pH, dapar (buffer)

Program Studi S2 Biomedik

Program Pascasarjana Unand
November 2010
 Calculating the pH

pH = - log [H+]
(Remember that the [ ] mean Molarity)

Example: If [H+] = 1 X 10-10

pH = - log 1 X 10-10
pH = - (- 10)
pH = 10
Example: If [H+] = 1.8 X 10-5
pH = - log 1.8 X 10-5
pH = - (- 4.74)
pH = 4.74
Acid/Base definitions
 Definition #1: Arrhenius (traditional)

Acids – produce H+ ions (or hydronium ions

H3O+)

Bases – produce OH- ions

(problem: some bases don’t have hydroxide

ions!)
Arrhenius acid is a substance that produces H+ (H3O+) in water

Arrhenius base is a substance that produces OH- in water

 Acid/Base Definitions
 Definition #2: Brønsted – Lowry

Acids – proton donor

Bases – proton acceptor

A “proton” is really just a hydrogen

atom that has lost it’s electron!
 A Brønsted-Lowry acid is a proton donor
A Brønsted-Lowry base is a proton acceptor

conjugate conjugate
base acid
acid base
ACID-BASE THEORIES

The Brønsted
definition means
NH 3 3is a
NH + BASE
H2O in NH4+ + OH-
water
Base — and Acid Acid Base
water is itself an
ACID
Dapar, pH dan Peran
dalam ikatan kimia
Biokimia terkait dengan sifat dan reaksi
senyawa organik
Dalam sel keberadaan senyawa dan
reaksi terjadi pada suasana berair/larutan
Air penting dalam reaksi biokimia dan
penentu sifat makromolekul, misal;
protein
 More About Water
H2O can function as both an ACID and a BASE.
In pure water there can be AUTOIONIZATION

Equilibrium constant for water = Kw

Kw = [H3O+] [OH-] = 1.00 x 10-14 at 25 oC
 More About Water
Autoionization OH-

H3O+

Kw = [H3O+] [OH-] = 1.00 x 10-14 at 25 oC

In a neutral solution [H3O+] = [OH-]
so Kw = [H3O+]2 = [OH-]2
and so [H3O+] = [OH-] = 1.00 x 10-7 M
Disosiasi air

 Air terdisosiasi jadi OH - dan H+

 pH digunakan untuk menentukan
konsentrasi H + dalam sel dan cairan
tubuh
 Gugus fungsi spt karboksil dan amino
terurai pada pH tertentu
Struktur Molekul Air
Jumlah air tubuh

 Pria : 55 - 65 % dari berat badan

 Wanita 10% lebih kecil
 Intra sel 55%
 Ekstra sel 45%
– Plasma 7.5 %
– Interstitial / limfe 22.5%
– Jaringan, tulang 15%
 Kebutuhan air
 Dewasa untuk kesehatan minum 2000 ml air

 Masukan
sebagai cairan 900 ml
dalam makanan 800 ml
Hasil oksidasi makanan 300 ml

Kehilangan
urin 1050 ml
tinja 100 ml
kulit/paru 850 ml
Sifat air berbeda dengan pelarut lain
Air : 0 0C 100 0C 540 kal/g
H2S: -85 -60 132
NH3: -78 -33 327
menandakan kekuatan intermolekuler yg kuat
adanya sifat elektrik dipolar
atom oksigen yg elektronegatif menarik elektron H
daerah positif skllg H
daerah negatif skllg O
tarikan elektrostatik antara molekul air; ik. hidrogen
Calculating fluid requirements

