Highly sensitive analysis of low-molecular-mass aldehydes in beverages using a hydroxylamine reagent by high-performance liquid chromatography with fluorescence detection.

In this study, a fluorescent reagent, 4-((aminooxy)methyl)-7-hydroxycoumarin (AOHC), was for the first time applied to label the low-molecular-mass aldehydes (LMMAs) through reductive oxyamination reaction to afford single N,O-substituted oxyamine derivatives at room temperatures with derivatization efficiencies as high as 96.8%. In the following high-performance liquid chromatography with fluorescence detection analysis, 12 LMMAs, including furfurals, aromatic aldehydes, and aliphatic aldehydes, were baseline-separated on an ODS column and detected with low limits of detection (LODs) (0.2-50 nM), and good precisions (intraday relative standard deviations [RSDs] were 2.40-4.68%, and interday RSDs were 4.65-8.91%). This approach was then adopted to analyze six alcoholic beverages and five dairy products, and nine LMMAs with concentrations in the 0.28-798.16 µM range were successfully detected with excellent accuracies (recoveries were 92.2-106.2%). Finally, the results were statistically analyzed and discussed. The proposed method has several advantages, including high sensitivity, room-temperature labeling, and the avoidance of further extraction and/or enrichment procedures, demonstrating its great utility for monitoring LMMAs in various complex matrices.

Nitrone formation: A new strategy for the derivatization of aldehydes and its application on the determination of furfurals in foods by high performance liquid chromatography with fluorescence detection.

A nitrone formation method using a novel fluorescent reagent 4-hydroxylaminopropyl-7-methoxylcoumarin (HAMC) has been developed and applied for the determination of furfurals in foods using high performance liquid chromatography with fluorescence detection (HPLC-FLD). Furfurals samples were derivatized by HAMC in aqueous condition at room temperature in less than 30min and were subjected to direct HPLC-FLD analysis without extra purification or extraction. Compared with other conventional methods, nitrone formation method produced single chromatographic peak for each analyte, leading to simpler chromatograms and enhancing the overall sensitivity. Low detection limits at sub-nM level (0.08-0.1nM) and high repeatabilities (intra-day RSD ≤ 4.2% and inter-day RSD ≤ 6.7%) were achieved. The linear range of the calibration curve was 0.2-4000nM with good correlation coefficients (R ≥ 0.9989). This method was successfully applied for furfurals determination in food samples, including honey and coffee. The accuracy was satisfactory with recoveries ranging from 89.7% to 106.7% Above all, this pre-column derivatization method is simple, reliable and highly sensitive, providing a promising way for future studies of aldehydes from various sources.

Reductive oxyamination: a method for the qualitative and quantitative analysis of monosaccharides with a new aminooxy reagent using high-performance liquid chromatography with fluorescence detection.

Derivatization of carbohydrates with aminooxy agents to form oximes can be used for qualitative and quantitative analysis of carbohydrates; however, the formation of isomeric products limits its application. A new reductive oxyamination procedure developed for the analysis of monosaccharides with a novel fluorescent O-substituted aminooxy reagent, 4-((aminooxy)methyl)-6-chloro-7-hydroxycoumarin (AOCC), is reported. In this procedure, monosaccharides undergo an oxime formation reaction with AOCC and are then readily reduced with 2-picoline-borane, followed by analysis with high-performance liquid chromatography with fluorescence detection. Good separation of five monosaccharide derivatives was achieved within 40 min with acetonitrile-water-tetrahydrofuran as the mobile phase. The detection limits were on the order of femtomoles. The linear range was 0.2-4000 nM, with a good correlation coefficient (R ≥ 0.9985). Furthermore, the method was applied for analysis of real samples, such as bovine milk powder, without complicated and tedious sample treatment. This reductive oxyamination method circumvents the problem caused by oxime isomers and can be used for the highly sensitive and selective analysis of monosaccharides with high accuracy, providing an effective and promising method for the analysis of carbonyls with aminooxy agents.

Highly sensitive method for aldehydes detection: Application to furfurals analysis in raisin and bovine milk powder.

A pre-column fluorescent derivatization method based on nitrone formation has been applied to determine furfurals (e.g. furfural (F), 5-methyfurfural (5-MF) and 5-hydroxymethylfurfural (5-HMF)) in food samples for the first time. An N-substituted hydroxylamine reagent 4-((hydroxyamino)butyl)-7-hydroxycoumarin (HAHC) was used to react with the aldehyde group of furfurals to form stable nitrone derivatives with high fluorescence intensities. The reactions proceeded under mild conditions in 30 min with high derivatization yields (>93%). A baseline-separation of three furfurals derivatives was subsequently achieved within 25 min on a reversed-phase column. The detection limits were at the low femtomol level (S/N = 3, 20 µL per injection). The linear range of the calibration curve was 0.4-4000 nM with good correlation coefficients (R2 ≥ 0.9991). The proposed method was further applied for food sample analysis, such as bovine milk powder and raisin. Satisfactory recoveries were obtained in the range of 94.7%-103.5%. Above all, this pre-column derivatization method is simple, fast and highly sensitive, providing an effective and promising way for future studies of aldehydes in different matrices.