Fluids and Electrolytes Functions of water

1. Transports nutrients to cells

(Sir Aldrin Autencio)
2. Removes waste products through urine
→ Found in serum, plasma, CSF and urine
3. Acts as the body’s coolant (perspiration)
Water 4. Determines the cell volume by its transport into and out of the cells
→ Maintains homeostasis
o Constant stability or balance inside and outside of each cell Fluid types
→ Is the primary fluid of the human body
 Isotonic solution
→ Is the solvent for all the processes in the human body
- Has the same solute concentration as another solution
→ Varies from 40 - 75% of total body weight with values declining by age and obesity
- No net fluid shifts occur between isotonic solutions
o Age of 60 and obesity: 40% ↓ of total body weight due to ↓ skeletal
- Example: NSS, blood (plasma = pH 7.4)
muscle mass with wide distribution of fats which displace water in the
body  Hypotonic solution
→ Normal values: - Has a lower solute concentration than another solution
o Males: 50-60% of the total body weight - Fluid shifts from the hypotonic solution into the more concentrated
compartment to equalize concentrations (net H2O gain)
 ↑ skeletal muscle, ↓ adipose tissue
- Example: half-normal saline (cells swell)
o Females: 45-50% of the body weight
 ↑ adipose tissue  Hypertonic solution
→ Responsible for most of the physiologic reactions of the human body: - Has a higher solute concentration than another solution (net H2O loss)
o Solvent - Example: 5% dextrose in NSS (cells shrink/shrivel/crenate)
o Lubricant
o Coolant Water input
o Cushion  Beverages (64%)
 Food (28%)
2 major compartments of total body water:  Metabolism (8%)
A. Intracellular fluid (ICF) – 66% - 24L – 2/3 (potassium)
→ Found inside the cells and if elevated may cause abnormality Water output
→ Main solvent for biochemical reaction (energy, metabolism and  Kidneys (59%)
respiration) → Principal organ of excretion of waste
B. Extracellular fluid (ECF) – 33% - 16L – 1/3 (sodium- where water goes, sodium  Skin (25%)
follows) → Released by sweat
→ Found in the circulation and may cause pulmonary edema o 50 mmol/L of Na+ lost
o Interstitial fluid (75%) – 11L o 5 mmol/L of K+ lost
 Provides or supplies nutrients to cell and body organs  Lungs (11%)
 Functions as a supply zone  Feces/Anus (5%)
o Intravascular fluid – plasma (25%) – 5L (capillaries – extend to deeper tissue to
deliver oxygen) Insensible liquids: Skin and lungs
Sensible liquids: Urine, stoll and blood (plasma)
The ICF is the fluid inside the cells while the ECF is the fluid found in the circulation like in the
interstitial fluid and in the intravascular fluid.
Plasma → expressed as osmoles/kg of water (w/w)
→ Contains 12% higher water content than whole blood/ serum → Plasma osmolality: 275 - 295
o 93% H20 → major contributors of osmolality:
o 7% solutes (amino acids, glucose, lipids, etc.) – Analytes  sodium
 chloride
Aquaporins → has 2 mechanisms:
→ Permeable membrane where water molecules can move easily  thirst
→ H2O passively enters the cell o ↑ EC osmolality
o Drying of the mouth (once the person has drank water:
Fluid movement ↑BV, ↓EC)

 Diffusion  ADH (Vasopressin/ Arginine vasopressin)

