Bioanalytical method by column-switching with direct injection of human plasma for determination of sulphonylureas

Authors

  • Juliana Veloso Ferreira Department of Pharmaceutical Products - Faculty of Pharmacy - Federal University of Minas Gerais image/svg+xml
  • Gerson Antônio Pianetti Department of Pharmaceutical Products - Faculty of Pharmacy - Federal University of Minas Gerais image/svg+xml
  • Christian Fernandes Department of Pharmaceutical Products - Faculty of Pharmacy - Federal University of Minas Gerais image/svg+xml

DOI:

https://doi.org/10.22456/2527-2616.91542

Keywords:

sulphonylureas, column-switching, restricted access media, fused core column, human plasma

Abstract

Sulphonylureas are widely used in the treatment of Diabetes mellitus, one of the main causes of death in human population. Their determination is essential in pharmacological research and in the development of new drugs. Generally, determination of sulphonylureas in biological matrices is performed using conventional sample preparation techniques, which frequently leads to an increase of analysis time and errors. In this context, a bioanalytical method for the simultaneous determination of sulphonylureas by direct injection of human plasma was developed and optimized. An automated column-switching high performance liquid chromatographic system with a restricted access media (RAM) column coupled to a fused-core column was employed. At the first dimension, a RAM column with mobile phase of ultrapure water pH 6.0 at a flow-rate of 1.0 mL min-1 was used. The valve switching time was 3 minutes. At the second dimension, a C18 guard-column coupled to a C18 fused core column with mobile phase of acetonitrile and 10 mM phosphate buffer pH 3.0 (54:46 v/v) at a flow-rate of 0.8 mL min-1 were employed. The column switching system was performed in backflush configuration with an analyte elution time of 1 minute. Flufenamic acid was used as the internal standard. The mean plasma protein exclusion percentage by the RAM-column was 104.5%. The developed and optimized method showed to be fast and simple, allowing the direct injection of biological sample into the chromatographic system and the simultaneous determination of three sulphonylureas in only 12 minutes, including the sample treatment, separation and detection.

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References

Queiroz MEC, Lanças FM. Análise de fármacos em material biológico: acoplamento microextração em fase sólida “no tubo” e cromatografia líquida de alta eficiência. Quim. Nova. 2005; 28(5):880-886.

Cassiano NM, Lima VV, Oliveira RV, De Pietro AC, Cass QB. Development of restricted-access media supports and their application to the direct analysis of biological fluid samples via high-performance liquid chromatography. Anal. Bioanal. Chem. 2006; 384:1462- 1469.

Mullet WM. Determination of drugs in biological fluids by direct injection of samples for liquidchromatographic analysis. J. Biochem. Biophys. Methods. 2007; 70:263–273.

Sadílek P, Satínský D, Solich P. Using restricted-access materials and column switching in high-performance liquid chromatography for direct analysis of biologically-active compounds in complex matrices. Trends Anal. Chem. 2007; 26(5):375-384.

Souverain S, Rudaz S, Veuthey JL. Restricted access materials and large particle supports for on-line sample treatment: an attractive approach for biological fluids analysis. J. Chromatogr., B. 2004; 801:141-156.

Chen Y, Guo Z, Wang X, Qiu C. Sample treatment. J. Chromatogr., A. 2008; 1184:191-219.

Desilets CP, Rounds MA, Regnier FE. Semipermeablesurface reversed-phase media for high-performance liquid chromatography. J. Chromatogr., A. 1991; 544:25-39.

Bakhtiar R, Majumdar TK. Tracking problems and possible solutions in the quantitative determination of small molecule drugs and metabolites in biological fluids using liquid chromatography–mass spectrometry. J Pharmacol Toxicol Methods. 2007; 55:227-243.

Lima VV, Cassiano NM, Cass QB. Desenvolvimento de colunas cromatográficas de meios de acesso restrito proteína imobilizada e suas avaliações para análise de fármacos com injeção direta de plasma humano. Quim. Nova. 2006; 29(1):72-78.

