Method for triglyceride measurement in small amounts of plasma

A method is described for determining triacylglyceride concentration in small amounts of plasma. After ethanolic-KOH digestion of diluted plasma aliquots, samples were neutralized with MgSO4 and glycerol was fluorimetrically assayed in supernatants by the coupling of glycerokinase, pyruvate kinase, and lactate dehydrogenase catalyzed reactions. Values were corrected by free glycerol present in the non-digested samples. Digestions were performed at different times and temperatures in order to establish optimum conditions for recoveries and reproducibility. Parallel determinations before and after phospholipid removal showed that their presence in plasma did not interfere with the obtained values. This method is especially useful for running many samples in parallel and for determinations in small experimental animals in which the amount of plasma is very limited.