→ Shift of solutes from an area higher concentration to an area of lower
concentration
→ Passive movement of ions across a membrane depends on the size and charge
of the ion being transported and on the nature of the membrane
→ Does not required energy (ATP)
 Active transport
→ Requires energy to move ions across the membrane
→ Requires ATP
→ Shift of solutes from an area of lower concentration to an area of higher
concentration
 Osmosis
→ Refers to the passive movement of fluid across a membrane from a lower
solute concentration and comparatively more fluid into an area of higher
solute concentration and comparatively less fluid
o Restore blood volume and reduce diuresis
B. Osmolarity
→ Measure of solute concentration
→ Measured in osmoles/L of solution (w/v)
→ Is inaccurate in cases of hyperlipidemia or hyperproteinemia
To maintain a normal plasma osmolality, osmoreceptors in the hypothalamus
respond quickly to small changes in osmolality. A 1 – 2% increase in osmolality causes a 4-
fold increase in circulating ADH (Antidiuretic Hormone) / Vasopressin. A 1 – 2% decrease in
osmolality shuts off ADH production. ADH- from hypothalamus and posterior pituitary-stores
and secretes ADH.
Homeostasis
↑ Plasma Osmolality = ↑ solutes, ↓ H2O --- hypothalamus  ↑ H2O, ↓ solute (dilute)

→ ↓ solute concentration , ↑ fluid  ↑ solute concentration, ↓ fluid ↓ Plasma Osmolarity = ↓ solutes, ↑ H2O – excreted  ↓ H2O, ↑ solute

Osmotic pressure Urine osmolality is decreased in:

→ Allows water to be distributed in the body 1. Diabetes insipidus - ↓ specific gravity

Hydrostatic pressure 2. Polydipsia

→ pressure of blood that pushes solutes out from capillaries 3. ↑ Syndrome of Inappropriate ADH Secretion (SIADH) & Hypovolemia
↓ solutes = ↑ hydrostatic pressure → Abnormal increase in ADH due to water retention

Albumin ↑ BV, ↑ Urine SG, ↑ ADH, ↓ Solute concentration

→ high molecular weight proteins; acts as a magnet (attracts water)
↑ albumin = ↑ reabsorption of solutes from interstitial fluid to capillaries
Osmolality and Osmolarity
A. Osmolality
→ Is a measure of the number of dissolved particles in a solution
Regulation of Blood Volume
Adequate blood volume is essential to maintain blood pressure and ensure good
perfusion to all tissues and organs. Sodium and water are interrelated and important in
maintaining the blood volume (Where sodium goes, water follows)
Sodium: main purpose is to maintain blood volume
If ↓ BV  ↓ BP, ↓ Perfusion of organs, ↓ solute
  Renin-Angiotensin-Aldosterone System (RAAS) a. Osmotic pressure b. Boiling point
- Responds primarily to a decreased blood volume ; ↑ plasma solute c. Freezing point
concentration → Most common due to its simplicity (↑ mole, ↓ temperature)
- Secretes aldosterone which increases sodium retention (distal convoluted d. Vapor pressure
tubules)
→ Not routinely used because alcohol tends to result to increase vapour
- Angiotensin II promotes vasoconstriction
→ Only detected when it decreases (↑ mole, ↓ vapor)
- Secretes Renin and Angiotensin
 Freezing point and Vapor Pressure: Principle of osmometer
- ↑ blood volume

Osmometry
Juxtaglomerular cells (glomerulus)  renin  angiotensinogen (hepatocytes) – renin 
→ Technique used to measure the concentration of dissolved solutes in a solution
angiotensin I –ACE  angiotensin II
A. Measured Osmolality
 Vasoconstriction of the efferent arteriole
→ Measured using an osmometer
 Stimulates adrenal cortex to produce aldosterone

