HLB固相萃取-高效液相色谱法同时测定花椒粉中碱性橙2、碱性橙21、碱性橙22染料的含量

Detection of basic orange 2,basic orange 21 and basic orange 22 in prickly ash powder by both HLB solid-phase extraction and high performance liquid chromatography

目的建立HLB固相萃取-高效液相色谱法测定花椒粉中碱性橙2、碱性橙21、碱性橙22的检测方法。方法花椒粉中碱性橙用乙腈超声提取后,经HLB固相萃取柱净化,采用Diamonsi L XDB-C_(18)柱色谱柱(4.6 mm×250 mm,5μm)分离,流速为1.0 ml/min,流动相为甲醇和20 mmol/L乙酸铵,梯度洗脱,柱温为30℃,紫外检测波长为449 nm。结果3种碱性橙在1.25μg/ml~20.0μg/ml浓度均呈良好的线性关系,相关系数(r)均>0.9998,检出限均为0.01 mg/kg,加标回收率为80.5%~93.4%,相对标准偏差(RSD)≤3.6%。结论该处理方法操作简便,能够有效提取、净化花椒粉中碱性橙2、碱性橙21、碱性橙22,杂质干扰明显减少,采用高效液相色谱法,重现性好、回收率满意。

Objective To establish a method for determining basic orange 2,basic orange 21 and basic orange 22 in prickly ash powder both by solid phase extraction-high performance liquid chromatography(HPLC).Methods Basic orange in prickly ash powder was extracted with acetonitrile ultrasound,purified through HLB solid phase extraction column.The separation was performed on Diamonsi L XDB-C18column(4.6 mm×250 mm,5μm)at a flow rate of 1.0 ml/min with mobile phases of methanol and 20 mmol/L ammonium acetate in gradient elution.The column temperature was 30℃,and the UV detection wavelength was 449 nm.Results Three basic orange showed a good linearity in the concentration range of 1.25μg/ml-20.0μg/ml,with correlation coefficient(r)>0.9998,and the limit of detection was 0.01 mg/kg.Standard recovery rate was within 80.5%-93.4%.The relative standard deviation(RSD)was≤3.6%.Conclusion The processing method is easy to operate,and can effectively extract and clean up orange from in prickly ash powder,less disturbance and interference peaks.The method had good reproducibility and satisfied recovery rate.