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The clean-up performed on solutions of PDIMs and cholesterol standards in petroleum ether show that, depending on the solvent mix and on the type of SPE used, the recovery of PDIMs is between 64 and 70%, whilst most of the cholesterol is removed from the system.
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The program satisfies the requirements of paragraph (f)(4)(iv) of this section because it makes available to all participants who do not meet the cholesterol standard a reasonable alternative standard to qualify for the reward.
Lipid pellets resuspended in 5 μl TN buffer and 5 μl of cholesterol standard solutions ranging from 0.3 to 7.8mM were supplemented with 500 μl of cholesterol liquid stable reagent provided by Thermo Electron Corporation.
For cholesterol analysis, aliquots of 4 µl of the different samples and cholesterol standard were applied to HP-K plates (Whatman, Clifton, NJ), developed in chloroform-acetone (95∶5v/v), stained with the CuSO4 reagent, and developed by charring at 170°C.
For cholesterol analysis, aliquots of 4 µl of the different extracts and cholesterol standard were applied to HP-K plates (Whatman, Clifton, NJ), developed in chloroform/acetone (95∶5 v/v), stained with the CuSO4 reagent, and developed by charring at 170°C.
The cholesterol standard curve was provided with the cholesterol assay kit.
The upper phase was collected, dried and analysed by thin layer chromatography using silica gel 60 plate (Merck, Darmstadt, Germany), which was developed with hexane/diethyl ether/acetic acid (87:20:1 by volume), with reference to internal cholesterol standard (Sigma, St Louis, MO).
Standardised procedures were used to determine fasting plasma glucose (hexokinase method), triacylglycerol and HDL-cholesterol (standard enzymatic methods) and serum insulin levels (1235 AutoDELFIA immunoassay system, DAKO, Ely, UK) [ 23].
The standard cholesterol tests measure quantity: A total cholesterol score below 200 is considered desirable, for example, and over 239 is high, a risk factor for heart disease.
Meanwhile, fasting blood samples were collected to measure LDL-C, HDL-C and total cholesterol using standard enzymatic procedures in the same clinical laboratory.
In the study, high cholesterol diet, standard atorvastatin, and high cholesterol diet with hydroalcoholic extract of gymnemic acid were fed to female rats for seven days.
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