Clinical Chemistry
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Clinical Chemistry 40: 2230-2234, 1994;
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Clinical Chemistry, Vol 40, 2230-2234, Copyright © 1994 by American Association for Clinical Chemistry

Detergent extraction and enzymatic analysis for fecal long-chain fatty acids, triglycerides, and cholesterol

MJ Lee, T Crook, C Noel and UM Levinson
Great Smokies Diagnostic Laboratory, Asheville, NC 28806.

We report a procedure for determining fecal long-chain fatty acids (LCFA), triglycerides, and cholesterol after detergent extraction with 10 mL/L Triton X-100, 6 mL/L Brij 30, and 0.1 mol/L HCl in isotonic saline by enzymatic analysis of the extraction supernates. Mean recoveries of calibrators ranged from 105% to 117%. Assays of fecal extracts were linear with concentration from 0 to 1000 mumol/L for LCFA, from 10 to 90 mumol/L for triglycerides, and from 0 to 300 mumol/L for cholesterol. Within-run CVs were < 3% for medium and high concentrations, and 3%, 14%, and 28% for low concentrations of LCFA, cholesterol, and triglycerides, respectively. Correlation with gravimetric determination resulted in coefficients (r2) of 0.72 for total fat, 0.63 for neutral fat, and 0.66 for nonesterified fat. Assays of extracts and residues prepared for gravimetric determination indicated that approximately 90% of triglycerides, cholesterol, and LCFA were extracted by organic solvents and that the extracts contain a significant amount of other fats. The proposed method appears accurate, precise, specific, and suitable for routine analysis.




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Copyright © 1994 by the American Association for Clinical Chemistry.