Chromatographic determination of low-molecular mass unsaturated aliphatic aldehydes with peroxyoxalate chemiluminescence detection after fluorescence labeling with 4-(N,N-dimethylaminosulfonyl)-7-hydrazino-2,1,3-benzoxadiazole

Abstract

A highly sensitive, selective and reproducible chromatographic method is described for determination of low-molecular mass unsaturated aliphatic aldehydes in human serum. The method combines fluorescent labeling using 4-(N,N-Dimethylaminosulfonyl)-7-hydrazino-2,1,3-benzoxadiazole with peroxyoxalate chemiluminescence. The derivatives were separated on a reversed-phase column C8 isocratically using a mixture of acetonitrile and 90mM imidazole-HNO3 buffer (pH 6.4, 1:1, % v/v). The calibration ranges were: 20-420nM for methylglyoxal, 16-320nM for acrolein, 15-360nM for crotonaldehyde and 20-320nM for trans-2-hexenal. The detection limits were ranged from 4.4 to 6.5nM (88-130fmol/injection), the recovery results were within the range of 87.4-103.8% and the intra and inter-day precision results were lower than 5.5%. The proposed validated method has been successfully applied to healthy, diabetic and rheumatic arthritis patients' sera with simple pretreatment method. In conclusion, this new method is suitable for routine analysis of large numbers of clinical samples for assessment of the oxidative stress state in patients.

Unsaturated Aldehyde Acrolein Promotes Retinal Glial Cell Migration

“…18 With respect to the involvement of acrolein in diabetes, acrolein was reportedly increased in the serum of patients with diabetes. 19 It was also shown that the urinary level of acrolein was higher in type 2 diabetes patients than in individuals without diabetes. 18 By contrast, little is known about the role of acrolein in ocular diseases, including DR.…”

Determination of acrolein in serum by high‐performance liquid chromatography with fluorescence detection after pre‐column fluorogenic derivatization using 1,2‐diamino‐4,5‐dimethoxybenzene

“…The regression equation (mean ± standard error, n = 3) was y = (0.895 ± 0.038) x + 0.066 ± 0.030, where y is the peak area ratio and x is the concentration of acrolein (μ M ). The sensitivity of the proposed method was 100 times greater compared with time‐resolved fluorescence assay (Togashi et al ., ), 15–20 times graeter compared with HPLC with electrochemical detection (Casella and Contursi, , ) and 4–10 times greater compared with HPLC with fluorescence detection (Ali et al ., ; Bohnenstengel et al ., ; Paci et al ., ). Additionally, the sensitivity was almost comparable to that of GC‐MS (Osório and de Lourdes Cardeal, ).…”

“…To date, several methods have been developed for the determination of acrolein using time-resolved fluorescence assay (Togashi et al, 2010), gas chromatography with mass spectrometry (GC-MS; Osório and de Lourdes Cardeal, 2011), high-performance liquid chromatography (HPLC) with fluorescence detection (Ali et al, 2014;Bohnenstengel et al, 1997;Paci et al, 2000), electrochemical detection Contursi, 2004, 2014) and liquid chromatography with tandem mass spectrometry (LC-MS/MS; Andreoli et al, 2003). In these methods, derivatization reactions were frequently adopted for the determination of acrolein in order to improve detectability and stability (Shibamoto, 2006).…”

“…In these methods, derivatization reactions were frequently adopted for the determination of acrolein in order to improve detectability and stability (Shibamoto, 2006). Several HPLC with fluor escence detection methods used fluorescent hydrazine reagents such as luminarin-3 (Paci et al, 2000) and 4-(N, N-dimethylaminosulfonyl)-7-hydrazino-2,1,3-benzoxadiazole (Ali et al, 2014). However, these derivatization reagents and by-products have strong fluorescence themselves and can interfere the detection of acrolein derivative.…”

All Eyes on Visible‐Light Peroxyoxalate Chemiluminescence Read‐Out Systems

“…Recently, a new chromatographic method for detecting low molecular mass aldehydes in human serum through a combination of fluorescence labelling and PO‐CL was published by Ali et al . Cellular oxidative stress can lead to lipid peroxidation of unsaturated fatty acids in cell membranes, which produces low molecular mass aldehydes which are cytotoxic and react with DNA .…”