基于HPLC-UV-MS技术的甘草和马钱子大鼠口服入血成分分析  

Characteristic Ingredients Profiling of Glycyrrhiza Radix and Semen Strychni in Rat Plasma Using HPLC-UV-MS

作　　者：张敏[1,2] 邓阳[1,2] 文雅洁[3] 方平飞[1,2] 罗世林[1,2] 王超[1,2] ZHANG Min DENG Yang WEN Ya-jie FANG Ping-fei LUO Shi-lin WANG Chao(Department of Pharmacy, The Second Xiangya Hospital, Central South University, Changsha 410011, China Institute of Clinical Pharmacy, Central South University, Changsha 410011, China Pharmacy School of Sun Yat-sen University, Guangzhou 510006, China)

机构地区：[1]中南大学湘雅二医院药学部,长沙410011 [2]中南大学临床药学研究所,长沙410011 [3]中山大学药学院,广州510006

出　　处：《中国药学杂志》2017年第9期731-737,共7页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(81473411;81573686)

摘　　要：目的建立解毒剂量甘草和中毒剂量马钱子药材提取液混合物的高效液相色谱-紫外-串联质谱(HPLC-UV-MS)分析方法,并运用于甘草和马钱子大鼠口服入血成分分析。方法采用Sino Chrome ODS-BP(4.6 mm×250 mm,5μm)色谱柱,水(含0.1%甲酸)(A)-乙腈(B)(V/V)为流动相,梯度洗脱:0.01 min 8%B,2 min 8%B,30 min 25%B,32 min 30%B,50 min45%B,53 min 85%B,60 min 85%B,61 min 8%B,77 min停止,流速0.3 mL·min^(-1)。UV检测波长254 nm,柱温40℃。质谱扫描采用正负离子模式同时检测的全扫描模式,正负离子模式锥孔电压分别30、25 V,毛细管电压分别为2.5、3.5 V。待测血浆用乙腈沉淀蛋白处理后测定。结果建立了同时测定甘草和马钱子药材提取物及入血成分的HPLC-UV-MS方法,比较了甘草和马钱子体内外物质变化。结论采用简便,可重复的HPLC-UV-MS方法对甘草和马钱子提取物体内外成分进行确证和分析,为甘草对马钱子解毒机制的物质基础研究提供实验依据。OBJECTIVE To establish a high performance liquid chromatography coupled with tandem mass spectrometry and ultraviolet detector （HPLC-UV-MS） method for determination of the mixture of Glycyrrhiza Radix extract at detoxification dosage and Semen Strychni extract at poisonous dosage, which will be applied to the characteristic ingredients profiling in rat plasma. METHODS Sino Chrome ODS-BP column （4. 6 mm ×250mm,5μm） was employed, which was maintained at 40 ℃ , and the mobile phase was composed of water （0. 1% formic aeid） （A） and aeetonitrile （B）. The gradient elution program was as follows： 0. 01 rain 8% B, 2 rain 8% B, 30 min 25% B, 32 rain 30% B, 50 min 45% B, 53 min 85% B, 60 min 85% B, 61 min 8% B, and terminated at 77 min, eluted at a flow rate of 0. 3 mL ·min^-1 A UV detector was equipped and the detection wavelength was 254 nm. ESI ^＋ mode was selected for strychnos alkaloids with cone voltage of 30 V and capillary voltage of 2. 5 V, while ESI- mode for glycyrrhiza with cone voltage of 25 V and capillary voltage of 3.5 V. Acetonitrile was used as protein precipitation agent to determine the characteristic ingredients in rat plasma. RESULTS An HPLC-UV-MS method for simultaneous determination of liquorice and Semen Stryehni extract was established and applied to measure the eomponents absorbed into blood. The material changes of liquorice and Semen Strychni in vivo and vitro were also compared. CONCLUSION A simple, repeatable HPLC-UV-MS method for analysis of Glyeyrrhiza Radix and Semen Stryehni in vivo and vitro was established. The related components are speculates, which provided the experimental basis to research the detoxification mechanism of Glycyrrhiza Radix against strychninization.

关 键 词：甘草 马钱子 解毒 高效液相色谱-紫外-串联质谱 入血成分 

分 类 号：R917[医药卫生—药物分析学]