An HPLC Method for the Analysis of Soybean- and Sunflower-Lecithin and their Enzymatic Hydrolysis Products. 4 th Euro Fed Lipids Congress

A.A. CARELLI; C. PENCI; CONSTENLA, D.

Madrid

Congreso; 4 th Euro Fed Lipids Congress; 2006

Universidad Complutense de Madrid (UCM)

An HPLC Method for the Analysis of Soybean- and Sunflower-Lecithin and their Enzymatic Hydrolysis Products Amalia A. CARELLI, Cecilia PENCI, Diana CONSTENLA Planta Piloto de Ingeniería Química (UNS-CONICET).  Bahía Blanca, Argentina. The enzymatic-hydrolysed lecithin has important technological applications in the food and pharmaceutical industries. The phospholipases are the enzymes commonly employed to hydrolyse lecithin, being required to develop methods that use traditional instruments in order to evaluate the products. With this purpose an HPLC method (High-Performance Liquid Chromatography) employing ultraviolet detection for the simultaneous analysis of phospholipids and lysophospholipids from soybean and sunflower oil was implemented. The separation of these compounds was achieved on an silica column (Alltech Lichrospher Si 60 5m, L=150 mm, i.d =4.6 mm) using a gradient elution method at room temperature. The mobile phase consisted in a ternary system of the following solvents: A (n-hexane); B (2-propanol/ acetic acid, 99.5:0.5 v/v); and C (methanol/ Milli Q water, 98:2 v/v). The detection and quantification by the external standard method of phospholipids and lysophospholipids were performed with an ultraviolet detector at 206 nm. This methodology allows an adequate resolution for the main phospholipids present in soybean- and sunflower-lecithin (phosphatidylcholine, phosphatidylethanolamine and phosphastidylinositol) and their respective lysophospholipids. Some hydrolyzed soybean- and sunflower-lecithin samples were evaluated by this method. In order to eliminate the free-fatty acids produced during the hydrolysis reaction, the hydrolyzed samples were eluted by solid phase extraction using diol-phase cartridge. The solvents were passed through in this elution order: chloroform, ethyl ether and methanol/ammonia solution. The methanol/ammonia fraction, containing lyso-phospholipids and phospholipids was collected, evaporated to dryness under nitrogen, suspended in n-hexane/2-propanol 50:50 v/v, and analyzed by HPLC. This method showed to be suitable for studying product distribution from enzymatic hydrolysis reactions of vegetable lecithin.