CALCULI QUANTITATIVE ANALYSIS

Presented by:-Damini. Moderator-Dr.Harjit kaur. BSc.-M.L.T 3rd year.

 It is sometimes necessary to obtain more precise information about the composition of a stone Physical techniques like X-ray diffraction optical crystallography Infra red spectroscopy have been employed but chemical analysis remains the most convenient procedure

Calculi analysis
Semi quantitative method (kit-method) gives the percentage of the component present in the stone Principle-Dissolve the sample of calculus as homogeneous as possible from this soln.various components are determined

Estimation carried out by this method in urinary calculi

Titrimetric method:Calcium Visual clour comparison:oxalate phosphate magnesium ammonium uric acid cystine

Sample preparation
Finely triturate the calculus to be analysed in a mortar Mix the resulting powder thoroughly To the amount of a level spatula,add 5 drops of sulphuric acid (95-97%) and stir to achieve complete dissolution (evolution of gas during dissolution indicates carbonate)

 Transfer the soln. to 100 ml graduate filled to 1/3rd with D/W ,make upto 50 ml mark with D/W

Mix well Transfer the soln. into reaction vessel

Carry out the estimations

CALCIUM
Principle-titrimetric determination using the reagent (ethylenedinitrilotetraacetic acid disodium salt) A trituration of calconcarboxylic acid is used as an indicator Procedure Sample 2 drops of 1 spatula of while mixing soln. NaOH soln. & the indicator add the reagent, (drop by drop)

colour changes from red to blue count the drops required for colour change to occur

no. of drops required multiplied by 5 gives the percentage calcium content of the calculus

Oxalate
Principle-the colour complex formed by iron (III) and sulfosalicylic acid is discharged by oxalate Procedure Sample add 2 drops of borate buffer , soln. 3 drops of iron (III)chloride soln., 3 drops of sulfosalicylic acid soln.

Allow to stand for 2 min.

 Compare with appropriate colour scale

Read off the percentage oxalate content of the calculus

Ammonium
Principle-on addition of nesslers reagent ammonium gives a yellow to brown soln. Procedure Sample add 3 drops of tetraiodomercurate soln. NaOH soln.

 Compare with appropriate colour scale

Read off the percentage oxalate content of the calculus

Phosphate
Principle-the molybdatophosphoric acid formed upon addition of ammonium molybdate is reduced to molybdenum blue by means of reducing agent Procedure Sample add ammonium molybdate soln. soln. Reduction soln. (4- methylaminophenol sulfate,sodium disulfite) Allow to stand for 5 min.

 Compare with appropriate colour scale

Read off the percentage oxalate content of the calculus

Magnesium
Principle-in a bufferd soln. magnesium reacts with a colour reagent(1-azo-2-hydroxy-3-(2,4dimethyl-carboxoanilido)-napthalene-1`-(2hydroxy-benzene-5-sodium sulfonate) soln. to form a red complex Procedure Sample add 10 drops of borate buffer soln. 10 drops of colour reagent

 After 1 min. compare with appropriate colour scale

Read off the percentage oxalate content of the calculus

Uric acid
Principle-in a buffered soln. molybdatophosphoric acid is reduced to form molybdenum blue by uric acid Procedure Sample add 3 drops of molybdatophosphoric soln. acid soln. allow to stand for 2 min. add 2 drops of borate buffer soln.

 Compare with appropriate colour scale

Read off the percentage oxalate content of the calculus

Cystine
Principle-cystine is reduced to cystine by sodium sulfite,in an alkaline medium cystine gives red colour with sodium nitroprusside Procedure Sample add 10 drops of ammonia soln.,a soln. Spoonful of sodium sulfite,dissolve after 1 min. add spoonful of sodium nitroprusside trituration

 Compare with appropriate colour scale

Read off the percentage oxalate content of the calculus

Advantages
This method gives almost identical results to the physical methods (infrared spectrometry and x-ray examination) Rapid diagnostic method

Disadvantages
Reagents used are harmful hence,have to used with great care

THANK YOU