CHEMICAL BIOLOGY III (BIOMOLECULES)

CHB 2203L | Jessa Marie D. Natividad | PLM College of Science

pH and Buffers
● A salt is produced in the reaction of an acid with a
OUTLINE base.
○ Examples include NaCl, Na₂SO₄, and
I.Acids and Bases NaOOCCH₃ (sodium acetate).
II.Conjugate Acids and Bases
III.pH and pOH: Measure of Acidity and Basicity
IV. Test for Acidity and Basicity
A. Using pH paper
V. Buffer Solution
VI. Henderson-Hasselbalch Equation
VII. Post-Lab
VIII. References

ACIDS AND BASES

● According to the Brønsted-Lowry theory, an acid is
a proton donor and a base is a proton acceptor.
○ We will be using the definition of Bronsted-Lowry
theory, which is more acceptable. He said that
acid is a proton donor. From the term
itself—proton—it donates hydrogen. Hydrogen
ion is the definition for proton. A base is a proton
acceptor or it accepts hydrogen. Of course,
typically your acid will contain hydrogen on its
molecule to be considered as an acid, kaya nga
siya tagabigay ng hydrogen, tagabigay ng
proton because it contains hydrogen, whereas
your base typically bears a hydroxyl group on its
structure. While your hydrogen bears the
positive (H⁺), you hydroxyl will bear the Figure 1.1 Examples of Strong Acids and Strong Bases
negatively charged (OH⁻). Syempre, for a
molecule, for a substance to behave as an acid,
CONJUGATE ACIDS AND BASES
it needs to encounter a base. So we will form
certain products. ACID + BASE → ___________ ● The product formed when an acid gives up a proton
So tatandaan, your solution can be considered is a potential proton acceptor and is called the
an acid if it encounters a base. Same as well conjugate base of the parent acid. Similarly, every
when a base encounters an acid, it can form base accepts a proton to produce a conjugate acid.
what we call conjugate acids and also conjugate ○ If an acid encounters a base, what will be your
bases. And also one of the characteristics, product form? If your acid encountered with a
special mention molecule, is water. So a base, of course acid is a proton donor, donates
molecule of water can be considered as what we hydrogen to your base. An acid that removes
call amphoteric. It can bear what we call a hydrogen on its structure will become a
hydronium ion and a hydroxyl part. (2𝐻2O conjugate base. A base that accepts hydrogen
from the acid will become a conjugate acid. The
+ −
→ 𝐻3𝑂 + 𝑂𝐻 ) It means water can be definition of conjugate base—it is the acid that
considered as an acid or a base (amphoteric). hydrogen is being removed. Conjugate acid is
Your water can be an acid depende sa partner the base that adds or accepts hydrogen from the
niya. Kung a partner ng water natin ay ang base, acid, therefore it becomes conjugate acid.
of course our water will become an acid, it will Tatandaan na there are only two products during
encounter ng acids and bases.
behave as an acid [𝐵𝐴𝑆𝐸 + 𝐻2𝑂 (𝑎𝑐𝑖𝑑)]. If ang
partner ng ating water is an acid, your water will
behave as a base [𝐴𝐶𝐼𝐷 + 𝐻2𝑂 (𝑏𝑎𝑠𝑒)]. Again,
your acid is a proton donor so typically, it will
contain hydrogen atom on its structure so it can
donate hydrogen ion sa base which will be a
proton acceptor—it accepts hydrogen and also
bears hydroxyl on its structure so ACID + BASE
→ conjugate base + conjugate acid.

COURSE CODE TRANSCRIBER/S: Gilbuena 1

 pH and Buffers TRANS # 3

TEST FOR ACIDITY AND BASICITY

● Done via red and blue litmus paper, or pH paper
○ When handling litmus or pH paper, handling with
bare hands is avoided. Why?
● Using red/blue litmus paper:
○ Red litmus turns blue in high pH
○ Blue litmus turns red in low pH
○ Both do not change color in neutral pH
○ It is necessary to test using both red and blue.
Why?

