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![Distribution of phosphate in blood
• Plasma contain both inorganic & organic phosphate.
• Phosphate: intracellular ion
• [Phosphate] of whole blood =40mg/dl
• Serum phosphate= 3-4 mg/dl
» Protein bound [10%]
» Complexed with Na, Ca, Mg [35%]
» Free [55%]](https://image.slidesharecdn.com/phosphoruspractical-200121114930/75/Phosphorus-practical-3-2048.jpg)
![Functions of phosphate
• Bone & teeth formation.
• ATP [energy currency of cell] [ATP → ADP + Pi]
– Involved in many energy intensive physiological
functions.
– Muscle contraction
– Neurological functions
– Electrolyte transport
• Essential for absorption & metabolism of carbohydrate.](https://image.slidesharecdn.com/phosphoruspractical-200121114930/75/Phosphorus-practical-4-2048.jpg)
![Functions of phosphate
• Acts as buffer. [H2PO4
- &HPO4
--]
H2PO4
- : HPO4
--
In acidosis 1:1
At pH 7.4 1:4
In alkalosis 1:9](https://image.slidesharecdn.com/phosphoruspractical-200121114930/75/Phosphorus-practical-6-2048.jpg)
![Regulation of phosphate level
• Role of PTH
– ↑es Phosphate excretion via kidney (phosphaturic
effect)
• Role of Calcitriol
– ↑es intestinal Phosphate absorption
• Role of calcitonin
– ↑es phosphate excretion via kidney.
– ↓es bone resorption &↑es bone mineralization
[phosphate deposition into bone]](https://image.slidesharecdn.com/phosphoruspractical-200121114930/75/Phosphorus-practical-7-2048.jpg)
![• HYPOPHOSPHATEMIA
– [Phosphate] : < 2.5 mg/dL (0.8 mmol/L).
– Mechanisms causing hypophosphatemia:
» Inadequate intake
» ↓ed intestinal absorption
» ↑ed excretion
» Shift from ECF to ICF](https://image.slidesharecdn.com/phosphoruspractical-200121114930/75/Phosphorus-practical-12-2048.jpg)
![• Hyperphosphatemia
[Phosphate] : > 4.0 mg/dL
– Mechanisms causing hyperphosphatemia:
• ↑ed phosphate intake
• ↓ed phosphate excretion
• Shift from ICF to ECF](https://image.slidesharecdn.com/phosphoruspractical-200121114930/75/Phosphorus-practical-17-2048.jpg)
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Phosphorus practical
- 1. Estimation of serumtotal phosphate by ANSA method Amit Jha Lecturer UCMS, Bhairahawa
- 2. Distribution of phosphatein body • Found in every cell of body. • Total body phosphate content= 700gm (25mole) – (both in organic & inorganic form) Mostly in combination with Ca in bone & teeth 10% is in association with protein, Lipid & Carbohydrate in muscle & blood Remaining is distributed in various chemical compound.
- 3. Distribution of phosphatein blood • Plasma contain both inorganic & organic phosphate. • Phosphate: intracellular ion • [Phosphate] of whole blood =40mg/dl • Serum phosphate= 3-4 mg/dl » Protein bound [10%] » Complexed with Na, Ca, Mg [35%] » Free [55%]
- 4. Functions of phosphate •Bone & teeth formation. • ATP [energy currency of cell] [ATP → ADP + Pi] – Involved in many energy intensive physiological functions. – Muscle contraction – Neurological functions – Electrolyte transport • Essential for absorption & metabolism of carbohydrate.
- 5. Functions of phosphate •Essential component of cyclic nucleotides. (cAMP, NADP) • Present in many coenzymes (TPP, NADP, PLP). – Thus req. for activity of many enzymes. • Phosphate is essential for formation of:- » Phosphoproteins » Phospholipids » Nucleic Acids
- 6. Functions of phosphate •Acts as buffer. [H2PO4 - &HPO4 --] H2PO4 - : HPO4 -- In acidosis 1:1 At pH 7.4 1:4 In alkalosis 1:9
- 7. Regulation of phosphatelevel • Role of PTH – ↑es Phosphate excretion via kidney (phosphaturic effect) • Role of Calcitriol – ↑es intestinal Phosphate absorption • Role of calcitonin – ↑es phosphate excretion via kidney. – ↓es bone resorption &↑es bone mineralization [phosphate deposition into bone]
- 8. ↓ in PlasmaPhosphate ↓ Stimulates 1-α-hydroxylase ↓ ↑es Calcitriol ↓ ↑es Intestinal absorption of Phosphate ↓ Plasma Phosphate ↑es towards normal
- 9. ↑ in PlasmaPhosphate ↓ Stimulates PTH ↓ ↑es Phosphate excretion via kidney ↓ Plasma Phosphate ↓es towards Normal
- 10. Principle: Inorganic Phosphorusby (ANSA Method) Phosphorus + Ammonium molybdate Acidic pH Ammonium Phosphomolybdate Heteropoly molybdenum Blue (680 nm) 1-Amino 2-Naphthol, 4- Sulfonic acid (ANSA)
- 11. Procedure: PPF: 1 mlSerum + 4 ml 10 % TCA, mixed and centrifuge Blank Standard Test Filtrate ---- ---- 2.5 ml Standard (4 mg/dl) ---- 2.5 ml ---- D/W 2.5 ml ---- ---- Molybdate solution 0.5 ml 0.5 ml 0.5 ml ANSA 0.2 ml 0.2 ml 0.2 ml Mix well and allow to stand at 10 minutes and add 1.5 ml of D/W to each tube. Mix and read OD at 680 nm (Red filter) Serum Phosphorus = OD T – OD B X 4 mg/dl OD S – OD B
- 12. • HYPOPHOSPHATEMIA – [Phosphate]: < 2.5 mg/dL (0.8 mmol/L). – Mechanisms causing hypophosphatemia: » Inadequate intake » ↓ed intestinal absorption » ↑ed excretion » Shift from ECF to ICF
- 13. • Inadequate intake& ↓ed intestinal absorption 13 Long-term low phosphate intake in setting of sudden ↑ in intracellular phosphate requirements eg: refeeding syndrome Intestinal malabsorption Vit. D def. ↓ Calbindin ↓ intestinal absorption Ingestion of Al/Mg antacids Binds to dietry phosphate ↓ absorption
- 14. 14 • ↑ed excretion Hyperparathyroidism↑ in PTH inhibit phosphate reabsorption in PCT Forced saline diuresis Inhibitory effect on PCT transport processes Fanconi syndrome
- 15. • Renal Phosphatewasting 15
- 16. • Shift fromECF to ICF 16 Carbohydrate diet Entry of PO4 & Glucose into cell Insulin therapy Entry of PO4 & Glucose into cell Short-term ↑ in cellular demand eg: Hungry bone syndrome Acute respiratory alkalosis Activation of PFK ↑ed glycolysis
- 17. • Hyperphosphatemia [Phosphate]: > 4.0 mg/dL – Mechanisms causing hyperphosphatemia: • ↑ed phosphate intake • ↓ed phosphate excretion • Shift from ICF to ECF
- 18. • ↑ed intake –IV or oral administration – Phosphate containing laxative
- 19. • ↓ed renalphosphate excretion 19 Acute or chronic renal failure ↓ GFR → ↓ excretion Hypoparathyroidism ↓ PTH → ↓ excretion
- 20. • Shifts fromICF to ECF 20 Transcellular shift Cell lysis •Lactic acidosis •Rhabdomyolysis •DKA •Leukemia •Respiratory acidosis •Lymphoma •Cytotoxic therapy