反相高效液相色谱法测定5种藏羌五加菜中金丝桃苷的含量

目的:建立5种藏羌五加菜中金丝桃苷的反相高效液相色谱法含量测定方法,并比较其金丝桃苷的含量差异。方法:采用液相色谱-质谱定性鉴别红毛五加、糙叶藤五加和蜀五加中金丝桃苷。金丝桃苷提取物通过60%乙醇溶液回流提取105min制得;Kromasil C18柱(4.6mm×250mm,5μm)、柱温30℃,流动相甲醇(A)-0.5%磷酸溶液(B),梯度洗脱;检测波长355nm,流速1mL/min。结果:金丝桃苷在0.17～2.55mg范围内峰面积与进样量具有良好的线性关系,回归方程Y=2×106X+1.94883×106(r=0.9990,n=6)。平均回收率100.31%,RSD为0.95%(n=9)。结论:本法简便、准确、重复性好,可用于五加菜质量的控制方法研究。四川阿坝藏族羌族自治州、四川甘孜藏族自治州的五加属植物中金丝桃苷含量:老叶远低于嫩叶;糙叶藤五加嫩叶(低温烘干品)≈蜀五加嫩叶(低温烘干品)>红毛五加嫩叶(低温烘干品)>无梗五加嫩叶(低温烘干品);糙叶藤五加的不同加工工艺:阴干>茶叶>烘干>热烫≈微波干燥≈晒干>>盐渍>冻藏1个月≈热烫后冻藏1个月。

Determination of Hyperoside in Tibetan Acanthopanax Leaves from Five Species by RP-HPLC

Objective: To establish a reversed-phase high-performance liquid chromatography(RP-HPLC) method for the determination of hyperoside in Tibetan Acanthopanax leaves from 5 different species.Methods: Hyperoside was identified by liquid chromatography-mass spectrometry(LC-MS).Hyperoside in samples were extracted by refluxing with 60% ethanol for 105 min.The chromatographic separation was achieved on a Kromasil C18 column at 30 ℃ through gradient elution using methanol as mobile phase A and 0.5% phosphoric acid at a flow rate of 1 mL/min.The detection wavelength was set at 355 nm.Results: A calibration curve equation describing the relationship between hyperoside concentration(Y) and peak area(X) was established as follows: Y=2 × 106X + 1.94883(r=0.990,n=6).The calibration curve was linear over the concentration range of 0.17-2.55.The average spike recovery rate of hyperoside was 100.31% with a RSD of 0.95%(n=9).Conclusion: A simple,accurate and repeatable method to determine hyperoside in Tibetan Acanthopanax leaves has been developed.Tender leaves of Tibetan Acanthopanax from the same species grown in Tibetan and Qiang Autonomous Prefecture and Ganzi Tibetan Autonomous Prefecture,Sichuan Province were found to be much richer in hyperoside than old ones.Low-temperature dried tender leaves of A.leucorrhizus(Oliv.) Harms var.fulvescens Harms & Rehd and A.setchuenensis Harms ex Diels had approximately equal hyperoside levels followed by tender leaves of A.giraldii Harms;tender leaves of A.sessiliforus Rupr had the lowest hyperoside level.Different drying methods were ranked in descending order from the highest to the lowest hyperoside content in dried A.leucorrhizus(Oliv.) Harms var.fulvescens Harms & Rehd as follows: shade drying > processing into tea > oven drying > blanching ≈ microwave drying ≈ sun drying >> salting > 1-month frozen storage ≈ 1-month frozen storage after blanching.