Validasi Metode HPLC untuk Penetapan Aspirin dan Asam Salisilat dalam Plasma Kelinci (Lepus curpaeums) secara Simultan

Agus Siswanto; Achmad Fudholi; Akhmad Kharis Nugroho; Sudibyo Martono

doi:10.22435/jki.v6i2.2922

Agus Siswanto Fakultas Farmasi Universitas Muhammadiyah Purwokerto, Dukuh Waluh, Purwokerto, Indonesia
Achmad Fudholi Fakultas Farmasi Universitas Gadjah Mada, Sekip Utara, Jogjakarta, Indonesia
Akhmad Kharis Nugroho Fakultas Farmasi Universitas Gadjah Mada, Sekip Utara, Jogjakarta, Indonesia
Sudibyo Martono Fakultas Farmasi Universitas Gadjah Mada, Sekip Utara, Jogjakarta, Indonesia

DOI: https://doi.org/10.22435/jki.v6i2.2922

Keywords: Aspirin, Salicylic Acid, Method validation, HPLC

Abstract

Aspirin is a nonsteroidal anti-inflammatory drug which also has the effect of antiplatelet for stroke prevention. Aspirin inside human body is very easy to break down into salicylic acid as the main metabolite. The aim of this study is to develop and validate the method for determinating aspirin and salicylic acid concentration in plasma by HPLC. Method validation including system suitability test, linearity test, determination of LOD and LOQ, recovery, accuracy and precision. Concentration of analytes in blood is measured by HPLC using benzoic acid as internal standard, with condition Purospher column Endcapped Star RP-18 (250 x 4.6 mm id, 5 m), acetonitrile : buffer phosphate 20 mM pH 2.5 (30:70 v/v) as mobile phase, injection volume 20 mL, flow rate 1.5 mL/minute, and UVVis detector λ 230 nm. The results showed that the proposed method meets the requirements of system suitability and good linearity (r > 0,990) with LOQ (aspirin = 0.024 mg/mL, salicylic acid = 0.336 mg/mL) and LOD (aspirin = 0.007 mg/mL, salicylic acid = 0.101 mg/mL). The method of analysis provides recovery of 85-115 %, accuracy and precision in accordance with the requirements for bioanalytical with CV < 5 %. Therefore, the proposed method is applicable to determine of aspirin and salicylic acid concentration in plasma.

References

Patrono C, Rocca B. Aspirin: promise and resistance in the new millennium. Arteriosclerosis, Thrombosis, and Vascular Biology. 2008;28(1):25–32.

Kannan S, Manivannan R, Balasubramaniam A, Kumar NS. Formulation and evaluation of aspirin delayed release tablet. International Journal of Comprehensive Pharmacy. 2010;1(4):1-3.

Mullangi R, Sharma K, Srinivas NR. Review of hplc methods and hplc methods with mass spectrometric detection for direct determination of aspirin with its metabolite(s) in various biological matrices. Biomed. Chromatogr. 2012;26(8):906-41.

Sweetman SC, Editor. Martindale the complete drug reference. Thirty-sixth ed. London: Pharmaceutical Press; 2009.

Dressman JB, Nair A, Abrahamsson B, Barends, DM, Groot DW, Kopp S, et al. Biowaiver monograph for immediaterelease solid oral dosage forms: acetylsalicylic acid. Journal of Pharmaceutical Sciences. 2012;101(8):2653-67.

Hakim L. Farmakokinetika. Yogyakarta: Bursa Ilmu; 2011.

Rojeab Y, Martin S, White R, Montenery S, Mcwilliams M, Walker M, Kisor D. In vitro dissolution and pilot pharmacokinetic studies of acetylsalicylic acid from an orally disintegrating tablet formulation of lowdose aspirin. IJMCR. 2011;2(2):72-7.

Sankar R, Dachinamoorthi, Shekar C. A Comparative Pharmacokinetic study of Aspirin Suppositories and Aspirin Nanoparticles Loaded Suppositories. Clinic Pharmacol Biopharm. 2012;1(3):1-5.

Broome TA, Brown MP, Gronwall RR, Casey MF, Meritt KA. Pharmacokinetics and plasma concentrations of acetylsalicylic acid after intravenous, rectal, and intragastric administration to horses. The Canadian Journal of Veterinary Research. 2003;67:297-302.

Kumar S, Jamadar LD, Bhat K, Musmade PB, Vasantharaju SG, Udupa N. Analytical method development and validation for aspirin. Int.J.ChemTech Res. 2010;2(1):389-99.

Bamigbola, E.A., Ibrahim, M.A., Attama, A.A., dan Uzondu, A.L. In Vitro-in Vivo Correlation of Four Commercial Brands of Aspirin Tablets Marketed in Nigeria. African Journal of Pharmacy and Pharmacology. 2009;5:1648–54.

Yamamoto E, Takakuwa S, Kato T, Asakawa N. Sensitive determination of aspirin and its metabolites in plasma by LCUV using on-line solid-phase extraction with methycellulose-immobilized anionexchange restricted access media. Journal of Chromatography. 2007;846:132-38.

Moffat AC, Osselton MD, Widdop B, Watts J, editor. Clarke’s analysis of drugs and poisons in pharmaceuticals, body fluids and postmortem material. Fourth edition. London-Chicago: Pharmaceutical Press; 2011.

Yuwono M, Indrayanto G. Validation of chromatographic methods of analysis. Profiles of Drug Substances Excipiens and Related Methodology. 2005;32:243-59.

FDA. Guidance for industry bioanalytical method validation. Rockville: United States Pharmacopoeial Convention Inc; 2001.

Sugiyono. Statistika Untuk Penelitian. CV Alfabeta: Bandung; 2009.

Lindholm J. Development and validation of hplc method for analytical and preparative purposes [disertasi]. Uppsala: Universitatis Upsaliensis; 2004.

Ramjith US, Sunith DK, Radhakrishnan Sand Sameer PA. HPLC study of aspirin and aspirin derivatives. International Journal Of Research In Pharmacy And Chemistry . 2013;3(1):1-5.

Harmita. Petunjuk pelaksanaan validasi metode dan cara perhitungannya. Majalah Ilmu Kefarmasian. 2004;I:117-35.