DOI: https://doi.org/10.24959/ophcj.19.964

Development and validation of HPLC/UV-procedures of efavirenz determination in biological fluids

O. I. Slabiak, I. M. Ivanchuk, L. Yu. Klimenko, Iu. S. Kolisnyk

Abstract


Efavirenz is a non-nucleoside reverse transcriptase inhibitor with a number of side effects manifested by psychiatric symptoms. There are cases of acute poisoning due to administration of efavirenz, including cases of suicide attempts; therefore, efavirenz may be approved as a toxic compound in forensic toxicology.

Aim. To apply the MiLiChrome® A-02 HPLC-analyzer system for efavirenz quantitative determination in biological fluids and perform validation of the procedures developed.

Results and discussion. Three HPLC/UV-procedures of efavirenz determination in blood and urine have been proposed. Validation of all procedures developed has been performed by such parameters as specificity, recovery, linearity, accuracy and precision in the variants of the methods of calibration curve and standard. The results of analysis have shown the absence of peaks with the retention time, which is coincident with the efavirenz retention time, on the chromatograms of blank-samples for all variants of procedures of the analyte isolation. All procedures of sample preparation show the high efficiency of efavirenz isolation both for blood and urine (at the level of 90 %). All procedures studied are characterized by the acceptable parameters of linearity, within-run and between-run accuracy and precision.

Experimental part. Sample preparation of blood and urine was carried out in three ways – 1) liquid-liquid extraction with the mixture of chloroform and 2-propanol (80 : 20); 2) 2-propanol extraction and salting-out with ammonium sulfate; 3) acetonitrile extraction and salting-out with ammonium sulfate. The chromatographic conditions were as follows: the column – Ø2 × 75 mm, ProntoSIL 120-5-C18 AQ, 5 μm; temperature – 40 °С; the flow rate – 100 μl/min; Eluent A – 0.2 M LiClO4 – 0.005 M HClO4; Eluent B – acetonitrile; the elution mode – linear gradient; detection – UV, 247 nm; the volume of injection – 2 μl.

Conclusions. The set of HPLC-procedures of efavirenz quantitative determination in blood and urine has been developed. Validation of the procedures developed has been performed.


Keywords


efavirenz; high-performance liquid chromatography; blood; urine; sample preparation; validation

Full Text:

PDF

References


Bastos, M. M., Costa, C. C. P., Bezerra, T. C., da Silva, F. C., Boechat, N. (2016). Efavirenz a nonnucleoside reverse transcriptase inhibitor of first-generation: approaches based on its medicinal chemistry. European Journal of Medicinal Chemistry, 108, 455–465. https://doi.org/10.1016/j.ejmech.2015.11.025

Waters, L., John, L., Nelson, M. (2007). Non-nucleoside reverse transcriptase inhibitors: a review. International Journal of Clinical Practice, 61 (1), 105–118. https://doi.org/10.1111/j.1742-1241.2006.01146.x

Porche, D. J. (2000). Efavirenz. Journal of the Association of Nurses in Aids Care, 11 (3), 95–98. https://doi.org/10.1016/s1055-3290(06)60280-x

Andany, N., Gold, W. L. (2016). Single–tablet antiretroviral treatment (once daily). Canadian Medical Association Journal, 188 (13), 971. https://doi.org/10.1503/cmaj.151412

Kenedi, C. A., Goforth, H. W. (2011). A systematic review of the psychiatric side-effects of efavirenz. AIDS and Behavior, 15 (8), 1803–1818. https://doi.org/10.1007/s10461-011-9939-5

Decloedt, E. H., Maartens, G. (2013). Neuronal toxicity of efavirenz: a systematic review. Expert Opinion on Drug Safety, 12 (6), 841–846. https://doi.org/10.1517/14740338.2013.823396

Abers, M. S., Shandera, W. X., Kass, J. S. (2014). Neurological and psychiatric adverse effects of antiretroviral drugs. CNS Drugs, 28 (2), 131–145. https://doi.org/10.1007/s40263-013-0132-4

Mutwa, P. R., Fillekes, Q., Malgaz, M., Tuyishimire, D., Kraats, R., Boer, K. R., Burger, D. M., van Schaik, R. H., Muganga, N., Geelen, S. P. (2012). Mid-dosing interval efavirenz plasma concentrations in HIV–1–infected children in Rwanda: treatment efficacy, tolerability, adherence, and the influence of CYP2B6 polymorphisms. Journal of Acquired Immune Deficiency Syndromes, 60 (4), 400–404. https://doi.org/10.1097/qai.0b013e3182569f57

Fillekes, Q., Natukunda, E., Balungi, J., Kendall, L., Bwakura-Dangarembizi, M., Keishanyu, R., Ferrier, A., Lutakome, J., Gibb, D. M., Burger, D. M., Walker, A. S. (2011). Pediatric underdosing of efavirenz: a pharmacokinetic study in Uganda. Journal of Acquired Immune Deficiency Syndromes, 58 (4), 392–398. https://doi.org/10.1097/qai.0b013e318235e560

Mollan, K. R., Smurzynski, M., Eron, J. J., Daar, E. S., Campbell, T. B., Sax, P. E., Gulick, R. M., Na, L., O’Keefe, L., Robertson, K. R., Tierney, C. (2014). Association between efavirenz as initial therapy for HIV-1 infection and increased risk for suicidal ideation or attempted or completed suicide: an analysis of trial data. Annals of Internal Medicine, 161 (1), 1–10. https://doi.org/10.7326/m14-0293

Efavirenz: suicides. (2015). Prescrire International, 24 (161), 156.

