Abstract
Citrulline is a non-protein building amino acid that plays important roles in nitric oxide production, vasodilation and ammonia detoxification. It is structurally related to arginine and is most abundant in melons and other cucurbit species. Most current methods for the quantification of citrulline and arginine require time-consuming chemical derivatization steps, and often use more expensive detection methods such as fluorescence detection. Hence, this study aimed to develop a simple and rapid reversed-phase high-performance liquid chromatography method using ultraviolet detection (RP-HPLC–UV), without requiring any analyte derivatization. The method was used to quantify the citrulline and arginine content of 13 Australian fruit and vegetable cultivars, comprising 11 species from 7 genera and 5 families. The highest citrulline contents were found in mature citron melon (Citrullus lanatus var. citroides) fruit (39.2 mg/g dry weight basis), which also had the highest citrulline: arginine ratio of 17.8. Red watermelon (Citrullus lanatus) flesh also contained high levels of citrulline (22.9 mg/g), with lower concentrations in other melons, pumpkin and squash (1–3 mg/g). Arginine levels were highest in red watermelon flesh (8.1 mg/g). This extraction protocol and HPLC method may be suitable for screening large number of samples for citrulline and arginine contents.
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Acknowledgements
The authors wish to thank Tania Collins for the assistance with the HPLC instrumentation.
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This work was funded in part by a New Staff Grant (RSH/5343) awarded by CQUniversity to one of the authors (MN). One of the authors (JJ) acknowledges support from the Australian Government in the form of a Research Training Program Scholarship.
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Joel Johnson: conceptualization, methodology, validation, formal analysis; investigation, data curation, writing—original draft, writing—review and editing. Browny Ohri: writing—review and editing. Kerry Walsh: writing—review and editing, supervision. Mani Naiker: conceptualization, methodology, writing—review and editing, supervision, funding acquisition.
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Joel B. Johnson declares that he has no conflict of interest. Browny Ohri declares that he has no conflict of interest. Kerry B. Walsh declares that he has no conflict of interest. Mani Naiker declares that he has no conflict of interest.
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Johnson, J.B., Ohri, B., Walsh, K.B. et al. A Simple Isocratic HPLC–UV Method for the Simultaneous Determination of Citrulline and Arginine in Australian Cucurbits and Other Fruits. Food Anal. Methods 15, 104–114 (2022). https://doi.org/10.1007/s12161-021-02110-4
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DOI: https://doi.org/10.1007/s12161-021-02110-4