LC—MS/MS法同时测定葛根药材中14种异黄酮类成分的含量

　　摘 要 目的：建立同时测定葛根药材中14 种异黄酮类成分含量的方法。方法：采用液相色谱串联质谱法同时测定14批葛根药材（野葛：PL-1～PL-7，粉葛：PT-1～PT-7）中14种异黄酮类成分的含量。色譜柱为 Extend C18，流动相为甲醇-水（梯度洗脱），流速为0.8 mL/min，柱温为22 ℃，进样量为5 μL；离子源为电喷雾离子源，检测方式为负离子检测，扫描方式为多级反应监测模式，喷射电压为-4 500 V，离子源温度为600 ℃，雾化气为60 psi ，辅助气为60 psi， 碰撞气为7 psi，气帘气为30 psi。采用SIMCA 13.0软件对上述14批药材进行聚类分析。结果： 14种待测异黄酮类成分在10～1 000 ng/mL（葛根素为10～5 000 ng/mL）范围内具有良好的线性关系（r均大于0.990 0），精密度、稳定性和重复性试验的RSD均小于5%，加样回收率范围为95%～105%，RSD范围为0.8%～4.5%（n=6）。野葛与粉葛中异黄酮类总含量差异较大，不同产地的葛根药材异黄酮含量差异较大；14种异黄酮类成分中，葛根素含量最高。聚类分析结果显示，14批葛根药材可聚为4类，PL-2为Ⅰ类，PL-3、PL-4为Ⅱ类，PL-5、PL-6、PL-7为Ⅲ类，其余为Ⅳ类。结论：该方法操作简单、重复性良好，可用于检测葛根药材中14 种异黄酮成分的含量，适用于该药材的整体质量控制。
　　关键词 葛根；粉葛；异黄酮类；液相色谱-串联质谱法；含量测定
　　Simultaneous Determination of 14 Kinds of Isoflavones in Pueraria radix by LC-MS/MS
　　WU Wenjie1，2，LIU Lianghong3，HUANG Ying1（1. School of Pharmacy，Hunan University of TCM，Changsha 410208，China； 2. School of Chemistry， Hong Kong Baptist University， Hong Kong， China； 3. School of Pharmacy， Hunan Pharmaceutical University， Hunan Huaihua 418000， China）
　　ABSTRACT OBJECTIVE： To establish the method for simultaneous determination of 14 kinds of isoflavones in Pueraria radix. METHODS： LC-MS/MS was adopted to detect 14 kinds of isaflavones in 14 batches of P. radix （P. radix：PL-1 to PL-7， P. thomsonii： PT-1 to PT-7）. The determination was performed on Extend C18 with mobile phase consisted of methanol-water （gradient elution） at the flow rate of 0.8 mL/min. The column temperature was set at 22 ℃. The sample size was 5 μL. Ion source was ESI source. The detection mode was negative ion detection. Scanning mode was MRM with jet voltage of -4 500 V； ion source temperature was set at 600 ℃， and atomization gas was 60 psi. The auxiliary gas was 60 psi， collision gas was 7 psi， air curtain gas was 30 psi. SIMCA 13.0 software wasused for cluster analysis of above batches. RESULTS： The linear range of 14 kinds of isoflavones ranged 10-1 000 ng/mL（puerarin of 10-5 000 ng/mL，r>0.990 0）. RSDs of precision， stability and reproducibility tests were all lower than 5%. The recoveries were 95%-105%（RSD were 0.8%-4.5%，n=6）. The total content of isoflavones were different significantly between P. radix and P. thomsonii. The contents of isoflavones in P. radix from different origins were different significantly. Among 14 kinds of isoflavones， the content of puerarin was the highest. Results of cluster analysis showed that 14 batches of P. radix could be clustered into 4 categories， including PL-2 as Ⅰ category， PL-3 and PL-4 as Ⅱ category， PL-5， PL-6 and PL-7 as Ⅲ category， other as Ⅳcategory. CONCLUSIONS： The method is simple and reproducible. It can be used for content determination of 14 kinds of isoflavones in P. radix and quality control.

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