盐处理下植物组织中甜菜碱和脯氨酸的HPLC-ESI-MS分析

Simultaneous Determination of Betaines and Proline in Plant Tissue under Salt Stress by HPLC-ESI-MS

通过高效液相色谱 -电喷雾质谱(HPLC -ESI-MS)联用技术 ,优化了 pH、流速、温度、鞘气和辅气等相关影响因素 ,建立了一个有选择性的、高灵敏度的分析方法 ,鉴定盐处理下植物组织提取物中的甜菜碱和脯氨酸 ,一次性快速地分析了甘氨酸甜菜碱、β_丙氨酸甜菜碱、葫芦巴碱、脯氨酸甜菜碱、4_羟脯氨酸甜菜碱和脯氨酸等6种化合物。其标准曲线 (50～1000ng/mL)呈直线 ,线性相关系数大于0.999 ;相对标准偏差(RSD)为0.5%～15%,回收率在94.0%～97.0 %之间。运用这一方法已经分析了大麦、小麦、大豆、甜菜和碱蓬等植物组织中的甜菜碱和脯氨酸 ,获得了较理想的实验数据 ,为揭示甜菜碱和脯氨酸与植物的耐盐性生理机制。

Glycine betaine(GB),β-alanine betaine(AB),trigonelline(Trg),proline betaine(PB),4-hyˉdroxyproline betaine(HPB)and proline in plant tissues under salt stress were selectively and sensitively deterˉmined by a rapid HPLC-ESI-MS method,after experimental parameters such as pH,flowrate,temperaˉture,and supplementary gases were optimized.The calibration curves equations of six compounds were linˉear from50to1000ng/mL,and the correlation was more than0.999.The relative standard deviations(RSDs)and the recovery varied from0.5%to15%and from94.0%to97.0%,respectively.This method was used to assay betaines and proline in the leaf and roots of wheat,barley,soybean,beet and Suaeda heteroptera,and satisfactory results were obtained.These results should be helpful for understandˉing the relationship between betaines and proline and the plant resistance to the salt stress,and provide sciˉentific basis for salt tolerance identification and breeding of varieties.