CN113156040A - 一种蟾蜍提取液中吲哚总碱的检验方法 - Google Patents
一种蟾蜍提取液中吲哚总碱的检验方法 Download PDFInfo
- Publication number
- CN113156040A CN113156040A CN202110532452.4A CN202110532452A CN113156040A CN 113156040 A CN113156040 A CN 113156040A CN 202110532452 A CN202110532452 A CN 202110532452A CN 113156040 A CN113156040 A CN 113156040A
- Authority
- CN
- China
- Prior art keywords
- taking
- product
- solution
- sulfuric acid
- ethanol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 18
- 239000003513 alkali Substances 0.000 title claims abstract description 15
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 title claims abstract description 15
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 title claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 40
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 40
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 36
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims abstract description 30
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000013558 reference substance Substances 0.000 claims abstract description 23
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 20
- SCULJPGYOQQXTK-OLRINKBESA-N Cinobufagin Chemical compound C=1([C@@H]2[C@@]3(C)CC[C@@H]4[C@@]5(C)CC[C@H](O)C[C@H]5CC[C@H]4[C@@]43O[C@@H]4[C@@H]2OC(=O)C)C=CC(=O)OC=1 SCULJPGYOQQXTK-OLRINKBESA-N 0.000 claims abstract description 16
- SCULJPGYOQQXTK-UHFFFAOYSA-N Cinobufagin Natural products CC(=O)OC1C2OC22C3CCC4CC(O)CCC4(C)C3CCC2(C)C1C=1C=CC(=O)OC=1 SCULJPGYOQQXTK-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000001514 detection method Methods 0.000 claims abstract description 14
- BGNGWHSBYQYVRX-UHFFFAOYSA-N 4-(dimethylamino)benzaldehyde Chemical compound CN(C)C1=CC=C(C=O)C=C1 BGNGWHSBYQYVRX-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000007605 air drying Methods 0.000 claims abstract description 10
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 10
- NLCIYDBRSJQKTO-UHFFFAOYSA-N chloroform;cyclohexane;propan-2-one Chemical compound CC(C)=O.ClC(Cl)Cl.C1CCCCC1 NLCIYDBRSJQKTO-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000007787 solid Substances 0.000 claims abstract description 10
- 238000005507 spraying Methods 0.000 claims abstract description 10
- 238000004809 thin layer chromatography Methods 0.000 claims abstract description 10
- QEEBRPGZBVVINN-UHFFFAOYSA-N Desacetyl-bufotalin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C1(O)CCC2C=1C=CC(=O)OC=1 QEEBRPGZBVVINN-UHFFFAOYSA-N 0.000 claims abstract description 8
- ATLJNLYIJOCWJE-CWMZOUAVSA-N bufogenin Chemical compound C=1([C@H]2C[C@H]3O[C@@]43[C@H]3[C@@H]([C@]5(CC[C@H](O)C[C@H]5CC3)C)CC[C@@]42C)C=CC(=O)OC=1 ATLJNLYIJOCWJE-CWMZOUAVSA-N 0.000 claims abstract description 6
- 229950006858 bufogenin Drugs 0.000 claims abstract description 6
- 238000001704 evaporation Methods 0.000 claims abstract description 6
- ATLJNLYIJOCWJE-UHFFFAOYSA-N resibufogenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C=1C=CC(=O)OC=1 ATLJNLYIJOCWJE-UHFFFAOYSA-N 0.000 claims abstract description 6
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 36
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 30
- 239000012088 reference solution Substances 0.000 claims description 19
- 239000012085 test solution Substances 0.000 claims description 19
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 14
- 239000000945 filler Substances 0.000 claims description 10
