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Detection of homocysteine in human saliva using protein precipitation and analysis by liquid chromatography and electrospray ionization quadrupole time-of-flight mass spectrometry: Profiles in healthy adults

Detection of homocysteine in human saliva using protein precipitation and analysis by liquid chromatography and electrospray ionization quadrupole time-of-flight mass spectrometry: Profiles in healthy adults

Abaye, Daniel A., Nielsen, Birthe and Boateng, Joshua S. ORCID: 0000-0002-6310-729X (2013) Detection of homocysteine in human saliva using protein precipitation and analysis by liquid chromatography and electrospray ionization quadrupole time-of-flight mass spectrometry: Profiles in healthy adults. Protein and Peptide Letters, 20 (12). pp. 1382-1389. ISSN 0929-8665 (Print), 1875-5305 (Online) (doi:10.2174/092986652012131112142531)

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Abstract

Homocysteine (Hcys) is a non-essential amino acid associated with a range of diseased and abnormal metabolic conditions. Hcys concentration in saliva is routinely determined by enzyme assays, which are broadly specific, but can be expensive and suffer from cross-reactivity. Total Hcys (tHcys) concentrations in eight healthy adults were determined to establish the inter-day variation during resting, normal and intensive physical activity, using the more sensitive analytical techniques of liquid chromatography and tandem mass spectrometry without prior derivatization. Saliva (~ 1.5 mL) was collected over four days; early morning (EA), normal activity (NA) and during physical activity (PA). Samples were processed by disulphide reduction, acetonitrile precipitation and then centrifugation-filtration. Extracts were chromatographically resolved and analysed on a quadrupole time- of-flight (QToF) mass spectrometer. The protonated [M+H]+, m/z 136.101 and product ([M+H]+- HCOOH) m/z 90.103 ions were then monitored against an internal standard (13CHcys) and external set of calibration standards. Mean tHcys concentration for the whole group, including exercise was 6.6 ± 8.0 (range 0.2 – 29.6 nmol/mL). Overall, concentration of tHcys was greater in males than the females but not significantly (p > 0.05). The mean EA concentration was significantly (p < 0.05) greater than NA for both males (p = 0340) and females (p = 0.0045). There were large within-subject variations (coefficient of variation; CV%; 24% to 103%). The limits of detection (LOD) and quantification (LOQ) were 0.07 and 0.22 nmol/mL, respectively. The procedure potentially provides a convenient means of analyzing salivary Hcys as a diagnostic disease marker.

Item Type: Article
Additional Information: [1] Copyright: © 2015 Bentham Science Publishers
Uncontrolled Keywords: acetonitrile precipitation, healthy adults, Homocysteine, liquid chromatography mass spectrometry, quadrupole time-of-flight, saliva
Subjects: Q Science > QD Chemistry
Q Science > QP Physiology
Faculty / Department / Research Group: Faculty of Engineering & Science
Faculty of Engineering & Science > Department of Pharmaceutical, Chemical & Environmental Sciences
Related URLs:
Last Modified: 17 Oct 2016 09:13
Selected for GREAT 2016: None
Selected for GREAT 2017: None
Selected for GREAT 2018: None
URI: http://gala.gre.ac.uk/id/eprint/12403

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