Application of Separation Techniques in Analytics of Biologically Relevant Sulfur Compounds

Głowacki, Rafał; Piechocka, J; Bald, Edward; Chwatko, Grażyna

doi:10.1007/978-3-030-63957-0_11-1

Cited by 1 publication

(2 citation statements)

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“…In this case, sample preparation involved treatment with 3 mol/L PCA followed by centrifugation to remove urinary proteins. This is one of the most commonly used techniques for effectively eliminating proteins from biological samples, by the addition of water-miscible organic solvent and (ultra)filtration [35][36][37][38] . Based on our earlier findings, sample acidification with a popular protein precipitating agent, namely PCA, was taken into account.…”

Section: Sample Preparationmentioning

confidence: 99%

“…Importantly, the approaches utilizing centrifugal concentrators or membrane filters were excluded to minimize plastic consumption and reduce the quantity of the samples. The use of ACN, typically recognized as the most effective protein precipitating agent among organic solvents, was also eliminated due to a large excess of ACN needed in relation to biological sample, comparing to 3 mol/L PCA which is needed for complete removal of proteins [35][36][37][38] . Taking into account the trace concentration of urinary proteins, the efficient protein precipitation is achieved by mixing the urine sample with 3 mol/L PCA and crashing at 10:1 ratio by volumes.…”

Section: Sample Preparationmentioning

confidence: 99%

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Application of the HPLC-ELSD technique for the determination of major metabolites of ibuprofen and creatinine in human urine

Piechocka,

Matwiej,

Gaweł

et al. 2023

Sci Rep

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The report presents robust and high throughput methods, based on liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD), for the simultaneous determination of major metabolites of ibuprofen (IBU), namely 2-hydroxyibuprofen and carboxyibuprofen (method A) as well as creatinine (Crn) (method B) in human urine. The assays primarily involve straightforward sample purification. For both methods, the chromatographic separation of the analytes is achieved within 8 min at room temperature on Poroshell 120 SB-C18 (75 × 4.6 mm, 2.7 µm) column using gradient elution. The eluents consisted of 0.1% formic acid in water and acetonitrile (method A) or water and methanol (method B) delivered at a flow rate of 1 or 0.5 mL/min, respectively. In relation to metabolites of IBU, the assay linearity was observed within 0.06–0.5 g/L in urine, while the Crn assay linearity was demonstrated within 0.5–30 mmol/L in urine. The limit of quantification for IBU metabolites was determined to be 0.06 g/L, and 0.5 mmol/L for Crn. These methods were successfully applied to urine samples delivered by ten apparently healthy donors showing that the HPLC-ELSD assays are suitable for human urine screening.

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Section: Sample Preparationmentioning

confidence: 99%