衍生化高效液相色谱法测定醋酸西曲瑞克中10种手性氨基酸的含量

目的衍生化高效液相色谱法(HPLC)测定醋酸西曲瑞克中10种手性氨基酸的含量。方法醋酸西曲瑞克水解后,采用Marfey法进行柱前衍生化反应,使用Hedera ODS-2 C18(4.6 mm×250 mm,5μm)色谱柱,流动相为0.065%甲酸溶液(A)-乙腈(B),按程序进行梯度洗脱,紫外检测器检测波长为340 nm。结果醋酸西曲瑞克水解后各氨基酸之间分离度良好,LOD和LOQ的浓度均分别为0.68μmol·L^(-1)和2.04μmol·L^(-1),组成醋酸西曲瑞克的10种手性氨基酸线性范围为10.20~24.49μmol·L^(-1)。结论本方法专属性强,准确度高,可用于同时测定醋酸西曲瑞克中10种手性氨基酸的含量。

Vaccination of Plasmid DNA Encoding Somatostatin Gene Fused with GP5 Gene of Porcine Reproductive and Respiratory Syndrome Virus Induces Anti-GP5 Antibodies and Promotes Growth Performance in Immunized Pigs

Somatostatin （SS） is a hormone that inhibits the secretion of growth hormone. Immunization against SS can promote the growth of animals. This paper described the effects of DNA immunization on the growth and antibody response in mice and pigs immunized with a plasmid DNA encoding SS fused with GP5 of porcine reproductive and respiratory syndrome virus （PRRSV）. A fragment of 180 bp encoding partial SS gene was amplified by PCR from the genomic DNA of peripheral blood mononuclear cells of pigs, and cloned as a fusion gene with PRRSV GP5 in plasmid pISGRTK3. Three times of immunization with the resulting plasmid pISG-SS/GP5 induced anti-GP5 antibodies in BALB/c mice and pigs, as demonstrated by GP5-specific ELISA and immunoblotting. Compared with pigs immunized with empty vector pISGRTK3, the growth performance of pigs immunized with pISG-SS/GP5 was increased by 11.1% on the 13th week after the last vaccination. The results indicated the plasmid DNA encoding SS and PRRSV GP5 fusion gene elicited anti-GP5 antibodies and improved the growth performance of immunized pigs.

HPLC法测定D-色氨酸甲酯盐酸盐中的左旋异构体

建立了测定D-色氨酸甲酯盐酸盐中左旋异构体的手性高效液相色谱方法。采用EnantioPak Y5(4.6 mm×250 mm, 5μm)色谱柱,以正己烷(含0.1%二乙胺)-乙醇(80:20)为流动相,检测波长280 nm。D-色氨酸甲酯盐酸盐与其左旋异构体之间分离度良好,L-色氨酸甲酯盐酸盐在0.9927~14.8905μg·mL^(-1)浓度范围内线性关系良好;D-色氨酸甲酯盐酸盐在0.9839~14.7588μg·mL^(-1)浓度范围内线性关系良好,R^(2)均为0.9998。本方法准确、简便,可以用于D-色氨酸甲酯盐酸盐中左旋异构体的质量研究。