Abstract
The coupling of anion exchange high-pressure liquid chromatography (HPLC) with electrospray ionization mass spectrometry (ESI-MS) allows for the simultaneous detection of the six forms of inositol phosphate (InsP). Here we describe a rapid quantitative analysis of InsPs by HPLC-ESI-MS, which can be applied to a wide array of sample types. With this method, InsPs could be separated and detected within 20 min of sample injection. The detection limit was as low as 25 pmol (i.e., ca. 2 nmol/g sample) for each type of InsP, which is particularly important for analytes that are often present at low abundance in nature.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cao S, Dong N, Chen J (2011) Synchronous fluorescence determination of phytic acid in foodstuffs and urine based on replacement reaction. Phytochem Anal 22:119–123
Chen Y, Chen J, Ma K, Cao S, Chen X (2007) Fluorimetric determination of phytic acid in urine based on replacement reaction. Anal Chim Acta 605:185–191
Irth H, Lamoree M, de Jong GJ, Brinkman UATh, Frei RW, Kornfeldt RA et al (1990) Determination of D-myo-1,2,6-inositol trisphosphate by ion-pair reversed-phase liquid chromatography with post-column ligand exchange and fluorescence detection. J Chromatogr A 499:617–625
March JG, Simonet BM, Grases F (1999) Fluorimetric determination of phytic acid based on the activation of the oxidation of 2,2′-dipyridyl ketone hydrazone catalysed by Cu(II). Analyst 124:897–900
Rounds MA, Nielsen SS (1993) Anion-exchange high-performance liquid chromatography with post-column detection for the analysis of phytic acid and other inositol phosphates. J Chromatogr A 653:148–152
Guse AH, Greiner E, Emmrich F, Brand K (1993) Mass changes of inositol 1,3,4,5,6-pentakisphosphate and inositol hexakisphosphate during cell cycle progression in rat thymocytes. J Biol Chem 268:7129–7133
Sjöberg PJR, Thelin P, Rydin E (2016) Separation of inositol phosphate isomers in environmental samples by ion-exchange chromatography coupled with electrospray ionization tandem mass spectrometry. Talanta 161:392–397
Guo J, Shi Y, Xu C, Zhong R, Zhang F, Zhang T et al (2016) Quantification of plasma myo-inositol using gas chromatography–mass spectrometry. Clin Chim Acta 460:88–92
Zhang S, Yang W, Zhao Q, Zhou X, Fan Y, Chen R (2017) Rapid method for simultaneous determination of inositol phosphates by IPC-ESI–MS/MS and its application in nutrition and genetic research. Chromatographia 80:275–286
Liu X, Villalta PW, Sturla SJ (2009) Simultaneous determination of inositol and inositol phosphates in complex biological matrices: quantitative ion-exchange chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 23:705–712
Duong QH, Clark KD, Lapsley KG, Pegg RB (2017) Determination of myo-inositol phosphates in tree nuts and grain fractions by HPLC-ESI-MS. J Food Compos Anal 59:74–82
Duong QH, Clark KD, Lapsley KG, Pegg RB (2017) Quantification of inositol phosphates in almond meal and almond brown skins by HPLC/ESI/MS. Food Chem 229:84–92
Acknowledgments
Financial supports for this research by The Graduate School of UGA and the Almond Board of California are gratefully acknowledged. Use of the Proteomics and Mass Spectrometry (PAMS) core facility with the assistance of Dr. Kevin D. Clark in UGA’s Department of Chemistry, Athens, GA, is greatly appreciated.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Duong, Q.H., Pegg, R.B. (2020). Quantitation of Inositol Phosphates by HPLC-ESI-MS. In: Miller, G. (eds) Inositol Phosphates. Methods in Molecular Biology, vol 2091. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0167-9_2
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0167-9_2
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0166-2
Online ISBN: 978-1-0716-0167-9
eBook Packages: Springer Protocols