B. Calculated Plasma Osmolality

Four other factors that affect blood volume:
1. Atrial Natriuretic Peptide (ANP)
Formula:
→ Promotes sodium excretion in the kidney and acts in regulating blood pressure
and fluid balance Posm = 2(Na+) + Glucose mg/dl + BUN mg/dl  Routinely used
→ Endogenous anti – hypertensive agent released by the cardiac atria 20 3
→ ↓ blood volume ; counteracts RAAS Posm = 1.86(Na+) + Glucose mg/dl + BUN mg/dl + 9
2. ADH (Vasopressin) 18 2.8
→ Conserves water by renal absorption Normal value = 275-295 mOsm/kg
→ Reabsorbs water
→ Produced by the posterior pituitary gland during dehydration Hypoosmolality
→ Activated by plasma osmolality (not always produced) → More water, less solutes; both ADH and thirst are suppressed
→ Signals the collection of H2O in kidneys and stops its excretion → Seen in cases of polydipsia and in patients with Diabetes insipidus
3. GFR increases with the volume expansion and decreases with volume depletion
4. An increase in plasma sodium will increase urinary sodium excretion Hyperosmolality
→ 90% - Na – osmotic activity in plasma → More solutes and less water
→ Determination of plasma sodium content is a determinant of the extracellular → Seen in SIADH (Syndrome of Inappropriate ADH secretion) and hypovolemia
volume content
 If there is ↑ Osmolality, homeostasis must be returned to normal by:
Determination of Osmolality  Sensitive Osmoles
Specimen: serum or urine  ECF Analytes which are restricted or found in the ECF (Na and
Instrument: Osmometer glucose)
→ Specimen: Serum (Plasma is not accepted because it contains anticoagulant that  H2O from the cells is efluxed to the ECF
stimulate osmotically active substances)  Insensitive Osmoles
→ Principle: Uses Sodium Chloride (NaCl) standard solution  Example: Alcohol in which it passively passes through the cell
→ Colligative properties – these are properties influenced by the number of solutes membrane (Alcohol automatically enters the cell
(particles or molecules) present and not its composition
Osmolal Gap SODIUM
→ Formula: Measured osmolality – calculated osmolality Electron Neutrality: body requires substitute if there is a loss of ion and it must have the
→ Indicates the presence of osmotically active particles other than sodium, glucose same charge
and BUN Sodium – Potassium ATPase Pump
→ Osmolal gap measures: Methanol, Ethanol, Lactate and β - hydroxybutyrate
→ NV: <10 mOsm/kg
Clinical Significance of increased Osmolal Gap:
o >10 mOsm/kg: presence of ingested alcohols
o >30 mOsm/kg: presence of alcohols in high quantity to render grave
prognosis

