Development and validation of HPLC/UV-procedures of secnidazole determination in blood and urine
Keywords:secnidazole, high-performance liquid chromatography, blood, urine, sample preparation, validation, method of standard
Aim. The system of HPLC-analyzer MiLiChrome® A-02 is widely used in Ukrainian laboratories of forensic toxicology. The purpose is to apply the HPLC-analyzer system for secnidazole quantitative determination in biological liquids and carry out validation of the developed procedures.
Methods. Sample preparation of blood and urine was carried out in three ways – 1) liquid-liquid extraction with organic solvents immiscible with water, 2) amphiphylic solvents extraction and salting-out with ammonium sulphate, 3) complex application of liquid-liquid extraction with organic solvents immiscible with water and amphiphylic solvents extraction with salting-out. Chromatographic conditions: column – Æ2´75 mm, ProntoSIL 120-5-C18 AQ, 5 μm; temperature – 40°С; flow rate – 100 μl/min; Eluent A – 0.2 M LiClO4 – 0.005 M HClO4; Eluent B – acetonitrile; elution mode – linear gradient; detection – UV, 277 nm; volume of injection – 2 μl.
Results. Validation of all developed procedures has been carried out by such parameters as specificity, recovery, linearity, accuracy and precision in the variant of the method of standard. The results of analysis have shown the absence of peaks with the retention time, which is coincident with the secnidazole retention time, on the chromatograms of blank-samples for all variants of procedures of analyte isolation. All procedures of sample preparation show the high efficiency of secnidazole isolation both for blood and urine (at the level of 90 %). All examined procedures are characterized by the acceptable parameters of linearity, within-run and between-run accuracy and precision.
Conclusions. The set of HPLC-procedures of secnidazole quantitative determination in blood and urine has been developed. Validation of the developed procedures has been carried out; isopropanol application in the acid medium is optimal for biological liquids sample preparation
- Brook, I. (2016). Spectrum and treatment of anaerobic infections. Journal of Infection and Chemotherapy, 22 (1), 1–13. doi: http://doi.org/10.1016/j.jiac.2015.10.010
- Upcroft, P., Upcroft, J. A. (2001). Drug Targets and Mechanisms of Resistance in the Anaerobic Protozoa. Clinical Microbiology Reviews, 14 (1), 150–164. doi: http://doi.org/10.1128/cmr.14.1.150-164.2001
- Rose, M. D., Bygrave, J., Sharman, M. (1999). Effect of cooking on veterinary drug residues in food Part 9 Nitroimidazoles. The Analyst, 124 (3), 289–294. doi: http://doi.org/10.1039/a809062i
- Sobel, R., Sobel, J. D. (2015). Metronidazole for the treatment of vaginal infections. Expert Opinion on Pharmacotherapy, 16 (7), 1109–1115. doi: http://doi.org/10.1517/14656566.2015.1035255
- Samuelson, J. (1999). Why Metronidazole Is Active against both Bacteria and Parasites. Antimicrobial Agents and Chemotherapy, 43 (7), 1533–1541. doi: http://doi.org/10.1128/aac.43.7.1533
- Freeman, C. D., Klutman, N. E., Lamp, K. C. (1997). Metronidazole. A therapeutic review and update. Drugs, 54, 679–708. doi: http://doi.org/10.2165/00003495-199754050-00003
- Commission Regulation (EU) No. 37/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin (2010). Official Journal of the European Union, L15, 1–72.
