Result Part A: Uric Acid test Concentration Absorbance (710nm) 0 0 Uric Acid Standard ( g/ml) 10 20 30 0.114 0.237 0.

413 40 0.534

Urine Samples Average Absorbance Concentration (g/ml) Part B: creatinine test Concentration Absorbance (710nm) 0 0

X 1.278 X1 1.196 1.237 94

Y 0.893 Y1 0.877 0.885 68

Z 0.889 Z1 0.886 0.888 69

Creatinine Standard ( g/ml) 10 20 30 0.061 0.093 0.110

40 0.205

Urine Samples Average Absorbance Concentration (g/ml) 0.204 45

X 0.163 X1 0.244 0.146 31

Y 0.152 Y1 0.140 0.131 29

Z 0.152 Z1 0.110

Part C: Urea test Concentration Absorbance (710nm) 0 0 20 0.0031 Urea Standard ( g/ml) 40 60 0.118 0.067 80 0.095 100 0.122

Urine Samples Average absorbance Concentration (g/ml)

X -0.006 X2 0.048 0.021 18

Y 0.055 Y2 0.067 0.061 51

Z 0.045 Z2 0.026 0.0355 30

Part D; total analyte for sample x, y and z Sample Total uric acid (g/ml/24hour) Total creatinine (g/ml/24hour) Total urea (g/ml/24hour) x 18.8 9.0 9 Y 13.6 6.2 25.5 Z 13.8 5.8 15

Part E: uric acid nitrogen, creatinine nitrogen and urea nitrogen for sample x, y, and z Sample uric acid nitrogen (g/ml/24hour) creatinine nitrogen (g/ml/24hour) urea nitrogen (g/ml/24hour) x 6.26 3.34 4.2 Y 4.53 2.30 11.9 Z 4.60 2.15 7

Calculation Part A: i) total uric acid 1) Total uric acid in 24 hour urine sample x Amount uric acid x dilution factor x urine volume in 24hour = 94g/ml x 100 x 2000ml/24hour =18.8g/ml/24hour 2) Total uric acid in 24 hour urine sample y Amount uric acid x dilution factor x urine volume in 24hour = 68g/ml x 100 x 2000ml/24hour =13.6g/ml/24hour 3) Total uric acid in 24 hour urine sample z Amount uric acid x dilution factor x urine volume in 24hour = 69 g/ml x 100 x 2000ml/24hour =13.8g/ml/24hour ii) uric acid nitrogen 1) Amount uric acid nitrogen in urine sample x = Amount of uric acid in 24hour x molar mass of nitrogen = 18.8g/24hour x ( = 6.26g/24 hour )

2)

Amount uric acid nitrogen in urine sample y = Amount of uric acid in 24hour x molar mass of nitrogen = 13.6g/24hour x ( = 4.53/24 hour )

3)

Amount uric acid nitrogen in urine sample z = Amount of uric acid in 24hour x molar mass of nitrogen = 39.5g/24hour x ( = 4.60/24 hour )

Part B: i) total creatinine 1) Total creatinine in 24 hour urine sample x Amount creatinine x dilution factor x urine volume in 24hour = 45g/ml x 100 x 2000ml/24hour =9g/ml/24hour 2) Total creatinine in 24 hour urine sample y Amount creatinine x dilution factor x urine volume in 24hour = 31g/ml x 100 x 2000ml/24hour =6.2g/ml/24hour 3) Total creatinine in 24 hour urine sample z Amount creatinine x dilution factor x urine volume in 24hour = 29 g/ml x 100 x 2000ml/24hour =5.8g/ml/24hour ii) creatinine nitrogen 1) Amount creatinine nitrogen in urine sample x = Amount of creatinine in 24hour x molar mass of nitrogen = 5.8g/24hour x ( = 3.34g/24 hour

2)

Amount creatinine nitrogen in urine sample y = Amount of creatinine in 24hour x molar mass of nitrogen = 6.2g/24hour x ( = 2.30/24 hour )

3)

Amount creatinine nitrogen in urine sample z = Amount of creatinine in 24hour x molar mass of nitrogen = 5.8g/24hour x ( = 2.15g/24 hour )

