Quantitative determination of 0.05 % chlorhexidine solution by capillary electrophoresis


  • A. M. Sampiiev Kuban State Medical University, Krasnodar, Russian Federation,
  • N. A. Davitavian Kuban State Medical University, Krasnodar, Russian Federation,
  • Ye. B. Nikiforova Kuban State Medical University, Krasnodar, Russian Federation,
  • Yu. F. Yakuba North Caucasian Federal Scientific Centre of Horticulture, Viticulture, Winemaking, Krasnodar, Russian Federation,




chlorhexidine, capillary zone electrophoresis, biguanides, amidines, antiseptics



The aim of the work was to estimate the quantitative content of 0.05 % chlorhexidine solution by capillary electrophoresis.

Materials and methods. The object of the study was the drug 0.05 % solution of chlorhexidine for local and external use of pharmacopoeia quality. The study was carried out on a Kapel-104T device, equipped with an ultraviolet photometric detector and personal computer with the Multichrom software.

Electrophoretic determination of the quantitative content of chlorhexidine was carried out in the UV region of the spectrum at a wavelength of 254 nm with a positive voltage on a capillary of 16 kV and a recommended capillary thermostating from 20 °C to 30 °C. The sample was introduced in pneumatical mode in the 30 mbar for 5 seconds. The analysis time was 5 minutes. As the leading electrolyte an aqueous solution consisting of 0.6 % solution of imidazole, 0.6 % solution of sodium tetraborate and 0.5 % solution of tartaric acid were used. The determination of chlorhexidine in the test solution was carried out in the UV spectral region at a wavelength of 254 nm, since at this wavelength one of the absorption maxima of the indicated drug was observed.

Results. The validation analysis showed that the technique is characterized by specificity, linearity in the concentration range from 100 μg/ml to 700 μg/ml (y = 0.3438х - 12.4789; r = 0.9913), accuracy for chlorhexidine levels of 80–120 % (R = 98.82–100.44 %), precision (Sr = 0.44–0.61 %). The relative error of the result of separate determination of chlorhexidine in 0.05 % solution using capillary electrophoresis was 1.28 %.

Conclusions. Quantitative determination of 0.05 % chlorhexidine solution by capillary electrophoresis was carried out and the parameters of the validation of the electrophoretic technique were established. The proposed electrophoretic conditions can serve as the basis for the development of methods for the quantitative determination of chlorhexidine bigluconate as part of this drug.



Zverkov, A. V., & Zouzova, A. P. (2013) Khlorgeksidin: nastoyashchee i budushchee odnogo iz osnovnykh antiseptikov [Chlorhexidine: Past, Present, and Future of the Famous Antiseptic Agent]. Klinicheskaya mikrobiologiya i antimikrobnaya khimioterapiya, 15(4), 279–285. [in Russian].

Gerke, A. N. (2015) Osnovnye principy mestnoj antimikrobnoj terapii v dermatologii [The basis of topical antimicrobial therapy in dermatology]. VetPharma, 1(23), 66–75. [in Russian].

Kasikhina, E. I. (2013) Hlorgeksidin: obzor lechebnykh vozmozhnostej i potencial'nykh klinicheskikh pokazanij v praktike akushera-ginekologa i venerologa [Chlorhexidine: a review of treatment options and potential clinical indications in the practice of an obstetrician/gynecologist and a venereologist]. Akurshestvo i ginekologiya, 4, 4–9. [in Russian].

Pogosyan, M.A. (2015) Hlorgeksidin – antiseptik, ne privodyashchij k bakteriorezistentstnosti [Chlorhexidine is an antiseptic that does not lead to bacterioresistance]. Byulleten' mededicinskih internet-konferencij, 5(10), 1234–1235. [in Russian].

Kouzmina, E., Lapatina, A., & Smirnova, T. (2014) Opolaskivateli polosti rta s khlorgeksidinom: e'ffektivnost' i bezopasnost' primeneniya (obzor literatury) [Efficacy and safety of chlorhexidine mouthrinses (literature review)]. Dental Forum, 2(53), 34–39. [in Russian].

Kvashnina, D. V., & Kovalishena, O. V. (2016) Ocenka primeneniya khlorgeksidina kak antisepticheskogo sredstva [Evaluation of chlorhexidine application as an antiseptic]. Medicinskij al'manakh, 3(43), 62–66. [in Russian].

Dounar, H. G., & Rzheuski, S. E. (2017) Antimikrobnaya aktivnost' gelya hlorgeksidina biglyukonata, prednaznachennogo dlya lecheniya kandidoza polosti rta [Antimicrobial activity of the gel with chlorhexidine digluconate intended for the treatment of oral candidiasis]. Vestnik Vitebskogo gosudarstvennogo

medicinskogo universiteta, 16(3), 91–97. [in Russian].

