First shift extrawork:

-methods of determination
Ex: AMS
method A. – principle
Substrate -> end product
Clinical Significance of each substrate measured
@ 1wholesheet pad paper

Second shift scope:

- electrolytes
- acid base balance
Extrawork:
-physiologic uses of various electrolytes and trace elements
-methods for determination of each electrolyte and trace elements
ex: Sodium
method A. – principle involved
-What diseases (table form) wherein electrolytes are increased/high or decreased
ELECTROLYTES
1. Electrolytes are charged particles. They are either cations of anions. The cations are attracted to
the cathode while the anions are attracted to the anode
a) Cations – positive ion, carries + charge
i. Attracted to cathode (negative electrode)
b) Anions – negative ion, carries – charge
i. Attracted to anode (positive electrode)
2. The cations in the serum are Na, K, Ca and Mg
a) Most abundant: Na
Least abundant: Mg
3. The anions in the serum are: Cl, HCO3, iPO4
a) Most abundant: Cl
Lease abundant: iPO4
4. Electrolyte profile is used in the calculation of anion gap. Anion gap value is high in metabolic
disorder and low in cancer
a) Electrolyte profile (4 tests): used in computation of anion gap
basis of selecting the 4 tests: 2 most abundant cations and anions
i. Determination of Na+ - average value: 140mEq/L 140mmol/L (equal only if
valence is = 1)
ii. Determination of K+ - average value: 4mEq/L or 4mmol/L
iii. Determination of Cl- - average value: 100mEq/L or 100mmol/L
iv. Determination of HCO3- - aveage value: 24-27 mEq/L or 24-27mmol/L
b) Sx used: serum
unit expressing conc elec: mEq or mmol/L
 c) Anion gap = (Na+ + K+) – (Cl- + HCO3-)
(140+4) – (100+24)
Anion gap = 20 = N = electrolyte order/balance
~ if anion gap = 30: metabolic acidosis, electrolyte disorder, advance diabetes
~ if anion gap = 4: accumulation of cancer proteins (basic proteins)
5. Serum osmolality - total number of moles of solutes dissolved in serum
a) Osmolality: total number of moles (gr solute / MW solute) of solutes per kg of solvent
b) NaCl - #1 solute in the serum
c) Glucose - #2 solute in the serum
d) BUN - #3 solute in the serum
e) 2 kinds of osmolality:
i. Actual osmolality – measured using osmometer
ii. Computed osmolality/Calculated osmolality – approximation of osmolality
 Base value from the conc of NaCl, glucose and BUN
 Osmolality based on 3 tests: NaCl, glucose and BUN
f) Na is ionized Na + Cl
i. Cl coexists with Na
ii. Na can partner with Cl and others
g) Serum osmolality – may be calculated or computed

𝑔𝑙𝑢𝑐𝑜𝑠𝑒 (𝑚𝑔 %) 𝐵𝑈𝑁

Serum osmolality Eq1= (𝑁𝑎 𝑥 2) + 𝑚𝑚𝑜𝑙 + 𝑚𝑚𝑜𝑙
18 2.8
𝐿 𝐿

𝑔𝑙𝑢𝑐𝑜𝑠𝑒 𝐵𝑈𝑁
Serum osmolality Eq2 = (𝑁𝑎 𝑥 1.86) + + +9
18 2.8
18 = MW of glucose (C6H12O6)
2.8 = measured from urea
6. Sodium determination is done using the ZUA method, EFP method and ISE method

outside: extracellular fluid (plasma, serum)

Cell Na+ most abundant cation in the ECF
K+ > Mg+ Cl- most abundant anion in the ECF

