Development and Validation of an Analytical Method for Volatiles with Endogenous Production in Putrefaction and Submersion Situations

A group of 16 volatile substances (ethyl acetate, 2-propanol, 1-propanol, methanol, acetone, ethanol, acetaldehyde, diethyl ether, methyl ethyl ketone, 1-butanol, 2-butanol, t-butanol, isobutanol, 2-methyl-1-butanol, 3-methyl-1-butanol and 1-pentanol) were qualitatively and quantitatively analyzed through a method developed for volatiles with endogenous production in putrefaction and submersion situations. The method was validated for blood, urine and vitreous humor, using a gas chromatograph (Varian 450-GC) with a flame ionization detector coupled to a headspace injector (HS-GC-FID). The vials were prepared by diluting 100 µL of the sample of interest in 1 mL of internal standard (acetonitrile 100 mg/L), using two capillary columns (VF-624ms and VF-5ms) with different polarities to ensure that all test compounds would be properly identified and undoubtedly distinguished from the rest. All volatiles were studied in a range of 50 to 2,000 mg/L in terms of selectivity/specificity, detection and quantification limits, linearity and calibration model, precision, accuracy, bias, robustness and stability according to the Scientific Working Group for Forensic Toxicology. Detection and quantification limits were between 1 to 8 mg/L and 4 to 24 mg/L, respectively, with coefficient of variation values under 10% in bias studies and in intermediate precision studies for most substances. The developed method was applied to real cases to test the method.