High-performance liquid chromatography with evaporative light-scattering detection method for determination of astragaloside IV in Radix astragali
- Author:
Xiang LI
1
Author Information
1. Department of Pharamaceutical Analysis
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(3):331-333
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To determine the content of astragaloside IV in Radix astragali by high-performance liquid chromatography with evaporative light-scattering detection (HPLC-ELSD). Methods: Radix astragali was extracted with MeOH-NH3 · H2O (9 : 1) for 60 min. The experimental condition was confirmed and the validation of the method was also tested. The chromatography condition was as follows: Hypersil ODS 2 column (4.6 mm × 250 mm, 5 μm); mobile phase was A: ACN, B: H2O, gradient elution; flow speed was 1.0 ml/min; temperature of column was room temperature; and the injection volume was 20 μl. The ELSD conditions were as follows: the temperature of drift tube was 40°C, the gas pressure was 3.5 bar (1 bar = 105 Pa), and the value of gain was 7. Results: The theoretical plate number of astragaloside IV was 214 798, the resolution was 1.858 and the tailing factor was 1.011. The calibration curves were linear in the range of 6.930-693.0 μg · ml-1, Y = 1.415 X + 7.509, r = 0.9994. The intra-day and inter-day precision (RSD) were all less than 2.0%. The stability (RSD) was 1.24% in 48 h and the recurrence (RSD) was 1.26% (n = 5). The limit of detection was 0.6930 μg · ml-1 and the recovery was 97.05% (RSD = 0.17%, n = 3) for astragaloside IV. The contents of astragaloside IV in 10 different batch of Astragalus membranaceus (Fisch.) Bge. were consistent to the standard of Chinese Pharmacopoeia. Conclusion: HPLC-ELSD can shorten the process and time of Radix astragali pretreatment. It is a simple and reliable method to determine the content of astragaloside IV in Radix astragali.
