Ascorbic Acid Saturation Test & Determination of Ascorbic Acid Content of Some Fruit Juices.

376 BCH Miss Tahani Al-shehri

OBJECTIVES • To measure the concentration of ascorbic acid urine sample after administration of a dose of ascorbic acid. .

• Water-soluble vitamins dissolve in water. The body cannot store them. Leftover amounts of the vitamin leave the body through the urine.Introduction • Vitamin C is a water-soluble vitamin that is necessary for normal growth and development. That means you need a continuous supply of such vitamins in your diet. .

Introduction Ascorbic acid Dehydroscorbic acid .

Functions of Vitamin C • Vitamin C is required for the growth and repair of tissues in all parts of your body. • It is necessary to form collagen • Vitamin C is one of many antioxidants in the body * The body does not manufacture vitamin C on its own ( Why ? ) .

Deficiency of Ascorbic acid • “ Scurvy “ .

.ascorbic acid saturation test • Administered ascorbic acid is taken up by the tissues until they are saturated. after which there is a sharp rise in the urinary ascorbic acid excretion.

dichloro-phenolindophenol DCPIP and the amount that is decolorized is a measure of the amount of the vitamin C present.Principle • In a protein-free filtrate of plasma or urine. ascorbic acid has the property of decolorizing a blue dye. .

Principle .

. • Glacial acetic acid is corrosive and any spilt on the skin should immediately be washed off with a large volume of water.Notes • Estimation of urinary ascorbic acid when the specimen arrives in the laboratory must carried out immediately . • Ascorbic acid is unstable in urine and a special bottle containing 20 ml of glacial acetic acid must be used for the urine collection.

2. . Pipette 5 ml of urine into a 100 ml volumetric flask and make up to the mark with 5% acetic acid. Pipette 5 ml of DCPIP dye solution into a conical flask.METHOD 1. Titrate with standard ascorbic acid solution (0. 4. 3. Measure the volume of the urine.02 mg/ ml) until the solution just becomes colourless.

Titrate with the dilute urine sample. 6.METHOD cont. Repeat the titrate and take the average . Pipette 5 ml of dye into a conical flask. Calculate how many milligrams of ascorbic acid are equivalent to 5 ml of the dye solution.. The titration must be done very rapidly. 5. 7. otherwise slow-reacting reducing substances present in the urine will contribute to the reduction of the dye 8.

9.. Calculate milligrams of ascorbic acid in total urine collected . Calculate how much ascorbic acid present in sample urine diluted 10.METHOD cont. .

Of ascorbic acid present in sample urine diluted =…………………………………… • Mg of ascorbic acid in total urine collected =………………………………………………………… . • Mg of ascorbic acid equivalent to 5 ml of the dye solution =………………………………………… • The conc.Results • Volume of urine =…………………………….