Determination of adenosines in muscle and intestinal digesta of Landrace pigs by high preformance liquid chromatography
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Graphical Abstract
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Abstract
Adenosines are the vector of energy metabolism. The aim of this work was to establish a method for the differences of energy metabolism from the pig muscle and intestinal digesta by high preformance liquid chromatography(HPLC).When the analytic condition of HPLC in feed was a reverse phase Waters C18 column (250 mm×4.6 mm,5 μm),mobile phase was methanol(V)/ 50mmol/L KH2PO4 buffer solution(pH6.5) (V 9∶91) with the flow rate of 0.5 mL/min,the column temperature was at 30(±1)℃,the detection wavelength was operated at 259 nm,and the injection volume was 10 μL,the calibration curves had good linearity over concentration range of 0.1-100 mg/L. The linear relative coefficient for ATP,ADP and AMP over concentration range of 0.1-100 mg/L were 0.999,0.998 and 0.996,respectively. The limit of detections(LOD, S/N ≥ 3)of ATP,ADP and AMP were 0.0001,0.004 and 0.004 mg/L,and their limit of quantifications (LOQ,S/N ≥ 10)were 0.002,0.02 and 0.02 mg/L,respectively. The mean recovery ratio of ATP,ADP and AMP were 92.00%-99.05%,100.77%-104.48% and 110.84%-113.72% with RSDs of 0.29%-1.02%, 0.33%-1.65% and 0.69%-1.92%,respectively. The whole time of analysis lasted for 10 min.This method has properties of rapidity,accuracy,sensitivity and repeatability,which can be applied in detecting the concentration of ATP,ADP and AMP and analyzing the energy metabolism.
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