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

Al-Shehri, Saad S.; Henman, Michael G.; Charles, Bruce G.; Cowley, David; Shaw, P. Nicholas; Liley, Helen; Tomarchio, A.; Punyadeera, Chamindie; Duley, John A.

doi:10.1016/j.jchromb.2013.05.001

Cited by 19 publications

(14 citation statements)

References 36 publications

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“…Since we intended primarily to estimate the concentration levels of free nucleobases in samples, we did not employ any rigorous sample pretreatment or acid hydrolysis of samples as usually applied when total amounts of nucleobases in samples were determined. 3,7,9,[12][13]15,[20][21][22] For the HPLC analysis beer samples (alcoholic and nonalcoholic samples of local producers) were degassed to eliminate Markelj et al: Optimization of High Performance Liquid ... dissolved CO 2 and diluted with mobile phase MP2 at a ratio 1:20.…”

Section: Sample Preparationmentioning

confidence: 99%

“…Acetate buffers are also preferred when mass spectrometric detection was combined with the LC separation. 20,22 Markelj et al: Optimization of High Performance Liquid ...…”

Section: Optimization Of Chromatographic Conditions For Separationmentioning

confidence: 99%

“…[16][17][18] For both techniques UV/VIS, 5,18 electrochemical 19 or MS detection is used. [13][14]18,[20][21][22] Since nucleobases are electroactive, in some papers voltammetric determination with static mercury drop electrode, by modified graphite/carbon electrodes, [23][24] and on the boron-doped carbon nanotubes are also reported. 27 In this study HPLC with isocratic elution with UV and MS detection was employed for the separation and determination of nine purine and pyrimidine bases.…”

Section: Introductionmentioning

confidence: 99%

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Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Markelj

Župančič

Pihlar

2016

ACSi

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Nine purine and pyrimidine bases were separated and determined simultaneously using reversed phase (RP) high performance liquid chromatography (HPLC) in some food samples and biological fluids. Chromatographic behavior of these ionizable compounds highly depends on the interactions with the solvent as confirmed experimentally and by calculation of distribution of this species as a function of pH. Chromatograms show the optimal separation of five purine (uric acid, hypoxanthine, xanthine, adenine, and guanosine), and four pyrimidine (cytosine, uracil, cytidine and tymine) bases at pH around four. Accordingly, acetate buffer was selected due to high buffer capacity in this region. By variation of pH, concentration of buffer and volume ratio between buffer and methanol, we found that a mixture of 50 mM acetate buffer of pH 4.0 ± 0.1 with 3% of methanol ensures reproducibility, complete separation in less than 15 minutes and compatibility with UV and MS detection. Developed screening method was validated and applied for the analysis of complex clinical and beverage samples.

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Section: Sample Preparationmentioning

confidence: 99%

“…Acetate buffers are also preferred when mass spectrometric detection was combined with the LC separation. 20,22 Markelj et al: Optimization of High Performance Liquid ...…”

Section: Optimization Of Chromatographic Conditions For Separationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Markelj

Župančič

Pihlar

2016

ACSi

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“…The diagnosis based on saliva provides information about the physiological condition, which has some advantages like low contamination risk and non-invasive procedure (Al-Shehri et al, 2013;Chiappin et al, 2007). Thus, several studies have utilized saliva analysis as a tool for monitoring steroid, peptide, and immune markers of sports training (Papacosta and Nassis, 2011;Papacosta et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

“…Thus, several studies have utilized saliva analysis as a tool for monitoring steroid, peptide, and immune markers of sports training (Papacosta and Nassis, 2011;Papacosta et al, 2013). However, the clinical trials used to analyze salivary biomarkers (Al-Shehri et al, 2013;Moreira et al, 2013;Novaković et al, 2013;Salazar et al, 2013) are relatively expensive and laborious, which decreases the number of teams using this technology.…”

Section: Introductionmentioning

confidence: 99%

Application of FT-IR spectroscopy to assess physiological stress in rugby players during fatigue test

et al. 2016

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Introduction: The diagnosis based on salivary biomarkers provides information about the physiological condition. However, the clinical trials used to analyze these biomarkers are relatively expensive and laborious. Thus, the purpose of this study was to identify the physiological stress in players using Fourier transform infrared spectroscopy (FT-IR). Methods: Thirteen male rugby players were submitted to the treadmill fatigue test and saliva collections were performed before and immediately after test. The FT-IR spectra of saliva samples were analyzed by the second derivative and cluster analysis. Results: From the results of cluster analysis were possible to discriminate the spectra of saliva before and after physical effort using the spectral region between 1490 to 1420 cm -1. Only the saliva spectra from two players were not discriminated in pre-exercise group and post-exercise group, which are in agreement with lowest value of heart rates. Conclusion: The second derivative showed differences between the average spectra of saliva samples collected pre and post-test, which explain the spectra discrimination by the cluster analysis using a specific infrared region for the identification of physiological stress.

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The determination of salivary oxypurines before and after exercise by combined liquid chromatography-field asymmetric waveform ion mobility spectrometry-time-of-flight mass spectrometry

Arthur

Wilson

Turner

et al. 2018

Int. J. Ion Mobil. Spec.

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A method combining field asymmetric waveform ion mobility spectrometry with liquid chromatography-mass spectrometry (LC-FAIMS-MS) has been developed for the analysis of the oxypurine compounds hypoxanthine (HX) and xanthine (XA) in saliva. Separation of the oxypurines from interfering matrix components was investigated using FAIMS-MS. The selected FAIMS parameters were then applied to the rapid LC-FAIMS-MS analysis of HX and XA using a short chromatographic separation method (7 min). A comparison of the LC-MS method with and without FAIMS applied, resulted in improved discrimination from saliva matrix interferences and improved chromatographic peak integration for both HX and XA using a FAIMS separation. A quantitative evaluation of the LC-FAIMS-MS method was performed giving limits of detection of 2.0 ng mL-1 for HX and 1.8 ng mL-1 for XA, and limits of quantification of 6.6 ng mL-1 for HX and 6.0 ng mL-1 for XA. The developed LC-FAIMS-MS method was applied to the targeted analysis of the oxypurine metabolites in saliva collected from healthy male athletes (n=11) before and after exercise designed to induce oxidative stress; post-exercise collection time-points included immediately after exercise, one hour and twenty-four hours' post-exercise. The salivary concentrations of both HX and XA were lower after physical exercise, compared to the pre-exercise (rest) concentrations and returned to approximately pre-exercise levels after twenty-four hours. The method reported has the potential for monitoring the salivary oxypurines, HX and XA, as biomarkers of oxidative stress and in other clinical applications.

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

Cited by 19 publications

References 36 publications

Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases

Application of FT-IR spectroscopy to assess physiological stress in rugby players during fatigue test

The determination of salivary oxypurines before and after exercise by combined liquid chromatography-field asymmetric waveform ion mobility spectrometry-time-of-flight mass spectrometry

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