TYPES OF HYPONATREMIA
Electrolytes A. Depletional Hyponatremia
→ These are ions capable of carrying an electrical charge → True loss of total body sodium
→ Principle: Ions either positively or negatively charged o Renal losses – diuretics, primary or secondary hypoaldosteronism,
Addison’s disease
→ Responsible for homeostasis
 Diuretics: Inhibits or blocks sodium and chloride absorption
→ Classified as: (promotes excretion of Na)
o Cations – positively charged (cathode)  Hypoaldosteronism: Decrease production of aldosterone ( ↓
o Anions – negatively charged (anode) Na2+ reabsorption)
o Non-renal losses – diarrhea, vomiting, burns, trauma
→ IV fluids promote Na absorption
ECF ICF ECF ICF
Cations Na+ K+ Anions Cl- PO4- B. DilutionalHyponatremia
→ Refers to low sodium concentration due to the effects of overhydration
Ca+2 Mg+2 HCO3-
→ Examples:
o SIADH – promotes water absorption in DCT and collecting duct
o Hyperglycemia – Na+ decreases by 1.6mmol/L for each 100mg/dL
Functions of Electrolytes: increment in blood glucose
 Glucose is osmotically active
1. Volume and osmotic regulation (Na, Cl, K)
 Glucose (ECF) allows diffusion of water from the cell (to dilute
2. Myocardial rhythm (K, Mg, Ca) gl;ucose) to the ECF thus ↑ water in the plasma
3. Regulation of ATPase ions pumps (Mg)  Promote Na excretion from the extracellular fluid into
4. Acid – base balance (HCO3) intracellular fluid
 Promote excretion of H2O from cells to extracellular fluids
5. Blood coagulation (Ca, Mg)
 Ruled in when blood glucose is <400 mg/dl
6. Neuromuscular excitability (K, Ca, Mg)
7. Production and use of ATP from glucose (Mg, PO4) C. Artifactual/ Pseudohyponatremia
8. Co-factors in enzyme activation (Mg, Ca) → Most common cause of hyponatremia but is not widely known
→ Are analytical errors that lead to false low sodium
→ Water is displaced by increase of lipids and proteins
Electrolyte determination: Na, K, Cl, HCO3 → Examples:
Potassium determination - can be collected by serum tubes provided that it has been o Hyperlipidemia – displaces water (diabetes mellitus, nephrotic
transferred within 30 mins (if not: increase of K) syndrome, cirrhosis)
o Hyperproteinemia
  In vitro hemolysis (During cell lysis, Hgb, a protein is expressed - It is directly proportional to the selective ion concentration. Makes use of glass
out to the sample and displaces water) membrane which selectively exchanges sodium 300X as rapidly as potassium.
 Hemolysis affects Na2+ if it is MARKED HEMOLYSIS - 2 methods of ISE:
- Sodium is not affected by hemolysis I. Direct ISE – sample is undiluted, results are higher than indirect ISE
II. Indirect ISE – sample is diluted. The activity of the ions is the same as its
EXAMPLES OF HYPERNATREMIA concentration
A. Excess water loss
o DI POTASSIUM
o Fever Hypomagnesemia
o Hyperventilation → ↓ Mg2+: promotes aldosterone production
o Severe burns
B. Increase intake or retention Potassium is 6 – 7 mmol/ L: alters the ECG
o Hyperaldosteronism / Conn's Syndrome Potassium is >10 mmol/L: cardiac arrest
→ ↑ production of aldosterone
→ Defect in adrenal cortex Traditional Methods:
o Ingestion of sea water Sodium Cobaltinitrite

Analytical Procedures
3 factors that influence distribution of K+
A. Chemical methods (Albanese Lein) – obsolete method
B. Flame Emission Spectrophotometry – measures unique light emitted by sodium 1. K+ losses occur when Na-K ATPase is inhibited by hypoxia hypomagnesemia and
C. Atomic Absorption Spectrophotometry digoxin intake
- The analyte under investigation are excited by light 2. Insulin promotes entry of K into the skeletal muscle and liver by increasing the Na-K
- 2 types: ATPase pump.
I. Flame Emission Spectrophotometry – measures light emitted by 3. Catecholamine
excited atoms. With a sodium filter (590nm), light emitted is directly  Epinephrine promotes K+ absorption in cell.
proportional to sodium concentration FEP has no need for light source.  Propanolon inhibits entry of K in cells.
The temperature of the flame is 2000-30000C. The monochromator
selects the suitable wavelength of the emitted light. 3 major mechanisms of diminished renal K excretion:
 Inert gases: Helium and Argon A. Reduced aldosterone- most common cause of chronic hyperkalemia
 Na(Yellow light); K(Violet), Li (Red), Mg(Blue) B. Renal Failure – reduced GFR and decreased tubular secretion
 Internal standard: Lithium and cesium a) Acute or chronic renal failure
II. Atomic Absorption Spectrophotometry (AAS) b) Addison’s disease
→ Principle: ground state atoms absorb light at different c) Severe dehydration
wavelengths C. Reduced distal delivery of sodium
→ Is used to measure the concentration by detecting absorption of a) Pseudohyperkalemia
electromagnetic radiation by atoms rather than molecules b) Thrombocytosis
→ the amount of light detected is inversely proportional to the c) In vitro hemolysis
amount of light leaving the hollow cathode lamp d) Recentrifugation of SST
→ Hallow Cathode Lamp - Symptoms: muscle weakness (8mmol/L), cardiac arrhythmias
 specific electrolyte determination
 Light source  If K is depleted, it is buffered by H+ to maintain electroneutrality
 1 HCl per electrolyte  If Na+ and H+ are seen in RBC, it is detected as decrease H+ levels
 Specific but expensive  Extracellular fluid concentrations detected in the lab: HCO- and K+
Special Specimen Consideration:
D. ISE’s (Glass Aluminum Silicate) 1. No hemolysis
- Most common analytical procedure, Newer method 2. No excessive fist clenching
- Directly proportional with FEP 3. No prolonged tourniquet application
- Measures the activity of ions in a solution and not its mass concentration
 4. Do not store on ice since it promotes extracellular shift of K + thereby whole blood  Rare
samples are stored at room temperature  Monoclonal protein present in the sample ( 1 – 3 mmol/L)
Chloride o ↓ plasma albumin concentration
Chloride shift mechanism o Refers to plasma cell dyscresias
 Hypercalcemia – 1% of Ca is found in plasma; 99% in bones
 Hypermagnesemia - intracellular/extracellular fluid

