage of an electrospray ionization interface is +2.5 to 3.5 kV, the curtain gas is 5 to 30L/min of nitrogen, the collision gas is 3 to 10L/min of nitrogen, the cluster removing voltage is 90 to 110V, the collision energy is 25 to 45eV, and the rest mass spectrum detection parameters of each component are shown in Table 1.
The method for measuring the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in the corydalis extracting solution is characterized in that the concentration of hydroxypropyl-beta-cyclodextrin in the background electrolyte solution is 5 to 40mmol/L, the concentration of ammonium formate is 5 to 50mmol/L, and the volume fraction of formic acid is 5 to 15%.
The method for measuring the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in the corydalis extracting solution is characterized in that the volume concentration of formic acid in the capillary electrophoresis-mass spectrum interface chemical modifier solution is 0.01-1%, and the volume fraction of methanol is 50-90%.
The invention has the following beneficial effects:
(1) The invention provides a method for directly measuring the content of chiral enantiomers of tetrahydropalmatine and tetrahydroberberine in a complex sample system of corydalis extracting solution.
(2) The invention carries out a large amount of screening on parameters such as the composition of a background electrolyte solution, the filling length of a chiral selector in a capillary before analysis, separation voltage and the like, determines to adopt optimal analysis conditions, can simultaneously ensure that two pairs of chiral enantiomers can be well separated, and avoids the problems of ionization inhibition and sensitivity reduction caused by the fact that the chiral selector reaches a mass spectrum detector.
(3) The method has the advantages of simple operation, high sensitivity and low detection cost (the consumption of the sample, the chiral selector and the organic solvent is less), and has good application prospect and use value.
(4) The methodological verification of the analysis method provided by the invention shows that the method has good precision, repeatability, accuracy and sensitivity, and can accurately determine each component.
Drawings
FIG. 1 is a capillary electrophoresis image of a control solution of example 1, wherein Peak 1 is an internal standard, peak 2 is l-tetrahydropalmatine, peak 3 is d-tetrahydropalmatine, peak 4 is(R)-Tetrahydroberberine with peak 5 of(S)-Tetrahydroberberine;
FIG. 2 is a capillary electrophoresis chart of the sample solution of example 1, in which Peak 1 is an internal standard, peak 2 is l-tetrahydropalmatine, peak 3 is d-tetrahydropalmatine, peak 4 is(R)-Tetrahydroberberine with peak 5 of(S)-Tetrahydroberberine is used.
Detailed Description
The present invention is further illustrated by the following examples, which are intended to be purely exemplary and are not intended to limit the scope of the invention, as various equivalent modifications of the invention will occur to those skilled in the art upon reading the present disclosure and fall within the scope of the appended claims.
Example 1:
a method for measuring the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis extract comprises the following steps:
(1) Preparation of control solutions
Respectively and precisely weighing l-tetrahydropalmatine d-tetrahydropalmatine, (/), (ii) tetrahydropalmatine, and (iii)R) -tetrahydroberberine and: (S) A proper amount of a reference substance containing 4 tetrahydroberberine components is dissolved by a formic acid aqueous solution with the volume fraction of 0.2 percent respectively and the volume is fixed, so as to prepare a single reference substance stock solution with each component with known concentration.
Precisely weighing appropriate amount of papaverine hydrochloride reference substance, dissolving with water and fixing volume to obtain internal standard reference substance solution with known concentration.
Then respectively and precisely absorbing the l-tetrahydropalmatine, the d-tetrahydropalmatine and (c)R) -tetrahydroberberine and: (S) A proper amount of single reference substance stock solution of the tetrahydroberberine is mixed, formic acid water solution with volume fraction of 0.2% is added to dilute by different times to obtain a series of mixed solutions with different concentrations, and then internal standard reference substance solution is respectively added to ensure that the concentration of the papaverine hydrochloride is 40 mug/mL, so that a series of reference substance solutions with different concentrations are obtained for later use.
(2) Preparation of test solution
Grinding rhizoma corydalis decoction pieces (processed with vinegar) into small granules, collecting 2.5 g, soaking in 70% ethanol water solution 20mL for 1 hr, ultrasonically extracting for 30 min, centrifuging at 12000 r/min for 5min, and collecting supernatant to obtain rhizoma corydalis extractive solution.
