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D6584_10 (17) 07-2017

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Set the instrument operating variables to the values specified in table one of the written standard. Weigh to the nearest 0.1 milligrams, approximately 100 milligrams of sample directly into a 10 milliliter septa vial. Using microliter syringes, add exactly 100 microliters of each internal standard an MSTFA. Shake the vials and allow it to set for 15 minutes to 20 minutes at room temperature. Add approximately eight milliliters of n-Heptane to the vial and shake. Inject 1 microliter leader of the reaction mixture into the cool on-column injection port, and start the analysis. Obtain a chromatogram and peak integration report. Identify peaks by comparison of retention times to the standards. For identification of additional peaks, use the relative retention times given in table four of the written standard and the reference chromatograms given in figure one. Mono, di, and triglycerides are separated according to carbon numbers, or CN. Monoglyceride consists of the four overlapping peaks with relative retention times, or RRT, of 0.76 and 0.83 to 0.86 with respect to the internal standard tricaprin. A pair of peaks, methyl esters with at carbon number of 24 may appear with the relative retention time of 0.80 to 0.82 and should not be included in the calculation of monoglyceride. Diglyceride is also primarily separated according to carbon number, but due to a varying double bonds in the molecules, baseline resolution of the peaks does not occur. The grouping of three to four peaks with relative retention time of 1.05 to 1.09, carbon number 34, 36, and 38, shall be attributed to diglyceride. Carbon number also separates triglyceride. Peaks with relative retention time of 1.16 to 1.31, CN 52, 54, 56, and 58, should be included in the calculation.

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Posted by: abuckmaster on Sep 14, 2018

D6584_10 (17) 07-2017

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