Infants and young children have high nutritional demands in order to support adequate growth and development, particularly during the transition from complementary feeding at 12 months of age to a mixed and varied diet at 36 months of age. Omega-3 long chain polyunsaturated fatty acid (n-3 LC-PUFA) such as docosahexaenoic acid (DHA; 22:6n-3) is the important constituents of the maturing brain, especially for visual and cognitive development. However only certain foods, such as fatty fish, contain n-3 LC-PUFA at concentrations sufficient for their needs. Additionally, these foods that are known to be rich in DHA may not be regularly provided to infants and toddlers due to concerns about potential food allergies or methylmercury exposure.

Although the importance of consuming n-3 LC-PUFA and essential fatty acids at the level of dietary recommendations in late infancy and early childhood (6-24 months) are highlighted, the current median n-3 LC-PUFA and estimated DHA intakes in toddlers in most countries are lower than the recommended levels. The European Food Safety Authority (EFSA) recommend that infants and young children (< 24 months) should consume 100 mg of DHA per day, while for older children (2-18 years), they recommend a daily intake of 250 mg. In order to bridge the gap between the current intake and recommended levels of n-3 LC-PUFA, general foods, especially infant and toddler formulas, should be enriched with n-3 LC-PUFA particularly DHA. With a growing body of research, the challenge is to find an ideal formula that is nutritionally balanced and human milk-like, especially with respect to the ratio between omega-6 and omega-3 fatty acids and DHA level.

In adding DHA into the toddler formula, there are several methods, among others are in the form of wet mix and dry blending. The aim of this study is to compare the bioavailability of different methods of adding DHA (dry blend versus wet mix) into formulas in healthy Indonesian toddlers age 2-3 years old for a period of one month. Furthermore, to evaluate the stability of milk-based formulas that are supplemented with DHA under same storage conditions, so as to monitor the stability of infant formula.

This study will use two phase approaches Phase 1, screening habitual n3 intake. Mothers of healthy toddlers, non-fish or non-n3 supplement babies (aged 2-3 years old) would be invited to complete a Food Frequency Questionnaire (FFQ) to determine their habitual dietary intake of long chain n3 fatty acids. The 120 babies with the lowest long chain n3 intake would be invited to proceed to phase 2 of the trial.

Phase 2, double blinded, randomized trial. Based on the outcome of the FFQ in phase 1 of the proposed trial, the 120 babies with the lowest habitual dietary intake will be allocated to diet in a random order for one month. Blood samples will be collected via dry blood spot kits at the start, middle and end of the diet along with faecal collection every day.

Groups are Milk drink unfortified (negative control), Milk drink unfortified plus fish oil (positive control), Milk drink fortified with DHA used wet mixing method and Milk drink fortified with DHA used dry blending method.

Population and subjects are Indonesian children aged 2-3 years old living in Jakarta will be selected if they meet the study criteria.

For stage-1: to get 50% of children having habitual low DHA intake, with degree of significance 5% and degree of reliability 95%, then 385 children will be needed.

For stage-2: This study will include 30 subjects per-study group

Inclusion Criteria:

• Boys and girls aged 2-3 years old, apparently healthy, living in the study area for minimally one month and parents permit them to participate in the study by signing the informed consent.

Exclusion Criteria:

• Having milk allergy and/or lactose intolerance, high omega-3 intake habit