Collection and determination of nucleotide metabolites in neonatal and adult saliva by high performance liquid chromatography with tandem mass spectrometry
| Autor: | Bruce G. Charles, David Cowley, Chamindie Punyadeera, Saad Al-Shehri, Helen G. Liley, John A. Duley, Paul N. Shaw, M. Henman, A. Tomarchio |
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| Rok vydání: | 2013 |
| Předmět: |
Adult
Male Purine Saliva Analyte Clinical Biochemistry Tandem mass spectrometry Biochemistry High-performance liquid chromatography Analytical Chemistry chemistry.chemical_compound stomatognathic system Limit of Detection Tandem Mass Spectrometry Humans Chromatography High Pressure Liquid Hypoxanthine Detection limit Chromatography Nucleotides Infant Newborn Reproducibility of Results Cell Biology General Medicine Middle Aged Xanthine Pyrimidines chemistry Purines Linear Models Female |
| Zdroj: | Journal of Chromatography B. 931:140-147 |
| ISSN: | 1570-0232 |
| DOI: | 10.1016/j.jchromb.2013.05.001 |
| Popis: | 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 mu M. 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 mu M respectively) compared to adults (4.3 +/- 3.3; 4.6 +/- 4.5 mu M); 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 photo-diode 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. (C) 2013 Elsevier B.V. All rights reserved. |
| Databáze: | OpenAIRE |
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