In this method, 1.0g of each sample powder was extracted thrice by warming (40°C – 50°C) and stiring with a magnetic stirrer for 1 hour with 20cm3 of 0.3N HCl. The combined extracts were diluted to 100cm3 with water and used for the total oxalate estimation.
For oxalate estimation, 5.0 cm3 of each extract was made alkaline with 1.0 cm3 of 5.0N NH40H. This was then made acid to phenolphthalein (2 or 3 drops of this indicator added, excess decolourizes solution) by drop wise addition of glacial acetic acid. 1.0 cm3 of 5% calcium chloride was then added and the mixture allowed to stand for 3 hours after which it was centrifuged at 3000 revolutions per minute (rpm) for 15 minutes. The supernatant was discarded and precipitates washed thrice with hot water with thorough mixing and centrifuging each time. Then to each tube 2.0 cm3 of 3N H2S04 was added and the precipitate dissolved by warming in a water bath (70°C – 80°C). The content of each tube was then titrated with freshly prepared 0.01N potassium permanganate solution. Titration was carried on at room temperature (29°C) until the first pink colour appeared throughout the solution and allowed to stand until the solution was colourless. The solution was then warmed to 70°C – 80°C and titration continued until a pink colour persisted for at least 30 seconds.