FLUID, BUFFER, ACID-BASE BALANCE

Rondang Soegianto

2010
Fluid content in individuals vary due to

variability in amt of adipose tissue.

High body water  lean

Low body water  obese

 Plasma > 90% H2O
Skin, muscle, internal organs 70-80% H 2 O
Skeleton only 22%  dry tissue
Fat 10%  lowest
 Intracellular fluid (ICF)
Extracellular fluid (ECF)
ICF compartment ~ 2/3 total body water
ECF: plasma = 1/5 of ECF
Interstitial fl ~ 4/5 of ECF
Minor ECF compartments:

- Lymph
- Transcellular fluid:
Cerebrospinal fl
Interoccular fl
Synovial fl
Cardial, intrapleural, peritoneal fl
Digestive juices
Transcelr fl does not affect fl balance of cell
Exception: Vomiting, diarrhea
Metabolism and Acid-Base Balance
A-B balance related to
- Respiration
- Metabolism
Impaired A-B balance of ECF 
- Lactic acidosis
- Ketosis
Chemical Terms
+
Acidity = [H ] (proton)
+
Pure water [H ] = 10 -7
+
[H ] > 10 -7 = Acid
+
[H ] < 10 -7 = Alkaline
pH = 7  [H+] = 10-7 M = 0.0001 mM
pH = 7.4  0.00004 mM
Note:
1. Absolute conc’s of H+ in organism much <<
other solutes
Ex. [Na + ] inECF = 145 mM

2. pH units are logarithmic  not linear scale

Rise of one unit pH = 10 fold rise in [H +]
pH of ECF (incl plasma) = 7.35 – 7.45
Outside this range: Acidosis <----- > Alkalosis

Compatible with life: pH 7.0 - 7.8

Exceeding this range may inhibit brain

function
Source of Protons
1. Dietary
a. Phospholipids: hydrol. of phosph. acids
b. Sulphur containing amino acids
Cystein and metionine from proteins
Degrd’ion in liver  sulphates and H +
2. Respiratory ( ~ 12.5 moles/day)
-
CO 2 + H 2O  H 2 CO 3 + H + HCO 3
+
3. Metabolic products
- Lactic acid
- Ketone bodies
- ATP hydrolysis
- NAD+ reduction
Totally per day: ~150 moles proton
Reversal by body reactions  no net charge
in protons
Body pH raised due to:

- Ingestion of weak acids (citric acid) as

- Na or K salts (from fruits)
- Hyperventilation (loss of CO 2 )
- Vomiting (loss of HCl from stomach)
Mechanism for pH adjustments
A. Buffer
B. Respiration
C. Renal acid secretion
 A. Buffers
-
Weak acid (binds added OH )
and
Conjugate base (binds added proton)

# Resists changes in pH when acid or base

added

# Conjugate base = weak acid - proton

Conj acid = weak base + proton
-
Blood buffer H 2 CO 3 : HCO
3
Salts Can Change pH of Blood or Lab
Sol’n
- Acidic salts ex NH 4 NO 3
NH4+ is weak acid
- Basic salts ex CH 3 COO-Na
CH3COO- is weak base
- Neutral salts ex NaCL, KNO 3
No weak acids or weak bases
Buffer Capacity

BC = Ability to consume added H+ and

added OH -

BC depends on
- Buffer conc’n
- pKa
Useful buffering is at pH within pKa + 1
or pKa - 1
Examples of Buffers
a. Proteins. Have many ionizable groups
and pKa values.
Present in differing conc’ns.
H.Prot  H+ + Prot -
H.Prot +  H + + Prot
Ex. Hb in erythrocytes (large amt)
H+ can penetrate eryth membrane
This buffering action influences plasma pH
b. Phosphates
One ionizable group, pKa = 6.8

Inorganic phosphates
H 2 PO4-   H + HPO 4
+ 2-

Organic phosphates (not much)

c. Bicarbonates
-
H 2 CO 3   H + HCO , pKa ~ 3
+
3
Only slightly useful to buffering cap at
pH=7.4
-
Important, since conc of H 2 CO 3 and HCO 3
can be regulated in response to pH change

Dynamic Buffer System
Active in respiratory system
B. Respiration
H 2 CO 3   H 2 O + CO 2
Enzyme: Carbonic anhydrase in RBC
and other tissues
Henderson-Hasselbalch eqn:
[salt]
pH – pKa + log _______________
[undiss. acid]
 -
For carbonic acid: [HCO 3 ]
pH = 3.0 + log ____________
[H2 CO 3 ]
# Lung ventilation >>  loss of CO2 >>

