HPLC测定远志碱水解8个成分含量

HPLC Determination of Eight Hydrolytic Constituents of Polygalae Radix

目的:建立高效液相色谱法(HPLC)同时测定远志碱水解8个成分(对羟基苯甲酸、对羟基肉桂酸、芥子酸、阿魏酸、苯甲酸、3,4,5-三甲氧基肉桂酸、对甲氧基肉桂酸、细叶远志皂苷)含量的方法,并测定10批不同来源远志碱水解产物的含量。方法:采用YMC-Pack ODS-A(250 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.05%磷酸水溶液(B)为流动相,梯度洗脱(0~6 min,20%~22%A;6~8 min,22%A;8~10 min,22%~25%A;10~14 min,25%~28%A;14~30 min,28%~36%A;30~33 min,36%~38%A),流速为1 mL·min^-1,柱温为30℃,检测波长为210、310 nm。结果:远志碱水解8个成分在33 min内均达到基线分离,且8个成分线性范围良好(r>0.9992,n=6),平均回收率为94.74%~104.95%(RSD<2.75%,n=6);所测样品中8个成分含量分别为0.02%~0.09%、0.20%~0.41%、0.25%~0.70%、0.18%~0.39%、0.19%~0.40%、0.05%~0.68%、0.28%~0.36%、1.77%~4.07%,不同来源远志水解后各成分含量差异明显。结论:该方法准确性和重复性良好,可用于远志碱水解成分含量测定,为远志质量控制提供参考。

Objective:To establish a HPLC method for the simultaneous determination of 8 constituents(P-hydroxybenzoic acid,P-hydroxycinnamic acid,mustard acid,ferulic acid,benzoic acid,3,4,5-trimethoxycinnamic acid,p-methoxycinnamic acid and tenuifolin)in the alkaline hydrolysate of Polygalae Radix and to determine their contents of 10 samples from different sources.Methods:The chromatographic separation was performed on YMC-Pack ODS-A column(250 mm×4.6 mm,5μm)with mobile phase consisting of acetonitrile(A)-0.05%phosphoric acid aqueous solution(B)at a flow rate of 1 mL·min^-1 in a gradient elution mode(0-6 min,20%-22%A;6-8 min,22%A;8-10 min,22%-25%A;10-14 min,25%-28%A;14-30 min,28%-36%A;30-33 min,36%-38%A),the column temperature was maintained at 30℃,and the detection wavelength was set at 210 and 310 nm.Results:Eight components in the hydrolysate of Polygala tenuifolia roots were detected by the established method and all reached the baseline separation within 33 min,with a good linear range of 8 components(r>0.9992,n=6).The average recovery was 94.74%-104.95%(RSD<2.75%,n=6).The contents of 8 components measured in all samples were significantly different after hydrolysis,ranging from 0.02%-0.09%,0.20%-0.41%,0.25%-0.70%,0.18%-0.39%,0.19%-0.40%,0.05%-0.68%,0.28%-0.36%,1.77%-4.07%respectively.Conclusion:The established method is accurate and reproducible,which can be used for the determination of hydrolytic constituents of Polygalae Radix,and providing a potential way for the quality control of Polygalae Radix.