固相萃取-亲水作用色谱法测定香菇中硫杂脯氨酸

通过对色谱条件和样品前处理方法进行优化,建立固相萃取—亲水作用色谱法测定香菇(Lentinula edodes)中硫杂脯氨酸的检测方法。优化后的色谱条件为：Inertsil HILIC 亲水作用色谱柱(4.6×250 mm,5μm)对样品进行分离测定,流动相采用乙腈-0.1%磷酸水(85∶15,v/v)等度洗脱,流速0.8 mL/min,检测波长193 nm,柱温30℃。前处理方法为：香菇样品采用甲醇-水(70∶30,v/v)提取,MAX阴离子固相萃取小柱进行净化处理,甲醇-1.0%甲酸(90∶10,v/v)作为洗脱剂对目标物进行洗脱。检测结果表明,硫杂脯氨酸在2~100μg/mL范围内呈良好的线性关系,相关系数为0.9998,加标回收率为96.3%~101.7%,相对标准偏差在1.3%~4.8%之间,检出限为0.76μg/g,定量限为2.31μg/g。该方法回收率高、简单、快速、无需衍生化,适用于香菇子实体硫杂脯氨酸的定量检测分析。

A Method for Detecting Thioproline in Lentinula edodes Fruit Bodies Using Solid Phase Extraction-Hydrophilic Interaction Chromatography

A simple and sensitive method for the detection and accurate measurement of thioproline in Lentinula edodes fruit bodies using solid phase-hydrophilic interaction chromatography is described.Samples were first dissolved in methanol-water (70∶30,v/v)and purified on a MAX anionic solid phase extraction column using methanol-1.0% formic acid (90∶10,v/v)as eluant.After purification,samples were analyzed at 30 ℃ using an Inertsil HILIC column (4.6×250 mm,5μm),acetonitrile-0.1% phosphoric acid solution (85∶15,v/v)as the mobile phase (flow rate 0.8 mL/min)and a detection wavelength of 193 nm.A good linear relationship(r2>0.9998)was recorded within the range 2~100μg/mL,recovery rates of 96.3%~101.7% were obtained,and the relative standard deviations (RSDs)varied between 1.3% and 4.8% (n=3).The limits of detection and quantification were 0.76μg/g and 2.31μg/g,respectively.