Determination of hippuric acid and methylhippuric acid in urine by high performance liquid chromatography after extracted with acetonitrile
10.3760/cma.j.issn.1001-9391.2016.04.020
- VernacularTitle:尿中马尿酸、甲基马尿酸测定的乙腈萃取-高效液相色谱法
- Author:
Jingjing QIU
1
;
Xiaolin RUAN
;
Banghua WU
;
Weifeng RONG
;
Ruibo MENG
Author Information
1. 510300广州,广东省职业病防治院,广东省职业病防治重点实验室
- Keywords:
Chromatography,high pressure liquid;
Hippuric acid;
Methylhippuric acid;
Urine
- From:
Chinese Journal of Industrial Hygiene and Occupational Diseases
2016;34(4):304-307
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish pretreatment conditions of hippuric acid (HA) and methyl-hippuric acid (MHA) in urine and HPLC conditions.Methods HA and MHA in urine were extracted with acetonitrile under acid condition and determinated by HPLC-DAD.The operating conditions by HPLC were C18 column (150 mm× 4.6 mm,5 μm),methanol-0.2％ acetic acid (contained 6.5 mmol/L potassium dihydrogen phosphate) (25:75,V/V) as mobile phase,1 ml/min as flow rate and wavelength was at 254 nm.Results The standard curves for HA,2-MHA and 3-MHA (4-MHA) showed good linearity between 9.91～2 974.20 μg/ml (r=0.999 98),1.91～ 573.60 μg/ml (r=0.999 84) and 2.00～598.65 μg/ml (r=0.999 85),respectively.The mean recoveries were 96.38％～98.01％,83.17％～94.05％,103.22％～104.45％,respectively.The within-run precision were 0.50％～ 1.20％,0.51％～1.59％,0.49％～0.95％,respectively,and the between-run precision were 1.70％～3.20％,1.30％～ 2.67％,0.86％～2.74％,respectively.The detection limit of HA,2-MHA and 3-MHA (4-MHA) were 0.18 μg/ml,0.46 μg/ml and 0.12 μg/ml,and the low determination concentrations of the method were 0.36 μg/ml,0.92 μg/ml and 0.24 μg/ml (1 ml urine).The urine can be kept 15 days at 4 ℃ refrigerator without significantly loss.Conclusion This method with simply pretreatment conforms to the relevant requirements of guide for establishing occupational health standards-part 5:determination methods of chemicals in biological materials.It can be used to detect HA and MHA in urine for occupational population exposure to toluene and xylene.