 Method 1: (based on energy intake): 1 ml of fluid per kcal

 Method 2: (based on body weight)
Age / gender ml/kg
Infant and children 1-10 kg 100-150
11-20kg add 50 ml/kg over 10kg
≥21 kg add 25 ml/kg over 20kg
Adolescents 40-60
Young adult 16-30 yrs 35-40
Average adults 30-35
Adult 55-65 yrs 30
Adult >65 yrs 25
 Method 3(based on nitrogen and energy intake): 1ml/kcal+100ml/gN
 Method 4(based on body surface area): 1500 ml/m2
Ikatan Molekul Air
AIR sebagai pelarut
Senyawa ion, spt garam
Senyawa polar non ion, gula,
alkohol
Ggs hidroksil senyawa non polar
dg mudah mengikat H air.
Senyawa amfipatik, mis
garam dari as lemak
 Punya: ggs polar(hidrofilik)
 ggs non polar (hidrofobik)
 Dalam larutan encer mmbtk micelles
 Mol amfipatik terlibat dl pembentukan
dan struktur membran
Kalium Laurat
Konsentrasi H+ dl sistem
biologi
 Dipertahankankan konstan
 Reaksi biokimia peka thd perubahan
H+
 proses metabolisme selalu
menghasilkan H+
Skema Micelle
Konstanta keseimbangan
Keq
 H2O === H+ + OH-

 K = (H+) (OH-)/(H2O)
 K air pada 25 oC = 1.8 x 10 -16 M (mol/l)
 Konsentrasi H2O dalam air murni 55.5 M
 (H+)= 1 x 10 -7 M
 K adalah konstanta disosiasi , merupakan
bilangan aksponensial, dinyatakan sebagai
pK
pH

 - log (H+)
 pH 7. Netral
 pH > 7, alkalis
 pH < 7, asam
 perubahan satu unit pH (7 jadi 6)
berarti penambahan konsentrasi H+
sepuluh kali lipat
Asam dan Basa Bronsted-
Lowry
 Asam , senyawa yg donasi proton (H+)
 Basa, senyawa yg terima proton ( H+)
 HA ----- H+ + A-
 HB+ ----- H+ + B:
 HB- ----- H+ + B 2-
 A, B disebut basa konjugat
Asam

 Asam kuat
– H2SO4, HCl
– Affinitas kecil thd proton, disosiasi
sempurna
Asam lemah
- As. Asetat, propionat, piruvat , air
- Affinitas tinggi thd proton, tidak mudah
berdisosiasi
Ionisasi Asam lemah

H3CCOOH==== H+ + H3CCOO -)
K = (H+) x (H3CCOO-)/(H3CCOOH)
K asam asetat pada 25 oC= 1.8 x 10 -5 M
(H+) = 4.2 x 10 -3 M
Persamaan Henderson-
Hasselbalch
 (H+) = K (HA)/(A-)
 log (H+) = log K + log (HA)/(A-)
 -log (H+) = -log K- log (HA)/(A-)
 -log (HA)/(A-) = + log (A-)/(HA)
 pH = pK + log (A-)/(HA)
 Titrasi asam lemah oleh
basa kuat
 Bila asam asetat dititrasi dg NaOH,
perubahan pH yang besar terjadi hanya pada
awal dan akhir titrasi
 Perubahan terkecil tdp pd titik tengah titrasi
 Pada titik ini (H3CCOOH atau HA) sama dg
(H3CCOO- atau A-)
 pH = pKa
 Kemampuan asam asetat tahan thd prbh pH
disebut kapasitas pendaparan
 Pendaparan ( buffering ) :
– Kecendrungan larutan untuk bertahan
secara lebih efektif terhadap perubahan
pH setelah penambahan asam atau basa
dibandingkan air dengan volume yang
sama
Banyak metabolit antara seperti gula fosfat
merupakan asam lemah
 Pendaparan penting untuk
Semua reaksi intrasel karena reaksi
akan melepaskan dan pengambilan
proton
Titrasi dengan Asam
Lemah
Berapa pH bila

 1. Rasio A-/HA = 100

 2. Rasio A-/HA = 0.1

 3. Konsentrasi H+ dari larutan 3.2 x

10-4 M

 4. Larutan KOH 2 x 10-6 M

Sistem dapar fisiologi

 pH berbagai cairan tubuh beragam

– cairan lambung = 1-2
– cairan usus = 8-9
– plasma = 7.40
Perubahan pH darah 0.2-0.4, medis serius,
tanpa koreksi , kematian
Dapar penting dalam darah adalah protein
(hemoglobin dalam eritrosit)
 Sistem dapar
ekstraseluler
 Sistem bikarbonat yang perlu
mengeluarkan CO2 produk metabolisme
– CO2 + H2O == H2CO3 == H+ + HCOO-

