Development and Validation of a Micellar Electrokinetic Capillary Chromatographic Method for the Assessment of Nucleosides, Potential Biomarkers, in Blood Serum

Buzatto, Adriana Zardini; Guedes, Sumaya F.; Vidotto, Lucas T.; Oliveira, Laurione Cândido de; Simionato, Ana Valéria Colnaghi

doi:10.5935/0103-5053.20130116

Cited by 1 publication

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“…The initial conditions for extraction of the analytes from serum samples were based on a previous work . Nevertheless, extraction procedure was reevaluated in order to improve analytes recoveries.…”

Section: Resultsmentioning

confidence: 99%

“…Serum samples were centrifuged with Amicon Ultra ultrafiltration membranes (3000 Da) from Millipore (Billerica, USA) for 60 min under 4°C (6000 rpm) and extracted by SPE with PBA resin . Such material has affinity for cis‐diol groups, present in most of the studied nucleosides (except T), under basic medium, whereas elution occurs under acidic medium (Supporting Information Fig.…”

Section: Methodsmentioning

confidence: 99%

“…160–300 mmol/L) are necessary in order to achieve good separation resolution, which often results in clogged capillaries and damaged equipment due to salt deposition. Furthermore, preconcentration strategies have not been widely studied for nucleosides analyses by CE and better detectabilities are still necessary .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Higher detectability method for the analysis of nucleosides, putative tumor biomarkers, in blood serum samples by CE‐UV with reversed EOF

Buzatto¹,

Zardini²,

Guedes³

et al. 2015

Electrophoresis

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The development and validation of methodologies for the analysis of biological samples is of outcome importance in order to obtain trustworthy results. This work reports a novel CE-UV method for the assessment of nucleosides, putative tumor biomarkers, in blood serum. The separation of seven nucleosides within c.a. 20 min has been achieved with: BGE 30 mmol/L borate at pH 9.90, 50 mmol/L CTAB, and 10% methanol; V = -10 kV; T = 20°C; and capillary dimensions of 56 cm × 50 μm. The sample plug was concentrated by a modified large volume sample stacking strategy that provided better detectability. Validation showed that the method is suitable for bioanalytical purposes and initial applications in serum samples from healthy subjects are also presented. Finally, statistical methods were applied to verify the effect of characteristics such as age, smoking habits, and alcohol consumption on nucleoside concentrations in blood serum. Univariate statistical analysis tests emphasized the need for age matching, which was confirmed by PCA-DA and PLS-DA. Cancer history in the nearby family may also interfere in nucleoside levels in blood serum, since adenosine concentrations were statistically higher for volunteers who declared having diseased relatives.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%