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UNIT: pH and Blood Gases

UNIT: pH and Blood Gases

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MLAB 2401- Clinical Chemistry Lab Manual
C
D 87
UNIT:pH and Blood Gases
11pH.wpd
Purpose
To become acquainted with theory and methods of measuring pH and blood gases.
Objectives
Upon completion of this exercise, the student will be able to:1.Review classroom notes on pH, Henderson-Hasselbach equation, normal values andexpected ratios, and pH electrodes.2.
22
Discuss the basic principles involved in pH, pO and pCO determinations on whole blood.3.Interpret the basic clinical significance of blood gas values.4.Discuss the basic theory behind operation of the pH electrode.5.Discuss the operation of the pH meter and the blood gas machine.
Principle I
Determination of blood gases enables the evaluation of a patient's acid-base balance. Blood gasinstruments in the laboratory are designed to measure the partial pressures of carbon dioxide
22
(pCO) and of oxygen (pO) as well as blood pH. Specialized electrodes designed for each gasdetermination are placed within the instrument so that one small blood sample suffices for allmeasurements.
22
 After measuring pH and pCO directly, it is possible to obtain bicarbonate and CO content values
2
by calculation or with the use of a nomograph. From a direct measurement of pH and pO, bloodoxygen % saturation can be determined.
Principle II
The
carbonic acid-bicarbonate buffer system
is the most important buffer system in the regulationof hydrogen ion balance in the body. In plasma, the relationship between pH and the bicarbonate-carbonic acid buffer system is expressed by the
Henderson-Hasselbach equation
:With this mathematical expression of the relationship between the
bicarbonate ion
and
carbonic acid 
, it is possible to calculate pH.
Principle III
323
The ratio of HCO (salt) to HCO (acid) is normally 20:1. With this ratio, the blood pH is 7.40.
 –
The pH falls (acidosis) as bicarbonate decreases in relation to carbonic acid. The pH rises(alkalosis) as bicarbonate increases in relation to carbonic acid.
Principle IV
Four categories of acid-base imbalance may be encountered:
metabolic acidosis
;
metabolic alkalosis
;
respiratory acidosis
; and
respiratory alkalosis
. In this context, “metabolic” refers to the
 
UNIT: pH and Blood Gases (continued)
D 88
C
MLAB 2401 - Clinical Chemistry Lab Manual
bicarbonate concentration in the Henderson-Hasselbach equation and the “respiratory” to thecarbonic acid.
Principle V
Blood gases and pH are generally performed on
arterial 
blood to provide acid-base and respiratoryinformation on a sample that is a mixture of blood from all parts of the body to tell how well thelungs are oxygenating the body.1.pH is the only way to determine if the body is too acid or too alkaline.a.
 Acidemia
and
alkalemia
refer to a condition of the blood.b.
 Acidosis
and
alkalosis
refer to the
 process
occurring in the patient which caused thecondition.
22
2.The
respiratory parameter 
is the pCO, or pressure/ tension exerted by dissolved CO gas
2
in the blood. pCO is influenced only by the function of the lungs.
2
a.CO gas should be considered an acid substance because in water carbonic acid
23
(HCO) is formed.
2
b.Transport forms of CO
2
1)dissolved CO gas
2233
2)combined with HO
6
HCO
6
HCO + H
 –+
3)as carboxyhemoglobin
222
3.Under normal conditions, dissolved CO gas has a pCO of approximately 40 mmHg. pCO
32
of 40 mm = 1.2 mEq/L. Normal HCO = 24 mEq/L. Total CO content = 25.2 mEq/L (or 
 –
mmol/L).
233
4.The H produced by the breakdown of HCO
6
HCO + H
++
a.is loosely carried and thus buffered with plasma proteins
4
b.H is excreted by the kidney as NH
++2
5.CO is removed by the lungs
22
High PCO – hypoventilation – respiratory acidosis by decreased elimination of CO bylungs.
22
Low PCO – hyperventilation – respiratory alkalosis by increased elimination of CO bylungs.
3
Non-Respiratory Parameters:
HCO and Base Excess (BE)
 –
 
UNIT: pH and Blood Gases (continued)
MLAB 2401- Clinical Chemistry Lab Manual
C
D 89
Bicarbonate ion and BE are
influenced 
by metabolic causes. Metabolic acid-base is under thecontrol of the kidneys.
3
1.HCO is an alkaline (base) substance excreted or conserved by the kidneys.
 –3
2.A high HCO or positive BE indicates metabolic alkalosis caused by loss of non-volatile acid
 –3
or gain of HCO
 –
a.excess vomitingb.excess diuretic therapyc.excess bicarbonate intake
33
3.A low HCO or negative BE indicates metabolic acidosis caused by loss of HCO or its
 –
neutralization by a non-volatile acid.a.seen in conditions causing the accumulation of organic acids as in diabeticketoacidosis, lactic acidosis, renal failure, etc.b.deficit of bicarbonate due to diarrhea, renal tubular acidosis, ammonium chloridetreatment.
Oxygen
222
The relationship between pO and hemoglobin O (HbO) is an s-shaped curve, the hemoglobin-oxygen dissociation curve.To fulfill its function as respiratory pigment, hemoglobin must specifically bind with high affinity tolarge quantities of oxygen, transport them, and release them in appropriate tissues. It is thetetrameric structure of hemoglobin that provides its unique oxygen-binding capacity and makesit superior.Each molecule of hemoglobin contains four (ferrous) iron-containing heme groups, each capable
222422
of binding one molecule of O (
Hb + 4 O
º
Hb(O)), each O added depends on the (pO)).The position of the oxygen-dissociation curve isdetermined by a number of factors in addition to thepreviously mentionedpartial pressure of oxygen;body temperature, red cell2, 3, DPG concentration,and pH (Bohr effect).Top left, oxygendissociation curves for human blood with different
2
pH(S) but constant pCO(40 mm Hg). DPGconcentration in

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