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CLINICAL CHEMISTRY

(CHEMICAL PATHOLOGY)

• SPECTRO
TOPIC
PHOTOMETRY
OF PRACTICAL • SYSTEM
IV YEAR MBBS INTERNATIONAL
UNITS
CLASS
(SI UNITS)
CASE STUDY
HISTORY & EXAMINATION_

20 YEARS OLD WOMEN BROUGHT TO EMERGENCY ROOM IN A COMATOSE STATE


DEEP AND RAPID BREATHING,FRUITY ODOR,SKIN & MUCUS MEMBRANES DRY
PATIENTS OLDER BROTHER HAD TYPE I DIABETES MELLITUS

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)

• READOUT DIGITAL DEVICE


 A METER / A DIGITAL READOUT DEVICE / RECORDER / LIGHT
EMITTING DIODE ( LED ) / LIQUID CRSTAL DISPLAY ( LCD )
CALCULATION

 TRUE ABS. OF TEST (A)


 TRUE ABS. OF STD (B)
 CONCENTRATION OF
STANDARD

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

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