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(CHEMICAL PATHOLOGY)
• SPECTRO
TOPIC
PHOTOMETRY
OF PRACTICAL • SYSTEM
IV YEAR MBBS INTERNATIONAL
UNITS
CLASS
(SI UNITS)
CASE STUDY
HISTORY & EXAMINATION_
LABORATORY DATA
SERUM RANDOM GLUCOSE ( 40 m mol / l ) , URINE GLUCOSE ( 4+)
pH ( 7.12 )
SERUM ACETONE ( 3+ )
SERUM SODIUM ( 128 m mol / l ). CHLORIDE ( 88 m mol / l ) AND
BICARBONATE ( 09 m mol / l )
POTASSIUM ( 5.7 m mol / l )
RAISED HEMATOCRIT,HEMOGLOBIN AND SERUM BUN
SPECTROPHOTOMETRY
PRINCIPAL OF SPECTROPHOTOMERRY
IT USES THE CHARACTERISTIC
PATTERN OF ELECROMAGNETIC
RADIATION ABSORBANCE FOR THE
IDENTIFICATION OF ANALYTES OF
INTEREST
IT MEASURES THE TRANSMITTED
RADIATION BY DETECTORS
CHEMICAL TESTS DONE ON A
SPECTROPHOTOMETER
• GLUCOSE, HbA1c, LIPID PROFILE AND LIPO-
PROTEINS , PROTEINS INCLUDING
IMMUNOGLOBULINS
• EXCRETIONS LIKE BILIRUBIN, UREA, CREATININE,
URIC ACID
• ENZYMES LIKE AST, ALT, ALP, CPK, LDH, AMYLASE,
ACID PHOSPHATASE, LIPASE, CK-MB AND MANY
OTHERS
• IONS LIKE CALCIUM, PHOSPHORUS, MAGNESSIUM,
• ELECTROLYTES LIKE SODIUM, POTASSIUM,
CHLORIDE, BICARBONATES
• HEMOGLOBIN, IRON, TIBC
COMPONENTS OF A
SPECTROPHOTOMETR
• LIGHT SOURCE
A BEAM OF SUFFICIENT POWER PROVIDING A
CONTINUUM OF WAVELENGTHS OVER THE
REGION OF INTEREST
TUNGSTEN LAMPS, TUNGSTEN HALOGEN LAMPS,
HYDROGEN AND DEUTERIUM DISCHARGE LAMPS
• MONOCHROMATORS
FOR ISOLATION OF THE REQUIRED RANGE OF
WAVELENGTH FOR THE SPECIFIC ANALYTES OF
INTEREST
FILTERS, DIFFRACTION GRATINGS OR PRISMS
COMPONENTS OF A
SPECTROPHOTOMETR
• SAMLPE HOLDER (CUVETTE OR CELL)
SQUARE CROSS SECTIONAL / CIRCULAR CROSS
SECTIONAL
TUBE WITH A PATH LENGTH OF 01 CM
• DETECTOR
PHOTOMULTIPLIER TUBE / PHOTOMULTIPLIER PLATE
( TO CONVERT LIGHT ENERGY INTO ELECTRICAL ENERGY)
FORMULA
( A / B ) MULTIPLIED BY
( CONC. OF STD)
RECOMMENDATIONS
• UNIT OF QUANTITY
(mol) / one can add a prefix
NO of moles= Weight in g / MW
• UNIT OF VOLUME
liter
• UNIT OF CONCENTRATION
mole = MW of a substance in g
dissolved in a liter
m mol / l
EXCEPTIONS
• Measurements of Proteins ( g / l )
• Measurements of Enzymes ( IU / l)
The amount of enzyme which will catalyze the
transformation of 01 micromole of substrate per
minute under standard conditions
• Measurements of Pressure of gases
( mmHg )
• Measurements of some hormones
CONV. U SI UNITS FACTOR
Glucose mg / dl m mol / l d 18
Cholesterol mg / dl m mol / l d 39
Proteins g / dl g/l m w 10
Bilirubin mg / dl u mol / l d 0.058
Urea mg / dl m mol / l d 6
Creatinine mg / dl u mol / l d 0.011
Uric Acid mg / dl m mol / l d 17
Calcium mg / dl m mol / l d 4
Phosphorus mg / dl m mol / l d 3