Simple and sensitive determination of malondialdehyde in human urine and saliva using UHPLC–MS/MS after derivatization with 3,4‐diaminobenzophenone

Abstract

Malondialdehyde has been used as a biomarker for lipid peroxidation in biological samples. An ultra-high performance liquid chromatography with tandem mass spectrometry method was developed to determine the levels of malondialdehyde in human urine and saliva samples. To select the optimum derivatization reagent from four diamino compounds, the reactivity and sensitivity of their derivatives were compared, and 3,4-diaminobenzophenone was selected. The optimum reaction conditions for malondialdehyde with 3,4-diaminobenzophenone were as follows: a reagent dosage of 50 mg/L, pH of 4, and reaction for 30 min at 50°C. The formed derivative product was analyzed using ultra-high performance liquid chromatography with tandem mass spectrometry without additional extraction or concentration steps. In the optimal conditions, the method was used to determine malondialdehyde concentration in human urine and saliva samples. The limits of quantification for malondialdehyde in biological samples were over a concentration range of 0.1-0.3 μg/L. Additionally, the calibration curve showed a linearity greater than r = 0.997. The method was used to analyze 14 human urine and saliva samples from healthy volunteers. Malondialdehyde was detected in the concentration range of 1.7-33.6 μg/g creatinine in all human urine samples and 0.1-1.3 μg/L in all human saliva samples.

Validated analytical method to determine new salivary lipid peroxidation compounds as potential neurodegenerative biomarkers

“…15-F 2t -IsoP) [10]. In general, these methods are based on colorimetric assays and immunoassays [4,[17][18][19], but also liquid chromatography (LC) coupled to mass spectrometry (MS) [10,20], or fluorescence detection methods [18] are used. Nevertheless, they have not been validated, so there is a lack of reliable analytical methods.…”

Bioanalytical and Mass Spectrometric Methods for Aldehyde Profiling in Biological Fluids

“…Liquid chromatography–mass spectrometry-based approaches have been used extensively to quantify derivatized carbonyl compounds, and recently for screening of unknown carbonyl compounds. Aldehyde derivatizations using 2,4-DNPH [143,197,198,199,200,201,202], dansylhydrazine (DnsHz) [203,204], N -(1-chloroalkyl)pyridinium [205], o -phenyldiamine [206], D-cysteine [207], 9,10-phenanthrenequinone (PQ) [208], 3-nitrophenylhydrazine [209], and 3,4-diaminobenzophenone [210] have been used to provide chromatographic retention and separation, efficient MS ionization, and MS/MS detectability. Typically, LC-MS analysis has been performed using selected reaction monitoring (SRM) with either atmospheric pressure chemical ionization (APCI), atmospheric pressure photoionization (APPI) or electrospray ionization (ESI).…”

Nuevos biomarcadores usados en el diagnóstico del daño renal en condiciones de estrés oxidativo

“…Esta técnica también se puede realizar utilizando diaminonaftaleno, hidralazina, ácido tiobarbitúrico, para la determinación de malondialdehído. 42 Proteínas con actividad antioxidante: PON-1, GST y NQO1…”