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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. (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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