饲料研究2024，Vol.47Issue(14)：128-133,6.DOI:10.13557/j.cnki.issn1002-2813.2024.14.022
猪小肠内容物中苯甲酸和马尿酸高效液相色谱检测方法的建立
Establishment of HPLC method for determination of benzoic acid and hippuric acid in pig small intestine contents
摘要
Abstract
In order to establish a high performance liquid chromatography(HPLC)method for the determination of benzoic acid and its metabolite hippuric acid in the contents of pig small intestine,ethyl acetate was used as extraction agent,potassium ferrocyanide zinc acetate was used as protein precipitator to extract benzoic acid from the sample,and the mobile phase was methanol:0.02％phosphoric acid water.Hippuric acid was extracted from samples with water,potassium ferrocyanide and zinc acetate,and the mobile phase was methanol:20 mmol/L ammonium acetate solution.The detection wavelength was 230 nm.The results showed that the linear relationship between the concentration of benzoic acid in the small intestine contents of pigs was good(R2=0.999 4).The limit of detection was 0.05 μg/g,and the limit of quantification was 0.10 μg/g.At 0.10,0.50,1.00,5.00,10.00 μg/g,the recovery was(101.62±4.49)％,and the coefficient of variation was lower than 4.80％and 9.96％,respectively.The addition concentration of hippuric acid in the contents of pig small intestine had a good linear relationship between 0.30 and 10.00 μg/g(R2=1.000 0).The limits of detection and quantification were 0.1 μg/g and 0.3 μg/g,respectively.The recovery of hippuric acid was(90.3±4.00)％at 0.50,1.00,5.00,20.00 μg/g,and the coefficient of variation within and between batches was lower than 5.93％and 5.26％,respectively.The results show that the pretreatment method of benzoic acid and hippuric acid is simple and time-saving,without solid phase extraction and use of toxic and harmful solvents,and saves cost.It can provide a reference for the detection of benzoic acid and its metabolite hippuric acid in the contents of pig small intestine.关键词
猪/苯甲酸/马尿酸/HPLC/小肠内容物Key words
pig/benzoic acid/hippuric acid/HPLC/small intestinal contents分类
农业科技引用本文复制引用
张寒,蒋永嘉,刘为,王志强,肖霞..猪小肠内容物中苯甲酸和马尿酸高效液相色谱检测方法的建立[J].饲料研究,2024,47(14):128-133,6.基金项目
江苏省人兽共患病学重点实验室资助项目(项目编号:R2303) （项目编号:R2303）
