HPLC-DAD法同时测定肝康复水丸剂中7个成分的含量

Simultaneous Determination of Seven Components in Gankangfu Water Pill by HPLC-DAD

目的:建立同时测定肝康复水丸剂中没食子酸、栀子苷、芍药内酯苷、芍药苷、丹酚酸B、丹皮酚和丹参酮Ⅱ_A含量的方法。方法:采用高效液相色谱(HPLC)法,以二极管阵列检测器(DAD),色谱柱为Agilent ZORBAX SB-C_18;流动相A为乙腈,流动相B为0.1%磷酸溶液,梯度洗脱;流速为1.0 m L/min;检测波长为267 nm(没食子酸)、235 nm(栀子苷、芍药内酯苷、芍药苷)、286nm(丹酚酸B)、274 nm(丹皮酚)、270 nm(丹参酮Ⅱ_A);柱温为30℃;进样量为10μL。结果:没食子酸、栀子苷、芍药内酯苷、芍药苷、丹酚酸B、丹皮酚、丹参酮Ⅱ_A检测质量浓度的线性范围分别为4.09～51.17μg/mL(r=0.999 5)、8.35～104.35μg/mL(r=0.999 1)、7.78～97.25μg/mL(r=0.999 9)、7.56～94.54μg/m L(r=0.999 3)、16.70～208.80μg/mL(r=0.999 5)、3.46～43.20μg/m L(r=0.999 7)、23.13～289.10μg/mL(r=0.999 9),检测限分别为0.09、0.15、0.13、0.12、0.16、0.04、0.24μg/mL,定量限分别为0.26、0.52、0.48、0.47、0.88、0.18、0.98μg/mL,平均回收率分别为91.83%、96.11%、96.83%、95.82%、96.93%、93.95%、94.79%(n=6),精密度试验的RSD均≤1.00%(n=6),供试品溶液48 h内稳定性试验的RSD均≤0.96%(n=6),重复性试验的RSD均≤0.99%(n=6)。结论:本方法准确、可靠,可用于肝康复水丸剂中上述7个成分的含量测定。

OBJECTIVE:To establish a method for simultaneous determination of the contents of gallic acid,jasminoidin,albiflorin,paeoniflorin,danshinolic acid B,paeonol and tanshinoneⅡA in Gankangfu water pill.METHODS:HPLC method with DAD was adopted.The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase A consisted of acetonitrile and mobile phase B consisted of 0.1%phosphate solution(gradient elution)at the flow rate of 1.0 mL/min.The detection wavelengths were set at 267 nm(gallic acid),235 nm(jasminoidin,albiflorin,paeoniflorin),286 nm(danshinolic acid B),274 nm(paeonol)and 270 nm(tanshinoneⅡA).The column temperature was set at 30℃and sample size was 10μL.RESULTS:The linear range of gallic acid,jasminoidin,albiflorin,paeoniflorin,danshinolic acid B,paeonol and tanshinoneⅡA were 4.09-51.17μg/mL(r=0.999 5),8.35-104.35μg/mL(r=0.999 1),7.78-97.25μg/mL(r=0.999 9),7.56-94.54μg/mL(r=0.999 3),16.70-208.80μg/mL(r=0.999 5),3.46-43.20μg/mL(r=0.999 7),23.13-289.10μg/mL(r=0.999 9),respectively.The detection limits were 0.09,0.15,0.13,0.12,0.16,0.04 and 0.24μg/mL;quantitative limits were 0.26,0.52,0.48,0.47,0.88,0.18 and 0.98μg/mL,respectively.The average recovery rates were 91.83%,96.11%,96.83%,95.82%,96.93%,93.95%and 94.79%,respectively(n=6).RSDs of precision test were not more than 1.00%(n=6).RSDs of stability test were not more than 0.96%within 48 h(n=6),and RSDs of repeatability test were than 0.99%(n=6).CONCLUSIONS:The method is accurate and reliable,and can be used for content determination of above 7 components in Gankangfu water pill.