phosphorus
TRANSCRIPT
الرحــمـــــن الله الرحــمـــــن بـســـــم الله بـســـــمالرحيـــمالرحيـــم
PhosphorusPhosphorus
PhosphorusPhosphorus
99 PrinciplePrinciple SampleSample ReagentsReagents ProceduresProcedures WavelengthWavelength ReadingReading CalculationCalculation Normal valuesNormal values InterpretationInterpretation
PhosphorusPhosphorusPrinciplePrinciple
Inorgaic phosphorus in serum
- ↑ - P- ↑ colour intensity
- absorbed at 640 nm
+ Ammonium molybdate
Phosphomolybdate complexStannous
chloride
Reduction byBlue
colour
Phosphorus Phosphorus SampleSample
Serum or Plasma(Not hemolyzed)
PhosphorusPhosphorusReagentsReagents
Reagent 1 (Colour reagent)Reagent 1 (Colour reagent) Ammonium molybdateAmmonium molybdate Formic acidFormic acid GlycerolGlycerol
Reagent 2 (Reducing reagent)Reagent 2 (Reducing reagent) Stannous chlorideStannous chloride
Standard Phosphorus reagent (4 Standard Phosphorus reagent (4 mg/dl)mg/dl)
PhosphorusPhosphorusProceduresProcedures
Blank Standard Test
Reagent 1(Colour reagent)
1 ml 1 ml 1 ml
Standard - 0.025 ml(25 µl) -
Sample - - 0.025 ml(25 µl)
Mix
Reagent 2(Reducing reagent)
0.1 ml(100 µl)
0.1 ml(100 µl)
0.1 ml(100 µl)
MixIncubate for 10 minutes at 37° c
Blank Standard Test
Blank
Reagent 1 (Colour reagent)
0 -1 –
2 –3 –
4 –5 –
0 -
1 –
2 –
3 –
4 –
5 –
0 -
1 –
2 –
3 –
4 –
5 –
0 -
1 –
2 –
3 –
4 –
5 –
0 -0.1 –
0.2 –0.3 –
0.4 –0.5 –
0.6 –0.7 –
0.8 –0.9 -
BlankReagent 2 (Reducing reagent)
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
BlankBlank
Standard
Reagent 1 (Colour reagent)
0 -1 –
2 –3 –
4 –5 –
0 -
1 –
2 –
3 –
4 –
5 –
0 -
1 –
2 –
3 –
4 –
5 –
0 -
1 –
2 –
3 –
4 –
5 –
Standard
StandardStandard
0 -
0.01 –
0.02 –
0.03 –
0.04 –
0.05 –
0.06 –
0.07 –
0.08 –
0.09 -
0 -
0.01 –
0.02 –
0.03 –
0.04 –
0.05 –
0.06 –
0.07 –
0.08 –
0.09 -
0 -
0.01 –
0.02 –
0.03 –
0.04 –
0.05 –
0.06 –
0.07 –
0.08 –
0.09 -
0 -0.1 –
0.2 –0.3 –
0.4 –0.5 –
0.6 –0.7 –
0.8 –0.9 -
StandardReagent 2 (Reducing reagent)
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
StandardStandard
Test
Reagent 1 (Colour reagent)
0 -1 –
2 –3 –
4 –5 –
0 -
1 –
2 –
3 –
4 –
5 –
0 -
1 –
2 –
3 –
4 –
5 –
0 -
1 –
2 –
3 –
4 –
5 –
Test
0 -
0.01 –
0.02 –
0.03 –
0.04 –
0.05 –
0.06 –
0.07 –
0.08 –
0.09 -
0 -
0.01 –
0.02 –
0.03 –
0.04 –
0.05 –
0.06 –
0.07 –
0.08 –
0.09 -
Serum
0 -
0.01 –
0.02 –
0.03 –
0.04 –
0.05 –
0.06 –
0.07 –
0.08 –
0.09 -
0 -0.1 –
0.2 –0.3 –
0.4 –0.5 –
0.6 –0.7 –
0.8 –0.9 -
TestReagent 2 (Reducing reagent)
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
0 -
0.1 –
0.2 –
0.3 –
0.4 –
0.5 –
0.6 –
0.7 –
0.8 –
0.9 -
TestTest
TestStandardBlank
PhosphorusPhosphorusWavelengthWavelength
640 nm640 nm
PhosphorusPhosphorusReadingReading
Read OD of the standard and the test Read OD of the standard and the test against the blank at 640 nmagainst the blank at 640 nm
OD standard = ……….? ……….OD standard = ……….? ……….
OD test = ……….? ……….OD test = ……….? ……….
PhosphorusPhosphorusCalculationCalculation
According to Beer-According to Beer-Lambert lawLambert law
OD standard → Known conc.OD standard → Known conc. (4 (4 mg/dl)mg/dl)
OD test → -P- conc. In the OD test → -P- conc. In the sample sample
(Unknown)(Unknown)
OD test x Standard conc.-P- conc. in sample = _____________________ OD standard
PhosphorusPhosphorusInterpretationInterpretation
↑ ↑ Increased values Increased values (Hyperphosphatemia)(Hyperphosphatemia)
Normal valueNormal value
↓ ↓ Decreased values Decreased values (Hypophosphatemia)(Hypophosphatemia)
PhosphorusPhosphorusInterpretationInterpretation
↑ ↑ Increased values Increased values (Hyperphosphatemia) (Hyperphosphatemia) False (Spurious):False (Spurious):
Hemolysis:Hemolysis: movement of –P- from movement of –P- from inside the RBCs into the serum.inside the RBCs into the serum.
TrueTrueDecreased renal excretionDecreased renal excretionVitamin D toxicityVitamin D toxicityIncreased –P- intakeIncreased –P- intake
PhosphorusPhosphorusInterpretationInterpretation
↓↓ Decreased values Decreased values (Hypophosphatemia)(Hypophosphatemia) Increased renal excretionIncreased renal excretion
Shifting to inside the cells as in Shifting to inside the cells as in
Insulin adminstrationInsulin adminstration
THANKS FOR ATTENTION
ALL THE BEST