Nicotinamide, one kind of Vitamin B, plays an important role for humans. To ensure quality control of nicotinamide in infant formula, a Certified Reference Material for nicotinamide in infant formula is being prepared in China for the first time. Studying on the procedure of sample preparation and HPLC analysis with different detectors like UV and MS, HPLC-UV and HPLC-IDMS are selected as the value assignment methods. And HPLC-IDMS method was validated in the international comparison CCQM-P78 and CCQM-K62. Homogeneity and stability are discussed in this paper as well.