It is very important for food chemistry and the food industry to precisely determine the contents of food components because the results are associated with the quality and safety of foods. Chromatography such as HPLC/photodiode array (HPLC/PDA) enables the separation and quantification of components in complex mixtures such as natural food additives and processed foods. This technique is widely used in the field of food science. The sensitivity of chromatography detectors varies for different analytes because each analyte has unique physical properties. Therefore, analytical standards corresponding to analyte are needed to perform calibrations. However, analytical standards, particularly for natural components, are not widely available. Recently, 1H-quantitative NMR (1H-qNMR) has become a powerful tool for quantifying analytes without having to rely on identical standards; however, the resolution of this technique is inferior to that of chromatography. Therefore, it is difficult to employ it directly to analyze natural food additives and processed foods. Our study has focused on method to remove the disadvantages of chromatography and 1H-qNMR. This has led to designing an off-line combination of HPLC/PDA and 1H-qNMR for estimating chromatographic relative molar sensitivity (RMS) of analytes with respect to a reference standard. The RMSs are calculated as follows: (1) artificial mixtures of the analyte and reference are analyzed using 1H-qNMR and HPLC/PDA; (2) the response ratios of each analyte and its reference, obtained by HPLC/PDA, are corrected using their molar ratios, as obtained by 1H-qNMR; (3) Using HPLC/PDA, analyte contents can be determined from the RMS values, peak area of analytes and reference, and the amount of reference standard precisely added to the sample solution. The accuracy of the calculated RMSs has been verified, and the applicability of our proposed method to determine the contents of components in natural food additives and processed foods has been evaluated.

Development of a HPLC/PDA method for quantitative analysis of food components without the need for analytical standards

254th National Meeting (2017)

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