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 241000212749 Zesius chrysomallus Species 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 9
- 239000000377 silicon dioxide Substances 0.000 claims description 9
- 238000001228 spectrum Methods 0.000 claims description 9
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 7
- KIWBPDUYBMNFTB-UHFFFAOYSA-N Ethyl hydrogen sulfate Chemical compound CCOS(O)(=O)=O KIWBPDUYBMNFTB-UHFFFAOYSA-N 0.000 claims description 6
- 239000012488 sample solution Substances 0.000 abstract description 2
- KBKUJJFDSHBPPA-UHFFFAOYSA-N 5beta-hydroxylcinobufagin Natural products CC(=O)OC1C2OC22C3CCC4(O)CC(O)CCC4(C)C3CCC2(C)C1C=1C=CC(=O)OC=1 KBKUJJFDSHBPPA-UHFFFAOYSA-N 0.000 description 6
- KBKUJJFDSHBPPA-ZNCGZLKOSA-N cinobufotalin Chemical compound C=1([C@@H]2[C@@]3(C)CC[C@@H]4[C@@]5(C)CC[C@H](O)C[C@@]5(O)CC[C@H]4[C@@]43O[C@@H]4[C@@H]2OC(=O)C)C=CC(=O)OC=1 KBKUJJFDSHBPPA-ZNCGZLKOSA-N 0.000 description 6
- 241000269417 Bufo Species 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 241001415440 Bufo gargarizans Species 0.000 description 3
- 241001415432 Duttaphrynus melanostictus Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- QEEBRPGZBVVINN-BMPKRDENSA-N bufalin Chemical compound C=1([C@H]2CC[C@]3(O)[C@H]4[C@@H]([C@]5(CC[C@H](O)C[C@H]5CC4)C)CC[C@@]32C)C=CC(=O)OC=1 QEEBRPGZBVVINN-BMPKRDENSA-N 0.000 description 2
- VTTONGPRPXSUTJ-UHFFFAOYSA-N bufotenin Chemical compound C1=C(O)C=C2C(CCN(C)C)=CNC2=C1 VTTONGPRPXSUTJ-UHFFFAOYSA-N 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 229930005303 indole alkaloid Natural products 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 description 2
- -1 sterol compounds Chemical class 0.000 description 2
- 230000008961 swelling Effects 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 206010000060 Abdominal distension Diseases 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 206010007247 Carbuncle Diseases 0.000 description 1
- 208000000419 Chronic Hepatitis B Diseases 0.000 description 1
- 208000031361 Hiccup Diseases 0.000 description 1
- 208000015817 Infant Nutrition disease Diseases 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 206010044302 Tracheitis Diseases 0.000 description 1
- 208000000260 Warts Diseases 0.000 description 1
- 206010000269 abscess Diseases 0.000 description 1
- WYTGDNHDOZPMIW-RCBQFDQVSA-N alstonine Chemical compound C1=CC2=C3C=CC=CC3=NC2=C2N1C[C@H]1[C@H](C)OC=C(C(=O)OC)[C@H]1C2 WYTGDNHDOZPMIW-RCBQFDQVSA-N 0.000 description 1
- 150000001412 amines Chemical group 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 150000002475 indoles Chemical class 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 201000007227 lymph node tuberculosis Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000004911 serous fluid Anatomy 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 201000010153 skin papilloma Diseases 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
-
- 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
- G01N1/30—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
- 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
- G01N30/34—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
- G01N30/48—Sorbent materials therefor
- G01N30/482—Solid sorbents
-
- 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
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/94—Development
-
- 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
- G01N30/90—Plate chromatography, e.g. thin layer or paper chromatography
