Circulating Choline and Phosphocholine Measurement by a Hydrophilic Interaction Liquid Chromatography-tandem Mass Spectrometry

Overview

Journal Heliyon

Specialty Social Sciences

Date 2023 Nov 29

PMID 38027764

Authors

Giulia Guerra

Francesco Segrado

Patrizia Pasanisi

Eleonora Bruno

Salvatore Lopez

Francesco Raspagliesi

Michela Bianchi

Elisabetta Venturelli

Affiliations

Soon will be listed here.

Abstract

Background: Given the growing interest in studying the role of choline and phosphocholine in the development and progression of tumor pathology, in this study we describe the development and validation of a fast and robust method for the simultaneous analysis of choline and phosphocholine in human plasma.

Methods: Choline and phosphocholine quantification in human plasma was obtained using a hydrophilic interaction liquid chromatography-tandem mass spectrometry technique. Assay performance parameters were evaluated using EMA guidelines.

Results: Calibration curve ranged from 0.60 to 38.40 μmol/L (R = 0.999) and 0.08-5.43 μmol/L (R = 0.998) for choline and phosphocholine, respectively. The Limit Of Detection of the method was 0.06 μmol/L for choline and 0.04 μmol/L for phosphocholine. The coefficient of variation range for intra-assay precision is 2.2-4.1 % (choline) and 3.2-15 % (phosphocholine), and the inter-assay precision range is < 1-6.5 % (choline) and 6.2-20 % (phosphocholine). The accuracy of the method was below the ±20 % benchmarks at all the metabolites concentration levels. In-house plasma pool of apparently healthy adults was tested, and a mean concentration of 15.97 μmol/L for Choline and 0.34 μmol/L for Phosphocholine was quantified.

Conclusions: The developed method shows good reliability in quantifying Choline and Phosphocholine in human plasma for clinical purposes.

References

Kirsch S, Herrmann W, Rabagny Y, Obeid R . Quantification of acetylcholine, choline, betaine, and dimethylglycine in human plasma and urine using stable-isotope dilution ultra performance liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2010; 878(32):3338-44. DOI: 10.1016/j.jchromb.2010.10.016. View

Otoki Y, Kato S, Kimura F, Furukawa K, Yamashita S, Arai H . Accurate quantitation of choline and ethanolamine plasmalogen molecular species in human plasma by liquid chromatography-tandem mass spectrometry. J Pharm Biomed Anal. 2016; 134:77-85. DOI: 10.1016/j.jpba.2016.11.019. View

Nitter M, Norgard B, de Vogel S, Eussen S, Meyer K, Ulvik A . Plasma methionine, choline, betaine, and dimethylglycine in relation to colorectal cancer risk in the European Prospective Investigation into Cancer and Nutrition (EPIC). Ann Oncol. 2014; 25(8):1609-15. DOI: 10.1093/annonc/mdu185. View

Knittelfelder O, Weberhofer B, Eichmann T, Kohlwein S, Rechberger G . A versatile ultra-high performance LC-MS method for lipid profiling. J Chromatogr B Analyt Technol Biomed Life Sci. 2014; 951-952:119-28. PMC: 3946075. DOI: 10.1016/j.jchromb.2014.01.011. View

Shunova A, Bockmann K, Minarski M, Franz A, Wiechers C, Poets C . Choline Content of Term and Preterm Infant Formulae Compared to Expressed Breast Milk-How Do We Justify the Discrepancies?. Nutrients. 2020; 12(12). PMC: 7763895. DOI: 10.3390/nu12123815. View

Garcia E, Shalaurova I, Matyus S, Wolak-Dinsmore J, Oskardmay D, Connelly M . Quantification of choline in serum and plasma using a clinical nuclear magnetic resonance analyzer. Clin Chim Acta. 2021; 524:106-112. DOI: 10.1016/j.cca.2021.11.031. View

