Rapid Analysis of Lewisite Metabolites in Urine by High-Performance Liquid Chromatography–Inductively Coupled Plasma-Mass Spectrometry

Rayman D. Stanelle¹, William J. McShane², Elena N. Dodova³, R. Steven Pappas¹, and Robert J. Kobelski^1
1Centers for Disease Control and Prevention, 4770 Buford Hwy., MS F-44, Atlanta, Georgia 30341; ²Battelle—Battelle Eastern Science and Technology Center, 1204 Technology Dr., Aberdeen, Maryland 21001; and ³Battelle—Centers for Disease Control and Prevention, 4770 Buford Hwy., MS F-50, Atlanta, Geogria 30341

A high-throughput method has been developed for determining Lewisite [dichloro(2-chlorovinyl)arsine] exposure by measuring the urine metabolite 2-chlorovinylarsonous acid (CVAA) and the oxidized metabolite 2-chlorovinylarsonic acid (CVAOA). The rapid sample preparation included a simple dilution of 400 µL of urine with 40 µL of water and 1 mL of buffer containing an internal standard and brief centrifugation prior to analysis by high-performance liquid chromatography–inductively coupled plasma-mass spectrometry (ICP-MS). CVAOA and CVAA were eluted isocratically with retention factors of approximately 3.0 and 4.2, respectively, from a reversed-phase polar embedded column with a cycle time of 5 min per sample. The dynamic reaction cell, typically used to remove polyatomic isobaric interferences, was not required for ICP-MS analysis because of the resolution of chloride from arsenical peaks of interest. This method was used to detect CVAA and CVAOA in the urine of a Lewisite-administered rat up to 24 h after exposure. The method demonstrated linearity over at least three orders of magnitude and had a method detection limit of 1.3 µg/L as CVAA (1.4 µg/L CVAOA). The relative standard deviations for quality control samples ranged from 3 to 6%. The method was sensitive and selective with no false-positives in 100 different urine samples collected from individuals with no known exposure to Lewisite. Ninety-six samples could be analyzed in an 8-h day.

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