ESTABLISH PROCEDURE DETERMINATION OF 2 - PHENOXYETHANOL CONTENT IN VACCINES BY REVERSED PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Abstract

Currently, 2 - phenoxyethanol (ethylene glycol monophenylete) with the chemical formula C₈H₁₀O₂ is used as a preservative in cosmetic products, drugs and vaccines. Commercial products such as cosmetics, drugs and vaccines must be tested for 2 - phenoxyethanol content before use in accordance with the regulations of the Pharmacopoeia and the National Regulatory Agency. This study was conducted to develop a procedure to determine 2 - phenoxyethanol content in vaccines according to the guidelines of ICH and the actual conditions of the Department. This study uses experimental describing method in the laboratory. A procedure for determination of 2 - phenoxyethanol content in havax and tetraxim vaccines on Thermo Scientific chromatography system, column C18 (150 x 4.6 mm), mobile phase of acetonitrile/water solution at the ratio 50/50 (v/v), DAD detector, wavelength 270 nm has been established and validated with detail results: system suitability (use 2 - phenoxyethanol solution 6 mg/ml), %RSD of the peak area is ​​0,39%; %RSD of retention time is 0,02%; tail drag coefficient is 1,22; number of theoretical plates is 13182. The repeatability on tetraxim and havax sample are respective: 0,37%; 0,42%. The reproducibility on tetraxim and havax samples are respective: 0,71%,; 0,48%. The recovery (R%) of the 2 - phenoxyethanol solution 5 µl/ml added to the tetraxim test sample are from 96,99% to 100,04%; of 2 - phenoxyethanol solution 6 mg/ml was added to the havax sample are from 96,80% to 100,43%. Robustness is determined through a small change in flow rate are respective: 0,9; 1,0; 1,1 ml/min, acetonitrile/water mobile phase ratio are respective: 52/48; 50/50; 48/52 (v/v); column temperature are respective: 20, 25, 30ºC; %RSD results of the 2 - phenoxyethanol content at the standard points are from 0,28 to 2,98%; The bias Δi of the standard points are from 0,01 to 5,31%. LOQ = 0,0026%. Linearity: %RSD at standard points from 1,15 - 3,19%; The bias of the standard points are from 0,16 to 6,68%. The specificity studied on degradation and forced conditions resulted in the secondary peak separating the main peak. The uncertainty of tetraxim and havax samples are respective 0,052 µl/dose; 0,070 mg/ml. We have succeeded in this study, achieving the purposes of optimizing and validating the procedure of determining 2 - phenoxyethanol content in vaccines in accordance with the practical conditions at the Department, in accordance with the vaccine containing 2 - phenoxethanol has different matrices from simple to complex. The uncertainty of the two test samples is consistent with the results for 2 -phenoxyethanol content in the sample.