HPLC一测多评法同时测定胃疡灵颗粒中8种成分的含量

目的:建立HPLC一测多评法同时测定胃疡灵颗粒中肉桂酸、桂皮醛、毛蕊异黄酮葡萄糖苷、芒柄花苷、芒柄花素、6-姜辣素、8-姜酚和10-姜酚的含量。方法:采用HPLC法,以Agilent SB-C₁₈柱(250 mm×4.6 mm,5μm)为色谱柱,流动相A为乙腈-甲醇(9∶1),流动相B为0.1%磷酸溶液,梯度洗脱(0~13 min,28%A;13~24 min,28%→35%A;24~39 min,35%→56%A;39~53 min,56%→85%A;53~60 min,85%→28%A),流速为1.1 ml/min,检测波长为280 nm,柱温为25℃。以毛蕊异黄酮葡萄糖苷为内参物,建立其他7个成分的相对校正因子,计算各成分含量。结果:肉桂酸、桂皮醛、毛蕊异黄酮葡萄糖苷、芒柄花苷、芒柄花素、6-姜辣素、8-姜酚和10-姜酚分别在0.59~11.80、7.47~149.40、2.18~43.60、1.91~38.20、4.49~89.80、3.87~77.40、0.94~18.80、1.46~29.20μg/ml(r≥0.9991)范围内线性关系良好;平均加样回收率分别为(96.89±1.04)%、(100.05±0.64)%、(99.52±1.18)%、(98.51±1.30)%、(99.38±0.77)%、(98.29±0.88)%、(97.72±1.26)%和(97.93±1.01)%(n=9);肉桂酸、桂皮醛、芒柄花苷、芒柄花素、6-姜辣素、8-姜酚和10-姜酚的相对校正因子分别为0.8263、0.8490、0.9720、1.2005、1.2766、0.7202和0.8749,一测多评法计算值和外标法实测值无明显差异。结论:本研究建立的HPLC一测多评法操作便捷、结果准确,可用于胃疡灵颗粒的质量控制。

Simultaneous determination of eight components in Weiyangling granules by HPLC-quantitative analysis of multi-components by single marker

Objective:To establish a high performance liquid chromatography quantitative analysis of multi-components by single marker(HPLC-QAMS)for simultaneous determination of cinnamic acid,cinnamaldehyde,calycosin-7-O-β-D-glucopyranoside,ononin,formononetin,6-gingerol,8-gingerol and 10-gingerol in Weiyangling granules.Methods:An HPLC method was established with Agilent SB-C₁₈(250 mm×4.6 mm,5μm)as a separate column.Acetonitrile-methanol(9∶1,phase A)and 0.1%phosphoric acid(phase B)were used as a mobile phase,and the flow rate was 1.1 ml/min with a gradient elution(0-13 min,28%A;13-24 min,28%→35%A;24-39 min,35%→56%A;39-53 min,56%→85%A;53-60 min,85%→28%A).The detection wavelength was set at 280 nm and the column temperature was 25℃.With calycosin-7-O-β-D-glucopyranoside as an internal standard,the relative correction factors of the other seven components were established and the contents of various components were calculated.Results:Cinnamic acid,cinnamaldehyde,calycosi-7-O-β-D-glucopyranoside,ononin,formononetin,6-gingerol,8-gingerol and 10-gingerol showed good linear relationships within the ranges of 0.59-11.80,7.47-149.40,2.18-43.60,1.91-38.20,4.49-89.80,3.87-77.40,0.94-18.80 and 1.46-29.20μg/ml(r≥0.9991),with average recovery rates of(96.89±1.04)%,(100.05±0.64)%,(99.52±1.18)%,(98.51±1.30)%,(99.38±0.77)%,(98.29±0.88)%,(97.72±1.26)%and(97.93±1.01)%(n=9)respectively.The relative correction factors of cinnamic acid,cinnamaldehyde,ononin,formononetin,6-gingerol,8-gingerol and 10-gingerol were respectively 0.8263,0.8490,0.9720,1.2005,1.2766,0.7202 and 0.8749.There were no significant differences between the calculated values by HPLC-QAMS method and the measured values by external standard method.Conclusion:The HPLC-QAMS method established by our study is simple and accurate,which could be used for the quality control of Weiyangling granules.