24-hrs. milk load test
Fasting blood of other 41 healthy participants was collected in EDTA vacutainers on day-15. A portion of blood was processed for hematological parameters within 30 minutes of collection and the rest was centrifuged at 1500 rpm for 20 minutes and the plasma was separated and distributed in four storage vials and stored at -20°C. All of them (51 participants) were instructed to drink 3×200 ml of milk along with their usual diet during the day (in the morning after breakfast, evening at 4 p.m. and at night after dinner) and come to the Laboratory next morning-9 am. On day-16, 10 ml. of fasting blood was collected and the separated plasma was stored as above (Figure 1: Flow sheet).
thumbnailFigure 1. Flow chart: 51 Participants were registered on day-0. Fasting blood was collected of 10 participants on day-1*. They were instructed to continue their usual diet for next 14 days. On day-15 fasting blood was collected of all 51 participants. They consumed 3 × 200 ml of milk during the day. On day-16 fasting blood was collected. They were divided into normal and vitamin B-12 deficient groups as per pl. vitamin B-12 concentrations. Deficient participants consumed 2 × 200 ml. milk every day for next 14 days and their fasting blood was collected on day-30. Dietary history was taken on day 1, 15, 30. Participant’s milk drinking time was in the morning after breakfast, 4 p.m. and at night after dinner.