RAPID ESTIMATION OF PHOSPHOLIPED BY IMMOBILIZED PHOSPHOLIPASE D FROM PISTACHIA KHINJUK IN FIA SYSTEM

Blue green gains of Pistachia khinjuk was collected from area of Harnai Balochistan and crude Phospholipase D was isolated by simple protean precipitation. This enzyme was covalently bounded on the surface of controlled porosity glass beads and packed in small glass columns. Immobilized PLD was used for quantitative determination of Phosphatidylcholine (Lecithin) and Phosphorylcholine in flow - injection analysis system. Immobilized columns of Phospholipase C from rice and Choline o xidase from chicken intestine were also prepared in same manner. The phosphatidylcholine was estimated by incorporation PLD column fallowed by choline oxidase column in flow stream. The resultant hydrogen peroxide produced as byproduct in flow injection s ystem was detected amperometrically by electrochemical hydrogen per oxide detector. The Phosphorylcholine was estimated by insertion of PLC column in FIA system and final product was estimated using spectrophotometer at λ Max 640 nm. The isolated PLD showe d rapid accuracy for estimation of lecithin and using immobilized PLC fallowed by Alkaline phosphatase found best for Phosphorylcholine estimation.