A series of process in achieving fortificant powder of natural folic acid was performed through fermentation of beans (tempeh), fermentation of vegetables and nixtamalization of corn. Experiment treatment was conducted on two types of fortificant based on nixtamal of corn (Zea mays var indentata) type (yellow corn and white corn) added to mixed pasta of soy tempeh or mung bean tempeh and fermented broccoli or fermented spinach at ratio of 1 : 1, 1 : 2,1 : 3, 1 : 4 and 1 : 5, homogenized and dried at 50 °C for 24 hours, respectively. Based on the best recovery of folic acid, optimization result of powder fortificant formulation was achieved at combination between mixed pasta of mung bean tempeh and fermented broccoli, and nixtamalized yellow corn at the ratio of 1 : 3, meanwhile combination between mixed pasta of soy tempeh and fermented spinach, and nixtamalized white corn at the ratio of 1 : 3. On both these treatments of folic acid recovery gave folic acid 248.74 µg/mL and 239.41 µg/mL, dissolved protein 1.03 mg/mL and 1.68 mg/mL, total sugar 686.55 and 482.88 mg/mL, total solids 93.90 % and 94.24 %, and reducing sugars 29.23 mg/mL and 13.31 mg/mL, respectively. Identification of folic acid monomer on each fortificant powder was found 9 folic acid monomers dominated by folic acid monomer with molecular weight (MW) 442.57 Dalton (Da.) and 442.74 Da. and relative intensities 9.74 % and 9.74 %, and 7 folic acid monomers dominated by folic acid monomer with MW of 442.68 Da. and relative intensities 10.1 %. Distribution of particles on both type of fortificant powders displayed polydisperse index of 1.289 and 0.925 with particles size 3990.5 nm and 724.3 nm, respectively.

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