Stimulation quantitation of saponins and sterones from Achyranthis Bidentatae Radix by high-performance liquid chromatography with double external standards calibration method
10.16438/j.0513-4870.2022-0014
- VernacularTitle:双外标校准高效液相色谱法同时测定中药牛膝中皂苷类成分和甾酮类成分
- Author:
Bing-xiao WU
1
;
Moo-seob KIM
1
;
Liu-ji ZHANG
2
;
Li-hua GU
3
;
Lin-nan LI
3
;
Li YANG
3
;
Zheng-tao WANG
3
Author Information
1. The MOE Key Laboratory for Standardization of Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
2. Henan Engineering Center for Development of Genuine Medicinal Materials, Henan Academy of Traditional Chinese Medicine, Zhengzhou 450004, China
3. The MOE Key Laboratory for Standardization of Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Shanghai R&D Center for Standardization of Chinese Medicines, Shanghai 201203, China
- Publication Type:Research Article
- Keywords:
Achyranthis Bidentatae Radix;
achyranthoside D;
italic>β-ecdysterone;
ouble external standards calibration;
HPLC;
quality evaluation
- From:
Acta Pharmaceutica Sinica
2022;57(6):1868-1873
- CountryChina
- Language:Chinese
-
Abstract:
Saponins and sterones are two main characteristic components in Achyranthis Bidentatae Radix. In order to control the quality of Achyranthis Bidentatae Radix more effectively, a high-performance liquid chromatography (HPLC) method was established by using double external standards calibration method (DESCM) for simultaneous determination of the contents of achyranthoside C, achyranthoside D, β-ecdysterone, 25R-inokosterone and 25S-inokosterone in Achyranthis Bidentatae Radix. Chromatographic separation was achieved on an Agilent Poroshell 120 EC-C18 (150 mm × 4.6 mm, 2.7 µm) using 0.1% phosphoric acid in water and 0.1% phosphoric acid in acetonitrile as mobile phase. The flow rate was 0.8 mL·min-1 and the column temperature was set as 35 ℃, the injection volume was 5 μL and the total analytical time was 30 min. β-Ecdysterone was used as the reference to calculate the relative correction factors (RCF) and relative retention time (RRT) of 25R-inokosterone and 25S-inokosterone, achyranthoside D was used for achyranthoside C. The RCFs of 25R-inokosterone, 25S-inokosterone, and achyranthoside C were 1.116, 1.056, and 0.888 1, respectively. The double external standards calibration method (DESCM) and external standard method (ESM) were used to calculate the contents of five ingredients in Achyranthis Bidentatae Radix samples from different sources and the variation between the results was within acceptable limits (RE ≤ 5%). The results showed that the contents of two saponins and three sterones of Achyranthis Bidentatae Radix were 0.597%-1.916% and 0.044%-0.150% respectively. The total content of saponins was about 10 times that of sterones. In conclusion, the established DESCM allowed simultaneous determination of five ingredients (achyranthoside C, achyranthoside D, β-ecdysterone, 25R-inokosterone, and 25S-inokosterone) in Achyranthis Bidentatae Radix, providing a scientific and feasible overall quality evaluation method for Achyranthis Bidentatae Radix.
