摘要
利用高效液相色谱-串联质谱（LC-MS/MS）方法直接分析植物酵素中多种未衍生化的氨基酸。样品用甲醇稀释5倍,超声提取30 min,离心10 min（速度为10 000 r/min）,取上清液分析。采用色谱柱Venusil ASB C18（250mm×4.6 mm,5μm）分离,以甲醇-乙酸-水混合溶液为流动相体系进行梯度洗脱,流速为0.5 mL/min,质谱喷雾电压为3 kV,离子源温度为150℃,去溶剂温度350℃,去溶剂气体流速为800 L/h。碰撞气氩气的流速保持压力为0.17 Pa。以被测物的提取离子流图峰面积进行定量,该分析方法的线性范围为0.5~200μmol/L（r2〉0.99）,回收率为86%~110%,可对植物酵素中16种氨基酸成分进行定量分析。该方法操作简便快速、准确可靠,还适用于食品、药品及天然产物中多种氨基酸成分的定量分析。
A method was established for underivatized amino acid determination in fermented beverage of plant extract. Samples were diluted with methanol for five times,extracted by ultrasonic vibration for 30 min,and high-speed centrifuged for 15 min at 10 000 r / min. The supernatant was separated and detected by high performance liquid chromatography-tandem mass spectrometry（HPLC-MS / MS）. The chromatographic column was Venusil ASB C18（250 mm×4. 6 mm,5 μm）. The elution was performed at a flow rate of 0. 5 mL / min using the mobile phases of methanol-acetic acid-water mixture. The MS detector was set as follows：ion source voltage 3 kV,ion source temperature 150 ℃,solvent temperature 350℃,gas flow rate 800 L / h. The collision gas was argon with a pressure of 0. 17 Pa. The quantitation analysis was carried out with peak area in extracted ion chromatograms. Good linearities were acquired in the range of 0. 5-200 μmol / L（r20. 99）for the amino acids. The recoveries were between 86% and110%. There were 16 amino acids in the fermented beverage of plant extract quantitatively analyzed.The method is simple,rapid,accurate and reliable in quantitative analysis of amino acid samples in the fields of pharmaceutical,food and natural products.
出处
《色谱》
CAS
CSCD
北大核心
2015年第3期309-313,共5页
Chinese Journal of Chromatography
基金
吉林省重点科技发展计划项目(20120901)
吉林省与中国科学院科技合作高技术产业化专项资金项目(2013SYHZ0008)
关键词
高效液相色谱-串联质谱
未衍生化氨基酸
植物酵素
high performance liquid chromatography-tandem mass spectrometry（HPLC-MS / MS）
un-derivatized amino acid
fermented beverage of plant extract
