In a comparative study of the Karl Fischer method with oven methods for determination of water in forages and animal feeds, oven methods yielded the following relative recoveries (expressed as a percentage of the recovery obtained by the Karl Fischer method) for hay, haylage, and corn silage, respectively: (1) drying at 135°C for 2 h (AOAC 930.15), 113,162, and 133%; (2) drying at 104°C for 3 h (AOAC 935.29), 96,122, and 113%; and (3) drying at 104°C for 6 h, 97, 129, and 117%. Relative recoveries for nonurea-containing and urea-containing feed, respectively, were as fol lows: (1) drying at 135°C for 2 h (AOAC 930.15), 116 and 2746% (2) drying at 104°C for 3 h (AOAC 935.29), 88 and 239%; (3) drying at 95°C for 5 h under vacuum (AOAC 934.01), 83 and 727% (4) drying at 104°C for 6 h, 90 and 427%; and (5) drying at 110°C for 3 h, 94 and 425%. Preliminary near-infrared reflectance calibrations for water (moisture) based on the Karl Fischer method were promising (r2 = 0.98; standard error of calibration = 0.20).
A Karl Fischer method for determining water (dry matter) in animal feed and forages was collaboratively studied. Water was extracted from animal feed or forage material into methanol–formamide (1 + 1) directly in the Karl Fischer titration vessel by high-speed homogenization. The water was titrated at 50°C with one-component Karl Fischer reagent based on imidazole. Ten blind samples were sent to 9 collaborators in the United States, Canada, and Germany. The within-laboratory relative standard deviation (repeatability) ranged from 1.14 to 6.99% for water or from 0.09 to 0.56% for dry matter. Among-laboratory (including within-) relative standard deviation (reproducibility) ranged from 5.35 to 10.73%, or from 0.44 to 0.77% for dry matter. The authors recommend that the method be adopted as Official First Action by AOAC INTERNATIONAL. A comparable alternative extraction procedure using boiling methanol is also recommended for Official First Action.
Oven methods for determining moisture (volatiles) in forages and other animal feeds are empirical. The moisture concentration obtained depends upon the time and temperature the sample was dried and is influenced by the presence of other volatiles than water. A validated reference method to measure water in forages and animal feeds could be used to evaluate the appropriateness of oven methods for various types of animal feeds and forages. Karl Fischer titration is a well-established method for determining water. However, thorough extraction of water from forages and feeds is a challenge because they often contain cellular structures that release water slowly. Water was successfully extracted into methanol-formamide (50 + 50) by high-speed homogenization and then titrated directly at 50 C with a one-component Karl Fischer reagent based on imidazole. The method is described in detail, results of day-to-day repeatability and laboratory-to-laboratory reproducibility are reported, and preliminary comparison data between oven methods are provided.
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