Determination of choline, betaine, and dimethylglycine in plasma by a high-throughput method based on normal-phase chromatography-tandem mass spectrometry
- PMID: 12560353
- DOI: 10.1373/49.2.286
Determination of choline, betaine, and dimethylglycine in plasma by a high-throughput method based on normal-phase chromatography-tandem mass spectrometry
Abstract
Background: The quaternary ammonium compounds, choline and betaine, and dimethylglycine (DMG) reside along a metabolic pathway linked to the synthesis of neurotransmitters and membrane phospholipids and to homocysteine remethylation and, therefore, folate status. Lack of a convenient, high-throughput method for the determination of these compounds has prevented population-based studies of their possible associations with lifestyle, nutrition, and chronic diseases.
Methods: Serum or plasma samples were deproteinized by mixing with three volumes of acetonitrile that contained d(9)-choline and d(9)-betaine as internal standards. We used a normal-phase silica column for the separation of choline (retention time, 2.8 min), betaine (1.3 min), DMG (1.15 min), and internal standards, which were detected as positive ions by tandem mass spectroscopy in the multiple-reaction monitoring mode, using the molecular transitions m/z 104-->60 (choline), m/z 113-->69 (d(9)-choline), m/z 118-->59 (betaine), m/z 127-->68 (d(9)-betaine), and m/z 104-->58 (DMG).
Results: For all three metabolites, the assay was linear in the range 0.4-400 micromol/L, and the lower limit of the detection (signal-to-noise ratio = 5) was < or =0.3 micromol/L. The within- and between-day imprecision (CVs) was 2.1-7.2% and 3.5-8.8%, respectively. The analytical recovery was 87-105%. The fasting plasma concentrations (median, 25th-75th percentiles) were 8.0 (7.0-9.3) micromol/L for choline, 31.7 (27.0-41.1) micromol/L for betaine, and 1.66 (1.30-2.02) micromol/L for DMG in 60 healthy blood donors. In individuals who had eaten a light breakfast, plasma concentrations of all three metabolites were significantly (25-30%) higher than in fasting individuals.
Conclusion: This is the first method for the combined measurement of choline, betaine, and DMG in human plasma or serum. The assay is characterized by simple sample preparation, no derivatization, high throughput, imprecision (CV) <10%, detection limits below the values seen in volunteers, and the high specificity provided by tandem mass spectroscopy.
Similar articles
-
An isotope dilution capillary electrophoresis/tandem mass spectrometry (CE-MS/MS) method for the simultaneous measurement of choline, betaine, and dimethylglycine concentrations in human plasma.Anal Bioanal Chem. 2016 Oct;408(26):7505-12. doi: 10.1007/s00216-016-9848-6. Epub 2016 Aug 8. Anal Bioanal Chem. 2016. PMID: 27503542
-
Measurement of concentrations of whole blood levels of choline, betaine, and dimethylglycine and their relations to plasma levels.J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Apr 15;957:41-5. doi: 10.1016/j.jchromb.2014.02.030. Epub 2014 Feb 28. J Chromatogr B Analyt Technol Biomed Life Sci. 2014. PMID: 24674988
-
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 Dec 15;878(32):3338-44. doi: 10.1016/j.jchromb.2010.10.016. Epub 2010 Oct 23. J Chromatogr B Analyt Technol Biomed Life Sci. 2010. PMID: 21074502
-
Betaine: a key modulator of one-carbon metabolism and homocysteine status.Clin Chem Lab Med. 2005;43(10):1069-75. doi: 10.1515/CCLM.2005.187. Clin Chem Lab Med. 2005. PMID: 16197300 Review.
-
Simple method for the routine determination of betaine and N,N-dimethylglycine in blood and urine.Clin Chem. 1998 Sep;44(9):1937-41. Clin Chem. 1998. PMID: 9732980
Cited by
-
Rumen-Protected Choline Improves Metabolism and Lactation Performance in Dairy Cows.Animals (Basel). 2024 Mar 27;14(7):1016. doi: 10.3390/ani14071016. Animals (Basel). 2024. PMID: 38612255 Free PMC article.
-
Profiling of primary and phytonutrients in edible mahlab cherry (Prunus mahaleb L.) seeds in the context of its different cultivars and roasting as analyzed using molecular networking and chemometric tools.PeerJ. 2023 Aug 30;11:e15908. doi: 10.7717/peerj.15908. eCollection 2023. PeerJ. 2023. PMID: 37663279 Free PMC article.
-
Serum 5-Methyltetrahydrofolate Status Is Associated with One-Carbon Metabolism-Related Metabolite Concentrations and Enzyme Activity Indicators in Young Women.Int J Mol Sci. 2023 Jul 1;24(13):10993. doi: 10.3390/ijms241310993. Int J Mol Sci. 2023. PMID: 37446171 Free PMC article.
-
The distribution of dietary choline intake and serum choline levels in Australian women during pregnancy and associated early life factors.Eur J Nutr. 2023 Oct;62(7):2855-2872. doi: 10.1007/s00394-023-03186-w. Epub 2023 Jun 28. Eur J Nutr. 2023. PMID: 37378694 Free PMC article.
-
Maternal dietary deficiencies in folic acid or choline worsen stroke outcomes in adult male and female mouse offspring.Neural Regen Res. 2023 Nov;18(11):2443-2448. doi: 10.4103/1673-5374.371375. Neural Regen Res. 2023. PMID: 37282475 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources