摘要:采用UPLC-MS/MS法檢測青蒿中青蒿素的含量,色譜柱Waters ACQUITY BEH C18(50 mm×2.5 mm,1.9 μm),以甲醇和水梯度洗脫,甲醇-水體積比為40%(0 min)-70%(4.5 min)-40%(5.0 min)-40%(5.5 min),流速0.30 mL/min,柱溫30 ℃,進樣量3.0 μL,離子模式(ESI+)分析,青蒿素母離子為m/z=283.3。子離子m/z=151.2,m/z=209.4。結果表明,在上述條件下,青蒿素與雜質分離完全,無明顯干擾,檢出限達到0.174 ng,平均回收率為99.38%,精密度良好,青蒿中青蒿素含量為(1 045.19±27.55) mg/kg,該法適用于青蒿素分析。
關鍵詞:UPLC-MS/MS;青蒿;青蒿素
中圖分類號:O657.6;Q591.1 文獻標識碼:A 文章編號:0439-8114(2013)20-5041-03
Detection of Artemisinin in Artemisia annua L. by UPLC-MS/MS
TENG Ming-gui1,XIE Cheng-cheng1,WANG Ye2,QIANG Yu-gang1,WANG Tao1,ZHAO Yang-yang1,
WANG Zhan-yong1,HOU Tian-gao2,JIANG Hai1
(1. School of Biological Science Engineering, Shaanxi University of Technology, Hanzhong 723000, Shaanxi, China;
2. Hanzhong Senior High School, Hanzhong 723000,Shaanxi, China)
Abstract: The detection of artemisinin in Artemisia annua L. was analysed by UPLC-MS/MS. The chromatographic column waters ACQUITY BEH C18(50 mm×2.5 mm,1.9 μm) was used in the analysis. The elution program was 40% (methanol/water, V/V, 0 min)-70%(4.5 min)-40%(5 min)-40%(5.5 min). The isolation was preformed with 0.3 mL/min at 30 ℃, and the injection volume was 3.0 μL in ESI(+) scan. The precursor ion of artemisinin was m/z=283.3, daughter ion were m/z=151.2,m/z=209.4. The results showed that artemisinin could be detected in these conditions. The detection limit was 0.174 ng with recovery of 99.38% and good precision. The content of artemisinin in Artemisia annua L. reached(1 045.19±27.55) mg/kg. It was suitable for detecting artemisinin in Artemisia annua L..
Key words: UPLC-MS/MS; Artemisia annua L.; artemisinin
青蒿(Artemisia annua L.),又名黃花蒿,菊科,一年生草本,高1.5 m,全株黃綠色,有濃烈揮發性香氣。氣香特異,味微苦,在全世界均有分布,其干燥地上部分具有清熱解暑、除蒸、截瘧等功效[1],主要含有倍半萜、黃酮、香豆素等成分[2-4]。青蒿素(Artemisinin)是青蒿的主要有效成分,青篙素為青蒿中含過氧基團的倍半萜內酯藥物,具有解熱、免疫、抗瘧,抗菌,抗寄生蟲作用,是我國惟一得到國際承認的抗瘧藥物[5]。2011年9月,我國藥學家屠呦呦因創制青蒿素和雙氫青蒿素新型抗瘧藥獲得拉斯克獎[6]。另外,青蒿素在抗腫瘤、腫瘤輔助化療、矽肺治療等方面也有療效[7]。目前,青蒿素已引起國際上的高度重視,原料藥材已實現規范化栽培,因此準確測定青蒿中青篙素含量對青蒿選種培育、青蒿素藥物生產及其藥理藥效研究都具有重要意義。
目前,青蒿素的測定方法包括:重量法(中華人民共和國藥典)[1],紫外分光光度法[8,9],熒光分析法[10,11],氣相色譜法[12],超臨界色譜分析法[13],高效液相色譜法[14]等。高效液相色譜法因重復性和穩定性好已被廣泛應用。但另一方面,由于青蒿素是含過氧橋的倍半萜類化合物,紫外吸收較弱,采用液相分離,紫外檢測器定量分析較困難?!?br>