Simultaneous determination of main phytoecdysones and triterpenoids in radix achyranthis bidentatae by high-performance liquid chromatography with diode array-evaporative light scattering detectors and mass spectrometry.
A liquid chromatographic method was developed for simultaneous determination of two main types of bioactive compounds: four phytoecdysones and eight triterpenoids in radix achyranthis bidentatae (RAB), i.e., polypodine B (1), ecdysterone (2), 25-R inokosterone (3), 25-S inokosterone (4), ginsenoside Ro (5), chikusetsusaponin IVa (6), zingibroside R1 (7), chikusetsusaponin IVa ethyl ester (8), 28-deglucosyl-chikusetsusaponin IVa (9), PJS-1 (10), 28-deglucosyl-chikusetsusaponin IVa butyl ester (11), and oleanolic acid (12). Optimum separations were obtained with a Zorbax C18 column, using a gradient elution with 0.08% aqueous formic acid (containing 5% isopropyl alcohol) and acetonitrile as mobile phase. Phytoecdysones were detected by diode array detector (DAD) at 242 nm, whereas triterpenoids were monitored by evaporative light scattering detector (ELSD) connected in series with DAD, temperature for the drift tube was 110 degrees C and the nitrogen flow rate was 3.2 L min(-1). The identity of the analytes was confirmed using retention times, ultraviolet absorbance and mass spectral data in comparison with reference compounds. The method was validated for acceptable precision (intra- and inter-day variation < or = 4.87%), accuracy (recovery > or = 88.9%) and sensitivity (LOD < or = 0.43 microg mL(-1) (DAD) and 26.0 microg mL(-1) (ELSD), LOQ < or = 0.97 microg mL(-1) (DAD) and 46.5 microg mL(-1) (ELSD), respectively). This rapid and reliable method was applied for the analysis of four cultivated and ten commercial samples. The results demonstrated that the method is suitable for routine analysis and quality control of RAB.
Li
Juan
J
Key Laboratory of Modern Chinese Medicines, China Pharmaceutical University, Ministry of Education, Nanjing, 210009, People's Republic of China.
Qi
Hui
H
Qi
Lian-Wen
LW
Yi
Ling
L
Li
Ping
P
D029044:Achyranthes / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004440:Ecdysone / Q000032:analysis* / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D010840:Phytosterols / Q000032:analysis* / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D012542:Scattering, Radiation; D014315:Triterpenes / Q000032:analysis* / Q000737:chemistry
D010840:Phytosterols; D014315:Triterpenes; C021238:phytoecdysones; D004440:Ecdysone