OUR LADY OF GUADALUPE COLLEGES

DEPARTMENT OF MEDICAL TECHNOLOGY

LABORATORY MANUAL IN CLINICAL CHEMISTRY 2

Name: KATE NICOLE C. PORLEY Date: FEBRUARY 18, 2023

Year and section: BSMT-3 Score:

ACTIVITY NO. 2
ELECTROLYTES

1.) In tabulated form, enumerate the reference range (serum, plasma, urine) of the following
electrolytes:
- Sodium
- Potassium
- Chloride
- Bicarbonate
- Magnesium
- Calcium
- Phosphate

Electrolytes Serum Plasma Urine

Sodium 135-145 mmol/L 135-145 mmol/L 120-240 mmol/d, varies with diet

Potassium 3.5–5.1 mmol/L 0.5 mEq/L 33–86 mmol/d

Chloride 98–107 mmol/L 98–107 mmol/L 110–250 mmol/d, varies with

diet

Bicarbonate 22-28 mmol/L 22-28 mmol/L

Magnesium 0.66–1.07 mmol/L 0.75 to 1.0 mmol/l 6-10 mEq/day (3-5 mmol/day)
(1.7–2.4 mg/dL) (1.8 to 2.4 mg/dl)
(colorimetric)
Calcium Total Calcium Child- 2.13–2.63 Calcium in the urine should be
Child- 2.13–2.63 mmol/L (8.5–10.5 100 to 300 milligrams per day
mmol/L (8.5–10.5 mg/dL) (mg/day) or 2.50 to 7.50
mg/dL) Adult- 2.24–2.53 millimoles per 24 hours
Adult- 2.24–2.53 mmol/L (9.0–10.1 (mmol/24 hours).
mmol/L (9.0–10.1 mg/dL) If the person is eating a diet low
mg/dL) in calcium, the amount of
calcium in the urine will be 50 to
Ionized Calcium 150 mg/day or 1.25 to 3.75
1.15–1.33 mmol/L mmol/24 hours
(4.6–5.3 mg/dL)

Phosphate Neonate- 1.45–2.91 2.8 to 4.5 mg/dL 13–42 mmol/d (0.4–1.3 g/d)
mmol/L (4.5–9.0
mg/dL)
Child ≤15 y-
1.29–2.26 mmol/L
(4.0–7.0 mg/dL)
Adult- 0.81–1.45
mmol/L (2.5–4.5
mg/dL)

2.) Illustrate and explain the effects of blood volume and osmolality on Na+ and water
metabolism.
The following are the effects of blood volume and osmolality on sodium and water metabolism:

● The kidneys respond by decreasing the GFR and secreting renin near the renal glomeruli
in feedback to decrease the blood volume and blood pressure. Renin will convert
angiotensinogen to angiotensin I, and it will change to angiotensin II.
● The angiotensin II provokes thirst, resulting in vasoconstriction to quickly increase the
blood pressure, it will stimulate the adrenal cortex to secrete aldosterone and increase
renal reabsorption of Na.
● Distal tubular reabsorption of Na+ and HCO3– in exchange for K+ and H+ will be
promoted by Aldosterone.
● Both thirst and ADH are stimulated by low blood volume, independent of osmolality.
● Epinephrine and norepinephrine are secreted in response to decreased blood volume.

Whenever a sodium ion is reabsorbed, it will be followed by water molecules, due to the renal
retention of sodium that has a profound effect on blood volume. The renal tubules reabsorbs 98%
to 99% of sodium along with most of the water while the large amounts of sodium are filtered in
the 150L of glomerular filtrate produced daily. By several liters per day, even 1% to 2%
reduction in tubular reabsorption of Na+ can increase water loss. It always depends on the water
intake and the circumstances of collection when it comes to urine osmolality values. Yet, there is
a general decrease in diabetes insipidus and polydipsia and increased in situations where ADH
secretion is increased by hypovolemia or hyperosmolality or in circumstances like inappropriate
ADH.

