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Estimation of Citric Acid from Aspergillus sp.

AIM To estimate the production of citric acid from Aspergillus sp.

INTRODUCTION Citric acid is one of the most important organic acid. Initially citric acid was extracted from fruits but now it is largely produced by microbial fermentation. Aspergillus niger and Aspergillus mentouis are widely used for commercial production of citric acid. It is widely used in food, soft drinks, pharmaceuticals, manufacturing of ink and dyes, leather industry, electroplating etc. Various carbohydrate rich raw materials such as corn, starch, sugar-beet and molasses are used as some material for sugar.

PRINCIPLE Many strains of Aspergillus niger which are mutants cannot oxidize citric acid and hence accumulate in the culture medium. The composition of the fermentation medium is critical for obtaining high yield of citric acid and hence it is essential to limit the growth of fungus so that high level of citric acid can accumulate in the medium. This can be accomplished by having a deficiency of trace metals in the medium. Artificial medium for the production of citric acid contains NH4NO3, MgSO4, KH2PO4 acid is added to achieve a low pH of 3.5. Sucrose serves as the carbon source for the production of citric acid. NH4NO3 is used to avoid formulations of oxalic acid and Glutamic acid fermentations. This experiment makes sure of simple acid base titration. The given sample is titrated against alkali of known strength, using Phenolphthalein as an indicator. The end point is pale pink colour. The volume of alkali used for titration is known and is used to calculate the normality and the percentage of acid in the filtrate.

REQUIREMENTS Conical flask, test tube, distilled water, inoculation loop, culture, shaker etc.

PROCEDURE The media was prepared pH obtained to 3.5 and sterilized. The spores of the culture were taken and were suspended in test tubes containing sterile water. 0.1 ml of spore suspension is inoculated into the medium under aseptic conditions, then flasks are incubated in a chemical shaker at 27 28 C for 10 days. The flasks were checked for fungal grown after 10 days of incubation. After the incubation period, the media containing the organisms were filtered using double layer. The filtrate was used for estimation. 10 ml of filtrate, add 2-3 drops of phenolphthalein. The solution is titrated against 0.1N NaOH solution taken in a burette.

RESULT The percentage of citric acid is found to be 0.532%

OBSERVATIONS Solution in the burette: 0.1N NaOH Solution in conical flask: 10ml of citric acid + 2-3 drops of phenolphthalein

Burette Reading Volume Initial 10 0 Final 7.6 7.6 Rundown

10

7.6

15.2

7.6

N1V1 (citric acid) = N2V2 (NaOH) 10 * N1 = 0.1 * 7.6 N1 = 0.1 * 7.6 / 10 = 0.076

Weight of citric acid in 1000 ml solution = Normality of citric acid *Equivalent weight of citric acid

= N1 * 70 = 5.32

Weight of citric acid in 1000 ml of solution = 5.32/10 = 0.532 g

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