Taking rhizoma corydalis extract, adding internal standard control solution to make papaverine hydrochloride concentration be 40 μ g/mL, and using as test solution.
(3) Determination of the Standard Curve
Respectively taking the mixed reference substance solutions with different concentrations prepared in the step (1), analyzing by a capillary electrophoresis-mass spectrometer to obtain a capillary electrophoresis chart (shown in figure 1), and obtaining the peak area of each reference substance in the reference substance solution from the chartA x Peak area of internal standard controlA s From the formulaf x/s = A x / A s The relative peak areas are respectively calculated. And (4) calculating to obtain a standard curve of each component by using the least square regression method respectively by using the relative peak area as an independent variable and the concentration of each reference substance as a dependent variable.
(4) Analysis of samples
Analyzing the sample solution prepared in step (2) with a capillary electrophoresis-mass spectrometer to obtain a capillary electrophoresis chart (shown in FIG. 2), and obtaining peak areas of the components to be detected in the sample solutionA x ', and peak area of internal standard controlA s ', from the formulaf x/s ’ = A x ’ / A s ' relative peak areas were calculated separately. And (4) respectively calculating the concentration of each component to be measured according to the standard curve of each component obtained in the step (3), wherein the result is as follows: l-tetrahydropalmatine 27.5. Mu.g/mL, d-tetrahydropalmatine 59.8. Mu.g/mL, ((ii))R) 1.28. Mu.g/mL of tetrahydroberberine, (b) and (d)S) -tetrahydroberberine 14.4 μ g/mL.
(5) In the determination method in the step (3) and the step (4), the conditions for the capillary electrophoresis-mass spectrometry combined determination are as follows:
the instrument comprises: a Beckman Coulter PA 800+ capillary electrophoresis instrument, an AB SCIEX API 4000 triple quadrupole mass spectrometer, and a flow distribution capillary electrophoresis-mass spectrometry interface;
capillary tube: a fused quartz capillary tube having a length of 100 cm, an inner diameter of 50 μm and an outer diameter of 360 μm;
background electrolyte solution: a 10 volume percent formic acid aqueous solution containing hydroxypropyl-beta-cyclodextrin with the concentration of 20mmol/L and ammonium formate with the concentration of 20 mmol/L;
capillary rinse and fill procedure prior to analysis: sequentially, deionized water is 0.5min (pressure 40 psi), 0.1 mol/L sodium hydroxide aqueous solution is 4 min (pressure 40 psi), deionized water is 0.5min (pressure 40 psi), formic acid aqueous solution with the concentration of 20mmol/L ammonium formate and the volume fraction of 10 percent is 4 min (pressure 40 psi), and background electrolyte solution is 1.68 min (pressure 10 psi), namely the filling length is 52.5 cm;
sample introduction conditions are as follows: injecting sample under pressure (0.5 psi) for 12 s;
separation voltage: +24 kV;
column temperature: 25. DEG C;
capillary electrophoresis-mass spectrometry interface chemical modifier solution: methanol aqueous solution with the volume fraction of 80 percent and the volume concentration of 0.1 percent of formic acid, and the flow rate is 2 mu L/min;
mass spectrum detection parameters: the positive ion multi-reaction monitoring mode comprises an electrospray ionization interface voltage of +3 kV, curtain gas (nitrogen gas) of 10L/min, collision gas (nitrogen gas) of 4L/min, cluster removing voltage of 98V, collision energy of 35 eV, and the rest detection parameters of each component are shown in Table 2.
TABLE 2 Mass spectrometric parameters
(6) The methodology of the method of the invention is examined as follows:
(6.1) examination of Linear relationship
The regression equation and linear range for the 4 components obtained by the operation of step (3) are shown in Table 3. The results show that the 4 components are in good linear relationship in the concentration range under investigation.
TABLE 3 results of linear relationship examination
(6.2) sensitivity examination
And (4) gradually diluting the reference substance solution by using a formic acid aqueous solution with the volume fraction of 0.2%, analyzing according to the method in the step (5), and taking the concentration of each component as a detection limit and a quantification limit when the signal-to-noise ratio of a mass spectrum signal is 3 and 10 respectively, wherein the results are shown in a table 4. Considering that the sample injection volume is only about 6nL, the detection method has high sensitivity.