HYPERCHLOREMIA
Cystic Fibrosis
→ Failure to excrete sodium and water
→ Hyperviscous secretion
Chloride generated by cellular metabolism within the tissue diffuses out into both plasma → Is an exocrine gland disorder characterized by excessive mucus secretion leading to
and RBCs. In the RBCs, CO2 forms carbonic acid (H2CO3) which splits into H+ and HCO3- . obstruction of the lungs and Upper respiratory tract due to precipitation of mucus
Deoxyhemoglobin buffers H+ whereas HCO3- diffuses out into the plasma and Cl- diffuses out → Patients have an increased sodium and chloride in sweat because of a defect of a
into the RBC to maintain homeostasis. gene known as CF transmembranous conductance regulator
→ Sweat Chloride Test – specific tool to identify patients suffering from Cystic Fibrosis
- Deoxyhemoglobin – buffers H+ ion → Individuals die within 40yrs.Old
- 1:20 ratio
PilocarpineIontopheresis by Gibson and Cooke
- 7.35-7.45 – blood pH
→ Sweat is collected in a gauze in which Chloride is measured directly with ISE
- ↑ Cl = ↑ Na reabsorbed → Allow the patient to sweat for 30 minutes
- ↓ Cl = ↓ Na reabsorbed → > 65 mmol/ l
Anion Gap → Normal: 5 – 40 mmol/l
→ Interference: Bromide, Cyanide, Cysteine
→ used in acid-base disorders
→ measures extracellular electrolytes Sweat Chloride Test: (Chloridometer)
→ is the difference between unmeasured anion and unmeasured cations (sulfate, → Most reliable parameter and most often measured
lactate, phosphate and organic acid) → Principle: Coloumetric
→ NV: <40mmol/L
→ Monitor severity of diarrhea
→ CF: 60-160mmol/L
→ Urine: 40 mmol/ 24 hours
→ >50 mg of sweat is collected within 30 minutes
→ used as a means of quality control
→ Formula:
Analytical Procedures:
1) AG = (Na + K) – (Cl + HCO3)
A. Mercuric Titration of Schales, Schales and Rice
NV: 12-20 mmol/L
→ Earliest method
2) AG = Na – (Cl + HCO3)
→ Uses mercuric nitrate as the reagent with diphenylcarbazone (DPC) as an
NV: 8-16 mmol/L indicator. Free mercuric ions combine with chloride to form insoluble HgCl 2.
 K+ may be excluded because it is an intracellular cation After the complete titration of the Cl-, excess Hg combines with DPC to form
Causes of Increased Anion Gap insoluble blue-violet complex indicating the end-point of titration. Absorbance
is read at 590nm.
 There is presence of unmeasured anions
 Commonly occurring
→ Interferences: Protein, Bilirubin, Hemolysis, lipemia
 Uremia, renal failure, lactic acidosis, salicylate poisoning, ethylene glycol, methanol
Causes of a Decreased Anion Gap
 B. Colorimetric – Amperometric Technique Bicarbonate
→ For back up analyzers 1. Bicarbonate-carbonic acid buffer system
→ Uses Cotlove chloridometer CO2 + H2O H2CO3 H+ + HCO3-
→ 2 separate electrical circuit employed:  In the glomerulus, H ions combine with HCO3 forming carbonic acid.
a) Colourimetric circuit H2CO3 dissociates to H+ and CO2, CO2 is reabsorbed back into the
→ Silver ions from silver electrode circulation
→ will react with Cl to form AgCl  Only a semi- permeable electrolyte
→ will stop measuring if there is an increase of Ag  HCO3 is filtered in the glomerulus but not reabsorbed
→ Halide ion – cyanide, thiocyanate
Sodium – Hydrogen Exchanger 3
Ag+2 + Cl -  AgCl2
Analytical procedures:
b) Amperometric circuit
a) Natelson Microgasometer Method
→ Detects increased current as free Ag accumulates after
complete titration of the Cl- in the solution. The amount of Cl- in - Reference method ; tedious
a solution is directly proportional to the time required to - All 3 forms of TCO2 are converted to CO2 by acidification of the sample.
generate silver to titrate the chloride This method measures as pressured of CO2 is liberated
b) Colorimetric method
C. Colorimetric methods - Uses cresol red (acidic) as pH indicator. A color change is produced from
→ Use of mercuric or ferric thiocyanate (FERRIC THIOCYANATE METHOD) red to yellow as CO2 gas diffuses across a silicone membrane, decreasing
the pH of the recipient buffer solution

