大蒜辣素提取物及其降解产物的HPLC-MSMS研究01.doc

大蒜及的LC-MS/MS研究 摘要：对大蒜提取物其进行研究。采用。大蒜提取液中主要含有硫代亚磺酸酯类成分。关键词：大蒜；大蒜辣素；稳定性；HPLC-MS/MS -mass spectrometric analysis of the chemical components and decomposition products in allicin extract of garlic WANG LI (, Song Min, HANG Tai-jun (Department of Pharmaceutical Analysis, China Pharmaceutical University, NanJing, 210009，China Abstract: To analyze the chemical components and decomposition products in allicin extract of garlic. The chemical components screening and identification were made with HPLC-MS/MS method by full scan TIC MS, HPLC retention time, product MS spectra and/or chemical reference standards. The stability of the extract in water and alcoholic solutions was also investigated. There were five major components in allicin extract which were all identified as thiosulfinates. The extract was stable for at least 3 months when stored at -20 oC as water solution. But obvious decomposition was observed with the increase of alcoholic concentration. The decomposition products were also identified by HPLC-MS/MS. Key words: Garlic; Allicin extract; Stability; HPLC-MS/MS 大蒜是历史悠久的药食两用佳品，大量研究证明，大蒜具有杀菌、降血脂、抗血栓、预防中风和高血压、保护肝脏和提高机体免疫力、抗癌等功能[1,2]。蒜氨酸及蒜酶催化裂解产生的大蒜辣素Allicin，二烯丙基硫代亚磺酸酯等化合物[3]是公认功效成分。但大蒜辣素，有强烈的蒜臭味，有时会出现绿褐变。为了充分发挥大蒜辣素的效果，大蒜辣素的稳定性问题，国内外室温下纯蒜辣素半衰期2-16小时，在大蒜汁或破碎大蒜中其半衰期2.4天，在11大蒜汁和水的稀释液中其半衰期22 天，冷冻干燥可以使蒜辣素的寿命延长0天[5]。有研究采用GC-MS对大蒜提取物的成份进行了分析，但由于本类成份的热不稳定性，所获得信息量有限[6]。本文采用高效液相色谱-质谱法HPLC-MS/MS)对大蒜辣素。 材料与方法 仪器 Agilent 1100系列高效液相色谱仪包括真空脱气泵、四元梯度泵、自动进样器、柱温箱和DAD检测器，美国Agilent公司；Thermo-FinniganTSQ Quantum Ultra AM型LC-MS/MS联用仪Xcalibur 1.2数据处理系统美国Thermo-Finnigan公司。 药品与试剂 精制蒜氨酸、蒜酶由新疆医科大学提纯度99.0；羟苯乙酯购自；新鲜白皮蒜市售, 产；甲醇为色谱纯；水为高纯水；其他试剂为分析纯。 色谱与质谱条件 色谱条件 ODS-2色谱柱4.6 mm×250 mm, 5 μm)，甲醇-1%甲酸50:50)流动相流速1.0 mL·min- 1柱温35 ℃检测波长242 nm进样量20 μL。 质谱条件 mL·min- 1进入质谱检测供试液的制备 大蒜辣素提取 大蒜经挑选、去皮、洗净后，用捣碎机破碎浆，35 ℃下酶解2 h，滤过收集，二氯甲烷萃取，无水硫酸钠干燥，减压浓缩定容至 mL，20 μL进行HPLC-MS/MS分析 大蒜辣素对照品 精密称取蒜氨酸蒜酶适量，用水超声溶解制成分别2 mg·mL- 11 mg·mL- 1的溶液，精密量各1mL，混合，在35 ℃水浴温孵酶解15 min，稀释10 mL，滤过20 μL进行HPLC-MS/MS分析[]。 结果与讨论 1 大蒜辣素提取成分大蒜辣素图