An Improved Method for Acetaldehyde Determination in Blood by High-Performance Liquid Chromatography and Solid-Phase Extraction

Abstract

This study reports on an improved method for acetaldehyde (ACH) determination in blood by high-performance liquid chromatography (HPLC). In the case of HPLC analysis, ACH is generally converted to derivatives for ultraviolet detection (for example 2,4-dinitrophenylhydrazine [DNPH] derivative). Nevertheless, elevation of the background during protein precipitation, hydrazone synthesis, or both frequently results in a serious loss of accuracy and precision of the analysis. The method in this study is developed to minimize the increase in nonspecific ACH-DNPH with a view to optimize mainly the synthetic condition of ACH-DNPH. The background is decreased dramatically by gentle deproteination, optimization of the DNPH amount and reaction pH, and reversed-phase solid extraction for the elimination of excess DNPH reagent. The standard curves show good linearity between 0 and 100 microM and minimal background is observed, indicating that the method is useful for monitoring the ACH concentration in blood.

An Optimized Method for the Measurement of Acetaldehyde by High‐Performance Liquid Chromatography

“…While most DNPH was removed from the cartridge with 1.5 ml of 70% methanol, AcH-DNP co-eluted in this washing step (data not shown),. This carryover effect resulted in either low efficiency of extraction or contamination with DNPH, as observed by other investigators (Kozutsumi et al, 2002; Ma and Klemm, 1997). Alternatively,, both AcH-DNP and DNPH were extracted by liquid-liquid extraction into an organic solvent (acetonitrile or isooctane) with negligible amounts remaining in the aqueous phase (data not shown).…”

“…The acid-catalyzed reaction had an optimum pH 4.0, since the rate-limiting step is the protonation of carbonyl oxygen, followed by the loss of water to form hydrazone. An optimum pH of 4.0 for the derivatization reaction was also found by Kozutsumi et al (2002), though stronger acidic conditions have been also reported (Bicking et al, 1988; Cotsaris and Nicholson, 1993). The reaction at the optimum pH was completed within 40 min at ambient temperature or with longer incubation at 4°C (see Fig.…”

“…In addition, RP-HPLC methodology has been developed and employed for evaluation of acetaldehyde content in biological fluids of experimental animals and human subjects (e.g., Akubue et al, 1994; Arolfo et al, 2009; Garver et al, 2000; 2001; Hamby-Mason et al, 1997; Nagy et al, 2004; Rouach et al, 2005; Di Padova et al, 1986; Kozutsumi et al, 2002; Lucas et al, 1986; Lynch et al, 1983; Zhang et al, 2006). The RP-HPLC method is based on selective labeling of acetaldehyde with DNPH, which produces a stable product, with visible/UV absorbance.…”

Determination of Acetaldehyde Content in Candesartan Cilexetil by HPLC

“…The most widely used derivatization agent is 2,4-dinitrophenylhydrazine (DNPH). In acidic media, DNPH reacted with carbonyl group of ACD to form Schiff base derivatization product DNPH-ACD [11,12] . The derivatization reaction leads to an orange colored mixture of DNPH-ACD as shown in fig.…”

Imaging acetaldehyde formation during ethanol metabolism in living cells using a hydrazinyl naphthalimide fluorescent probe

“…This is mainly due to the furtive nature of acetaldehyde, necessitating careful sample processing and detection methods such as spectrophotometry, nuclear magnetic resonance imaging (NMR), or gas chromatography mass-spectrometry (GC-MS), which typically result in processing and destruction of the sample. 14 The use of carbonyl-responsive fluorescent probes 15–21 could potentially circumvent these problems by allowing for the real-time, unobtrusive visualization of acetaldehyde as it is produced in living cells.…”