- G01N30/95—Detectors specially adapted therefor; Signal analysis
-
- 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
- G01N2030/3007—Control of physical parameters of the fluid carrier of temperature same temperature for whole column
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/48—Sorbent materials therefor
- G01N2030/484—Solid sorbents
Abstract
本发明涉及一种蟾蜍提取液中吲哚总碱的检验方法，包括如下步骤，步骤S1：取本品0.5‑2ml，加甲醇3‑6ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色，步骤S2：取本品0.5‑2ml，加氯仿3‑6ml，震摇5‑15分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色，步骤S3：取本品0.1‑0.3ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷‑氯仿‑丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液；本发明具有检测准确、操作简单的优点。
Description
技术领域
本发明属于检测吲哚总碱的技术领域，具体涉及一种蟾蜍提取液中吲哚总碱的检验方法。
背景技术
蟾皮为蟾蜍科动物中华大蟾蜍或黑眶蟾蜍的干燥全皮，蟾皮的化学成分主要有华蟾酥毒基，脂蟾毒配基， 蟾毒灵等，以及由它们与琥珀酸，丁二酸，精氨酸等形成的酯类，如蟾毒灵-3-丁二酰精 氨酸酯，蟾毒它灵-3-丁二酰精氨酸酯，华蟾毒精-3-丁二酰精氨酸酯，脂蟾毒配基-3 -丁二酰精氨酸酯等，还含有吲哚类生物碱，如5-羟色胺，蟾蜍色胺，蟾蜍季胺，蟾蜍噻宁 和脱氢蟾蜍色胺等，另外尚含氨基酸、多肽及甾醇类化合物，蟾皮具有清热解毒，利水清胀功效，用于治疗痈疽、肿毒、瘰疬、肿瘤，疳积腹胀，慢性气管炎等。目前，蟾皮提取物制剂主要有华蟾素注射液、华蟾素片剂、华蟾素口服液等，其中华蟾素注射液更是一种传统的生物药制剂、广泛用于临床，具有解毒、消肿、止痛的功效，临床上主要用于治疗中晚期肿瘤、慢性乙型肝炎，还用于治疗顽固性呃逆、扁平疣及银屑病等症，现有技术中还没有对于蟾蜍吲哚总碱含量检测的方法；针对这些不足之处，有必要开发一种检测准确、操作简单的蟾蜍提取液中吲哚总碱的检验方法。
发明内容
本发明的目的是为了克服现有技术的不足，而提供一种检测准确、操作简单的蟾蜍提取液中吲哚总碱的检验方法。
本发明的目的是这样实现的：一种蟾蜍提取液中吲哚总碱的检验方法，包括如下步骤，
步骤S1：取本品0.5-2ml，加甲醇3-6ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色； 步骤S2：取本品0.5-2ml，加氯仿3-6ml，震摇5-15分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色； 步骤S3：取本品0.1-0.3ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰；供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2的三次水为流动相A相,乙腈为流动相B相；检测波长为296nm；柱温20℃。
包括如下步骤，
步骤S1：取本品1ml，加甲醇5ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色； 步骤S2：取本品1ml，加氯仿5ml，震摇10分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色； 步骤S3：取本品0.2ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰；供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2(用磷酸调节pH值为3.2)的三次水为流动相A相,乙腈为流动相B相；检测波长为296nm；柱温20℃。
包括如下步骤，
步骤S1：取本品2ml，加甲醇6ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色； 步骤S2：取本品2ml，加氯仿6ml，震摇15分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色； 步骤S3：取本品0.3ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰；供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2(用磷酸调节pH值为3.2)的三次水为流动相A相，乙腈为流动相B相；检测波长为296nm；柱温20℃。
本发明的有益效果：本发明的优点在于利用色谱法，准确的检测处蟾蜍中吲哚总碱的含量，其他有益效果在具体实施例中详细说明。
附图说明
图1是本发明一种蟾蜍提取液中吲哚总碱的检验方法中的梯度洗脱的模式图。
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例，基于本发明中的实施例，本领域技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
除非另有定义，本说明书所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同，本说明书中所使用的术语只是为了描述具体的实施方式的目的，不是用于限制本发明，本说明书所使用的术语如“和/或”包括一个或多个相关的所列项目的任意的和所有的组合；此外，下面所描述的本发明不同实施方式中所涉及的技术特征只要彼此之间未构成冲突就可以相互结合。
实施例1
如图1所示，一种蟾蜍提取液中吲哚总碱的检验方法，包括如下步骤，
步骤S1：取本品0.5-2ml，加甲醇3-6ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色。 步骤S2：取本品0.5-2ml，加氯仿3-6ml，震摇5-15分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色。 步骤S3：取本品0.1-0.3ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰。供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2的三次水为流动相A相,乙腈为流动相B相；检测波长为296nm；柱温20℃。
本实施例中来源是蟾蜍科动物中华大蟾蜍或黑眶蟾蜍的干燥分泌物，多于夏、秋二季捕捉蟾蜍，洗净，挤取耳后腺及皮肤腺的白色浆液，专业提取而成的以十八烷基硅烷键合硅胶为填充剂；色谱法：以pH为3.2(用磷酸调节pH值为3.2)的三次水为流动相A相,乙腈为流动相B相；检测波长为296nm；柱温20℃，采用梯度洗脱的模式进行分离，梯度模式如图1所示，理论板数按华蟾酥毒基峰、脂蟾毒配基峰计算应分别不低于4000。
对照品溶液的制备：取华蟾酥毒基对照品、脂蟾毒配基对照品、蟾毒灵对照品适量，精密称定，加甲醇分别制成每1ml各含华蟾酥毒基、脂蟾毒配基5mg的溶液，即得。
供试品溶液的制备：精确量取本品2ml，精密加入甲醇10ml，用0.45μm滤膜过滤即得。
测定法：分别精密吸取上述两种对照品溶液与供试品溶液各5 μl，注入液相色谱仪，测定，即得。
