STABILIZATION OF VITAMIN A USING VITAMIN E AS ANTIOXIDANT IN LYOPHILIZED AUTOLOGOUS SERUM AND ITS ANTIBACTERIAL PROPERTIES

Abstract

Dry eye syndrome is defined as an ocular surface disease caused by a lack of tear production or increased tear evaporation. Dry eye treatment with conventional drugs were less effective due to the nature and composition of the drugs that less akin to tears. Autologous serum is used as eyedrops in dry eye disease because it is natural and does not cause allergic reactions, as well as having biomechanical and biochemical properties that resemble normal tears. In liquid form, the content of serum didn’t last long, so the storage method of the serum was developed by using lyophilization with sucrose as lyoprotectant. The use of sucrose in lyophilization can only stabilize the protein content in serum so that the content of vitamin A in serum still decreased. In an effort to increase the stability of vitamin A in serum, this research was conducted to prepare freeze-dried autologous serum with the addition of vitamin E as an antioxidant of vitamin A. The purpose of this study was to determine the effect of vitamin E in stabilizing the concentration of vitamin A in freeze-dried autologous serum and checking its antibacterial properties by total plate count method. In this method, the separated blood serum was diluted with 0.9% sodium chloride followed by adding sucrose 60mM and vitamin E in different concentrations 0.5; 1; 2.5; 5% then this preparation was lyophilized. After that the serum was stored at 4°C and the content of vitamin A was analyzed every month by HPLC. Freeze-dried autologous serum with 1% addition of vitamin E showed the best results in stabilizing the vitamin A content up to 2 months. The content of vitamin A left in the 2-month freeze-dried serum was 0.0191-0.0761 ppm.