Quantitative anaylsis of acetone in blood via headspace gas chromatography/flame ionization detection

Abstract

The purpose of this research project was to develop and validate a method for the quantitation of acetone in blood using Headspace Gas Chromatography with Flame Ionization Detection. The following items were included in the development: preparation of standards and positive controls, testing the necessity of using salts, establishment of a suitable internal standard concentration, determination of an appropriate sample size, limit of detection calculation, establishing the linear dynamic range, testing for carryover effects, method validation including case samples, and checking the stability of samples containing acetone. The blood-acetone quantitation method was successfully developed and validated. Standards and controls were accurate and precise and yielded linear results. The addition of salt to sample vials was deemed unnecessary. An internal standard concentration of 0.05% (v/v) n-propanol in water and a sample size of 0.25 mL were found to be suitable. The limit of detection was calculated to be 0.333 mg/dL while the limit of quantitation was calculated to be 0.816 mg/dL. However, quantitations were inaccurate below 1.0 mg/dL and blood-acetone concentrations above 250.0 mg/dL are not generally encountered. Therefore, an administrative linear dynamic range was set from 1.0 to 250.0 mg/dL. No carryover between samples occurred. Interday and interindividual variations were 6.93 and 0.77%, respectively. When checking the stability of samples containing acetone, time accounted for 97.7% variation while temperature only accounted for 0.22% variation.