LC—MS/MS法同時(shí)測定葛根藥材中14種異黃酮類成分的含量

　　摘 要 目的：建立同時(shí)測定葛根藥材中14 種異黃酮類成分含量的方法。方法：采用液相色譜串聯(lián)質(zhì)譜法同時(shí)測定14批葛根藥材（野葛：PL-1～PL-7，粉葛：PT-1～PT-7）中14種異黃酮類成分的含量。色譜柱為 Extend C18，流動(dòng)相為甲醇-水（梯度洗脫），流速為0.8 mL/min，柱溫為22 ℃，進(jìn)樣量為5 μL；離子源為電噴霧離子源，檢測方式為負(fù)離子檢測，掃描方式為多級反應(yīng)監(jiān)測模式，噴射電壓為-4 500 V，離子源溫度為600 ℃，霧化氣為60 psi ，輔助氣為60 psi， 碰撞氣為7 psi，氣簾氣為30 psi。采用SIMCA 13.0軟件對上述14批藥材進(jìn)行聚類分析。結(jié)果： 14種待測異黃酮類成分在10～1 000 ng/mL（葛根素為10～5 000 ng/mL）范圍內(nèi)具有良好的線性關(guān)系（r均大于0.990 0），精密度、穩(wěn)定性和重復(fù)性試驗(yàn)的RSD均小于5%，加樣回收率范圍為95%～105%，RSD范圍為0.8%～4.5%（n=6）。野葛與粉葛中異黃酮類總含量差異較大，不同產(chǎn)地的葛根藥材異黃酮含量差異較大；14種異黃酮類成分中，葛根素含量最高。聚類分析結(jié)果顯示，14批葛根藥材可聚為4類，PL-2為Ⅰ類，PL-3、PL-4為Ⅱ類，PL-5、PL-6、PL-7為Ⅲ類，其余為Ⅳ類。結(jié)論：該方法操作簡單、重復(fù)性良好，可用于檢測葛根藥材中14 種異黃酮成分的含量，適用于該藥材的整體質(zhì)量控制。
　　關(guān)鍵詞 葛根；粉葛；異黃酮類；液相色譜-串聯(lián)質(zhì)譜法；含量測定
　　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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