2Cl- + Hg (SCN) 2  HgCl + 2(SCN) - will dissociate Hg binded by SCN c) Enzymatic method

3(SCN) - + Fe+3  Fe (SCN) 3 what is measured - Alkalinizes the sample to convert all forms of CO2 to HCO3-. HCO3- is used
to carboxylate phosphoenolpyruvate (PEP) in the presence of PEP
carboxylase, which catalyzes the formation of oxaloacetate
→ Principle: Thiocyanate ions are displaced from mercury by chloride ions. Free 𝑃𝐸𝑃 𝑐𝑎𝑟𝑏𝑜𝑥𝑦𝑙𝑎𝑠𝑒
thiocyanate combines with ferric ions to form the ferric PEP + HCO3- → oxaloacetate + H2PO4
thiocyanate, a red complex that can be quantitated 𝑀𝐷𝐻
Oxaloacetate + NADH + H → malate + NAD+
spectrophotometrically at 525 nm.
The rate of change in absorbance of NADH at 340 nm is proportional to the
D. ISE
concentration of HCO3-.
→ Membrane made of silver chloride – silver sulphide
340 nm – maximal absorptivity of NADH; constant
→ Membrane: tri – n – actylpropylammoniachloride decanol
d) ISE
E. Spectrophotometric
- Uses silicone membrane
→ Mercuric thiocyanate/ Whitehorn titration technique – reddish complex (480
- Most commonly used (modern automation)
nm)
→ Ferric perchlorate – colored complex
Specimen considerations:
1. Arms must be free from scratches wound and rashes due to contamination of serous
fluid
2. Patient must be aged 48 hours

Hypergammaglobulinemic States
→ ↓ electrolytes because protein diffuse water
Magnesium → Active form of Vitamin D which increases calcium absorption in the intestine
and it enhances the effect of PTH on bone resorption
Analytical procedures:
C. Calcitonin
a) AAS
→ Promotes Ca excretion in the body
→ Reference method. Lanthanum and strontium are contained in the
diluents to bind with phosphate to prevent the formation of magnesium- → Synthesized in the medullary cells of the thyroid gland; secreted when the
phosphate compound which are not measured. concentration of calcium in the blood increases. It has a calcium-lowering
b) Fluorometric methods effect by inhibiting the actions of both PTH and Vitamin D

