GC-MS/SIM法测定不同产地杜仲叶中的氨基酸

Determination of amino acids in Eucommia ulmoides Oliv leaves by gas chromatography-mass spectrometry/selected ion monitor

目的建立杜仲叶中氨基酸含量测定的GC-MS/SIM方法,并用此方法测定不同产地杜仲叶中氨基酸含量。方法杜仲叶采用6mol.L-1盐酸提取,提取液经BSTFA+1%TMCS衍生化后进样,全扫描定性,选择离子监测内标法定量。色谱柱为HP-5MS石英毛细管柱(30m×0.25μm×0.25mm),初始柱温为100℃,保持3min,程序升温8℃.min-1至300℃,保持6min,分流比为10∶1。结果各氨基酸在相应的范围内线性关系良好(r>0.999),平均回收率在89.6%～106.3%(n=5),RSD在1.7%～6.0%。采用该法对4个不同产地的杜仲叶进行氨基酸的含量测定,结果表明杜仲叶中含有氨基酸15种,湖北恩施产杜仲叶氨基酸含量较高,为9.95%;重庆长寿产杜仲叶氨基酸含量较低,为5.80%。结论本法简便灵敏、重现性好、准确度高,适用于杜仲叶中氨基酸的分离检测。

Objective To develop a method to determine amino acid in Eucommia ulmoides Oliv leaves by gas chroma- tography-mass spectrometry/selected ion monitor （GC-MS/SIM）. Methods Amino acids as TMS derivatives were separated and identified on HP-5MS column （30 mN 0.25 μm× 0.25 ram）. The initial column temperature of 100 ℃ was held for 3 min, and then programmed at 8 ℃ min-1 to 300℃ for 6 rain. Identification was performed on the basis of the retention time and the mass peak intensity ratio. The quantity was determined hy the internal standard technique and the internal standard was sarcosine. Results All the 15 amino acids found in Eucomrnia ulmoides Oliv leave, showed good linearity in its corresponding linear range. The recovery was at 89.6 %-106.3% and the RSD was 1.7%-6.0%. Conclusion The method is simple, accurate and sensitive, which can determine the amino acids in Eucommia ulmoides Oliv leaves.