Abstract
A rapid and simple liquid chromatographic method with UV detection has been developed for the determination of hydrochlorothiazide (HCTZ), cilazapril (CL) and its active metabolite cilazaprilat (CLT) in urine. Sample preparation for urine consisted of solid-phase extraction using styrene-divinylbenzene (SDB-2) cartridges. The chromatographic system was a Zorbax Eclipse XDB-C18 column with a mixture of methanol and 10 mM phosphate buffer, pH 2.3 with gradient (20 to 60% of methanol) as mobile phase at a flow rate of 1.0 mL min−1. The detection was performed at the wavelength of 206 nm. Enalapril maleat was used as an internal standard. The detector response was linear in the range of 2.4–30.0, 1.6–15.0 and 1.8–20.0 μg mL−1 for HCTZ, CL and CLT, respectively. LOQ was determined to be 2.4, 1.6 and 1.8 μg mL−1 for HCTZ, CL and CLT, respectively. Both intra- and inter-day precision were within acceptable limits. The method has been applied to urine samples obtained from three hypertensive patients after intake of HCTZ and CL therapeutic dose.
References
Dollery C (1999) Therapeutic drugs, 2nd edn. Churchill Livingstone, Edinburgh, UK
Register of Medicines of Republic of Serbia (2008) BB-Soft, Belgrade, Serbia
Erk N (1999) J Pharm Biomed Anal 20:155–167. doi:10.1016/S0731-7085(99)00009-6
Erk N, Onur F (1996) Anal Lett 29:1963–1974. doi:10.1080/00032719608000802
Dinc E, Baleanu D (2002) J Pharm Biomed Anal 30:715–723. doi:10.1016/S0731-7085(02)00359-X
Atay O, Tamer U, Arikan D (2001) Anal Lett 34:1153–1161. doi:10.1081/AL-100104961
Cetin G, Sungur S (2005) Fresenius Environ Bull 14(5):353–356
Hillaert S, De Grauwe K, Van den Bossche W (2001) J Chromatogr A 924:439–449. doi:10.1016/S0021-9673(01)00714-2
Lee HW, Park WS, Cho SH, Kim MH, Seo JH, Kim YW, Kim SS, Lee YS, Lee KT (2007) Talanta 71:62–67. doi:10.1016/j.talanta.2006.03.020
Prieto JA, Akesolo U, Jiménez RM, Alonso RM (2001) J Chromatogr A 916:279–288. doi:10.1016/S0021-9673(01)00565-9
Prieto JA, Jiménez RM, Alonso RM, Ortiz E (2001) J Chromatogr B 754:23–34. doi:10.1016/S0378-4347(00)00580-6
Prieto JA, Jiménez RM, Alonso RM (1998) J Chromatogr B 714:285–292. doi:10.1016/S0378-4347(98)00226-6
ICH Harmonised tripartite guideline validation of analytical procedures: text and methodology Q2(R1). Parent Guideline dated October 27, 1994 (Complementary guideline on methodology dated 6 November 1996 incorporated in November 2005). http://www.ich.org/LOB/media/MEDIA417.pdf
US Food and Drug Administration, Center for Drug Evaluation and Research (CDER) (2001) Guidance for industry: bioanalytical method validation. http://www.fda.gov/CDER/GUIDANCE/4252fnl.pdf
Williams PEO, Brown AN, Rajaguru S, Francis RJ, Bell AJ, Dewland PM (1990) Eur J Drug Metab Pharmacokinet 15:63–67 PMID: 2166672
Acknowledgment
We gratefully acknowledge the financial support from the Ministry of Science and Environment of the Republic of Serbia, grants No. 142072 and 142047.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Vujić, Z., Crevar, M., Obradović, V. et al. Simultaneous Determination of Hydrochlorothiazide, Cilazapril and Its Active Metabolite Cilazaprilat in Urine by Gradient RP-LC. Chroma 70, 1221–1225 (2009). https://doi.org/10.1365/s10337-009-1286-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1365/s10337-009-1286-6