Development, validation, and comparison of four methods to simultaneously quantify L-arginine, citrulline, and ornithine in human plasma using hydrophilic interaction liquid chromatography and electrospray tandem mass spectrometry

dc.contributor.authorLai, Xianyin
dc.contributor.authorKline, Jeffrey A.
dc.contributor.authorWang, Mu
dc.contributor.departmentDepartment of Biochemistry and Molecular Biology, IU School of Medicineen_US
dc.date.accessioned2016-05-04T17:55:40Z
dc.date.available2016-05-04T17:55:40Z
dc.date.issued2015-11
dc.description.abstractTo understand the role of l-arginine depletion in impaired nitric oxide synthesis in disease, it is important to simultaneously quantify arginine, citrulline, and ornithine in the plasma. Because the three amino acids are endogenous analytes, true blank matrix for them is not available. It is necessary and valuable to compare the performance of different approaches due to lack of regulatory clarity for validation. A two-step sample preparation method using methanol as protein precipitation reagent was developed in this study is used for sample preparation. Because true blank matrix for endogenous analytes is not available, water as blank matrix, 1% BSA in PBS as blank matrix, surrogate analyte, and background subtraction were designed to establish successful quantification methods. Four methods to simultaneously quantify arginine, citrulline, and ornithine in human plasma using hydrophilic interaction liquid chromatography and electrospray tandem mass spectrometry were developed, validated, and compared. The developed two-step sample preparation method using methanol as protein precipitation reagent in this study needs less time and provides higher recovery comparing with other approaches. Three of the four methods, water as blank matrix, 1% BSA in PBS as blank matrix, and surrogate analyte, have been successful in fulfilling all the criteria, while background subtraction has failed. Results of the measured concentrations in 97 human plasma samples using the three methods show that the difference between any two methods or among the three methods presents 100% of samples with less than 20% for all the three amino acids and majority of them are under 10%. The developed two-step sample preparation method using methanol as protein precipitation reagent is simple and convenient. Three of the four methods are fully validated and the validation is successful. The BSA functioned effectively as a blank matrix for these three amino acids, considering cost, data quality, matrix similarity, and practicality.en_US
dc.eprint.versionAuthor's manuscripten_US
dc.identifier.citationLai, X., Kline, J. A., & Wang, M. (2015). Development, validation, and comparison of four methods to simultaneously quantify l-arginine, citrulline, and ornithine in human plasma using hydrophilic interaction liquid chromatography and electrospray tandem mass spectrometry. Journal of Chromatography B, 1005, 47–55. http://doi.org/10.1016/j.jchromb.2015.10.001en_US
dc.identifier.urihttps://hdl.handle.net/1805/9523
dc.language.isoenen_US
dc.publisherElsevieren_US
dc.relation.isversionof10.1016/j.jchromb.2015.10.001en_US
dc.relation.journalJournal of Chromatography Ben_US
dc.rightsPublisher Policyen_US
dc.sourceAuthoren_US
dc.subjectarginineen_US
dc.subjectcitrullineen_US
dc.subjectornithineen_US
dc.subjectplasmaen_US
dc.titleDevelopment, validation, and comparison of four methods to simultaneously quantify L-arginine, citrulline, and ornithine in human plasma using hydrophilic interaction liquid chromatography and electrospray tandem mass spectrometryen_US
dc.typeArticleen_US
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