Collection and determination of nucleotide metabolites in neonatal and adult saliva by high performance liquid chromatography with tandem mass spectrometry

Al-Shehri, S., Henman, M., Charles, B. G., Cowley, D., Shaw, P. N., Liley, H., Tomarchio, A., Punyadeera, C. and Duley, J. A. (2013) Collection and determination of nucleotide metabolites in neonatal and adult saliva by high performance liquid chromatography with tandem mass spectrometry. Journal of Chromatography B, 931 140-147. doi:10.1016/j.jchromb.2013.05.001


Author Al-Shehri, S.
Henman, M.
Charles, B. G.
Cowley, D.
Shaw, P. N.
Liley, H.
Tomarchio, A.
Punyadeera, C.
Duley, J. A.
Title Collection and determination of nucleotide metabolites in neonatal and adult saliva by high performance liquid chromatography with tandem mass spectrometry
Journal name Journal of Chromatography B   Check publisher's open access policy
ISSN 1570-0232
1873-376X
Publication date 2013-07-15
Sub-type Article (original research)
DOI 10.1016/j.jchromb.2013.05.001
Volume 931
Start page 140
End page 147
Total pages 8
Place of publication Netherlands
Publisher Elsevier
Collection year 2014
Language eng
Formatted abstract
Saliva contains a number of biochemical components which may be useful for diagnosis/monitoring of metabolic disorders, and as markers of cancer or heart disease. Saliva collection is attractive as a non-invasive sampling method for infants and elderly patients. We present a method suitable for saliva collection from neonates. We have applied this technique for the determination of salivary nucleotide metabolites. Saliva was collected from 10 healthy neonates using washed cotton swabs, and directly from 10 adults. Two methods for saliva extraction from oral swabs were evaluated. The analytes were then separated using high performance liquid chromatography (HPLC) with tandem mass spectrometry (MS/MS). The limits of detection for 14 purine/pyrimidine metabolites were variable, ranging from 0.01 to 1.0 μM. Recovery of hydrophobic purine/pyrimidine metabolites from cotton tips was consistently high using water/acetonitrile extraction (92.7–111%) compared with water extraction alone. The concentrations of these metabolites were significantly higher in neonatal saliva than in adults. Preliminary ranges for nucleotide metabolites in neonatal and adult saliva are reported. Hypoxanthine and xanthine were grossly raised in neonates (49.3 ± 25.4; 30.9 ± 19.5 μM respectively) compared to adults (4.3 ± 3.3; 4.6 ± 4.5 μM); nucleosides were also markedly raised in neonates. This study focuses on three essential details: contamination of oral swabs during manufacturing and how to overcome this; weighing swabs to accurately measure small saliva volumes; and methods for extracting saliva metabolites of interest from cotton swabs. A method is described for determining nucleotide metabolites using HPLC with photodiode array or MS/MS. The advantages of utilising saliva are highlighted. Nucleotide metabolites were not simply in equilibrium with plasma, but may be actively secreted into saliva, and this process is more active in neonates than adults.
Keyword Saliva
Neonates
Nucleotide metabolites
Purines
HPLC
Mass spectrometry
Pyrimidine Metabolism
Newborn-Infants
Plasma Creatinine
Inborn-Errors
Purine
Hypoxanthine
Xanthine
Disorders
Cortisol
Uridine
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 6 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 8 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Sun, 11 Aug 2013, 10:12:13 EST by System User on behalf of School of Pharmacy