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Deuterated vitamin D standards are used commonly as internal standards in LC-MS/MS analysis of vitamin D3 and 25-hydroxyvitamin D3 in food. However, the use of various eluent additives, such as methylamine, formic acid and ammonium formate, also contributes to matrix effects and the performance of analysis by affecting accuracy and robustness. For the first time, continuous post-column infusion experiments of isotopically labelled vitamin D3-[d6] were performed to evaluate ion-suppression in a wide variety of food (salmon, cheese, pork