Analysis of Ascorbic Acid - also Known as Vitamin C- using HPLC in Banana

In this video, we will talk about the analysis of ascorbic acid, also known as vitamin C using HPLC. Banana has long known to be a superfood It contains a lot essential nutrients that are beneficial to the human body, such as vitamins and minerals Ascorbic acid, also known as vitamin C, is abundant in banana containing about 10% of daily amount required Health benefits of ascorbic acid include prevention of infections, synthesis of collagen and production of iron Quantitative analysis of ascorbic acid in various food is important in health and nutrition sciences, food processing and quality labelling Some of the methods of analysis are AOAC 967.21 which is based on titrimetric method for vitamin C in preparations and juices, AOAC 2012.22 which is based on the measurement of vitamin C in infant and adult formula Let’s start off with the sample preparation Peel off the banana skin and cut the banana Weigh the cut banana into a centrifuge tube Add in a mixture of acetic acid, metaphosphoric acid and TCEP into the sample TCEP is important in the extraction process. Ascorbic acid is easily oxidised into dehydroascorbic acid. TCEP helps to reduce dehydroascorbic acid into ascorbic acid. Vortex the sample mixture Centrifuge the sample mixture Filter the supernatant with nylon filter Dilute the filtered solution Ascorbic acid stock solution at concentration of 1000 ppm was prepared using a diluent mixture of metaphosphoric acid, acetic acid and TCEP. The stock solution was diluted to 7 different concentrations as working standards using the same diluent as stock solution. Let’s move on to the HPLC analysis Load the samples into the HPLC system Load analysis and inject the samples A calibration curve is built with an ascorbic acid concentration ranging from 0.1 ppm to 10 ppm Finally, this calibration curve is then used to quantitate ascorbic acid in banana samples It is important for the ascorbic acid in sample to be stable and not oxidized during the analysis In order to evaluate the stability of ascorbic acid in sample, an extracted banana sample was injected consecutively for 10 hours The peak areas of the injections were compared The peak area of ascorbic acid remains consistent throughout the period, showing that it is stable in the sample #Shimadzu #HPLC #vitamin C #vitamin C content #ascorbic acid #analysis of vitamin C #analysis of ascorbic acid #analysis using HPLC #analysis vitamin C in banana #Shimadzu Asia pacific #GCMS Analytical instrument Liquid Chromatography