Improvement of Quality Control Standard of Bufonis Venenum in Shexiang Baoxin Pills
10.13422/j.cnki.syfjx.20211646
- VernacularTitle:麝香保心丸中蟾酥的质量控制标准提升
- Author:
Li-yao SHI
1
;
Yun-ge FANG
2
;
Liang-mian CHEN
2
;
Rui WANG
1
;
Zhi-min WANG
2
;
Hui-min GAO
2
;
Peng JIANG
3
;
Chang-sen ZHAN
3
Author Information
1. Shanxi University of Chinese Medicine，Jinzhong 030619，China
2. National Engineering Laboratory for Quality Control Technology of Chinese Herbal Medicines， Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences，Beijing 100700，China
3. Shanghai Hutchison Pharmaceuticals Co. Ltd.，Shanghai 200201，China
- Publication Type:Journal Article
- Keywords:
Bufonis Venenum;
Shexiang Baoxin pills;
cinobufagin;
bufalin;
resibufogenin;
quantitative analysis of multi-components by single marker （QAMS）;
high performance liquid chromatography （HPLC）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(2):159-165
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo develop a quantitative analysis of multi-components by single marker （QAMS） for determination of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills， and to provide a method for improving the national standard of the pills. MethodHigh performance liquid chromatography （HPLC） was developed for simultaneous determination of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills and the methodology validation was carried out. The chromatographic separation was performed on a Nucleosil 100-5 C18 column （4.6 mm×250 mm， 5 μm） with the mobile phase of acetonitrile -0.1% potassium dihydrogen phosphate aqueous solution （pH adjusted to 3.2 with phosphoric acid） （48∶52）， and the flow rate was 0.6 mL·min-1， the detection wavelength was set at 296 nm and the column temperature was 35 ℃. Taking cinobufagin as the internal standard， the relative correction factors （RCFs） of bufalin and resibufogenin were calculated， and the key influencing factors of RCFs were investigated. Relative retention time was used for the chromatographic peak location of the analyte， combining with the on-line ultraviolet spectroscopy and accurate relative molecular weight obtained by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-QTOF/MS）. The external standard method was used to verify the contents of three components obtained by QAMS. ResultQAMS was established for the determination of bufalin， cinobufagin and resibufogenin in the samples， and RCFs of cinobufagin to bufalin and resibufogenin were 0.922 and 1.01， respectively. The total content of the three marker compounds in 11 batches of Shexiang Baoxin pills was 33.7-36.0 µg per pill. There was no significant difference between the quantitative results of QAMS and external standard method. ConclusionThe established method can be used for the quality control of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills. It is suggested that bufalin should be considered as one of three marker compounds， and the sum of bufalin， cinobufagin and resibufogenin should be used for the content limit of this preparation.