TABLE 4 sensitivity examination results
(6.3) examination of precision, reproducibility and stability
Taking one sample solution prepared in the step (2), repeatedly injecting the sample solution for 6 times according to the analysis conditions in the step (5), and measuring the RSD of the concentrations of the 4 components according to the step (3) and the step (4) as shown in the table 6. The results show that the precision is good.
And (3) taking the same part of rhizoma corydalis extracting solution, respectively adding an internal standard reference substance solution according to the step (2), preparing 6 parts of test sample solution, respectively carrying out sample injection analysis according to the analysis conditions in the step (5), and measuring the RSD of the concentration of 4 components according to the step (3) and the step (4) as shown in the table 6. The results show good reproducibility.
And (3) taking the sample solution prepared in the step (2), standing at normal temperature for 0, 1, 2, 4, 8, 24, 48 and 72 hours, then injecting samples according to the analysis conditions in the step (5), measuring the concentrations of 4 components according to the step (3) and the step (4), wherein the RSD (red signal) is shown in Table 5, and the concentrations of all the components have no descending trend. The results show that 4 components are stable for 72 hours at room temperature.
TABLE 5 results of precision, repeatability, stability studies
(6.4) sample application recovery Rate examination
6 parts of test solution is prepared according to the step (2), and the concentration of the 4 components is measured according to the step (3) and the step (4). Respectively taking 1mL of 6 test sample solutions, precisely adding 1mL of the reference sample solution prepared in the step (1), uniformly mixing, measuring the concentrations of the 4 components according to the step (3) and the step (4), and calculating the sample adding recovery rate of each component. The measurement results are shown in table 6, the sample recovery rate is within the range of 89.1% -110%, and the method is good in accuracy.
TABLE 6 investigation results of sample recovery (%)
Example 2:
a method for measuring the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis extract comprises the following steps:
(1) Preparation of control solutions
Respectively and precisely weighing appropriate amount of reference substances of 4 components of l-tetrahydropalmatine, d-tetrahydropalmatine, (R) -tetrahydroberberine and (S) -tetrahydroberberine, respectively dissolving with 0.2% formic acid water solution by volume fraction, and fixing volume to obtain single reference substance stock solution with known concentration.
Precisely weighing appropriate amount of papaverine hydrochloride reference substance, dissolving with water and fixing volume to obtain internal standard reference substance solution with known concentration.
Respectively and precisely sucking appropriate amounts of single reference substance stock solutions of l-tetrahydropalmatine, d-tetrahydropalmatine, (R) -tetrahydroberberine and (S) -tetrahydroberberine, mixing, adding 0.2% formic acid water solution by volume fraction to dilute by different times to obtain a series of mixed solutions with different concentrations, respectively adding internal standard reference substance solution to make the concentration of papaverine hydrochloride be 40 mug/mL, and obtaining a series of reference substance solutions with different concentrations for later use.
(2) Preparation of test solution
Grinding rhizoma corydalis decoction pieces (processed with vinegar) into small particles, collecting 2.5 g, soaking in 20mL of water for 1 hr, ultrasonically extracting for 30 min, centrifuging at 12000 r/min for 5min, and collecting supernatant to obtain rhizoma corydalis extract.
Taking rhizoma corydalis extract, adding internal standard reference solution to make papaverine hydrochloride concentration be 40 μ g/mL, and using as test solution.
(3) Determination of the Standard Curve
And (2) respectively taking the mixed reference substance solutions with different concentrations prepared in the step (1), analyzing by using a capillary electrophoresis-mass spectrometer to obtain a capillary electrophoresis chart, obtaining the peak area Ax of each reference substance in the reference substance solution and the peak area As of the internal standard reference substance from the capillary electrophoresis-mass spectrometer, and respectively calculating to obtain relative peak areas according to the formula fx/s = Ax/As. And (4) calculating to obtain a standard curve of each component by using the least square regression method respectively by using the relative peak area as an independent variable and the concentration of each reference substance as a dependent variable.
