A high-performance liquid chromatographic assay for HI-6 oxime in plasma.
A high-performance liquid chromatographic (HPLC) assay was developed to determine HI-6 (1-(2-hydroxyiminomethyl-1-pyridinio-3-(4-carbamoyl-1-py ridiniol-2-oxapropane dichloride)) concentrations in small volumes of plasma. A 100-microL plasma sample added to 900 microL of distilled water was microfiltered. Filtrate (200 microL) was injected onto an HPLC instrument containing a 100-microL sample loop, a C18 column, and an ultraviolet (UV) wavelength detector. Limit of sensitivity for HI-6 was 2.5 micrograms/mL. Extraction of efficiency (n = 12) at 10 and 100 micrograms HI-6/mL plasma was 69.4 +/- 6.6% (SD) and 81.5 +/- 2.0% (SD), respectively. Protein-plasma binding of HI-6 did not occur. HI-6 was stable when frozen at -20 degrees C for up to 10 days (0.025 less than p less than 0.05). Correlation coefficients representing standard curve linearity ranged from 0.9986 to 0.9999 (n = 6). Within-day and between-day coefficients of variation (n = 6) for unknown samples ranged from 4.4 to 8.3% and 5.8 to 17.1%, respectively. Bias of unknown samples ranged from -10.5 to 5.7%. The method's sensitivity, accuracy, and precision indicate that it can be used to accurately measure HI-6 concentrations in 100 microL of plasma.
United States Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, Maryland 21010-5425.
M P
MP
McCluskey
C R
CR
Clark
B G
BG
Talbot
M D
MD
Green
B J
BJ
Lukey
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism; D002801:Cholinesterase Reactivators / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D010091:Oximes; D011485:Protein Binding; D011726:Pyridinium Compounds / Q000097:blood*; D011817:Rabbits; D015203:Reproducibility of Results
D001798:Blood Proteins; D002801:Cholinesterase Reactivators; D010091:Oximes; D011726:Pyridinium Compounds; C022870:asoxime chloride