In hemolysis: K+ is abnormally elevated

Na+ measurement
a) Reaction of Na in serum with Zinc Uranyl acetate
𝑁𝑎 + 𝑍𝑈𝐴 → 𝑦𝑒𝑙𝑙𝑜𝑤 𝑝𝑟𝑜𝑑𝑢𝑐𝑡 (𝑚𝑒𝑎𝑠𝑢𝑟𝑒 𝑖𝑛 𝑠𝑝𝑒𝑐𝑡𝑟𝑜)
b) EFP: Emission Flame Photometry
𝑏𝑢𝑟𝑛𝑠 𝑒𝑙𝑒𝑐𝑡𝑟𝑜𝑙𝑦𝑡𝑒𝑠 𝑖𝑛 𝑠𝑒𝑟𝑢𝑚 → 𝑦𝑒𝑙𝑙𝑜𝑤 𝑓𝑙𝑎𝑚𝑒 (𝑚𝑒𝑎𝑠𝑢𝑟𝑒𝑑 𝑏𝑦 𝑚𝑒𝑎𝑛𝑠 𝑜𝑓 𝐸𝐹𝑃)
i. Parts of EFP
 Atomizer – create fine spray of serum sx, bring spray to flame
 Flame – provided by gas in air flame, or gas in oxygen flame
 Monochromator – to isolate colored emission (isolate yellow flame from
violet/purple flame)
  Photocell – for color to turn into electrical current
 Galvanometer/ Potentiometer – to measure electrical current
c) ISE: Ion Sensitive Electrode (Ion Selective/Specific Electrode)
i. Electrode for Na+: glass ISE
7. Sodium maintains normal hydration of the body, forms osmotic pressure and the major influence
of serum osmolality
a) Human body = 70% water
b) Forms osmotic pressure – so water cannot go and clog into all areas due to the pressure
formed by Na
c) Na+ - with highest conc in serum among solutes
8. Sodium concentration in the blood may be either hypernatremia or hyponatremia
a) Hypernatremia – high levels of Na in blood
i. Ex: patient undergoing diuretic therapy: hyponatremia
Patient w/ congestive heart failure: hypernatremia
b) Hypokalemia – low levels of Na in blood
9. Potassium is important in the contraction of muscles (K+: banana)
a) Contraction of heart muscle
b) Contraction of skeletal muscle
*hyperkalemia: lead to cardia arrest
10. Potassium determination in the serum is done using:
a) Sodium cobaltinitrite
𝑁𝑎3𝐶𝑜 (𝑁𝑜2)6 → 𝑦𝑒𝑙𝑙𝑜𝑤 𝑝𝑟𝑜𝑑𝑢𝑐𝑡 (𝑠𝑝𝑒𝑐𝑡𝑟𝑜)
b) EFP
𝑓𝑙𝑎𝑚𝑒 𝑏𝑢𝑟𝑛 𝐾 (𝑠𝑒𝑟𝑢𝑚) → 𝑣𝑖𝑜𝑙𝑒𝑡 𝑓𝑙𝑎𝑚𝑒 → 𝑚𝑜𝑛𝑜𝑐ℎ𝑟𝑜𝑚𝑎𝑡𝑜𝑟 (𝑓𝑖𝑙𝑡𝑒𝑟) 400𝑛𝑚
c) ISE: antibiotic valinomycine electrode

11. Calcium is found in the bones and teeth

a) Intracellular fluid: K > Mg
b) Extracellular fluid: Na > Cl
c) Bones and teeth: Ca+2 >
d) Most abundant mineral in serum: Na
e) Most abundant mineral in human body: Ca+2
f) Most abundant mineral in the blood: Fe+3
g) Most abundant mineral inside RBC: K+ and Mg+
12. The three forms of calcium in the blood:
a) Ionized form – unbound, free, Ca+2, 50% of total
- physiologically active
b) Albumin bound form – bound to albumin, 40% of total
c) Complexed form – remaining 10%, Ca3(PO4)2
99% of body’s Ca: bones and teeth (cement)
Only 1% of body’s Ca is in blood
13. Factors affecting blood calcium levels:
a) Vitamin D – solar vitamin (skin is trapped in vit D from sunlight)
i. Promote absorption of Ca from the food at the small intestines
 ii. High Vit D = High Ca
b) Parathyroid hormone (PTH) – high PTH = high Ca
i. Major mineral in Ca metabolism
c) Calcitonin – protein hormone (with “-in)
i. High calcitonin = Low Ca
14. Calcium concentration in the serum is measured by:
a) EDTA titration – oldest method
𝐶𝑎 + 2 + 𝐸𝐷𝑇𝐴 → 𝑤ℎ𝑖𝑡𝑒 𝑝𝑝𝑡 (𝐶𝑎 𝑒𝑑𝑒𝑡𝑎𝑡𝑒)
b) Colorimetric method: Clark and Collip method

