HPLC测定秦艽配伍药对前后特征性成分的变化

Determination of Changes of Characteristic Components Before and After Different Gentiana Combinations by HPLC Method

摘要:

目的：探讨药对秦艽-威灵仙、秦艽-桑寄生、秦艽-防己配伍前后特征性成分变化的规律。建立HPLC测定方法。方法：Agilent A3000250×046 PurFuit 5 C₁₈色谱柱（4.6 mm×250 mm，5 μm），流动相0.04% H₃PO₄水溶液（A）-乙腈（B），梯度洗脱（0~15 min，5%~15% B；15~25 min，15%~25% B；25~35 min，25%~35% B；35~40 min，35%~70% B；40~55 min，70%~95% B），流速0.8 mL·min^-1，检测波长220 nm，柱温25 ℃。结果：药对秦艽-威灵仙中分离出16个特征峰（w1~w16），秦艽-桑寄生中分离出18个特征峰（s1~s18），秦艽-防己中分离出21个特征峰（f1~f21），其中峰号w1，w3，w4，w5，w6，w7，w8，w10，w11，w13，w16，s3，s4，s5，s7，s9，s10，s11，s12，s13，s14，f1，f4，f6，f8，f12，f14，f15，f16，f21峰面积升高，w2，w9，s1，s2，s6，s8，f2，f3，f5，f10，f11，f13，f17，f19峰面积降低，w14，w15，s16，s17，s18，f9，f20峰面积不变，其中w8，s7，f9为獐芽菜苦苷，w10，s8，f11为龙胆苦苷，s13，f16，f17分别为槲皮苷、防己诺林碱、粉防己碱。w12，s15，f7，f18为药对中新产生的成分。结论：秦艽配伍用药后所含的特征性成分含量发生了变化且有新成分的生成，本研究所建立的HPLC分析秦艽不同配伍药对前后特征性成分变化的方法简便、准确、重复性较好。

Abstract:

Objective: To establish an HPLC method for determination of changes of characteristic components before and after the compatibility of Gentianae Macrophyllae Radix-Clematidis Radix et Rhizoma,Gentianae Macrophyllae Radix-Taxilli Herba, and Gentianae Macrophyllae Radix-Stephaniae Teerandrae Radix. Method: The chromatographic column was Agilent A3000250×046 Pursuit 5 C₁₈ (4.6 mm×250 mm,5 μm), with 0.04% phosphoricacid solution (A)-acetonitrile (B) as the mobile phase for gradient elution (0-15 min,5% -15% B;15-25 min,15% -25% B;25-35 min,25% -35% B;35-40 min,35% -70% B;40-55 min,70% -95% B) at the flow rate of 0.8 mL·min^-1. The detection wavelength was set at 220 nm,and the column temperature was set at 25 ℃. Result: The 16 characteristic peaks (w1-w16) were isolated from Gentianae Macrophyllae Radix-Clematidis Radix et Rhizoma herbal pair;18 characteristic peaks (s1-s18) were isolated from Gentianae Macrophyllae Radix-Taxilli Herba herbal pair;and 21 characteristic peaks (f1-f21) were isolated from Gentianae Macrophyllae Radix-Stephaniae Teerandrae Radix herbal pair. Among them,the peak areas of w1,w3,w4,w5,w6,w7,w8,w10,w11,w13,w16,s3,s4,s5,s7,s9,s10,s11,s12,s13,s14,f1,f4,f6,f8,f12,f14,f15,f16 and f21 were increased;peak areas of w2,w9,s1,s2,s6,s8,f2,f3,f5,f10,f11,f13,f17 and f19 were decreased;and the peak areas of w14,w15,s16,s17,s18,f9 and f20 were not changed. w8,s7 and f9 were swertiamarin;w10,s8 and f11 were gentiamarin;s13,f16 and f17 were quercitin,fangchinoline and tetrandrine. w12,s15,f7 and f18 were the new components produced in the herbal pairs. Conclusion: After Gentianae Macrophyllae Radix compatibility,the contents of characteristic ingredient were changed and new components were generated. The established HPLC method was simple,accurate and repeatable to analyze the changes of characteristic components before and after different Gentianae Macrophyllae Radix drug combinations.