In order to establish an HPLC-MS/MS method for detecting progesterone in raw milk,raw milk was extracted with methanol,and then determined by HPLC-MS/MS after the purification with a C18 solid phase extraction column....In order to establish an HPLC-MS/MS method for detecting progesterone in raw milk,raw milk was extracted with methanol,and then determined by HPLC-MS/MS after the purification with a C18 solid phase extraction column. Qualitative determination was performed according to retention time and selective ion abundance ratio,and quantification was performed by external standard method. The results showed that progesterone had a good linear relation in the range of0. 1-5. 0 ng/ml,with a correlation coefficient of 0. 999 9. The detection limit of the established method was 0. 02 μg/kg,and the quantification limit was0. 04 μg/kg. In the standard addition range of 0. 1-5. 0 ng/ml,the recovery ranged from 79. 6% to 105. 1%,with relative standard deviation≤8. 9%. Compared with the industry standard,the detection limit and the quantification limit of the established method are significantly reduced,and the sensitivity of the method is enhanced,so this method is suitable for the detection of progesterone residue in fresh milk in pastures.展开更多
为了建立快速、准确检测生鲜乳中布鲁氏杆菌的方法,试验采用细菌基因组提取试剂盒提取生鲜乳中的布鲁氏杆菌基因组DNA,针对布鲁氏杆菌保守抗原Omp31、Omp25设计引物,并设计细菌16S r DNA基因序列作为内参引物,通过引物浓度和退火温度的...为了建立快速、准确检测生鲜乳中布鲁氏杆菌的方法,试验采用细菌基因组提取试剂盒提取生鲜乳中的布鲁氏杆菌基因组DNA,针对布鲁氏杆菌保守抗原Omp31、Omp25设计引物,并设计细菌16S r DNA基因序列作为内参引物,通过引物浓度和退火温度的调整优化,建立了生鲜乳中布鲁氏杆菌的多重PCR检测方法。结果表明：建立的多重PCR检测结果与预期结果一致,可以扩增出布鲁氏杆菌Omp31（472 bp）、Omp25（140 bp）特异性片段以及细菌16S r DNA的通用片段。而对于其他致病菌只能扩增出1 500 bp的细菌16S r DNA通用片段,具有较强的特异性。该多重PCR检测方法灵敏度可达1×10^5cfu/m L,较单一PCR检测方法灵敏度有所下降。说明本试验建立的检测方法可完成生鲜乳中布鲁氏杆菌的快速检测。展开更多
基金Supported by Science and Technology Open Cooperation Project of Henan Province(162106000017)Science and Technology People-benefiting Plan Project of Henan Province(152207110004)+1 种基金Major Science and Technology Program of Henan Province(16110051020)Puyang Science and Technology Plan Project(160215)
文摘In order to establish an HPLC-MS/MS method for detecting progesterone in raw milk,raw milk was extracted with methanol,and then determined by HPLC-MS/MS after the purification with a C18 solid phase extraction column. Qualitative determination was performed according to retention time and selective ion abundance ratio,and quantification was performed by external standard method. The results showed that progesterone had a good linear relation in the range of0. 1-5. 0 ng/ml,with a correlation coefficient of 0. 999 9. The detection limit of the established method was 0. 02 μg/kg,and the quantification limit was0. 04 μg/kg. In the standard addition range of 0. 1-5. 0 ng/ml,the recovery ranged from 79. 6% to 105. 1%,with relative standard deviation≤8. 9%. Compared with the industry standard,the detection limit and the quantification limit of the established method are significantly reduced,and the sensitivity of the method is enhanced,so this method is suitable for the detection of progesterone residue in fresh milk in pastures.
文摘为了建立快速、准确检测生鲜乳中布鲁氏杆菌的方法,试验采用细菌基因组提取试剂盒提取生鲜乳中的布鲁氏杆菌基因组DNA,针对布鲁氏杆菌保守抗原Omp31、Omp25设计引物,并设计细菌16S r DNA基因序列作为内参引物,通过引物浓度和退火温度的调整优化,建立了生鲜乳中布鲁氏杆菌的多重PCR检测方法。结果表明：建立的多重PCR检测结果与预期结果一致,可以扩增出布鲁氏杆菌Omp31（472 bp）、Omp25（140 bp）特异性片段以及细菌16S r DNA的通用片段。而对于其他致病菌只能扩增出1 500 bp的细菌16S r DNA通用片段,具有较强的特异性。该多重PCR检测方法灵敏度可达1×10^5cfu/m L,较单一PCR检测方法灵敏度有所下降。说明本试验建立的检测方法可完成生鲜乳中布鲁氏杆菌的快速检测。
