Quantitation of Acrolein-Derived (3-Hydroxypropyl)mercapturic Acid in Human Urine by Liquid Chromatography−Atmospheric Pressure Chemical Ionization Tandem Mass Spectrometry:  Effects of Cigarette Smoking

Abstract

Recently published data suggest that acrolein (1), a toxic but weakly carcinogenic constituent of cigarette smoke, may be involved as a causative factor for the mutations frequently observed in the p53 tumor suppressor gene in lung cancer in smokers. Biomarkers are needed to further assess the possible relationship between acrolein uptake and cancer. In this study, we analyzed 3-hydroxypropylmercapturic acid (3-HPMA, 2) in human urine. 3-HPMA is a major metabolite of acrolein in laboratory animals. The method employs [ 13 C 3 ]3-HPMA as internal standard, with analysis and quantitation by LC-APCI-MS/MS-SRM. Clean, readily quantifiable chromatograms were obtained. The method was accurate and precise and required only 0.1 mL of urine. Median levels of 3-HPMA were significantly higher (2900 pmol/mg creatinine, N = 35) in smokers than in non-smokers (683 pmol/mg creatinine, N = 21) (P = 0.0002). The effect of smoking was further assessed by determining levels of 3-HPMA before and after a 4 week smoking cessation period. There was a significant 78% decrease in median levels of urinary 3-HPMA after cessation (P < 0.0001). The relationship between levels of urinary 3-HPMA and those of acrolein-derived 1,N 2 -propanodeoxyguanosine (PdG) adducts in lung was investigated in 14 smokers. There was a significant inverse relationship between urinary 3-HPMA and α-hydroxy-PdG (3) but not γ-hydroxyPdG (4) or total adduct levels. The results of this study clearly demonstrate that acrolein uptake in smokers is significantly higher than in non-smokers, and underline the need for further investigation of the possible relationship of acrolein uptake to lung cancer.

Acrolein: Sources, metabolism, and biomolecular interactions relevant to human health and disease

“…This is readily recognized by the observation that the main urinary metabolite of acrolein, 3-hydroxypropyl mercapturic acid, is found at about twice the level in urine of smokers compared to nonsmokers [11]. In the same study, the levels of the acrolein metabolite showed a median decrease of 78% after 4 wk of smoking abstinence (P < 0.0001).…”

“…DHN-MA was found in human urine at concentrations of about 2.7 μg/L (8.4 nM) [34]. The analogous acrolein metabolite (S-(3-hydroxypropyl)mercapturic acid, HPMA) was reported to be present in the urine of a nonsmoker at a concentration of 422 μg/L (1.9 μM) [11]. Roethig et al [35] determined the excretion of HPMA for nonsmoking subjects to be about 250 μg/24 h, which corresponds to about 0.7 μM if the urine volume was 1.6 L/day.…”

“…The main pathway for elimination of acrolein is conjugation with glutathione (GSH) in the liver, followed by enzymatic cleavage of the γ-glutamic acid and glycine residues, respectively, in the liver and in the kidney [58,59], and N-acetylation of the resultant cysteine conjugate to form S-(3-oxopropyl)-N-acetylcysteine (OPMA) in the kidney. Reduction of this aldehyde yields S-(3-hydroxypropyl)-N-acetylcysteine (HPMA), the main metabolite of acrolein found in urine [11,60], while oxidation of the aldehyde group produces S-carboxyethyl-N-acetylcysteine (carboxyethyl mercapturic acid, CEMA) (Fig. 7).…”

“…Enzymemediated epoxidation of acrolein produces glycidaldehyde, an instable intermediate that can react with water to yield glyceraldehyde [61] or it can form a conjugate with GSH [62]. Additional metabolites identified in the 0-24-h urine after oral administration of [2,[3][4][5][6][7][8][9][10][11][12][13][14] C] acrolein to rats (2.5 mg/kg) include 3-hydroxpropionic acid (6-30% relative abundance), malonic acid (trace), and N-acetyl-S-(2-carboxy-2-hydroxyethyl)-cysteine (8-18%), an endproduct metabolite of GSH addition to glycidaldehyde [62] (Fig. 7).…”

“…In humans, the mean levels of HPMA in first-void morning urine are about 4.0 nmol/mg creatinine for smokers and 0.7 nmol/mg creatinine for nonsmokers [11]. The average concentration of 3-HPMA in the urine of the smoker subjects was 1095 ng/mL urine, which translates into HPMA excretion of about 1.7-2 mg/day [11] (= 7.7-9 μmol).…”