Rasio HCOO-/H2CO3 = 20/1 diperlukan

supaya pH plasma 7.40
Cairan intraseluler menggunakan ionisasi
kedua asam fosfat sebagai sistem dapar
H2PO4- ===H+ + HPO4 2- ,
pK=7.21
 Renal tubule acidosis（RTA）
 Urine acidification is impaired
 無法分泌H+以平衡過多的代謝酸（nonvolatile
acid），所以產生代謝性酸中毒
 Defect in PT H+-secretion（proximal RTA）
– Cystinosis胱胺酸症
– Fanconi’s syndrome
– Carbonic anhydrase inhibitors
 Defect in distal tubule H+-secretion（distal RTA）
– Medullary sponge kidney
– Amphotericin B
– Secondary to urinary obstruction
 Treatment: ingest alkali (HCO3-)
Response to acid-base disorders
 Respiratory defense
– Metabolic acidosis: ↑H+↑ventilation
rate
 Type-I-DM patient (keto acid製造增
加): Kussmaul respiration: 呼吸肌會
疲勞, 所以呼吸代償會受損, 酸中毒
更嚴重
– Metabolic alkalosis:↓H+  ↓ventilation
rate
Response to acid-base
disorders

 Renal defense
– Secretion of H+
– HCO3- reabsorbed
– Production and excretion of NH4+
DM patient: ketone bodies
Diarrhea:
Normal Acid-Base Balance

 Normal pH 7.35-7.45
 Narrow normal range

 Compatible with life 6.8 - 8.0

___/______/___/______/___
6.8 7.35 7.45 8.0
Acid Alkaline
Maintenance of Balance

Balance maintained by:

 Buffering systems

 Lungs

 Kidneys
 Buffer Systems

 Prevent major changes in pH

 Act as sponges… H+
 3 main systems H+
Bicarbonate-carbonic acid buffer
H+
Phosphate buffer
Protein buffer
Buffer Systems

 Bicarbonate buffer - most important

Active in ECF and ICF
 Phosphate buffer
Active in intracelluar (ICF) fluid
 Protein buffer - Largest buffer store
Albumins and globulins (ECF)
Hemoglobin (ICF)
 Bicarbonate-Carbonic Acid

 Body’s major buffer

 Carbonic acid - H2CO3 (Acid)
 Bicarbonate - HCO3 (Base)

1.2 mEq/L 24 mEq/L

1 20
H2CO3 ……………… HCO3
pH = 7.4
 Bicarbonate-Carbonic Acid

 Ratio important
 Not absolute values
 Person with COPD (CAL)

2.41mEq/L 20
48 mEq/L
H2CO3 ………………
7.4
HCO3
 Regulation

 Key concept
 Carbonic anhydrase equation

CO2 +H2O H2CO3 H+ + HCO3

Carbon Carbonic Bicarbonate
Dioxide Acid

(ACID) (BASE)
 Acid
Substance that contains H+ ions
that can be released (H2CO3)
Carbonic acid releases H+ ions
 Base
Substance that can accept H+ ions
(HCO3)
Bicarbonate accepts H+ ions
 As CO2 increases, carbonic acid
increases, H+ ions increase
 pH drops….. becomes more acidic

CO2 +H2O H2CO3 H+ + HCO3

Carbonic Bicarbonate
Acid
CO2 H2CO3 H+ HCO3
(pH Acidic <7.35)
 As HCO3 increases, H+ decreases
 pH rises, becomes more alkaline

CO2 +H2O H2CO3 H+ + HCO3

Carbonic Bicarbonate
Acid
CO2 H2CO3 H+ HCO3

(pH Basic >7.45)

