Study on Quality Control of Eight Components in Baohou Tablets Based on Quantitative Analysis of Multi-components by Single Marker

OBJECTIVE To develop a quantitative analysis of multi-components by single marker(QAMS) method for simultaneous determination of the content of calycosin7-O-β-D-glucopyranoside, ononin, formononetin, aucubin, harpagide, harpagoside, oroxin B and oroxin A in Baohou tablets. METHODS Using HPLC with harpagide as an internal reference substance, its relative correction factor with calycosin7-O-β-D-Glucopyranoside, ononin, formononetin, aucubin, harpagoside, oroxin B and oroxin A was established and its content was calculated. The Agilent TC-C₁₈ column (250 mm×4.6 mm, 5 μm) was adopted, the column temperature was set at 30℃. The mobile phase was acetonitrile-methanol (9︰1)(A) and 0.2% phosphoric acid solution(B) with gradient elution, the flow rate was 1.0 mL·min^-1. The detection wavelength were set at 254 nm for calycosin7-O-β-D-glucopyranoside, ononin and formononetin, 210 nm for aucubin, harpagide and harpagoside, 280 nm for oroxin B and oroxin A. The injection volume was 10 μL. RESULTS The linear range of calycosin7-O-β-D-glucopyranoside, ononin, formononetin, aucubin, harpagide, harpagoside, oroxin B and oroxin A were 0.61-15.25, 0.54-13.50, 1.26-31.50, 0.79-19.75, 3.38-84.50, 1.07-26.75, 7.78-194.50, 4.89-122.25 mg·L^-1(r=0.999 3-0.999 8) respectively. The average recovery were 98.02%, 96.99%, 97.91%, 98.32%, 99.30%, 97.83%, 100.06% and 99.62%(RSD=0.79%-1.42%, n=9), respectively. RCFs of calycosin7-O-β-D-glucopyranoside, ononin, formononetin, aucubin, harpagoside, oroxin B and oroxin A were 1.051 6, 1.209 9, 0.721 7, 0.875 0, 0.899 7, 1.127 0 and 0.760 4, respectively. There were no significant differences between the calculated values by QAMS and the measured values by external standard method. CONCLUSION The method is simple and accurate, and can be used for quality evaluation of Baohou tablets.