- Lamp, K. C., Freeman, C. D., Klutman, N. E., Lacy, M. K. (1999). Pharmacokinetics and Pharmacodynamics of the Nitroimidazole Antimicrobials. Clinical Pharmacokinetics, 36 (5), 353–373. doi: http://doi.org/10.2165/00003088-199936050-00004
- Gillis, J. C., Wiseman, L. R. (1996). Secnidazole. A review of its antimicrobial activity, pharmacokinetic properties and therapeutic use in the management of protozoal infections and bacterial vaginosis. Drugs, 51 (4), 621–638. doi: http://doi.org/10.2165/00003495-199651040-00007
- El Walily, A. F. M., Abdine, H. H., Razak, O. A., Zamel, S. (2000). Spectrophotometric and HPLC determination of secnidazole in pharmaceutical tablets. Journal of Pharmaceutical and Biomedical Analysis, 22 (6), 887–897. doi: http://doi.org/10.1016/s0731-7085(99)00290-3
- Sun, H. W., Wang, F. C., Ai, L. F. (2007). Simultaneous determination of seven nitroimidazole residues in meat by using HPLC-UV detection with solid-phase extraction. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 857 (2), 296–300. doi: http://doi.org/10.1016/j.jchromb.2007.07.039
- Ravi, S. K., Naidu, M. U. R., Sekhar, E. C., Rao, T. R. K., Shobha, J. C., Rani, P. U., Surya, K. J. (1997). Rapid and selective analysis of secnidazole in human plasma using high-performance liquid chromatography with ultraviolet detection. Journal of Chromatography B: Biomedical Sciences and Applications, 691 (1), 208–211. doi: http://doi.org/10.1016/s0378-4347(96)00419-7
- Li, X., Sun, J., Wang, G., Zheng, Y., Yan, B., Xie, H. et. al. (2007). Determination of secnidazole in human plasma by high-performance liquid chromatography with UV detection and its application to the bioequivalence studies. Biomedical Chromatography, 21 (3), 304–309. doi: http://doi.org/10.1002/bmc.758
- Zhu, D., Hu, K., Tao, W., Feng, L., Duan, H., Jiang, X., Chen, J. (2011). Evaluation of the Bioequivalence and Pharmacokinetics of Two Formulations of Secnidazole after Single Oral Administration in Healthy Volunteers. Arzneimittelforschung, 57 (11), 723–726. doi: http://doi.org/10.1055/s-0031-1296674
- Tenenbaum, H., Cuisinier, F. J. G., Liboux, A., Pichard, E., Montay, G., Frydman, A. (1993). Secnidazole concentrations in plasma and crevicular fluid after a single oral dose. Journal of Clinical Periodontology, 20 (7), 505–508. doi: http://doi.org/10.1111/j.1600-051x.1993.tb00398.x
- Mitrowska, K., Antczak, M. (2017). Development and validation of a liquid chromatography with tandem mass spectrometry method for the determination of nitroimidazole residues in beeswax. Journal of Separation Science, 40 (5), 1158–1166. doi: http://doi.org/10.1002/jssc.201600928
- Rúbies, A., Sans, G., Kumar, P., Granados, M., Companyó, R., Centrich, F. (2015). High-throughput method for the determination of nitroimidazoles in muscle samples by liquid chromatography coupled to mass spectrometry. Analytical and Bioanalytical Chemistry, 407 (15), 4411–4421. doi: http://doi.org/10.1007/s00216-014-8436-x
- Azarova, I. N., Baram, G. I. (2014). Primenenie perhlorata litiya v obraschenno-fazovoy vyisokoeffektivnoy zhidkostnoy hromatografii aminosoedineniy. Sorbtsionnyie i hromatograficheskie protsessyi, 14 (1), 858–867.
- Shovkova, O. V., Klimenko, L. Yu., Shovkova, Z. V., Kostina, T. A. (2018). Development and validation of HPLC/UV-procedure of secnidazole determination. Journal of Organic and Pharmaceutical Chemistry, 16 (3 (63)), 30–38. doi: http://doi.org/10.24959/ophcj.18.948
- Klimenko, L. Yu. (2016). The integrated approach to development and validation of the procedures of analytes quantification in biological fluids for chemical and toxicological analysis. Kharkiv, 816.
- Moffat, A. C., Osselton, M. D., Widdop, B. (2011). Clarke’s analysis of drugs and poisons in pharmaceuticals, body fluids and postmortem material: 4th ed. London: Pharmaceutical Press, 2609.
- Klimenko, L. Yu. (2014). Development of approaches to determination of linearity, accuracy and precision of UV-spectrophotometric methods of quantitative determination by the method of standard in forensic and toxicological analysis. Farmatsyia Kazakhstana, 4 (155), 31–35.
- Klimenko, L. Yu., Trut, S. M., Poluyan, S. М. (2014). Determination of validation characteristics of UV-spec-trophotometric method of doxylamine quantitative determination in blood in the variant of the method of standard. News of Pharmacy, 2 (78), 53–58. doi: http://doi.org/10.24959/nphj.14.1969
How to Cite
Copyright (c) 2018 Oksana Shovkova, Lina Klimenko, Zoia Shovkova, Olena Mykytenko
This work is licensed under a Creative Commons Attribution 4.0 International License.
Our journal abides by the Creative Commons CC BY copyright rights and permissions for open access journals.