高效液相色谱法测定胡椒粉中胡椒碱的含量

Determination of Piperine in Pepper Powder by HPLC Method

建立了高效液相色谱法测定胡椒粉中胡椒碱含量的分析方法。样品用乙醇提取,涡旋2min,超声30min,离心后,0.45μm滤膜过滤。采用Waters Atlantis T3色谱柱分离(250 mm×4.6 mm×5μm),以甲醇∶水(75∶25)作流动相,紫外检测器检测,检测波长343nm,进样量20μL,外标法定量。在此优化的实验条件下,胡椒碱的浓度在0.5~10μg/mL范围内与峰面积呈良好的线性关系,回归方程分别为Y=12777X-37170(r=0.9998),检出限为25mg/kg,回收率在96.20%~98.30%之间,相对标准偏差在1.85%~3.12%之间。该方法前处理简便、重现性好,可适用于胡椒粉中胡椒碱含量的测定。

To establish a high performance liquid chromatography method for the determination of piperine in pepper power.The samples are extracted with ethanol,vortexed for 2 min,sonicated for 30 min,centrifuged,and filtered through a 0.45μm filter.Using Waters Atlantis T3 column for separation（250 mm×4.6 mm×5μm）,methanol∶water（75∶25）as the mobile phase,detected with UV detector,the detection wavelength is 343 nm,injection volume：20μL,quantitate with external standard.Under the optimized conditions,there is a good linear relationship between the concentration of piperine and the peak area in the range of 0.5~10μg/mL.The regression equation is Y =12777 X-37170（r=0.9998）,the detection limit is 25 mg/kg,the recovery rate is between 96.20% and 98.30%,and the relative standard deviation is between 1.85% and 3.12%.The method has simple pretreatment and good reproducibility and can be applied to the determination of piperine in pepper powder.