Quantitative Clinical Diagnostic Analysis of Acetone in Human Blood by HPLC: A Metabolomic Search for Acetone as Indicator

Abstract

Using high-performance liquid chromatography (HPLC) and 2,4-dinitrophenylhydrazine (2,4-DNPH) as a derivatizing reagent, an analytical method was developed for the quantitative determination of acetone in human blood. The determination was carried out at 365 nm using an ultraviolet-visible (UV-Vis) diode array detector (DAD). For acetone as its 2,4-dinitrophenylhydrazone derivative, a good separation was achieved with a ThermoAcclaim C18 column (15 cm × 4.6 mm × 3 μm) at retention time (t R) 12.10 min and flowrate of 1 mL min−1 using a (methanol/acetonitrile) water elution gradient. The methodology is simple, rapid, sensitive, and of low cost, exhibits good reproducibility, and allows the analysis of acetone in biological fluids. A calibration curve was obtained for acetone using its standard solutions in acetonitrile. Quantitative analysis of acetone in human blood was successfully carried out using this calibration graph. The applied method was validated in parameters of linearity, limit of detection and quantification, accuracy, and precision. We also present acetone as a useful tool for the HPLC-based metabolomic investigation of endogenous metabolism and quantitative clinical diagnostic analysis.

Derivatization procedures and their analytical performances for HPLC determination in bioanalysis

“…formaldehyde, can be determined as a trace in various matrices, the resulting hydrazone having an absorbing maximum (λ max ) at 360 nm (Soman, Qiu, & Li, 2008). Other examples of determining simple carbonylic compounds in biomatrices by means of this derivatization reaction are malondialdehyde in human urine (Kim, Jung, & Kho, 2017), acetaldehyde in hepatoma cell culture medium, blood and plasma (Guan, Rubin, & Anni, 2012) and acetone in blood samples (Kalkan, Sahiner, Cakir, Alpaslan, & Yilmaz, 2016;Fujii et al, 2014).…”

Surface enhanced substrate simulation optimized coupled PLS model for quantitative detection of acetone in transformer oil

“…The content of acetone, as one of the aging characteristics of oil-paper insulation, can be used as a parameter to assess the aging condition of the insulation system [7]. Means of detection of dissolved acetone content in transformer oil mainly include high performance liquid chromatography (HPLC) [8], electrochemical method [9] and spectrophotometric method [10]. The most commonly used method is HPLC, but the complexity of the operation and the frequent replacement of columns determine that this method can only be completed in the laboratory and is not suitable for long-term monitoring [11].…”

Accidental Acetone Ingestion in Liver Transplant Patient With Alcohol Relapse: A Case Report

“…In healthy individuals, these ketone bodies can be naturally produced in the liver by the breakdown of fatty acids. This mainly occurs in fasting states when glucose stores and insulin levels are low, and glycogen breakdown cannot produce enough glucose [ 2 , 3 ]. The body continues to have normal glucagon and epinephrine levels, causing fatty acids to be released from fat cells.…”