Determination of 2,3-dimercaptosuccinic acid in mice blood and tissues by HPLC with fluorescence detection.
2,3-Dimercaptosuccinic acid (DMSA) is an orally effective chelating agent for the treatment of heavy metal poisoning. The increasing therapeutic use of DMSA has stimulated the need for sensitive and selective methods for its determination in biological samples, as well as study on pharmacokinetics and tissue distribution. According to the previously reported method, an improved method was established for the determination of DMSA in mice blood and tissues, in which oxidized DMSA was reduced by the disulfide-reducing agent, dithiothreitol (DTT), and DMSA was converted to a highly fluorescent and stable derivative by reaction with monobromobimane (mBBr) in alkaline solution. Acetonitrile was used for deproteinization and dichloromethane was used for condensation and purification, which significantly shortened the amount of time used to process the sample. Meanwhile isocratical elution was performed and excellent separation of the DMSA derivative was obtained, this enabled a run finish within 20 min. The limits of quantitation were 0.025 microg/ml in brain and 0.1 microg/ml in blood, lung, heart, intestine, liver, spleen and kidney, respectively. The calibration curves were linear in all samples (r(2)>0.992) with a range of 0.025-1.6 microg/ml for brain homogenate and 0.1-6.4 microg/ml for blood and homogenates of lung, heart, intestine, liver, spleen and kidney, respectively. Therefore, the method is simple, rapid and sensitive, and it could be applicable to the studies in an animal model to evaluate the distribution of DMSA in blood and tissues.
Ju
Xue Hai
XH
Centre for Research in Ecotoxicology and Environmental Remediation, Agro-Environmental Protection Institute, Ministry of Agriculture of the People's Republic of China, Tianjin 300191, China.
Shi
Ying
Y
Liu
Na
N
Guo
Rui Chen
RC
Wang
Ben Jie
BJ
Cui
Xi
X
D000818:Animals; D001774:Blood Chemical Analysis; D002614:Chelating Agents / Q000032:analysis* / Q000627:therapeutic use; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D007855:Lead Poisoning / Q000188:drug therapy; D008297:Male; D051379:Mice; D013050:Spectrometry, Fluorescence / Q000379:methods*; D004113:Succimer / Q000032:analysis* / Q000627:therapeutic use
D002614:Chelating Agents; D004113:Succimer