摘要
目的改进高效液相色谱法测定尿液中苯巯基尿酸的方法。方法尿样经过离心后,取上清液,加入水涡旋15 s,硫酸溶液(50%)涡旋30 s并需在10 min内加入氢氧化钾(7.8 mol/L)溶液涡旋30 s。以氯仿和异丙醇(V/V=5∶1)混合液进行液-液提取,提取液经氮气吹干后,甲醇定容注入液相色谱仪,经EC-C18反相色谱柱(150 mm×4.6 mm,4μm)分离,采用紫外检测器(VWD)检测,以保留时间定性,峰面积定量。结果本法苯巯基尿酸在200.0μg/L^1 000.0μg/L浓度内呈线性,相关系数为0.999 4,以3倍噪音值计算方法检出限为2.0μg/L。在200.0μg/L^1 000.0μg/L内添加低、中、高3个浓度水平,样品加标回收率为95.67%~97.74%;批内精密度为0.41%~1.56%(n=6),批间精密度为0.44%~2.52%(n=6)。结论本法简单实用,分析时间短,灵敏度高,定性定量准确,可以满足尿样中苯巯基尿酸的检测要求。
Objective To improve the high performance liquid chromatography method in the determination of S-phenylmercapturic acid in urine. Methods Urine sample was centrifugated,and the supernatant was vortexed for 15 seconds for diluting with water. Then the theulfuric acid and potassium hydroxide solution（ 7. 8 mol/L） were added one by one in 10 minutes and vortexed for 30 s. S-phenylmercapturic acid can be extracted from above urine solution with chloroform and isopropyl alcohol mixture（ V/V = 5∶1）,and redissolved in methanol after being dried under nitrogen blowing. Then the S-phenylmercapturic acid can be separated by EC-C18 column（ 150 mm × 4. 6 mm,4 μm） and detected by VWD. The retention time was used for qualitative analysis and the peak area was for quantification. Results In this method,good linearity was obtained when the concentration of phenylmercapturic acid was within 200. 0 μg/L-1 000. 0 μg/L,with a correlation coefficient（ r） of 0. 999 4.The detection limit was 2. 0 μg/L using triple noise method. Low,medium and high concentrations of S-phenylmercapturic acid were added in the range of 200. 0 μg/L-1 000. 0 μg/L. The spiked recoveries ranged from 95. 67% to 97. 74%. The intra-assay precision was within 0. 41%-1. 56%（ n = 6）,and the inter-assay precision was within 0. 44%-2. 52%（ n = 6）.Conclusion This method was more simple,practical,sensitive and accurate. It can meet the request for the determination of trichloroacetic acid in urine.
出处
《中国卫生检验杂志》
CAS
2018年第3期272-275,共4页
Chinese Journal of Health Laboratory Technology
基金
浙江省医药卫生科技计划项目(2016KYB241)
杭州市卫生科技计划重点项目(2015Z10)
关键词
高效液相色谱
尿
苯巯基尿酸
High performance liquid chromatography
Urine
S -phenylmercapturic acid
