摘要
本研究以多拉菌素为内标,建立了犬血浆中塞拉菌素的高效液相色谱-荧光检测方法。用乙腈作为血浆蛋白沉淀剂,用SampliQ C18型SPE固相萃取柱进行净化,用1-甲基咪唑和三氟乙酸酐衍生化处理。色谱条件为ODS-1色谱柱(250mm×4.6mm),流动相为甲醇∶乙腈∶1%1-庚烷磺酸∶0.4%乙酸=40.0∶57.6∶0.9∶1.5(v/v/v/v),流速1mL.min-1,进样量20μL,激发波长355nm,发射波长465nm,柱温30℃。方法的检测限为0.25ng.mL-1,线性范围0.5～50.0ng.mL-1,变异系数为1.47%～3.31%,方法的样品平均回收率为94.0%。结果表明,所建方法准确、灵敏度高和重复性好,可用于犬体内塞拉菌素血药含量的测定。
The aim of this study was to establish a method for detection of selamectin in dog plasma by using doramectin as an internal standard.Samples were measured by high-performance liquid chromatography with fluorescence detection after automated solid phase extraction.After proteins separated with acetonitrile,a solid phase extraction was performed to plasma samples.The eluate was evaporated to dryness under stream of nitrogen gas,and followed by chemical derivation using N-methylimidazole and trifluoroacetic anhydride.An aliquot sample（20 μL） was pumped into the chromatographic column connected with fluoresce detector that wavelength was set at 355 nm for excitation and 465 nm for emission at 30℃.The mobile phase composed of methanol-acetonitrile-sodium 1-heptanesulfonate（1%）-acetic acid（0.4%）（40.0：57.6：0.9：1.5,v/v/v/v） were used,with flow rate 1 mL·min-1.The method gave linear calibration graphs（R2=0.999 3） over the concentration range studied（0.5-50.0 ng·mL-1） with detection limit,variation coefficient and average recovery of samples were 0.25 ng·mL-1,1.47%-3.31% and 94.00% respectively.It indicated that the method we established was reliable,sensitive and with good repeatability and suitable for detection of selamectin in dog plasma.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2011年第9期1346-1350,共5页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家肉牛牦牛产业技术体系(CARS-38)
"十一五"国家科技支撑计划项目(2006BAD31B02
2008BADB4B05)
