Dietary fiber sometimes is defined chemically as nonstarch polysaccharides plus lignin or as other specific chemical entities. Analysis of dietary fiber according to a chemical definition typically involves gas chromatography, which allows separation and quantitation of chemical constituents that are added to arrive at a dietary fiber value. Other definitions of fiber are broader, defining it to be whatever is not digested in the alimentary tract. Analytically, this definition translates into the gravimetric sum of the material remaining after a series of enzymatic and chemical treatments that simulate in vivo digestion. Various methods reflect the gravimetric definition, which might include as dietary fiber some protein, resistant starch, and even lipids that are not digested by particular assay conditions. We used a recently proposed bile-enzymatic-gravimetric assay for total dietary fiber on commonly consumed seeds (hulled and unhulled sesame, caraway, and poppy) and visually found these seeds to be undigested. We then determined the impact of the undigested seeds on measured dietary fiber content by spiking homogenized daily menus with 5% by weight of these seeds and calculating recoveries with 2 assumptions: seeds are 100% fiber because they are not digested, and the fiber content of seeds is as determined by assay. Calculated recoveries were very different depending on which assumption was made (71-90% or 99109%, respectively), and the difference was closely related to the seed’s protein content.
A recently proposed bile-enzymatic-gravimetric total dietary fiber (TDF) method was modified and the new procedure was compared with the original method, the traditional AOAC enzymatic-gravimetric determination (AOAC Official Method 985.29), and another simplified AOAC procedure by analyzing several diet composites, including National Institute of Standards and Technology 1548 total diet reference material. The original and modified bile-enzymatic-gravimetric procedures also were compared by analyzing 9 food samples from a collaborative study of the original method. The modified method consistently yielded values about 10% lower than the original method but closer to reference values and to values from AOAC Offical Method 985.29, suggesting results that are more in line with accepted TDF standard methodology. Our modified method was used to analyze 180 fresh-frozen diet composites with TDF values ranging from 0.6 to 3.2 g/100 g wet weight. Samples were from 2 multicenter feeding studies sponsored by the National Heart, Lung and Blood Institute: DELTA (Dietary Effects on Lipoproteins and Thrombogenic Activity) and DASH (Dietary Approaches to Stop Hypertension). The mean relative standard deviation (RSD) for duplicate analyses was 1.1%. For 40 assays of a quality control diet composite over 9 months, the standard deviation was 0.1 g/100 g wet weight (4.9% RSD), indicating the method’s excellent precision for routine use.
We have evaluated the determination of lactate dehydrogenase (EC 1.1.1.27) isoenzyme 1 activity by chemical inhibition of the other isoenzymes with perchlorate and with 1,6-hexanediol. In the hexanediol method, we studied the effect of the duration of incubation with the inhibitor; a 5-min incubation yielded results closest to those of an immunochemical technique (Isomune-LD). The perchlorate method was the most precise, and the hexanediol method the least, although for none of the techniques did the coefficient of variation exceed the medically acceptable limit prescribed by the College of American Pathologists. Pairwise correlation among the immunoprecipitation, electrophoretic, and chemical inhibition methods was good (r greater than 0.991), although the differences between means were statistically significant (except for the comparison of the two chemical inhibition methods). Because of its ease, low cost, and precision, we recommend the perchlorate method for routine use.
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