3.) Enumerate and explain the methods of determination for the following electrolytes:

Sodium

As the years passed by, sodium has been measured in various ways, including chemical
methods, flame emission spectrophotometry (FES), atomic absorption spectrophotometry (AAS),
and ISEs. ISEs are the most routinely used method in clinical laboratories. To develop a potential
produced by having different ion concentrations on either side of the membrane, ISE method
preferred to use a semipermeable membrane. The two types of ISE measurement are the direct
and indirect. Direct measurements come up with an undiluted sample to communicate with the
ISE membrane, on the other hand, Indirect measurement, the dilute sample if for measurement.
The results will vary if the samples are hyperlipidemic or hyperproteinemic because it will lead
to a falsely decreased measurement of ionic activity in millimoles per liter of plasma, and in
direct method measures in plasma water only. Protein build up on the membrane through
continuous use is the one source of error with ISEs.

Potassium

Just like in Sodium, the current method of choice is ISE. A valinomycin membrane is used to
selectively link potassium, resulting in a resistance change which might correspond to potassium
concentration. Inner electrolyte solution is KCl.
Chloride

Usual method used in Chloride is the ISE. an ion-exchange membrane is used to selectively link
Cl ions for ISE measurement. Method using coulometric generation of silver ions is the
amperometric-coulometric titration that combines with Cl to measure the Cl concentration.
Excess or free Ag is used to designate the endpoint when all Cl in a patient is bound to Ag. The
coulometric generator and timer will be off as the Ag accumulates. The digital (Cotlove)
chloridometer (Labconco Corporation) uses this principle in Cl analysis.

Bicarbonate

In the presence of phosphoenolpyruvate (PEP) carboxylase, HCO3 is used to carboxylate

phosphoenolpyruvate that catalyzes the build up of oxaloacetate. The rate of change in
absorbance of NADH is proportional to the concentration of HCO3 .

Magnesium

For measuring the total serum Mg2 calmagite, formazen dye, and methylthymol blue are
involved. In calmagite, to form a reddish-violet complex that may be read at 532 nm, Mg2
should bind with calmagite. The formazen dye method, to form a colored complex, the Mg2
should bind with the dye. While in the methylthymol blue method, to form a colored complex,
Mg2 should bind with the chromogen. The reference method for measuring Mg2 is AAS.

Calcium

Either ortho-cresolphthalein complexone (CPC) or arsenzo III dye were used for the total Ca2
analysis to form a complex with Ca2. Ca2 will be released from its protein carrier and complexes
by acidification of the sample prior to dye-binding reaction. The CPC method uses 8
hydroxyquinoline to avoid interference in Mg2. Membranes impregnated with special molecules,
may be used by these systems but reversibly, link Ca2 ions. An electrical potential established
across the membrane that is proportional to the ionized Ca2 concentration when the Ca2 binds to
those membranes.

Phosphate

The formation of an ammonium phosphomolybdate complex is one of the usual methods for
phosphorus determination. It can be measured using ultraviolet absorption at 340 nm or can be
reduced to form a stable blue chromophore or molybdenum blue which is read between 600 and
700 nm.
 References

Bishop, M. L., Fody, E. P., & Schoeff, L. E. (Eds.). (2010). Clinical Chemistry: Techniques,

Principles, Correlations. Wolters Kluwer Health/Lippincott Williams & Wilkins.

Calcium - urine. (2019, May 6). UCSF Health. Retrieved February 15, 2023, from

[Link]

Cardiothoracic Critical Care. (n.d.). [Link]

Magnesium measurement, urine | Allina Health. (n.d.). Allina Health account. Retrieved

February 15, 2023, from [Link]

Phosphorus blood test - San Francisco. (2017, November 20). UCSF Health. Retrieved February

15, 2023, from [Link]

Rackley, C. R., & Toffaletti, J. G. (2021). Blood Gases and Critical Care Testing: Physiology,

Clinical Interpretations, and Laboratory Applications. Elsevier Science.

Serum Potassium - Clinical Methods. (n.d.). NCBI. Retrieved February 15, 2023, from

[Link]