本品按华中蟾浆原液计算，含去乙酰华蟾毒它灵、伪异沙蟾毒精、华蟾毒它灵、华蟾酥毒基(C26H34O6)和脂蟾毒配基(C24H32O4)的总量不得少于4.5％；梯度法，总吲哚生物碱计算总量不得少于6.0%。
实施例2
包括如下步骤，
步骤S1：取本品1ml，加甲醇5ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色。 步骤S2：取本品1ml，加氯仿5ml，震摇10分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色。 步骤S3：取本品0.2ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰。供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2(用磷酸调节pH值为3.2)的三次水为流动相A相,乙腈为流动相B相；检测波长为296nm；柱温20℃。
实施例3
包括如下步骤，
步骤S1：取本品2ml，加甲醇6ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色。 步骤S2：取本品2ml，加氯仿6ml，震摇15分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色。 步骤S3：取本品0.3ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰。供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2(用磷酸调节pH值为3.2)的三次水为流动相A相，乙腈为流动相B相；检测波长为296nm；柱温20℃。
具体实施方式是对本发明的进一步说明而非限制，对本领域普通技术人员来说在不脱离本发明实质内容的情况下对结构做进一步变换，而所有这些变换都应属于本发明所附权利要求的保护范围。
Claims (3)
1.一种蟾蜍提取液中吲哚总碱的检验方法，其特征在于：包括如下步骤，
步骤S1：取本品0.5-2ml，加甲醇3-6ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色； 步骤S2：取本品0.5-2ml，加氯仿3-6ml，震摇5-15分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色； 步骤S3：取本品0.1-0.3ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰；供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2的三次水为流动相A相,乙腈为流动相B相；检测波长为296nm；柱温20℃。
2.根据权利要求1所述的一种蟾蜍提取液中吲哚总碱的检验方法，其特征在于：包括如下步骤，
步骤S1：取本品1ml，加甲醇5ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色； 步骤S2：取本品1ml，加氯仿5ml，震摇10分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色； 步骤S3：取本品0.2ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰；供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2(用磷酸调节pH值为3.2)的三次水为流动相A相,乙腈为流动相B相；检测波长为296nm；柱温20℃。
3.根据权利要求1所述的一种蟾蜍提取液中吲哚总碱的检验方法，其特征在于：包括如下步骤，
步骤S1：取本品2ml，加甲醇6ml，加对二甲氨基苯甲醛固体少量，滴加硫酸数滴，即显蓝紫色； 步骤S2：取本品2ml，加氯仿6ml，震摇15分钟，蒸干，残渣加醋酐少量使溶解，滴加硫酸，初显蓝紫色，渐变为蓝绿色； 步骤S3：取本品0.3ml，置10ml量瓶中，加乙醇至刻度，作为供试品溶液，再取脂蟾毒配基及华蟾毒基对照品，加乙醇分别制成每1ml含1mg的溶液，作为对照品溶液，照薄层色谱法试验，吸取上述4种溶液各10μl，分别点于同一硅胶G薄层板上，以环己烷-氯仿-丙酮（4∶3∶3）为展开剂，展开，取出，晾干，喷以10%硫酸乙醇溶液，加热至斑点显色清晰；供试品色谱中，在与对照药材色谱相应的位置上，显相同颜色的斑点；在与对照品色谱相应的位置上，显相同的一个绿色及一个红色斑点；色谱条件：以十八烷基硅烷键合硅胶为填充剂；以pH为3.2(用磷酸调节pH值为3.2)的三次水为流动相A相,乙腈为流动相B相；检测波长为296nm；柱温20℃。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110532452.4A CN113156040A (zh) | 2021-05-17 | 2021-05-17 | 一种蟾蜍提取液中吲哚总碱的检验方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110532452.4A CN113156040A (zh) | 2021-05-17 | 2021-05-17 | 一种蟾蜍提取液中吲哚总碱的检验方法 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113156040A true CN113156040A (zh) | 2021-07-23 |
Family
ID=76876298
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110532452.4A Pending CN113156040A (zh) | 2021-05-17 | 2021-05-17 | 一种蟾蜍提取液中吲哚总碱的检验方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113156040A (zh) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101019891A (zh) * | 2007-02-14 | 2007-08-22 | 周亚球 | 蟾皮总碱及其制备和分析方法以及其制剂 |
| CN107589213A (zh) * | 2017-10-23 | 2018-01-16 | 安徽华润金蟾药业股份有限公司 | 一种蟾皮药材或饮片或包含蟾皮制剂的质量控制方法 |
-
2021
- 2021-05-17 CN CN202110532452.4A patent/CN113156040A/zh active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101019891A (zh) * | 2007-02-14 | 2007-08-22 | 周亚球 | 蟾皮总碱及其制备和分析方法以及其制剂 |
| CN107589213A (zh) * | 2017-10-23 | 2018-01-16 | 安徽华润金蟾药业股份有限公司 | 一种蟾皮药材或饮片或包含蟾皮制剂的质量控制方法 |
Non-Patent Citations (6)
| Title |
|---|
| 杨世高 等: "华蟾素分散片质量标准研究", 安徽农业大学学报, vol. 36, no. 04, pages 598 - 602 * |
| 汪元符: "HPLC同时测定干蟾皮中华蟾酥毒基和酯蟾毒配基成分的含量", 中国执业药师, vol. 12, no. 12, pages 25 - 30 * |
| 王鹏飞 等: "基于特征图谱和一测多评法的蟾酥药材质量控制研究", 中国中药杂志, vol. 43, no. 14, pages 2863 - 2871 * |