𝐶𝑎 + 2 + 𝐾2𝐶2𝑂4 → 𝑤ℎ𝑖𝑡𝑒 𝑝𝑝𝑡 (𝐶𝑎𝐶2𝑂4)

𝐶𝑎𝐶2𝑂4 + 𝐻2𝑆𝑂4 → 𝐻2𝐶2𝑂4 (𝑜𝑥𝑎𝑙𝑖𝑐 𝑎𝑐𝑖𝑑)
dissolve
𝐻2𝐶2𝑂4 + 𝐾𝑀𝑛𝑂4 (𝑡𝑖𝑡𝑟𝑎𝑡𝑒) → 𝑑𝑒𝑐𝑜𝑙𝑜𝑟𝑖𝑧𝑎𝑡𝑖𝑜𝑛(𝑒𝑛𝑑 𝑝𝑜𝑖𝑛𝑡)
𝑝𝑢𝑟𝑝𝑙𝑒 𝑠𝑜𝑙𝑛
c) Routine method: Autoanalyzer
𝐶𝑎 + 2 + 𝑂𝐶𝑃𝐶 → 𝑦𝑒𝑙𝑙𝑜𝑤
𝑠𝑒𝑟𝑢𝑚 𝑑𝑦𝑒
𝑜𝑟𝑡ℎ𝑜𝑐𝑟𝑒𝑠𝑜𝑙𝑝𝑡ℎ𝑎𝑙𝑒𝑖𝑛 𝑐𝑜𝑚𝑝𝑙𝑒𝑥 ?

d) AAS: Most preferred method

i. Atomic Absorption spectrophotometry
ii. Measure Ca and Mg
iii. Hollow cathode tube -> release radiation -> absorbed unk (ex: Ca+2) -> amt
radiation absorbed by unk meas AAS
iv. Mg +2: extracellular
15. *
16. Chloride determination in sweat is used in the diagnosis of cystic fibrosis of the pancreas. The
method employed is pilocarpine iontophoresis
a) Cl-: measure
i. Sweat
 sweat Cl det: Dx of cystic fibrosis (inability of person to reabsorb Cl
prior to sweating)
Cl goes out with sweat
ii. Serum Cl = dehydration
 Hyperviscoate body fluid*
 Pulmonary fluid viscous -> scarred lungs -> infection -> death
iii. Sweat Cl- determination
 Old: cellophane method
 New: Pilocarpine into pluresis
a. Alkaloid drug
b. Administered underneath the skin
 Effects:
 a. Sialogigic effect (salivation)
b. Diaphoretic effect

K2CrO5
𝑆𝑤𝑒𝑎𝑡 𝑆𝑥 + 𝐴𝑔𝑁𝑂3 →→→ 𝑟𝑒𝑑

1) Abnormally high Cl-: detected cystic fibrosis

2) Cl- sweat = 50-60 mmol/L
3) Serum Cl det:
a) Schales & Schales – Mercuric titrations method
i) Manual
𝑆𝑒𝑟𝑢𝑚 𝐶𝑙 − +𝐻𝑔2 − −→ 𝑒𝑛𝑑 𝑝𝑜𝑖𝑛𝑡 (𝑣𝑖𝑜𝑙𝑒𝑡 𝑏𝑙𝑢𝑒)
𝑖𝑛𝑑𝑖𝑐𝑎𝑡𝑜𝑟
b) Chlondometer – sample
i. Instrumental

17. Fe determionation in the serum is done by adsorption of various dyes

a) Iron: Fe+2: food: newly formed blood
b) Fe +3: blood (old)
c) Fe+2 oxidized Fe+3 predominantly n k blood -> Fe+3 formed
d) Steps in det of Fe+3
i. Remove protein transport by HCl
 Fe+2: transferrin
 Fe+3: ferritin
ii. Reduce all Fe+3 (inert) to Fe+2 (active)
iii. Dye Fe+2
 Ferrozine dye
 Tripyridyl triazine
 Bathophenanthroline

18. Cu determination is used in the diagnosis of Wilson’s disease

a) Copper deficiency: Menke’s syndrome “kinky hair”
b) Copper toxicity: Wilson’s disease
i. Elevated copper in the serum
ii. Copper is not circulated bc of the deficiency of transport protein (ceruloplasmin)
iii. Excess copper is deposited in liver tissue w/c causes hepatolenticular
degeneration (Wilson’s disease)