高效液相色谱法测定饲料中盐酸赛庚啶和盐酸可乐定含量

摘要

本实验建立了高效液相色谱法测定饲料中盐酸赛庚啶和盐酸可乐定含量的方法。取2 g样品以0.1%盐酸-乙腈沉淀蛋白，8000 r/min高速离心3 min后合并上清液，50℃减压旋转蒸发至干。5 mL 0.1%盐酸-乙腈（90∶10， V/V）溶液溶解后通过混合型阳离子交换柱净化，收集洗脱液于50℃减压旋转蒸发至干，用1 mL 0.1％盐酸-乙腈（90∶10，V/V）溶液溶解后，过0.22μm尼龙过滤膜，上机测定。液相色谱条件为：流动相A为0.1％HCl（pH＝2.0），流动相B为乙腈，进行梯度洗脱，色谱柱为十八烷基硅烷键合相，柱温30℃，流速1.0 mL/min，检测器为二极管阵列检测器，检测波长为210 nm。实验结果表明，该方法在1～100 mg/kg线性关系良好（r＞0.999），在预混合饲料、浓缩饲料、配合饲料中添加量为1、10、100 mg/kg时方法的准确性（回收率＞70%）和重现性（RSD＜10%）好，可用于饲料中盐酸赛庚啶和盐酸可乐定含量的检测。%In order to determine the residue of cyproheptadine hydrochloride and clonidine hydrochloride in feed staff,a method by high performance liquid chromatography (HPLC) was established.2 g sample was extracted by 0.1%HCl-acetonitrile(90∶10,V/V),and centrifuged at 8000 r/min for 3 minutes,then the liquid supernatant was merged.After it was vacuum rotary evaporated at 50 ℃,0.1% HCl-acetonitrile (90∶10,V/V) was used to solute the residue.Strong acid cation exchange SPE column was selected to purify the sample.After elution liquid was evaporated,0.1% HCl-acetonitril (90∶10,V/V) was used to solute the residue.After the sample was filtrated by a 0.22 μm nylon filtter,it was detected by HPLC.The mobile phase was 0.1%HCl(pH=2.0)and acetonitrile(gradient elution),and the solid phase was eighteen alkyl bonded phase.The column temperature was 30℃and the current speed was 1.0 mL/min.The separated sample was detect-ed by the diode array detector with a wave length of 210nm.The results showed that the method had a good linearity range of 1 ~ 100 mg/kg (r > 0.999).The recovery and the repeatability were good at the additive of 1,10,100 mg/kg (recovery rate>70%,RSD<10%)of premixed feed,concentrated feed and assorted feed.The method was adaptable to determine the concentration of cyproheptadine hydrochloride and clonidine hydrochloride in feed staff.