| 陈婷: "HPLC法同时测定蟾酥药材中华蟾酥毒基和酯蟾毒配基的含量", 西北药学杂志, vol. 30, no. 06, pages 690 - 693 * |
| 黄玉叶 等: "蟾皮的提取和含量测定及临床应用研究进展", 中华中医药学刊, vol. 29, no. 07, pages 1636 - 1638 * |
| 黄瑞松 等: "壮药蟾蜍皮质量控制方法的研究", 中国实验方剂学杂志, vol. 19, no. 05, pages 132 - 135 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Du et al. | A green ionic liquid-based vortex-forced MSPD method for the simultaneous determination of 5-HMF and iridoid glycosides from Fructus Corni by ultra-high performance liquid chromatography | |
| Sun et al. | Simultaneous rapid quantification of ginsenoside Rg1 and its secondary glycoside Rh1 and aglycone protopanaxatriol in rat plasma by liquid chromatography–mass spectrometry after solid-phase extraction | |
| Huo et al. | Recent applications in CEC | |
| Peng et al. | Simultaneous ligand fishing and identification of human serum albumin binders from Eucommia ulmoides bark using surface plasmon resonance-high performance liquid chromatography–tandem mass spectrometry | |
| Wang et al. | Simultaneous determination of nucleosides and their bases in Cordyceps sinensis and its substitutes by matrix solid‐phase dispersion extraction and HPLC | |
| Pang et al. | A metal organic framework polymer monolithic column as a novel adsorbent for on-line solid phase extraction and determination of ursolic acid in Chinese herbal medicine | |
| Luo et al. | Molecularly imprinted polymer solid‐phase microextraction coupled with ultra high performance liquid chromatography and tandem mass spectrometry for rapid analysis of pyrrolizidine alkaloids in herbal medicine | |
| Zhang et al. | Screening and isolation of potential neuraminidase inhibitors from leaves of Ligustrum lucidum Ait. based on ultrafiltration, LC/MS, and online extraction-separation methods | |
| CN105361185A (zh) | 一种用大孔树脂分离纯化核桃青皮中多酚类物质的方法 | |
| CN101028354B (zh) | 豨莶风湿片的检测方法 | |
| CN104849369B (zh) | 一种麻附甘药物质量检测方法 | |
| CN107315061A (zh) | 一种治疗妇女子宫出血的茜芷中药制剂的质量控制方法 | |
| Yan et al. | Comparative study on major bioactive components in natural, artificial and in-vitro cultured Calculus Bovis | |
| CN113156040A (zh) | 一种蟾蜍提取液中吲哚总碱的检验方法 | |
| Zhang et al. | Metal–organic framework assisted matrix solid‐phase dispersion microextraction of saponins using response surface methodology | |
| Jiang et al. | Miniaturized solid-phase extraction using a mesoporous molecular sieve SBA-15 as sorbent for the determination of triterpenoid saponins from Pulsatilla chinensis by ultrahigh-performance liquid chromatography-charged aerosol detection | |
| Qin et al. | Ultrasonic-assisted liquid–liquid extraction and HILIC–ELSD analysis of ginsenoside Rb1, astragaloside IV and dulcitol in sugar-free “Fufangfufangteng Heji” | |
| Pang et al. | Monolithic column prepared with UiO-66-2COOH MOF as monomer for enrichment and purification of ursolic acid in plants by online solid-phase extraction | |
| CN115728404B (zh) | 蓝芩口服液对照提取物及其制备方法和应用 | |
| CN109085258A (zh) | 一种琴叶榕的质量检测方法 | |
| CN112798713B (zh) | 一种中药复方抑菌凝胶剂的鉴定和含量测定方法 | |
| Liang et al. | Identification and quantification of C21 steroidal saponins from radix cynanchi atrati by high‐performance liquid chromatography with evaporative light scattering detection and electrospray mass spectrometric detection | |
| CN112051124A (zh) | 一种海龙胶口服液中海龙和/或甘草的快速薄层鉴别方法 | |
| CN112526057A (zh) | 妇炎净胶囊的薄层色谱鉴别方法 | |
| Matysik et al. | Computer-aided optimization of stepwise gradient profiles in thin-layer chromatography |
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 |