Lin, S.-C., Huang, C.-F., Shen, L.-J. Wang, H.-J., Lin, Ch.-Y., et al. (2015) Formulation and stability of an extemporaneous 0.02% chlorhexidine digluconate ophthalmic solution. J. Formos. Med. Assos., 114, 1162–1169. https://doi.org/10.1016/j.jfma.2014.08.003

George, J., Klika, A. K., & Higuera, C. A. (2017) Use of Chlorhexidine Preparations in Total Joint Arthroplasty. J. Bone Jt Infect., 2(1), 15–22. doi: 10.7150/jbji.16934

Fiorentino, F. A. M., Corrêa, M. A. & Salgado, H. R. N. (2010) Analytical Methods for the Determination of Chlorhexidine: A Review. Crit. Rev. Anal. Chem., 40, 89–101. doi: 10.1080/10408340903232020

Tyzhigirova, V. V., & Timofeeva, O. N. (2017) Analiz lekarstvennykh preparatov khlorgeksidina biglyukonata i miramistina metodami UF-spektrofotometrii i tonkoslojnoj khromatografii [Analysis of drugs chlorhexidine bigluconate and miramistine by UV- spectrophotometry and thin-layer chromatography]. Innovacionnye tekhnologii v farmacii Proceedings of the All-Russian Scientific and Practical Conference with international participation, (P. 115–121). Sumy: IGMU. [in Russian].

Mubeen, R. S., Mantri, A. P., Singh, S. K., et al. (2016) Simultaneous estimation of chlorhexidine digluconate and miconazole nitrate by RP- HPLC. European Journal of Biomedical and Pharmaceutical sciences, 3, 617–620.

Mohamme, T. G., & Abdel Aziz, E. M. (2017) Development and validation of a simple, fast, isocratic stability indicating RP-HPLC-UV method for the determination of chlorhexidine and its impurity para-chloroaniline in bulk and finished product. Journal of Pharmacy, 7, 01–08. doi: 10.9790/3013-0706010108

Bogdanovska, L., Saliu, S., Popovska, M., Dimitrovska, A., Ugrinova, L., & Petkovska, R. (2014) Development and validation of RP–HPLC assay of chlorhexidine in gingival crevicular fluid. Arh. farm., 64, 69–82. doi: 10.5937/arhfarm1402069B

Siddiqui, M. R., AlOthman, Z. A., & Rahman, N. (2017) Analytical techniques in pharmaceutical analysis: A review. Arabian Journal of Chemistry, 10(1), 1409–1421. https://doi.org/10.1016/j.arabjc.2013.04.016

Kumar, M., Bhatia, R., & Rawal, R. K. (2018) Applications of various analytical techniques in quality control of pharmaceutical excipients. J. Pharm Biomed Anal., 157, 122–136. https://doi.org/10.1016/j.jpba.2018.05.023

Ma, H., Bai, Y., Li, J., & Chang, Y. X. (2018) Screening bioactive compounds from natural product and its preparations using capillary electrophoresis. Electrophoresis, 39(1), 260–274. doi: 10.1002/elps.201700239

Hamdan, I. I. (2017) Capillary electrophoresis in the analysis of pharmaceuticals in environmental water: A critical review. J. Liq. Chromatogr. Relat. Technol., 40, 111–125. doi: 10.1080/10826076.2017.1293550

Abramova, E., Alekseeva, N., & Egorov, A. (2012) Attestaciya/kvalifikaciya (validaciya) oborudovaniya i analiticheskikh metodov v farmacevticheskom proizvodstve [Instrumentation and Analytical Methods Attestation/Qualification (Validation) when Carrying out Pharmaceutical Production]. Analitika, 1, 60–62. [in Russian].

(2015) Gosudarstvennaya farmakopeya Rossijskoj Federacii [State Pharmacopoeia of the Russian Federation] Retrieved from http://femb.ru/feml. [in Russian].

(2017). USP39-NF34. Rockville. Retrieved from http://www.usp.org/ sites/default/files/usp_pdf/EN/products/usp39-nf34-index-supplement1.pdf.


How to Cite

Sampiiev, A. M., Davitavian, N. A., Nikiforova, Y. B., & Yakuba, Y. F. (2019). Quantitative determination of 0.05 % chlorhexidine solution by capillary electrophoresis. Zaporozhye Medical Journal, (4). https://doi.org/10.14739/2310-1210.2019.4.173352



Basic research