Krishnam TR, Ibraham I. Solid-phase extraction technique for the analysis of biological samples. J. Pharm. Biomed. Anal. 1994; 12(3):287-294.

Fagundes VF, Leite CP, Pianetti GA, Fernandes C. Rapid and direct analysis of statins in human plasma by column-switching liquid chromatography with restricted-access material. J. Chromatogr. B. 2014; 947- 948:8-16.

World Health Organization. [citad 2015 Jun 17]. Available from: http://www.who.int/en/.

Goodman LS, Gilman A. As Bases Farmacológicas da Terapêutica. 12 ed. Rio de Janeiro: Mcgraw-Hill; 2012.

Ramalho ACR, Lima ML. Insulina e Antidiabéticos Orais. In: Silva P, editor. Farmacologia. 7ed. Rio de Janeiro: Guanabara Koogan; 2006. p. 805-823.

Moreau RLM, Siqueira MEPB. Toxicologia Analítica. 1ª ed. Rio de Janeiro: Guanabara Koogan; 2008.

Polagani SR, Pilli NR, Gajula R, Gandu V. Simultaneous determination of atorvastatin, metformin and glimepiride in human plasma by LC–MS/MS and its application to a human pharmacokinetic study. J. Pharm. Anal. 2013; 3(1): 9-19.

El-Enany NM, Abdelal AA, Belal FF, Itoh YI, Nakamura MN. Development and validation of a repharsed phase-HPLC method for simultaneous determination of rosiglitazone and glimepiride in combined dosage forms and human plasma. Chem Cent J. 2012; 6(9):1-10.

Sultana N, Arayne MS, Iftikhar B. Simultaneous determination of atenolol, rosuvastatin, spironolactone, glibenclamide and naproxen sodium in pharmaceutical formulations and human plasma by RP-HPLC. J. Chin. Chem. Soc. 2008; 55:1022-1029.

Chakradhar L, Kallem R, Karthik A, Sundari BT, Ramesh S, Mullangi R, et al. A rapid and highly sensitive method for the determination of glimepiride in human plasma by liquid chromatography–electrospray ionization tandem mass spectrometry: application to a pre-clinical pharmacokinetic study. Biomed. Chromatogr. 2008; 22:58-63.

Yuzuak N, Ozden T, Eren S, Ozilhan S. Determination of glimepiride in human plasma by LC-MS-MS. Chromatographia. 2007; 66:165-168.

Gedeon C, Kapur B, Aleksa K, Koren G. A simple and rapid HPLC method for the detection of glyburide in plasma original research communication (analytical). Clin. Biochem. 2008; 41:167-173.

Souza KC, Andrade GF, Vasconcelos I, Viana IMO, Fernandes C, Souza EMB. Magnetic solid-phase extraction based on mesoporous silica-coated magnetic nanoparticles for analysis of oral antidiabetic drugs in human plasma. Mater. Sci. Eng., C. 2014; 40:275-280.

Viana IMO, Lima PPR, Soares CDV, Fernandes C. Simultaneous determination of oral antidiabetic drugs in human plasma using microextraction by packed sorbent and high-performance liquid chromatography. J. Pharm. Biomed. Anal. 2014; 96:241-248.

Song Y, Maeng J, Hwang H, Park J, Kim B, Kim J, et al. Determination of glimepiride in human plasma using semi-microbore high performance liquid chromatography with column-switching. J. Chromatogr., B. 2004; 810:143–149.

Polson C, Sarkar P, Incledon B, Raguvaran V, Grant R. Optimization of protein precipitation based upon effectiveness of protein removal and ionization effect in liquid chromatography-tandem mass spectrometry. J. Chomatogr. B. 2003; 785:263-275.

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Published

17-07-2019 — Updated on 10-03-2026

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How to Cite

Veloso Ferreira, J., Antônio Pianetti, G., & Fernandes, C. (2026). Bioanalytical method by column-switching with direct injection of human plasma for determination of sulphonylureas. Drug Analytical Research, 3(1), 16–22. https://doi.org/10.22456/2527-2616.91542 (Original work published July 17, 2019)

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ORIGINAL ARTICLES