ANALYTICAL METHOD DEVELOPMENT FOR DETERMINATION OF RESIDUAL ERYTHROMYCIN IN ORAL POLIOMYELITIS VACCINE BY HIGH-PERFORMANCE LIQUID CHROMATORAPHY

• Main Author: Tri Wibowo, Lukman
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/78849
• Institution: Institut Teknologi Bandung

Vaccine is a preparation that contains antigenic substrance which can generate active and specific immunity in human. Oral poliomyelitis vaccine is suspension which contains poliomyelitis virus type 1, 2, or 3; or in combination. In vaccine preparation, any other components other than antigen are normally added as excipients; such as adjuvant, stabilizer, solvent, and preservative. Erythromycin is known to be added in vaccine and used as antibacterial agent to prevent contamination. Purpose of this study is to develop a determiJ1ation method of residual erythromycin in oral poliomyelitis vaccine using high-performance liquid chromatography (HPLC) with UV detection. System Of HPLC used in this study are column Inertsil@ C8, 4,6 i.d. x 250 mm, particle size 5 gm,; detection at 215 nm; mobile phase acetonitrile : phosphate buffer 0.03 M pH 5 = 30 : 70; flow rate 1mL/min; and volume of injection 100 PL. Extraction method is liquid-liquid extraction. The method was validated by testing linearity, limit of detection, limit of quantitation, accuracy, and precicion. Linear curve of calibration obtained from standard solution from 40-400 pg/mL with linear regression equation y 1239.7x 8901.5 and correlation coefficient r 0.9992. While the linear regression equation obtained from spiked matrix with analyte from 40-400 pg/mL is y 1137.1 x — 20986 and correlation coefficient r — 0.9995. Limit of detection of erythromycin in standard solution and in matrix are 14.80 ug/mL and 11.75 ug/mL, while limit of quantization of erythromycin in standard solution and in matrix are 49.32 pg/mL and 39.18 ug/mL. Recovery of erythromycin ranged from 89.49 — 93.51% based on standard solution linearity and 100.78 — 107.13% based on spiked matrix linearity. Method showed a good Intraday precicion with coefficient of variance CV: 1.16, 0.51, and 1.59% in day 1 to 3, and CV 0.54% for interday precicion. These results indicate that this method was validated and can be used for determination of erythromycin in oral poliomyelitis vaccine.