1. The determination of ascorbic acid in butter by the usual macrotitration method was found to be impossible because this substance rapidly became oxidized while the butter was being melted, so that only about half could be recovered.
2. A simple micro-titration procedure enabled more reliable results to be obtained, but the loss of ascorbic acid from smaller samples could not be reduced below about 10% under the most favourable conditions.
3. Kinetic experiments have been carried out on butter serum and buttermilk to which ascorbic acid had been added, and from the results it has been deduced that the oxidation of ascorbic acid takes place by three distinct concomitant reactions: one catalysed by the original copper, one catalysed by added copper, and one catalysed by added ferric iron.
4. It has been calculated that in an unsalted butter of normal metallic content half the ascorbic acid would be oxidized in about 12 hr. at cold storage temperature ( – 13° C). At this temperature, 0·1 p.p.m. of added copper is about twenty times as effective a catalyst as 1·0 p.p.m. of added iron. At room temperature the corresponding half-oxidation time would be about 2 hr., and here 0·1 p.p.m. of added copper is about four times as active as 1·0 p.p.m. of added iron. In salted butter the inhibitory effect of the chloride ion would increase these times somewhat.
5. The chemical state in which copper and iron exist in milk and related products has been discussed, especially in relation to the kinetic results obtained.