固相萃取-反相高效液相色谱法测定汤料中的碱性橙Ⅱ

摘要

A rapid and effective method was established for the determination of basic orange Ⅱ in soup stock by solid-phase extraction coupled with reversed-phase high performance liquid chromatography (SPE-RP-HPLC).The samples were extracted with acidified methanol solution.The supernatant was added to the HLB solid phase extraction cartridge,leaching with formic acid aqueous solution,eluting with ammoniated methanol.The solution was then filtrated by 0.22 μ m membrane before testing.The separation was performed by a Waters RP C18 chromatographic column (4.6mm*250mm,5 μ m) using methanol-ammonium acetate (20mmol/L) as mobile phase,with diode array detector and external standard method peak area quantification.The linear range of basic orange Ⅱ was in the range of 1.0 μ g/mL ～ 25.0 μ g/mL with a correlation coefficient of 0.9992.The detection limit was 0.04 μ g/mL and the quantitative limit was 0.12 μ g/mL.The recovery rate was 89.2％ ～ 91.5％.The method has the advantages of simple pretreatment,good purifying effect,high sensitivity and fast detection rate.It is suitable for the separation and quantitative detection of basic orange Ⅱ in soup stock.%建立一种快速、高效测定汤料中碱性橙Ⅱ的固相萃取-反相高效液相色谱(SPE-RP-HPLC)方法.用酸化甲醇溶液提取样品,上清液加入到HLB固相萃取小柱后,选择甲酸-水溶液淋洗,氨化甲醇洗脱,过0.22μm滤膜后上机检测.采用Waters RP C18色谱柱(4.6mm×250mm,5μm)分离,以甲醇-乙酸铵(20mmol/L)水溶液作为流动相,用二极管阵列检测器检测,外标法峰面积定量.结果表明,碱性橙Ⅱ在1.0μg/mL ～ 25.0μg/mL浓度范围内线性良好,相关系数r2为0.9992,检出限为0.04μ g/mL,定量限为0.12μg/mL,加标回收率达到89.2％ ～ 91.5％.该方法具有前处理简单、净化效果好、灵敏度高和检测速度快的优点,适用于汤料中碱性橙Ⅱ的分离和定量检测.