→ Use of calcein, a fluorescent dye that removes calcium to prevent Parathyroid gland
interferences. Use of 8-hydroxyquinoline binds with magnesium. secrets parathyroid
Separate serum ASAP to prevent leakage of Mg+2 into the serum hormone
c) Colorimetric methods
→ Most commonly used
I. Calagmite forms a complex with Mg+2 (reddish-violet) which is then Bone
Intestine Kidneys
measured spectrophotometrically at 532 nm, EGTA (ethylene glycol
tetraacetic acid) prevents calcium interference stimulates osteoclasts
to release stored Ca2+ 1. reabsorbs Ca
II. Formazan dye method - Mg+2 binds with the dye to form a colored
in the bone to the 2. promotes excretion
complex, 660 nm promotes Ca2+ and plasma (promote bone of phosphates
III. Methylthymol blue - Mg+2 binds with methylthymol blue phosphate absorption breakdown)
3. activates renal -1-
Alpha hydroxylase
 In lactation, there is ↓ Mg because it is used and consumed for milk production
 ↓ Mg (↓ ATP, which is needed for Na – K ATPase pump) = ↓ K+ intracellularly
PTHrP (PTH – related Peptide)
→ Associated with cancers
Calcium
→ Mimics the action of PTH
Regulators of serum Calcium:
→ Binds to PTH receptors yiel;ding ↑ calcium levels
A. Parathyroid hormone (PTH)
Analytical procedures:
→ Promotes Ca reabsorption
a) AAS
→ PTH secretion in blood is stimulated by a negative feedback mechanism. It
→ reference method; specimens are diluted with lanthanum HCl to reduce
will act on the bone to activate bone resorption, and the kidney to increase
viscosity and prevent interferences from protein and other ions. Not
phosphate secretion and calcium reabsorption returning Ica +2 back to
commonly used - expensive
normal. PTH will also activate Vitamin D for maximal reabsorption of calcium
in the SI and Vitamin D enhances the effect of PTH on bone resorption b) Dye-binding methods
→ 3 major effects of PTH: → Ca must be separated first from protein carrier

 Activates bone resorption – breakdown of osteoclast with subsequent Ortho-Cresolphtalein Complexone

release of calcium into the ECF  Dye: Arsenazo III
 In the kidney, PTH conserves calcium by increasing tubular  Mg+2 inhibitor: 8-hydroxyquinoline
reabsorption of calcium ions  570 nm, purple color
 PTH stimulates renal production of Vitamin D c) ISE – liquid membrane
B. 1, 25 Dihydroxycholecalciferol [1,25-(OH)2-D3)
→ Promotes Ca absorption Special Specimen Considerations:
1. Blood must be obtained anaerobically because alkalosis produce falsely low
calcium levels
Phosphorus
Analytical procedures:
a) Fiske-Subbarow Colorimetric method
- Formation of colorless AP (ammonium phosphomolybdate); measures
the absorbance at 340 nm. It can be reduced to form molybdenum blue
which is read at 600-700 nm. Use the following reducing agents:
1. ANSA (aminonaphthol-sulfonic acid)
2. Ascorbic acid
3. Methyl-para-aminophenol sulphate
b) Enzymatic method – neutral pH, NADPH is quantified
2, 3 BPG (Biphosphoglycerine)
→ Related to affinity of oxygen to haemoglobin
→ ↓ O2 to Hgb
→ Contains phosphorus
Specimens:
1. Serum
2. Plasma (Lithium heparin)
3. Urine (24hr) – should be 24 hr because of diurnal variation
Phosphate: ↑ morning; ↓ night

Relationships

Interrelated / Directly
proportional
Sodium and Water
Sodium and Chloride
NADH and Bicarbonate
Inversely proportional
Sodium and Potassium
Chloride and Bicarbonate
Calcium and Phosphorus