(4) Analysis of samples
And (3) analyzing the sample solution prepared in the step (2) by using a capillary electrophoresis-mass spectrometer to obtain a capillary electrophoresis chart, and obtaining peak areas Ax ' of each component to be detected in the sample solution and a peak area As ' of an internal standard reference substance from the chart, wherein the relative peak areas are respectively calculated by a formula fx/s ' = Ax '/As '. And (4) respectively calculating the concentration of each component to be measured according to the component standard curve obtained in the step (3), wherein the result is as follows: 23.7 mu g/mL of l-tetrahydropalmatine, 55.8 mu g/mL of d-tetrahydropalmatine, 0.706 mu g/mL of (R) -tetrahydroberberine, and 9.24 mu g/mL of (S) -tetrahydroberberine.
(5) In the determination method in the step (3) and the step (4), the conditions for the capillary electrophoresis-mass spectrometry combined determination are as follows:
the instrument comprises the following steps: the system comprises a Beckman Coulter PA 800+ capillary electrophoresis instrument, an AB SCIEX API 4000 triple four-stage rod mass spectrometer and a flow-through capillary electrophoresis-mass spectrum interface;
capillary tube: a fused silica capillary having a length of 100 cm, an inner diameter of 50 μm and an outer diameter of 360 μm;
background electrolyte solution: a 10 volume percent formic acid aqueous solution containing hydroxypropyl-beta-cyclodextrin with the concentration of 20mmol/L and ammonium formate with the concentration of 20 mmol/L;
capillary rinse and fill procedure prior to analysis: sequentially, deionized water is 0.5min (pressure 40 psi), 0.1 mol/L sodium hydroxide aqueous solution is 4 min (pressure 40 psi), deionized water is 0.5min (pressure 40 psi), formic acid aqueous solution with the concentration of 20mmol/L ammonium formate and the volume fraction of 10 percent is 4 min (pressure 40 psi), and background electrolyte solution is 1.68 min (pressure 10 psi), namely the filling length is 52.5 cm;
sample introduction conditions are as follows: injecting sample under pressure (0.5 psi) for 12 s;
separation voltage: +24 kV;
column temperature: 25. DEG C;
capillary electrophoresis-mass spectrometry interface chemical modifier solution: a methanol aqueous solution with the volume fraction of 80 percent and the formic acid volume concentration of 0.1 percent, and the flow rate is 2 mu L/min;
mass spectrum detection parameters: the positive ion multi-reaction monitoring mode comprises an electrospray ionization interface voltage of +3 kV, curtain gas (nitrogen gas) of 10L/min, collision gas (nitrogen gas) of 4L/min, cluster removing voltage of 98V, collision energy of 35 eV, and the rest detection parameters of each component are shown in Table 7.
TABLE 7 Mass spectrometric parameters
The foregoing is only a preferred embodiment of the present invention, and those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be construed as the protection scope of the present invention.
Claims (4)
1. A method for measuring the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis tuber extracting solution is characterized by comprising the following steps:
(1) Preparation of control solutions: respectively preparing l-tetrahydropalmatine, d-tetrahydropalmatine and (B)R) -tetrahydroberberine and (b)S) Four kinds of reference substance solutions of tetrahydroberberine and an internal standard reference substance solution of papaverine hydrochloride with known concentration, precisely sucking the four kinds of reference substance solutions, mixing, and adding formic acid with volume fraction of 0.01 to 10 percentDiluting the aqueous solution by different times to obtain mixed solutions with different concentrations, and adding the papaverine hydrochloride internal standard reference substance solution into the mixed solutions respectively to ensure that the concentrations of the papaverine hydrochloride in the mixed solutions are the same and are 10 to 100 mu g/mL, thus obtaining reference substance solutions with different concentrations for later use;
(2) Preparation of a test solution: adding the above papaverine hydrochloride internal standard reference solution into rhizoma corydalis extract to make the concentration of papaverine hydrochloride be the same as that of papaverine hydrochloride in the reference solution obtained in step (1), and making into test solution;
