基于液质联用技术对不同产地淡豆豉核苷和氨基酸类成分的分析

Determination of Amino Acids and Nucleosides in Semen Sojae Praeparatum from Different Regions by Using UFLC-MS/MS

目的：建立液质联用技术同时测定不同产地淡豆豉中38种核苷和氨基酸类含量的方法。方法：采用超快速液相色谱和三重四极杆-线性离子阱串联质谱以及亲水色谱柱（2.1 mm×100 mm,3.5μm）,流动相0.2%甲酸水-0.2%甲酸乙腈梯度洗脱,流速0.6 m L·min-1,采用电喷雾正离子化及多反应监测模式（MRM）,对安徽、江苏、河南3个产区不同采收期共8个批次淡豆豉中的22种氨基酸类和16种核苷类成分进行含量测定;并对结果进行PCA-DA聚类分析及t检验。结果：所建方法使淡豆豉中22种氨基酸类和16种核苷类成分得到较好分离,浓度与峰面积呈良好的线性关系,加样回收率在93.74%-104.32%,RSD在0.6%-3.5%;8个批次样品基本可以测得22种氨基酸类和16种核苷类成分,各批次间核苷与氨基酸的含量都有明显差异,总氨基酸质量分数在7.241-34.21 mg·g-1,总核苷质量分数在14.21-82.53 mg·g-1;不同产地样品的氨基酸含量高低排序为安徽产＆gt;江苏产＆gt;河南产,不同产地样品的核苷含量高低排序为江苏产与安徽产＆gt;河南产,同一产地贮存时间短的样品总氨基酸和总核苷含量也高于贮存时间长的。结论：该方法适用于38种核苷和氨基酸类成分含量的同时测定,比较了不同产地淡豆豉的差异性,在一定程度上反应了不同产区间淡豆豉的质量优劣,为后续药效活性研究提供参考。

Objective： To establish liquid chromatography-mass spectrometry（ LC-MS） method for content determination of amino acid and nucleoside in Semen Sojae Praeparatum from different origins. Method：The contents of 22 amino acids and 16 nucleosides in 8 bathes of Semen Sojae Praeparatum（ collected from Anhui,Jiangsu and Henan provinces） were determined by an ultra fast liquid chromatography coupled with triple quadrupole mass spectrometer. Waters XBridge Amide column（ 2. 1 mm × 100 mm,3. 5 μm） was used with0. 2% formic acid acetonitrile-0. 2% formic acid solution as the mobile phase for gradient elution at a flow rate of0. 6 m L·min-1. In addition,electrospray ionization and multiple reaction monitoring（ MRM） mode were used. The results were used for PCA-DA cluster analysis and t test. Result： The 22 amino acids and 16 nucleosides in Semen Sojae Praeparatum were well separated,and showed good linear relationship between concentration and peak area.The average recovery rate was between 93. 74% and 104. 32%,RSD 0. 6%-3. 5%. There were significant differences in the amino acids and nucleosides contents among different batches. The total contents of amino acids were at 7. 241-34. 21 mg·g-1,and the total contents of nucleosides were at 14. 21-82. 53 mg·g-1. The contents of amino acids were in the following order from high level to low level： Anhui 〉 Jiangsu〉 Henan,and nucleosides were in the following order from high level to low level： Jiangsu,Anhui 〉Henan. The total contents of amino acids and total contents of nucleosides in samples with shorter storage time were higher than those with longer storage time in the same region. Conclusion： This method is applicable for the simultaneous determination of contents of 38 kinds of amino acids and nucleosides. The differences in the study can fill up the blank in Semen Sojae Praeparatum from different origins reflect the quality of samples from different origins,and provide references for the further pharmacodynamic research.