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The general purpose of chromatography is to separate components in a mixture by moving this mixture through a support for which the different components have different affinity and hence get separated. (Marie, 2003). The application of Vitamin C for HPLC allows the determination of Vitamin C in an easy, fast, and precise method. The kit includes all reagents in ready to use for preparation and separation of the samples. From our observation we see that the sample that we analyze do not have Vitamin C. These happened because of the retention time for sample A, B and C which are 3.317, 3.311 and 3.317 µg/ml is not the same as the retention time of the standard which are 3.270, 3.272, and 3.270 µg/ml. The retention of vitamin C is often used as an estimate for the overall nutrient retention of food products because it is highly sensitive to oxidation and leaching into water-soluble media during storage (Davey et al.,2000).The concentration (µg/ml) of the Vitamic C in all samples are 0.104 and the area(mAU) of all samples are 0.These means there is no Vitamin C presence in this experiment. Vitamin C is a water soluble vitamin. It is unstable vitamin and can be lost easily. There are many factors that can might effect the results of Vitamin C in this experiment. One of the factor is because of the time of sample that is expose in the atmosphere. The sample is easy to oxidized in the air, so the time to expose the sample to the open atmosphere should be minimized. Besides that, The temperature also affect vitamin C level. Areas with cool nights produce citrus fruits with higher vitamin C levels. Hot areas produce fruit with lower levels of vitamin C (Padayatty et al., 2003). Also environment conditions that increase the acidity of citrus fruits also increase vitamin C levels. Furthermore, grinding and fractionation during milling of the sample cause major losses of Vitamin C. This can cause mechanical pressure to the sample that can lead to loss of Vitamin C....
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