(3) Determination of the standard curve: respectively taking the control solution with different concentrations prepared in the step (1), analyzing by using a capillary electrophoresis-mass spectrometer to obtain a capillary electrophoresis-mass spectrogram, obtaining peak areas Ax of four control substances in the control solution and peak areas As of an internal standard control substance from the diagram, respectively calculating by using a formula fx/s = Ax/As to obtain relative peak areas, and respectively calculating by using a least square regression method by using the relative peak areas As independent variables and the concentrations of the four control substances As dependent variables to obtain a standard curve;
(4) And (3) sample analysis: analyzing the sample solution obtained in the step (2) by using a capillary electrophoresis-mass spectrometer to obtain a capillary electrophoresis-mass spectrogram, obtaining peak areas Ax ' of four to-be-detected components in the sample solution and peak areas As ' of an internal standard reference substance from the diagram, respectively calculating relative peak areas by using a formula fx/S ' = Ax '/As ', and respectively calculating the concentrations of the four to-be-detected components of l-tetrahydropalmatine, d-tetrahydropalmatine, (R) -tetrahydroberberine and (S) -tetrahydroberberine according to the standard curve obtained in the step (3);
the conditions of the capillary electrophoresis-mass spectrometry combined determination are as follows: the capillary is a fused quartz capillary, the length is 75 to 120cm, the inner diameter is 50 to 75cm, and the outer diameter is 360 mu m; the background electrolyte solution is formic acid aqueous solution containing hydroxypropyl-beta-cyclodextrin and ammonium formate; before analysis, the capillary tube washing and filling procedures are that deionized water is used for 0.5min under the pressure of 40psi,0.1 mol/L sodium hydroxide aqueous solution is used for 4 min under the pressure of 40psi, deionized water is used for 0.5min under the pressure of 40psi, formic acid aqueous solution containing ammonium formate is used for 4 min under the pressure of 40psi, and the filling length of background electrolyte solution is 40-70 cm; the sample injection condition is that the pressure sample injection is 0.5 psi, the sample injection time is 3 to 20 s, the separation voltage is +15 to 30 kV, and the column temperature is 20 to 30 ℃; the capillary electrophoresis-mass spectrum interface chemical modifier solution is a methanol water solution containing formic acid, and the flow rate is 0.5 to 3 mu L/min; the mass spectrum detection parameters are a positive ion multi-reaction monitoring mode, the voltage of an electrospray ionization interface is plus 2.5 to 3.5 kV, curtain gas is 5 to 30L/min of nitrogen, collision gas is 3 to 10L/min of nitrogen, cluster removing voltage is 90 to 110V, collision energy is 25 to 45eV, and the rest mass spectrum detection parameters of each component are shown in the following table,
。
2. The method for determining the content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in rhizoma corydalis extract according to claim 1, wherein the L-tetrahydropalmatine, d-tetrahydropalmatine, and (C), (D), (C) and (D) in step (1) are used to determine the content of tetrahydropalmatine and tetrahydropalmatine chiral enantiomers in rhizoma corydalis extractR) -tetrahydroberberine, (ii) tetrahydroberberine: (iii)S) The preparation method of four reference substance solutions of the tetrahydroberberine comprises the following steps: precisely weighing l-tetrahydropalmatine, d-tetrahydropalmatine, and (A) and (B)R) -tetrahydroberberine and: (S) -4-component reference substances of tetrahydroberberine are respectively dissolved by formic acid water solution with volume fraction of 0.01 to 10 percent and the volume is determined, and each reference substance solution with known concentration is prepared; the preparation method of the papaverine hydrochloride internal standard reference substance solution comprises the following steps: accurately weighing papaverine hydrochloride reference substance, dissolving with water, and metering volume to obtain internal standard reference substance solution with known concentration.
3. The method for determining the content of chiral enantiomers of tetrahydropalmatine and tetrahydroberberine in rhizoma corydalis extract according to claim 1, wherein the concentration of hydroxypropyl- β -cyclodextrin in the background electrolyte solution is 5 to 40mmol/L, the concentration of ammonium formate is 5 to 50mmol/L, and the volume fraction of formic acid is 5 to 15%.
