HPLC-QAMS法同时测定榆槐片中11种有效成分的含量

Simultaneous Determination of 11 Active Components in Yuhuai Tablets by HPLC-QAMS

目的:建立同时测定榆槐片中栀子新苷、山栀子苷、羟异栀子苷、京尼平龙胆双糖苷、栀子苷、地榆皂苷Ⅰ、地榆皂苷Ⅱ、芸香柚皮苷、柚皮苷、橙皮苷和新橙皮苷等11种有效成分含量的方法。方法:采用高效液相色谱-一测多评(HPLC-QAMS)法。以Agilent TC-C_(18)为色谱柱,以乙腈-0.1%磷酸溶液为流动相进行梯度洗脱,流速为1.0 mL/min,柱温为30℃,检测波长为238 nm(栀子新苷、山栀子苷、羟异栀子苷、京尼平龙胆双糖苷和栀子苷)、203 nm(地榆皂苷Ⅰ和地榆皂苷Ⅱ)、283 nm(芸香柚皮苷、柚皮苷、橙皮苷和新橙皮苷)。以栀子苷为内参物,计算其余10种成分相对于该成分的相对校正因子,从而计算10批样品中各成分的含量,并与外标法所得结果进行比较。结果:栀子新苷、山栀子苷、羟异栀子苷、京尼平龙胆双糖苷、栀子苷、地榆皂苷Ⅰ、地榆皂苷Ⅱ、芸香柚皮苷、柚皮苷、橙皮苷和新橙皮苷检测质量浓度的线性范围分别为0.87~43.50、1.99~99.50、4.06~203.00、7.35~367.50、12.97~648.50、28.98~1449.00、3.79~189.50、1.57~78.50、18.05~902.50、0.66~33.00和14.38~719.00μg/mL(r均大于0.9990),精密度、重复性、稳定性(24 h)试验的RSD均小于2%(n=6),平均加样回收率为96.90%~100.10%,RSD为0.67%~1.74%(n=9)。HPLC-QAMS法与外标法所测得的10批榆槐片中栀子新苷等10种有效成分的含量比较,差异均无统计学意义(P>0.05)。结论:本研究所建立的HPLC-QMAS法操作简便、结果准确,可用于榆槐片中栀子新苷、山栀子苷、羟异栀子苷、京尼平龙胆双糖苷、栀子苷、地榆皂苷Ⅰ、地榆皂苷Ⅱ、芸香柚皮苷、柚皮苷、橙皮苷和新橙皮苷含量的同时测定。

OBJECTIVE:To establish the method for simultaneous determination of 11 active components in Yuhuai tablets,such as gardoside,shanzhiside,gardenoside,genipin 1-gentiobioside,geniposide,ziyuglycosideⅠ,ziyuglycosideⅡ,narirutin,naringin,hesperidin and neohesperidin.METHODS:HPLC-QAMS method was adopted.The determination was performed on Agilent TC-C_(18) column(250 mm×4.6 mm,5μm)with mobile phase consisted of acetonitrile(A)-0.1%phosphoric acid solution(B)(gradient elution)at the flow rate of 1.0 mL/min.The column temperature was set at 30℃.The detection wavelengths were set at 238 nm for gardoside,shanzhiside,gardenoside,genipin 1-gentiobioside and geniposide,203 nm for ziyuglycosideⅠand ziyuglycosideⅡ,and 283 nm for narirutin,naringin,hesperidin and neohesperidin.Using geniposide as an internal reference,the relative correction factors of other 10 components relative to this component were calculated,and the contents of each component in 10 batches of samples were calculated.The results obtained by HPLC-QAMS method were compared with those obtained by external standard method.RESULTS:The linear ranges of gardoside,shanzhiside,gardenoside,genipin 1-gentiobioside,geniposide,ziyuglycosideⅠ,ziyuglycosideⅡ,narirutin,naringin,hesperidin and neohesperidin were 0.87-43.50,1.99-99.50,4.06-203.00,7.35-367.50,12.97-648.50,28.98-1449.00,3.79-189.50,1.57-78.50,18.05-902.50,0.66-33.00 and 14.38-719.00μg/mL(all r>0.9990).RSDs of precision,repeatability and stability(24 h)tests were all less than 2%(n=6).The average recoveries were 96.90%-100.10%,and RSDs were 0.67%-1.74%(n=9).There was no significant difference in the contents of 10 active components as gardoside between HPLC-QAMS method and external standard method in 10 batches of Yuhuai tablets(P>0.05).CONCLUSIONS:The HPLC-QMAS method established in this study is convenient and accurate.It can be used for the simultaneous determination of gardoside,shanzhiside,gardenoside,genipin 1-gentiobioside,geniposide,ziyuglycosideⅠ,ziyuglycosideⅡ,narirutin,naringin,hesperidin and neohesperidin in Yuhuai tablets.