Validasi Metode Penetapan Kadar Lisinopril dalam Spiked Plasma Secara Ultra Performance Liquid Chromatography Melalui Derivatisasi dengan 1-Fluoro 2,4 Dinitrobenzen

Ririn Sumiyani, Sudibyo Martono, . Sugiyanto


ABSTRACT: A rapid, accurate, and sensitive method for determining lisinopril in spiked plasma was developed by means using an Ultra Performance Liquid Chromatography (UPLC) with 1-fluoro 2,4 dinitrobenzen (FDNB) derivatization. Lisinopril was precolumn derivatized with FDNB at optimum condition, i.e. room temperature and borate buffer at pH 11, subsequently analyzed with UPLC. Isocratic condition of acetate buffer (0.01 M, pH 3.50) : acetonitrile : metanol = 70 : 10 : 20 (v/v/v) as mobile phase, 0.3 mL/min of flow rate at λ 296 nm were applied at Acquity BEH C18 column, resulting a linearity of lisinopril at range of concentration of 5,0-100 ng/mL (Y = 410,59x + 211,91, r = 0.93). The accuration of the established method was achieved by 88,59±6,01 to 101,70± 2,56% recovery, while the precision was shown with RSD value of 2,57- 8,16 %, limit of detection (LOD) instrument of 0,73 ng/mL and limit of quatification (LOQ) 2,44 ng/mL, dwith R2 = 0,9987 dan r = 0,9993. In addition, the resulted LOD and LOQ more or less similar with the published HPLC-MS-MS method (1.03-10.0 ng/mL). Hence, it could be concluded that the developed UPLC method can be used as an alternative method for determining lisinopril in plasma.

Keywords: Lisinopril, FDNB, derivatization, UPLC

ABSTRAK: Penetapan kadar lisinopril dalam spiked plasma secara Ultra Performance Liquid Chromatography (UPLC) melalui derivatisasi dengan 1-fluoro 2,4 dinitrobenzen (FDNB) merupakan metode yang cepat, sensitif dan akurat. Derivatisasi precolumn lisinopril dan FDNB optimum pada suhu kamar, suasana dapar borat pH 11,0, dilanjutkan analisis secara UPLC isokratis menggunakan kolom Acquity BEH C dan fase gerak dapar asetat (0,01 M pH 3,50): asetonitril:metanol (70: 10: 20, v/v/v), laju alir 0,3 mL/menit pada λ 296 nm, menghasilkan linieritas kadar lisinopril dalam spiked plasma pada rentang 5,0 -100 ng/mL terhadap luas area lisinopril-DNB dengan persamaan Y = 410,59x + 211,91 dengan R2 = 0,9987 dan r = 0,9993 Akurasi metode ditunjukkan dengan nilai % rekoveri sebesar 88,59±6,01 smpai dengan 101,70± 2,56 %. Ketelitian ditunjukkan dengan nilai RSD 2,57- 8,16 %, sedangkan Batas Deteksi Instrumen = 0,73 ng/ mL dan Batas Kuantitasi = 2,44 ng/mL. Hasil Batas Deteksi penelitian ini relatif sama dengan Batas Deteksi penetapan kadar lisinopril secara HPLC-MS (1,03- 10,0 ng/mL). Dengan demikian dapat disimpulkan bahwa metode ini berpotensi dikembangkan sebagai metode alternatif pengganti HPLC-MS untuk penetapan lisinopril dalam plasma.

Kata kunci: Lisinopril, FDNB, derivatisasi, UPLC

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