Blood H 2 CO 3 <<  pH >>

= Respiratory alkalosis
Ventilation <<  Resp acidosis
Quantitative Values of Buffer on Acid Load
1. Plasma protein 1% of total buffering
of acid administration
2. Erythrocytes ~6% (H+ penetrate RBC)
3. ECF carbonic/bicarbonate system ~42%
4. Intracellular protein ~51%
EC H+ exchanges for IC Na + (36%) and
for IC K+ (15%)
Some Na+ derived from bone apatite crystals
In chronic acidosis  bone resorption 
Ca 2+ and phosphate released
Kidney in Acid-Base Balance Control
Maintains many solutes in plasma by adjusting
rate of excretion in urine thru

- reabsorption from glomerular filtrate

- secretion into urine

Glomer. Filtr.: high Na+ and bicarbonate

(equals conc in plasma0
 -
Loss of HCO 3  pH of blood <<

Recovery:
Protons secreted into lumen from
tubular cells

In the lumen:
HCO 3- + H +  H2 CO
H 2 CO 3  H 2 O + CO 2
CO2 diffuses back into tubuli
Importance of Various Buffer Systems

* Protein buffer system primarily important

intracellularly

* Hb buffer system buffers H+ generated

from carbonic acid (H 2 CO 3 )

* The phosphate buffer system is an

important urinary buffer
Line Order of Defense Mechanisms Against
Changes in [H+ ]

1. Chemical buffer systems

First line of defense. H + not eliminated but
incorporated into buffers

2. Respiratory system, second line of defense

a. Works thru pulmonary ventilation
Increase of arterial [H+ ] by metabolism
stimulates resp. Center in brain stem
 >> pulm. ventilation
 +
b. When arterial [H ] falls, pulm. Vent. <<
 shallow breathing. Metab.
CO2 diffuses from cell  blood,
faster than CO2 removed
from
blood  lungs. CO2 accuml in
+
blood  restores [H ]
3. Kidneys, third line of defense
Most potent A-B regulating mechanism
- vary removal of proton from any source
- conserve or eliminate bicarbonate ion

Ex. Renal compensation for acidosis

+ -
For each H  to urine, new HCO3
+
returns to plasma to buffer another H
still remaining in body fluid
Thus:
Kidneys are able to restore pH to normal
Respond continuously to pH change until
compensation is complete

Lungs can only adjust amt of CO 2 that forms

H + in the body
1. CHEMICAL
BUFFER

2. PHYSIOLOGICAL
BUFFER

MECHANCAL
RESPIRATION
EXCRETION OF CO2

RENAL MECHANISM
EXCRETION OF H+
Kidneys Secrete Ammonium

H+ transp. actively from tubular cells 

tubular plasma.
Capacity limited to urinary pH = 4.5
H + in tub fl must be buffered to << free H +
Important urinary buffers:

1. Filtered phosphate buffer

2. Secreted ammonia
Secreted H + first buffered by phosph buffer.
Phosph in tub fl comes from ingested phosph
= dietary excess
With high H + excretion, buff cap. of phosph
Exceeded. Kidney cannot respond by >> phos
excrt’n. Only phosph reabsorption subjected
to control mechanism.
Next: Tubular cells secrete NH3
NH3 + H +  NH 4  urine
NH3 synthesized from glutamine in tub. cells
Acid-Base Balance

Ratio of [HCO 3- ] : [CO2 ]

Normal = 20/1
Resp acd < 20/1 CO 2 >>
Resp alk > 20/1 CO 2 <<
-
Met ac < 20/1 [HCO 3 ] <<
Met alk > 20/1 [HCO -3 ] >>
Compensation:
-
Resp Ac. Kidneys conserve filtered HCO
Resp Alk: Kidneys conserve H + 3
Excrete more HCO 3-
Met Ac : Renal and chemical
- buffers take up H +
- lungs blow off CO 2
- kidneys excrete H
-
- conserve HCO 3

Met Alk: Liberation of H +

Ventilation <<, CO 2 retained in body fld
 Causes of A-B Imbalance

Resp Ac : Hypoventilation
Lung disease
Depression of resp centr by drug or disease

Resp Alk. Fever, Anxiety, Aspirin poisoning

Met Ac Severe diarrhea, DM

Strenuous exercise, Uremia (renal failure)

Met Alk Vomiting

Ingestion of alkaline drugs
References:
1. Biochemistry for the Medical Sciences
E.A. Newsholme and A.R. Leech
John Wiley & Sons, 1983
2. Biochemistry. A Foundation
Peck Ritter
Broks/Cole Publ Co, 1996
3. Human Physiology. From Cells to Systems
L. Sherwood
Brooks/Cole, 2004
4. Medical Biochemistry
A.C Brownie, J.C. Kernohan
Elsevier, 2005