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- Title
相対モル感度を用いたsingle-reference HPLC 法が定量値に影響を及ぼす 要因の検討と機能性表示食品中のルテイン定量への応用
- Authors
酒井有希; 増本直子; 西﨑雄三; 大槻 崇; 松藤 寛; 杉本直樹; 佐藤恭子
- Abstract
Chromatographic techniques, such as high-performance liquid chromatography (HPLC), are typically adopted as quantitative methods for the separation and identification of functional substances classed as Food with Function Claims (FFC). Although the quantitative values determined by chromatographic approaches are ensured by using high purity reference materials, for most functional substances, such references are not commercially available. In this study, we applied an alternative quantitative technique using the relative molar sensitivity (RMS), namely single-reference HPLC, to quantitatively analyze the content of lutein in FFC. In this method, RMS of the analyte to a single reference compound was used to determine the analyte content in FFC using HPLC/photodiode array detector (PDA). Notably, the approach did not require the use of any reference materials. Since there were no researches on factors affecting on quantitative value by single-reference HPLC method, the concentration ranges of the analyte and single reference, which yielded reliable RMS values, were comprehensively evaluated to obtain accurate quantitative values by single-reference HPLC. Consequently, when the signal to noise ratio of the peaks corresponding to the single reference and analyte was above 50, the differences in the quantitative values were within approximately 1.5%. Based on the obtained results, RMS of lutein to a single reference compound, specifically sudan I, which is a stable commercially available analytical standard, was established at 8.18. It was found that there was no significant difference between the lutein content of FFC determined utilizing the single-reference HPLC method and the absolute calibration curve approach. The calibration curve was generated using lutein with adjusted purity measured by 1H-qNMR. The outcomes of the study demonstrate that lutein in FFC can be indirectly quantified employing inexpensive and high-purity sudan I as a single reference.
- Publication
Japanese Journal of Food Chemistry & Safety, 2020, Vol 27, Issue 3, p123
- ISSN
1341-2094
- Publication type
Article