Zeisel S, da Costa K . Choline: an essential nutrient for public health. Nutr Rev. 2009; 67(11):615-23. PMC: 2782876. DOI: 10.1111/j.1753-4887.2009.00246.x. View

Sarkar A, Ghosh D, Haldar D, Sarkar P, Gupta B, Dastidar S . A rapid LC-ESI-MS/MS method for the quantitation of choline, an active metabolite of citicoline: Application to in vivo pharmacokinetic and bioequivalence study in Indian healthy male volunteers. J Pharm Biomed Anal. 2012; 71:144-7. DOI: 10.1016/j.jpba.2012.07.003. View

Buszewski B, Noga S . Hydrophilic interaction liquid chromatography (HILIC)--a powerful separation technique. Anal Bioanal Chem. 2011; 402(1):231-47. PMC: 3249561. DOI: 10.1007/s00216-011-5308-5. View

10.

Michel V, Yuan Z, Ramsubir S, Bakovic M . Choline transport for phospholipid synthesis. Exp Biol Med (Maywood). 2006; 231(5):490-504. DOI: 10.1177/153537020623100503. View

11.

Koc H, Mar M, Ranasinghe A, Swenberg J, Zeisel S . Quantitation of choline and its metabolites in tissues and foods by liquid chromatography/electrospray ionization-isotope dilution mass spectrometry. Anal Chem. 2002; 74(18):4734-40. DOI: 10.1021/ac025624x. View

12.

de Vogel S, Ulvik A, Meyer K, Ueland P, Nygard O, Vollset S . Sarcosine and other metabolites along the choline oxidation pathway in relation to prostate cancer--a large nested case-control study within the JANUS cohort in Norway. Int J Cancer. 2013; 134(1):197-206. DOI: 10.1002/ijc.28347. View

13.

Yue B, Pattison E, Roberts W, Rockwood A, Danne O, Lueders C . Choline in whole blood and plasma: sample preparation and stability. Clin Chem. 2008; 54(3):590-3. DOI: 10.1373/clinchem.2007.094201. View

14.

Ojiakor O, Rylett R . Modulation of sodium-coupled choline transporter CHT function in health and disease. Neurochem Int. 2020; 140:104810. DOI: 10.1016/j.neuint.2020.104810. View

15.

Guasch-Ferre M, Hu F, Ruiz-Canela M, Bullo M, Toledo E, Wang D . Plasma Metabolites From Choline Pathway and Risk of Cardiovascular Disease in the PREDIMED (Prevention With Mediterranean Diet) Study. J Am Heart Assoc. 2017; 6(11). PMC: 5721752. DOI: 10.1161/JAHA.117.006524. View

16.

Heaton J, McCalley D . Some factors that can lead to poor peak shape in hydrophilic interaction chromatography, and possibilities for their remediation. J Chromatogr A. 2015; 1427:37-44. DOI: 10.1016/j.chroma.2015.10.056. View

17.

Greco G, Letzel T . Main interactions and influences of the chromatographic parameters in HILIC separations. J Chromatogr Sci. 2013; 51(7):684-93. DOI: 10.1093/chromsci/bmt015. View

18.

Ohkawa R, Kurano M, Sakai N, Kishimoto T, Nojiri T, Igarashi K . Measurement of plasma choline in acute coronary syndrome: importance of suitable sampling conditions for this assay. Sci Rep. 2018; 8(1):4725. PMC: 5856837. DOI: 10.1038/s41598-018-23009-x. View

19.

Chandler T, White H . Choline and methionine differentially alter methyl carbon metabolism in bovine neonatal hepatocytes. PLoS One. 2017; 12(2):e0171080. PMC: 5289486. DOI: 10.1371/journal.pone.0171080. View

20.

Bernhard W, Raith M, Shunova A, Lorenz S, Bockmann K, Minarski M . Choline Kinetics in Neonatal Liver, Brain and Lung-Lessons from a Rodent Model for Neonatal Care. Nutrients. 2022; 14(3). PMC: 8837973. DOI: 10.3390/nu14030720. View