Curley, P., Siccardi, M., Moss, D. M., Owen, A. (2016). Development and validation of an LC-MS/MS assay for the quantification of efavirenz in different biological matrices. Bioanalysis, 8 (20), 2125–2134. https://doi.org/10.4155/bio-2016-0021

Montgomery, E. R., Edmanson, A. L., Cook, S. C., Hovsepian, P. K. (2001). Development and validation of a reverse-phase HPLC method for analysis of efavirenz and its related substances in the drug substance and in a capsule formulation. Journal of Pharmaceutical and Biomedical Analysis, 25 (2), 267–284. https://doi.org/10.1016/s0731-7085(00)00495-7

Saras-Nacenta, M., López-Púa, Y., Lípez-Cortés, L. F., Mallolas, J., Gatell, J. M., Carné, X. (2001). Determination of efavirenz in human plasma by high-performance liquid chromatography with ultraviolet detection. Journal of Chromatography B: Biomedical Sciences and Applications, 763 (1–2), 53–59. https://doi.org/10.1016/s0378-4347(01)00357-7

Ramachandran, G., Kumar, A. K., Swaminathan, S., Venkatesan, P., Kumaraswami, V., Greenblatt, D. J. (2006). Simple and rapid liquid chromatography method for determination of efavirenz in plasma. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 835 (1–2), 131–135. https://doi.org/10.1016/j.jchromb.2006.03.014

Barreiros, L., Cunha-Reis, C., Silva, E. M., Carvalho, J. R., das Neves, J., Sarmento, B., Segundo, M. A. (2017). Development and validation of a liquid chromatography-MS/MS method for simultaneous quantification of tenofovir and efavirenz in biological tissues and fluids. Journal of Pharmaceutical and Biomedical Analysis, 136, 120–125. https://doi.org/10.1016/j.jpba.2016.12.028

Azarova, I. N., Baram, G. I. (2014). Sorbtsionnye i Khromatograficheskie Protsessy, 14 (1), 858–867.

Slabiak, O. I., Ivanchuk, I. M., Klimenko, L. Yu., Tokaryk, G. V., Lebedynets, V. O., Yaremenko, V. D. (2018). Development and validation of HPLC/UV-procedure for efavirenz quantitative determination. Journal of Pharmaceutical Sciences and Research, 10 (11), 2829–2835.

Klymenko, L. Yu. (2015). Kompleksnyi pidkhid do rozrobky ta validatsii metodyk kilkisnoho vyznachennia analitiv u biolohichnykh ridynakh v khimiko-toksykolohichnomu analizi. Doctor’s thessis. Kharkiv, 816.

Guidance for the Validation of Analytical Methodology and Calibration of Equipment used for Testing of Illicit Drugs in Seized Materials and Biological Specimens (2009). United Nations Office on Drugs and Crime, Laboratory and Scientific Section,New York.

Slabiak, O. I., Ivanchuk, I.M., Klimenko, L. Yu., Mykytenko, O. Ye. (2018). Study of efavirenz extraction from aqueous solutions. Nauka i innovaciia, 1, 165–171.


GOST Style Citations


1.         Efavirenz a nonnucleoside reverse transcriptase inhibitor of first-generation: approaches based on its medicinal chemistry / M. M. Bastos, C. C. P. Costa, T. C. Bezerra et al. // Eur. J. Med. Chem. – 2016. – Vol. 108. – P. 455–465. https://doi.org/10.1016/j.ejmech.2015.11.025

2.         Waters, L. Non-nucleoside reverse transcriptase inhibitors: a review / L. Waters, L. John, M. Nelson // Int. J. Clin. Pract. – 2007. – Vol. 61, Issue 1. – P. 105–118. https://doi.org/10.1111/j.1742-1241.2006.01146.x 

3.         Porche, D. J. Efavirenz / D. J. Porche // JANAC. – 2000. – Vol. 11, Issue 3. – P. 95–98. https://doi.org/10.1016/s1055-3290(06)60280-x 

4.         Andany, N. Single-tablet antiretroviral treatment (once daily) /N. Andany, W. L. Gold // CMAJ. – 2016. – Vol. 188, Issue 13. – P. 971. https://doi.org/10.1503/cmaj.151412 

5.         Kenedi, C. A. A systematic review of the psychiatric side-effects of efavirenz / C. A. Kenedi, H. W. Goforth // AIDS Behav. – 2011. – Vol. 15, Issue 8. – P. 1803–1818. https://doi.org/10.1007/s10461-011-9939-5 