Part C: i) total urea 1) Total urea in 24 hour urine sample x Amount urea x dilution factor x urine volume in 24hour = 18 g/ml x 250 x 2000ml/24hour =9g/ml/24hour 2) Total urea in 24 hour urine sample y Amount urea x dilution factor x urine volume in 24hour = 51 g/ml x 250 x 2000ml/24hour =25.5g/ml/24hour 3) Total urea in 24 hour urine sample z Amount urea x dilution factor x urine volume in 24hour = 30 g/ml x 250 x 2000ml/24hour =15g/ml/24hour ii) Urea nitrogen 1) Amount urea nitrogen in urine sample x = Amount of urea in 24hour x molar mass of nitrogen = 9g/24hour x ( = 4.2g/24 hour 2) Amount urea nitrogen in urine sample y = Amount of urea in 24hour x molar mass of nitrogen = 25.5g/24hour x ( = 11.9g/24 hour 3) Amount urea nitrogen in urine sample z = Amount of urea in 24hour x molar mass of nitrogen = 15g/24hour x ( = 7g/24 hour ) ) )

Discussion

The folin reagent which are stable in the acidic form, however if want the reduction of the folin reagent to occur it have to increase the untill pH 10. Therefore in the experiment the folin solution was added with the NaOH which are help to increasing the pH of the reagent for the reduction. (Becker, Caldwell, & Zachgo, 1996) Uric acid causes phosphotungstic acid to reduce to blue tungsten oxides. Uric acid is oxidized to allantolin. In the experiment is show that the solution is become dark blue colour which indicate the uric acid was present in the urine and the phosphotungstic acid was reduced to tungsten oxides which appear in dark blue colour (Chawla, 2003). The increasing of the blue intensity indicate the higher concentration of allantolin which form from the uric acid. Piric acid is a substance that able to react with creatinine and giving a reddish yellow coloured compound in alkaline medium. This reaction is called Jaffes reaction, first the piric acids have to react with the alkaline medium first like sodium hydroxide to form a alkaline picrate. After that the alkaline picrate was react to the creatinine will form reddish yellow crystals which are creatinine picrate. The higher the creatinine present in the sample the higher the intensity of the colour measured at 520nm (Shanmugam, Kumar sathish, & Panneer Selvam, 2010). The urea test are also known as Berthelot reaction which are testing for the ammonium ion present in sample. The urease will broken down the urea into the ammonia ion and carbon dioxide. After that the hypochlorite will added to react with the urea form monochloramine which will react again with the phenol to form a indophenol dye (Ochei & Kolhatkar, 2000). The higher the concentration of urea will form the higher concentration of indophenol. Therefore the intensity of the solution will higher. The urea is the product of urea cycle which using for eliminate the excess ammonia which arise from the deamination of aminoacid our body. The normal range for the urea amount inside the urine is 12 gram to 20 gram per hour. The low level of urea is indicate that the patient malnutrition or kidney problem cause the urea cannot eliminate in the urine for example for elderly people. While for the patient who has a higher amount of urea in the urine means that the person taking too much of the

protein diet and the rate of the protein break down are much frequent or faster than the other for example like athelets. The uric acid is the major-end product of purine metabolism. The uric acid which is insoluble in water and because of it the high level of uric acid in urine may cause the kidney stone. The high level of uric acid on urine mostly are associated by the hyperuricemia which will cause the formation of gout on the joint. The reason that cause the patient high level of uric acid in the urine is the patient taking purine rich food for example mushroom, beans and spinach.

Bibliography
Becker, J. M., Caldwell, G. A., & Zachgo, E. A. (1996). Biotechnology: A Laboratory Course. san Diego: acedemic press. Chawla. (2003). Practical Clinical Biochemistry: Methods and Interpretations (3rd ed.). New Delhi: Jaypee Brothers Medical Publisher (p) Ltd. Ochei, J., & Kolhatkar, A. (2000). Medical Laboratory Science : Theory And Practice. New Dehli: Mc-Graw Hill Publishing Company Limited. Shanmugam, S., Kumar sathish, T., & Panneer Selvam, T. (2010). Laboratory Handbook on Biochemistry. New Dehli: PHI learning private limited.