Determination of lipoprotein lipase activity in post heparin plasma of streptozotocin‐induced diabetic rats by high‐performance liquid chromatography with fluorescence detection

Abstract

The activity of lipoprotein lipase (LPL), an enzyme responsible for lipoprotein metabolism, would vary in diseases and metabolic disorders. For determination of LPL activity, a highly sensitive high performance liquid chromatography (HPLC) method using a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ) was applied to determinate the oleic acid (OA) generated from triolein by LPL activity without multiple solvents extraction step. We studied the optimal conditions of the reaction including the effect of emulsifiers, deproteinizing solvents, and the concentration of bovine serum albumin (BSA). Ten millimolar concentrations of triolein, 5% of BSA, 1% of Gum arabic (GA), and acetonitrile showed the optimum conditions for measuring the LPL activity. The accuracy values for the determination of LPL activity in 10 microL of rat post heparin plasma were 108.73 approximately 114.36%, and the intra- and inter-day precision values were within 1.28% and 2.91%, respectively. The limit of detection was about 4.53 nM (signal-to-noise ratio 3). The proposed method was applied to determination of LPL activity in post heparin plasma of normal and streptozotocininduced diabetic rats associated with 52.3% reduction. The established assay system could be used for determining LPL activity in different physiological and pathological conditions to clarify the relationship between LPL activity and diabetes mellitus.

Depolarizing Actions of GABA in Immature Neurons Depend Neither on Ketone Bodies Nor on Pyruvate

“…In addition, the argument that 4 mM DL-BHB is equivalent to 2 mM active D-BHB (Rheims et al, 2009) is invalidated by the observations that L-BHB exerts complex biological actions (Moore et al, 1976;Webber and Edmond, 1977;Herzberg and Gad, 1984;Eaton et al, 2003;Tsai et al, 2006;Chou et al, 2008). Also, ketogenic diet reduces glucose utilization by 10% per millimolar plasma ketone bodies (Robinson and Williamson, 1980;Harding and Charlton, 1990;LaManna et al, 2009), hampering the interpretation and relevance of observations made using ACSF with glucose and DL-BHB.…”

Ameliorative Effect of Hexane Extract of Phalaris canariensis on High Fat Diet‐Induced Obese and Streptozotocin‐Induced Diabetic Mice

“…At the end of obesity and chronic diabetes experiments all mice were anesthetized with 1.0% pentobarbital sodium and blood was obtained from the retroorbital plexus of each animal following the injection of heparin (100 IU kg −1 body weight) into a tail vein for 10 min [20]. The liver and kidney were removed according to defined anatomical landmarks [21].…”

Cyanidin-3-O-β-glucoside improves obesity and triglyceride metabolism in KK-Aymice by regulating lipoprotein lipase activity

“…After 12 weeks of dietary intervention, all mice were anesthetized with 1.0% pentobarbital sodium and blood was obtained from the retro‐orbital plexus of each animal following the injection of heparin (100 IU kg −1 body weight) into a tail vein for 10 min 15. The blood samples were then centrifuged at 1600 × g for 15 min at 4 °C for the preparation of plasma.…”