This experiment was conducted to establish a method for simultaneous determination of several water-soluble vitamins in rumen fluid by high performance liquid chromatographic (HPLC). Rumen fluid sample was extracted with hexane and metaphosphoric acid, and the supernatant was evaporated in a nitrogen stream at 40 ℃. The concentrate was dissolved by mobile phase A. Chromatographic condition was the following: SBC8 column, particle side 5 μm, 4.6 mm×150 mm; mobile phase A, Vmethanol:V0.05 mol/L trisodium phosphate (pH=2.5)= 7:93; mobile phase B, Vmethanol:V0.05 mol/L trisodium phosphate (pH=2.5)=90:10; flow rate was 1.0 mL/ min; detection wavelength was 245 nm. The results showed as follows: 1) the minimum detecting concentrations of niacinamide, nicotinic acid, folic, riboflavin acid and cyanocobalamin were 12.90, 12.00, 74.11, 24.00 and 9.57 ng/mL (signal to noise ratio=3), and correlation coefficients of linear regression were 0.999 9, 0.999 3, 0.990 3, 0.999 0 and 0.999 5, respectively; 2) the average recoveries of adding standard niacinamide, nicotinic acid, folic acid, riboflavin and cyanocobalamin in rumen fluid were 97.5%, 96.8%, 73.2%, 84.4% and 89.1%, and the average relative standard deviations of those were 2.9%, 3.0%, 15.6%, 7.8% and 5.0%, respectively. The results indicate that the contents of niacinamide and nicotinic acid in rumen fluid can be simultaneously determined by this method.
ZHAO Yunjun
,
MENG Qingxiang
. Simultaneous Determination of Several Water-Soluble Vitamins in Rumen Fluid by High Performance Liquid Chromatography[J]. Chinese Journal of Animal Nutrition, 2013
, 25(2)
: 356
-363
.
DOI: 10.3969/j.issn.1006-267x.2013.02.017
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