4. The method for measuring the content of the chiral enantiomers of tetrahydropalmatine and tetrahydroberberine in the rhizoma corydalis extract as claimed in claim 1, wherein the volume concentration of formic acid in the capillary electrophoresis-mass spectrometry interface chemical modifier solution is 0.01 to 1%, and the volume fraction of methanol is 50 to 90%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010495542.6A CN111537593B (en) | 2020-06-03 | 2020-06-03 | Method for measuring content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis extracting solution |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010495542.6A CN111537593B (en) | 2020-06-03 | 2020-06-03 | Method for measuring content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis extracting solution |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111537593A CN111537593A (en) | 2020-08-14 |
| CN111537593B true CN111537593B (en) | 2022-12-02 |
Family
ID=71976566
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010495542.6A Active CN111537593B (en) | 2020-06-03 | 2020-06-03 | Method for measuring content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis extracting solution |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111537593B (en) |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2007230174B2 (en) * | 2006-03-28 | 2012-03-22 | Georgia State University Research Foundation | Polymeric Sulfated Surfactants for Capillary Electrophoresis (CE) and CE-Mass Spectrometry (CE-MS) |
| ES2320718B1 (en) * | 2007-11-26 | 2010-03-03 | Universidad De Alcala. | PROCEDURE FOR THE IDENTIFICATION AND QUANTIFICATION OF THE ENANTIOMERS OF THE CARNITINE NON-PROTEIC AMINO ACID IN FOOD BY CAPILLARY ELECTROPHORESIS COUPLED TO MASS SPECTROMETRY. |
| CN108872432A (en) * | 2018-07-11 | 2018-11-23 | 南京海昌中药集团有限公司 | The quality determining method of rhizoma corydalis health product and its processed product |
-
2020
- 2020-06-03 CN CN202010495542.6A patent/CN111537593B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN111537593A (en) | 2020-08-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | Global characterization of neutral saccharides in crude and processed Radix Rehmanniae by hydrophilic interaction liquid chromatography tandem electrospray ionization time-of-flight mass spectrometry | |
| CN109696499B (en) | High-sensitivity determination method for nitrosamine in water based on high-resolution mass spectrum | |
| CN111175394A (en) | Method for detecting plasma catecholamine and metabolites thereof by liquid chromatography-tandem mass spectrometry | |
| CN103983725B (en) | The rapid assay methods of cumarin and safrole in a kind of essence and flavoring agent | |
| CN108918711B (en) | Detection method of polyphenol compounds in tobacco leaves | |
| CN107941950B (en) | Method for simultaneously determining nine functional components in different parts of Changshan grapefruit by ultra-high performance liquid chromatography-tandem mass spectrometry | |
| CN112730706A (en) | Method for detecting biological small molecule marker by liquid chromatography-tandem mass spectrometry | |
| CN107356689B (en) | Method for determining 12 Amadori compounds in tobacco | |
| CN107677744B (en) | Method for detecting morphological mercury in animal tissue cells | |
| CN108152425B (en) | Method for detecting lignanoids in sesame oil by high performance liquid chromatography | |
| CN107345946B (en) | The method for preparing purified of methcathinone standard substance for forensic science illicit drugs inspection | |
| CN111798937B (en) | Method for establishing metabonomics database of wolfberry tissue and application thereof | |
| CN111537593B (en) | Method for measuring content of tetrahydropalmatine and tetrahydroberberine chiral enantiomers in corydalis extracting solution | |
| CN110609106B (en) | Method for detecting 11 organophosphorus pesticide residues in white paeony root by using ultra-performance liquid chromatography-mass spectrometry technology | |
| CN108956812B (en) | Method for detecting water-soluble organic chloride pesticide residue in soil | |
| CN108693286B (en) | Method for detecting genotoxic impurity diisopropyl sulfate in medicine | |
| CN114113403B (en) | Method for determining liquid chromatography-mass spectrometry (LC-MS) of Hedan tablets | |
| CN116642970A (en) | Sample pretreatment method for simultaneously detecting 6 drug concentrations in blood | |
| CN105158372A (en) | Method for determining urocanic acid and ethyl ester thereof in cosmetics | |
| CN107102087B (en) | Method for detecting contents of various organic acids in coptis chinensis by ion chromatography | |
| CN115728408A (en) | Method for simultaneously determining seven artemisinin related compounds in artemisia annua | |
| CN113866305A (en) | Method for rapidly and accurately analyzing theanine in fresh tea leaves based on liquid chromatography-mass spectrometry technology | |
| CN107515262B (en) | Liquid chromatography-mass spectrometry method for simultaneously determining lincomycin and gentamicin in animal plasma | |
| CN109283272B (en) | Detection method of 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and sodium salt thereof | |
| CN105717228A (en) | Method for detecting content of methylphenol substances in baijiu |
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 |