固相萃取-高效液相色谱-蒸发光散射法检测药食同源食用酵素中γ-氨基丁酸含量

Determination of γ-aminobutyric acid content in edible enzymes from medicinal and edible homologous sources by solid phase extraction-high performance liquid chromatography-evaporative light scattering detection

中文关键词:  药食同源  γ-氨基丁酸  食用酵素  SPE-HPLC-ELSD  固相萃取

英文关键词:medicinal and edible homologous sources  γ-aminobutyric acid  edible enzymes  solid phase extraction-high performance liquid chromatography-evaporative light scattering detection  solid phase extraction

中文摘要:

      目的 建立一种基于固相萃取-高效液相色谱-蒸发光散射法(solid phase extraction-high performance liquid chromatography-evaporative light scattering detection, SPE-HPLC-ELSD)快速检测药食同源食用酵素中γ-氨基丁酸(γ-aminobutyric acid, GABA)含量的分析方法。方法 选取西洋参发酵液为代表样品, 经离心(8000 r/min, 10 min)、超声提取(10 min, 25 ℃)后, 对比不净化、分散固相萃取净化、亲水-亲脂平衡(hydrophilic-lipophilic balance, HLB)通过式固相萃取净化3种不同的处理方式优化样品前处理过程, 开发优化HPLC-ELSD仪器方法实现含量准确测定, 并推广应用于其他多种药食同源食用酵素产品中的GABA含量测定。结果 选用优化后的SPE-HPLC-ELSD测定GABA, 在50~2000 μg/mL质量浓度范围内线性指标良好(r=0.999), 加标回收率在85.93%~96.30%, 重复性相对标准偏差为0.87% (n=6)。结论 本方法高效、准确, 适用于多种药食同源食用酵素中GABA定量分析, 也可为复杂基质下的GABA测定及强极性、低紫外吸收小分子化合物检测提供新的途径和参考。

英文摘要:

      Objective To establish a rapid analysis method based on solid phase extraction-high performance liquid chromatography-evaporative light scattering detection (SPE-HPLC-ELSD) for determining the γ-aminobutyric acid (GABA) content in edible enzymes from medicinal and edible homologous sources. Methods Fermented panacis quinquefolii radix liquid was selected as a representative sample. After centrifugation (8000 r/min, 10 min) and ultrasonic extraction (10 min, 25 ℃), 3 kinds of different sample pretreatment methods—non-purification, dispersive solid phase extraction purification, and hydrophilic-lipophilic balance (HLB)-based solid phase extraction purification were compared and optimized. An optimized HPLC-ELSD instrumental method was developed to achieve accurate content determination. The method was then applied to determine the GABA content in other medicinal and edible homologous enzyme products. Results GABA was determined by optimized SPE-HPLC-ELSD. GABA showed a good linear relationship (r=0.999) in the mass concentration range of 50–2000 μg/mL. The recovery rate ranged from 85.93% to 96.30%, with a repeatability relative standard deviation of 0.87% (n=6). Conclusion This method is efficient and accurate, suitable for the quantitative analysis of GABA in various medicinal and edible homologous enzymes. It also provides a new approach and reference for GABA determination in complex matrices and for detecting small molecules with strong polarity and low ultraviolet absorption.