Respiratory & Renal Regulation

 Lungs control CO2

 Kidneys control HCO3

kidneys (bicarbonate)
pH =
lungs (carbon dioxide)
 Respiratory Regulation
Mechanisms of control …
 Hyperventilation -- blow off CO2
 Hypoventilation -- retain CO2
Regulation rapid...
 Seconds to minutes
Measured by PaCO2 - Normal
35-45 mm Hg
Renal Regulation
Mechanism of control
 Excretion or retention of
H+ or HCO3
Regulation….. Slow
 Hours to days to change pH
Normal serum HCO3
 22-26 mEq/L
Acid-Base Imbalances
 Ratio of 20 to 1 out of
balance
 Acidosis (acidemia)
 pH falls below 7.35
 Increase in blood carbonic
acid or
 Decrease in bicarbonate
Acid-Base Imbalances

 Alkalosis(alkalemia)
 pH greater than 7.45

 Increase in bicarbonate

or
 Decrease in carbonic acid
Acid-Base Imbalances

Primary cause or origin:

 Metabolic

Changes brought about by

systemic alterations
(cellular level)
 Respiratory

Changes brought about by

respiratory alterations
Acid-Base Imbalances
Compensation
 Corrective response of kidneys
and/or lungs
Compensated
 Restoration of pH and 20 : 1 ratio
Uncompensated
 Inability to adjust pH or 20 : 1
ratio
Four Basic Types of
Imbalance
 Respiratory Acidosis
 Respiratory Alkalosis

 Metabolic Acidosis

 Metabolic Alkalosis
Respiratory Acidosis
 Carbonic acid excess
 Exhaling of CO2

inhibited H2CO3
 Carbonic acid builds up

 pH falls below 7.35

 Cause = Hypoventilation
(see chart)
 Acid-Base Imbalances
 Normal

1.2 mEq/L 24 mEq/L

H2CO3 ……………… HCO3
1 20

7.4
Respiratory Acidosis

1 13
7.21
Respiratory Acidosis
 Compensation: How?
 Opposite regulating mechanism
 Problem = depressed breathing, build up of
CO2 in blood
 Response - Kidney retains HCO3
(Response ….. Slow)
Respiratory Alkalosis
 Carbonic acid deficit
 Increased exhaling
of CO2
 Carbonic acid decreases
 pH rises above 7.45 H2CO3
 Cause = hyperventilation (see chart)
 Acid-Base Imbalances
 Normal

1.2 mEq/L 24 mEq/L

H2CO3 ……………… HCO3
1 20

7.4
Respiratory Alkalosis

1 40
7.70
Respiratory Alkalosis
 Compensation:
 Problem = excess “blowing off” of
CO2
 Result = decrease in carbonic acid
and increase in HCO3
 Response: Kidney excretes excess
bicarbonate
Metabolic Acidosis
 Base-bicarbonate deficit
 Low pH (< 7.35)
 Low plasma bicarbonate (base)
 Cause = relative gain in H+
(lactic acidosis, ketoacidosis)
or actual loss of HCO3
(renal failure, diarrhea)
 Acid-Base Imbalances
 Normal

1.2 mEq/L 24 mEq/L

H2CO3 ……………… HCO3
1 20

7.4
 Metabolic Acidosis
 Kidney failure (decrease in
bicarbonate)

1 10
7.10
 Metabolic Acidosis
 Lactic acidosis, keto acidosis
(increase acid… no change in
bicarbonate)

1 10
7.10
Metabolic Acidosis
 Compensation:
 Problem = low HCO3 (base) or high H+ ion
(acid)
 Response: Lungs hyperventilate
 Get rid of CO2
(decrease PaCO2 and therefore raise level of
HCO3)
 Metabolic Alkalosis
 Bicarbonate excess
 High pH (> 7.45)
 Loss of H+ ion or gain of HCO3
 Most common causes vomiting, gastric
suctioning (NG tube)
 Other: Abuse of antacids, K+
wasting diuretics
 Acid-Base Imbalances
 Normal

1.2 mEq/L 24 mEq/L

H2CO3 ……………… HCO3
1 20

7.4
Metabolic Alkalosis

1 30
7.58
Metabolic Alkalosis
 Compensation:
 Problem = too much base
 Response: Lungs compensate by
hypoventilating
 Retain CO2, increase PaCO2
 Increase acid level in blood
TERIMA KASIH