6.         Decloedt, E. H. Neuronal toxicity of efavirenz: a systematic review / E. H. Decloedt, G. Maartens // Expert Opin. Drug Saf. – 2013. – Vol. 12, Issue 6. – P. 841–846. https://doi.org/10.1517/14740338.2013.823396 

7.         Abers, M. S. Neurological and psychiatric adverse effects of antiretroviral drugs / M. S. Abers, W. X. Shandera, J. S. Kass // CNS Drugs. – 2014. – Vol. 28, Issue 2. – P. 131–145. https://doi.org/10.1007/s40263-013-0132-4 

8.         Mid-dosing interval efavirenz plasma concentrations in HIV-1-infected children inRwanda: treatment efficacy, tolerability, adherence, and the influence of CYP2B6 polymorphisms / P. R. Mutwa, Q. Fillekes, M. Malgaz et al. // J. Acquir. Immune Defic. Syndr. – 2012. – Vol. 60, Issue 4. – P. 400–404. https://doi.org/10.1097/qai.0b013e3182569f57 

9.         Pediatric underdosing of efavirenz: a pharmacokinetic study in Uganda / Q. Fillekes, E. Natukunda, J. Balungi et al. // J. Acquir. Immune Defic. Syndr. – 2011. – Vol. 58, Issue 4. – P. 392–398. https://doi.org/10.1097/qai.0b013e318235e560

10.       Association between efavirenz as initial therapy for HIV-1 infection and increased risk for suicidal ideation or attempted or completed suicide: an analysis of trial data / K. R. Mollan, M. Smurzynski, J. J. Eron et al. // Ann. Intern. Med. – 2014. – Vol. 161, Issue 1. – P. 1–10. https://doi.org/10.7326/m14-0293 

11.       Efavirenz: suicides // Prescrire Int. – 2015. – Vol. 24, Issue 161. – P. 156.

12.       Development and validation of an LC-MS/MS assay for the quantification of efavirenz in different biological matrices / P. Curley, M. Siccardi, D. M. Moss, A. Owen // Bioanalysis. – 2016. – Vol. 8, Issue 20. – P. 2125–2134. https://doi.org/10.4155/bio-2016-0021 

13.       Development and validation of a reverse-phase HPLC method for analysis of efavirenz and its related substances in the drug substance and in a capsule formulation / E. R. Montgomery, A. L. Edmanson, S. C. Cook, P. K. Hovsepian // J. Pharm. Biomed. Anal. – 2001. – Vol. 25, Issue 2. – P. 267–284. https://doi.org/10.1016/s0731-7085(00)00495-7 

14.       Determination of efavirenz in human plasma by high-performance liquid chromatography with ultraviolet detection / M. Saras-Nacenta, Y. López-Púa, L. F. Lípez-Cortés et al. // J. Chromatogr. B Biomed. Sci. Appl. – 2001. – Vol. 763, Issues 1–2. – P. 53–59. https://doi.org/10.1016/s0378-4347(01)00357-7 

15.      Simple and rapid liquid chromatography method for determination of efavirenz in plasma / G. Ramachandran, A. K. Kumar, S. Swaminathan et al. // J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. – 2006. – Vol. 835, Issues 1–2. – P. 131–135. https://doi.org/10.1016/j.jchromb.2006.03.014 

16.       Development and validation of a liquid chromatography-MS/MS method for simultaneous quantification of tenofovir and efavirenz in biological tissues and fluids / L. Barreiros, C. Cunha-Reis, E. M. Silva et al. // J. Pharm. Biomed. Anal. – 2017. – Vol. 136. – P. 120–125. https://doi.org/10.1016/j.jpba.2016.12.028 

17.       Азарова, И. Н. Применение перхлората лития в обращенно-фазовой высокоэффективной жидкостной хроматографии аминосоединений / И. Н. Азарова, Г. И. Барам // Сорбционные и хроматографические процессы. – 2014. – Т. 14, Вып. 1. – C. 858–867.

18.       Development and validation of HPLC/UV-procedure for efavirenz quantitative determination / O. I. Slabiak,I.M. Ivanchuk, L. Yu. Klimenko et al. // J. Pharm. Sci. & Res. – 2018. – Vol. 10 (11). – P. 2829–2835.

19.       Клименко, Л. Ю. Комплексний підхід до розробки та валідації методик кількісного визначення аналітів у біологічних рідинах в хіміко-токсикологічному аналізі: дис. … докт. фарм. наук / Л. Ю. Клименко. – Х., 2015. – 816 с.

20.       Guidance for the Validation of Analytical Methodology and Calibration of Equipment used for Testing of Illicit Drugs in Seized Materials and Biological Specimens / United Nations Office on Drugs and Crime, Laboratory and Scientific Section. –New York: United Nations, 2009. – 70 p.

21.      Study of efavirenz extraction from aqueous solutions / O. I. Slabiak, I. M. Ivanchuk, L. Yu. Klimenko, O. Ye. Mykytenko // Наука и инновация. – 2018. – №1. – С. 165–171.





Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

Abbreviated key title: Ž. org. farm. hìm.

ISSN 2518-1548 (Online), ISSN 2308-8303 (Print)