猪肝脏和肌肉组织中β-兴奋剂克伦特罗的高效液相色谱电喷雾质谱分析研究

Analysis of Clenbuterol in Pig Liver and Muscle by High Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometry

建立了猪肉和肝组织中克伦特罗的高效液相色谱电喷雾质谱 (HPLC ESI/MS)分析方法。在最佳色谱条件下克伦特罗的保留时间为 1 6min ,样品空白无干扰。定量分析的线性范围为 2～ 1 0 0mg/L ;最低检出限为 0 .75mg/L ;方法回收率为 95 %～ 98% ;RSD小于 2 .0 %。利用ESI/MS/MS对 β 兴奋剂克伦特罗进行了质谱解析 ,选择特征离子峰m/z 2 77、2 5 9和 2 0 3作为准确定性的依据。对实际生物样品猪瘦肉和肝进行检测结果表明 ,当样品中克伦特罗残留含量较高时 ,可以利用紫外检测数据对其进行准确定量。当残留含量低于最低检出限时 ,可以根据HPLC/ESI/MS结果中有无特征峰出现 。

For beta-agonist clenbuterol, its typical ion peaks at m/z 277, 259 and 203 in electrospray ionization mass spectrometry (ESI/MS) were found and elucidated. The high performance liquid chromatography (HPLC)-ESI/MS method for analyzing the residue of clenbuterol in pig liver and muscle was developed. With the optimal mobile phase of MeOH-10 mmol/L NH4Ac (45:55, V/V) at 0.9 mL/min and the separation column of 250 mm x 4.6 mm i.d. ODS, clenbuterol and the sample blanks can be completely separated with the retention time of 16 min. With 243 nm detection, the linear range was between 2 and 100 mg/L with the detection limit of 0.75 mg/L, the recovery of 95%-98% and the RSD less than 2.0%. Using the proposed HPLC-ESI/MS, clenbuterol can be accurately identified with both retention time and the typical ion peaks. The experimental results of the real biologic samples of pig liver and muscle show the residue content of clenbuterol can be accurately determined by ultraviolet detection when the residue of clenbuterol. was higher. For the residue of clenbuterol lower than its detection limit, the quantitative result was not able to obtain, however the accurately qualitative result can be made according to its typical ion peaks at corresponding elution time.