快速溶剂萃取-液质联用法同时测定6种白芷呋喃香豆素  

Simultaneous determination of six furancoumarins in Angelicae Dahuricae Radix using accelerated solvent extraction and LC-MS/MS

作　　者：魏宁[1,2] 原梅[2] 阳海鹰[2] 庄笑梅[2] 孙璐[1] 李桦[2] 

机构地区：[1]沈阳药科大学,沈阳110016 [2]军事医学科学院毒物药物研究所,北京100850

出　　处：《药物分析杂志》2015年第8期1385-1392,共8页Chinese Journal of Pharmaceutical Analysis

基　　金：国家"重大新药创制"科技重大专项(2012ZX09301003-001)

摘　　要：目的：通过利用液质联用方法同时测定川白芷提取液中6种主要呋喃香豆素成分含量并比较3种不同提取方法的提取量,建立一种更高效可靠的中药材质量控制方法。方法：建立液相色谱-质谱联用法（LC-MS/MS）定量测定白芷药材中的6种呋喃香豆素成分（欧前胡素、异欧前胡素、水合氧化前胡素、氧化前胡素、佛手柑内酯和花椒毒酚）,采用CAPCELL PAK MGⅡC18色谱柱（2.0 mm×100 mm,3.0μm）,流动相为含0.1%甲酸乙腈-含0.1%甲酸、5 mmol·L-1甲酸铵水,梯度洗脱,运行4.0 min,流速0.3 m L·min-1,柱温25℃,以ESI为离子源,在阳离子扫描多反应监测（MRM）模式下,对6种呋喃香豆素成分的离子对进行扫描检测,其中6种成分的定量离子对为欧前胡素m/z 271.1/147.0、异欧前胡素m/z 271.1/147.0、水合氧化前胡素m/z 305.0/203.0、氧化前胡素m/z 287.0/203.0、佛手柑内酯m/z 217.1/174.0、花椒毒酚m/z 203.0/147.0;运用正交设计试验优化快速溶剂萃取法（ASE）的提取工艺,并比较了ASE法、回流提取法和超声辅助提取法的萃取量。结果：欧前胡素、异欧前胡素、水合氧化前胡素、氧化前胡素、佛手柑内酯和花椒毒酚6种被测成分的质量浓度在0.05~1.6、0.05~1.6、0.02~0.64、0.02~0.64、0.02~0.64、0.02~0.64μg·m L-1范围内线性关系良好（相关系数〉0.997）;回收率在95.3%~102.4%之间,RSD均小于3.6%。正交试验设计优化ASE法,优化得到的萃取条件为萃取温度90℃,静态萃取时间6 min,保持循环次数为2次,萃取溶剂为95%乙醇溶液;其他参数设定为系统默认值。经条件优化后ASE萃取得到的白芷6种呋喃香豆素含量显著高于回流法和超声法,ASE萃取得到的主要成分（欧前胡素、异欧前胡素）的含量比其他2种提取方法高10%~20%,其他4种含量相对较低的成分萃取量提高显著,含量相比增加了1.4~2.2倍。结论：ASE法可快速高效萃取白芷香豆素成分,与液相�Objective：To compare the extraction capacity of three extraction methods [ accelerated solvent extraction （ ASE）, reflux extraction and ultrasonic extraction ] utilizing liquid chromatographymass spectrometry （ LC- MS/ MS） for simultaneous determination of six furancoumarins in Angelicae Dahuricae Radix, so as to establish an efficient and reliable quality control method for TCM. Methods：An LC-ESI-MS/MS method was originally established to quantify six furancoumarins （ imperatorin, isoimperatorin, oxypeucedanin hydrate, oxypeucedanin, bergapten and xanthotoxol） in Angelicae Dahuricae Radix. ASE, as a popular automated technology,was used to extract these furancoumarins from the herb product. The separation was performed on a CAPCELL PAK MGII Cls column（2. 0 nun× 100 mm,3.0 μm） under gradient elution with flow rate of 0. 3 mL min - 1. A binary mobile phase system adapted with mobile phase A of water containing 0. 1% formic acid and 5 mmol . L-1 ammoninn formate and mobile phase B of acetonitrile containing 0. 1% formic acid was adopted. The column temperature was set at 25 ℃ and the inject volume was 3 μL. The detection was accomplished by multiple-reaction monitoring （MRM）scanning via electrospray ionization（ESI） source operating in the positive ionization mode. The quantitative ion pairs of the six analytes were as follows： imperatorin m/z 271.1/147.0, isoimperatorin m/z 271.1/147.0, oxypeucedanin hydrate m/z 305.0/ 203.0, oxypeueedanin m/z 287.0/203.0, bergapten m/z 217.0/174.0, and xanthotoxol m/z 203.0/147.0. The to- tal runtime for one injection was 4 min. The primary ASE parameters, including extraction temperature, static time and extraction solvent were optimized by an orthogonal test, based on the yield of imperatorin and isoimperatorin. The extraction efficiency of the ASE was then compared with reflux extraction and ultrasonic-aided extraction. Resuits： Good linearity was achieved in the ranges of 0. 05 - 1.6,0. 05 - 1.6,0.02 - 0. 64,0.02 - 0. 64,0.02 - 0. 64, and

关 键 词：川白芷 呋喃香豆素 欧前胡素 异欧前胡素 水合氧化前胡素 氧化前胡素 佛手柑内酯 花椒毒酚 快速溶剂萃取 正交试验 液质联用 

分 类 号：R917[医药卫生—药物分析学]