How do we define and identify if our unknown solution is

an acid or a base? Of course, you have unknown acid
and base, hindi given yung pH value so we cannot
identify. We have different measurements, different
Figure 1.2 Acid and Base yields Conjugate base and techniques such as the first one—yung ginagamit nating
Conjugate acid red and blue litmus paper. Actually, it is made up of
paper, meron din siyang laman na lichens which is highly
pH AND pOH: sensitive to pH. Actually, may purple pa nga e but this is
MEASURE OF ACIDITY AND BASICITY not very usual. Ang usual na ginagamit ay red and blue
● pH (power of Hydrogen) litmus paper. When you’re handling your litmus or your
○ Concentration of [H⁺] pH paper, do not handle with bare hands because your
○ Lower pH, higher H⁺; more acidic hands, pawisin yan, it has moisture, of course. Acidic to
○ pH = -log[H⁺] or pH = -log[H₃O⁺] neutral ang ating moisture. Ang pawis natin ay acidic to
○ ‘Pag acid, concentration ng hydrogen ang neutral. Magkakaroon kayo ng false positive or false
+ negative result doon sa inyong test. So that’s why, if
hinahanap. ↓ 𝑝𝐻, ↑ 𝐻 you’re handling red and blue litmus paper, you need to
○ pH = -log[H₃O⁺] is used if we are encountering handle them nang naka gloves.
water acting as an acid but normally, pH =
-log[H⁺]
● pOH (Hydroxide ion)
○ Concentration of [OH⁻]
○ Lower pOH, higher OH, more basic
○ pOH = -log[OH⁻]
○ Tandaan, base solution contains hydroxide or
the Hydroxyl ion so therefore in base, we are
measuring the pOH or the hydroxide ion Figure 1.3 Biologist for Leni-Kiko
concentration.
So what is the idea of using red and blue litmus paper?
○ ‘Pag base, concentration ng hydroxyl or
− You have an unknown solution, you do not know if it is an
hydroxide ion ang hinahanap. ↓ 𝑝𝑂𝐻, ↑ 𝑂𝐻 acid or a base. What you should do is dip both of your
red and blue litmus paper doon sa ating solution. If your
Determine the [H+], [OH-], pH, and pOH given the ff: red litmus paper stays as red but your blue litmus paper
turns red, it is red–red, therefore the solution is acid.
[H+] [OH-] pH pOH Another scenario, if your red litmus paper turns blue and
your blue litmus paper stays as blue, you have two blue
2.3x10⁻⁴ −11 3.64 10.36 results, you have a base solution. Another scenario, if
4. 37 × 10 your red stays as red and blue stays as blue, you have a
neutral solution. HIndi lang basta red ang ididip mo, hindi
1. 58 × 10
−13 6.3x10⁻² 12.80 1.2 lang basta blue, sabay dapat dinidip ang red and blue
litmus paper so you can identify if both of them changed
color.
pH = -log [H+] pH>7 Basic
pOH = –log [OH-] pH<7 Acid USING pH PAPER
[H+] = 10[⁻ᵖᴴ] pH=7 Neutral Solution ● Using pH paper:
[OH-] = 10[⁻ᵖᴼᴴ] ○ Color change also occurs; each color
pH+pOH = 14 corresponds to a specific pH range
○ Some pH paper have multiple indicators on a
How do I identify if it is an acid or a base? Acid should
single strip, for more accurate pH estimation
have a lower pH. Base should have a higher pH. Kung
○ Has color chart for reference
pagbabasehan and pOH value, it is considered as an
acid if it has higher pOH but base kapag lower pOH.
Another one, which is more highly used, mas preferable
‘to bukod sa pH meter. Kapag wala tayong pH meter,
mas preferable ‘tong gamitin, ang pH paper. This one
could indicate the pH number. Same procedure: you

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 pH and Buffers TRANS # 3

have an unknown solution, then you have a pH strip. are doing thesis, in vitro studies, kagaya ng enzymes,
Once you dip on your unknown solution, after a few normal cells, cancer cells, bacteria—lahat ‘yan galing sa
second, there will be what we call bonds of color. So the organism. For example, sa katawan ng tao, kumuha ka
bonds of color, you will just check kung anong pH ng breast cancer cell, galing siya sa katawan ng tao,
number corresponds to that bonds of color. Kung ano meron siyang certain pH condition na kailangan niya
yung kaparehong color na nakita niyo. For example, para mabuhay. Of course, kung ilalabas mo yan,
kapareho niyo yung number 4 color, therefore its pH tatanggalin mo yan doon sa organism, for example,
number is around 4. So this is just an estimation of pH normal breast cells at cancer cells sa breast, tatanggalin
hanggang 1-14. But if you want to be precise, kunwari mo yan doon sa organism, kailangan nilang mabuhay.
may 4.14, 7.5, 7.7 ang kailangan niyo, you need to use They need what we call a buffer, where it has the best pH
now a pH meter. In a laboratory setup, usually ang condition na kagaya ng organism. Kaya nga siya in vitro
ginagamit lang ay red and blue litmus paper and pH studies, ilalabas mo yung enzyme or yung organism
meter. kagaya ng cells and bacteria pero gagayahin mo,
i-sisimulate mo yung condition doon sa loob ng
pinanggalingan nilang organism. Kung ang pH na
kailangan nila ay 7, outside the organism, kailangan
nilang pH is also 7. Beyond 7 or below 7, mamamatay
sila that’s why meron tayong tinatawag na buffer solution.
Ginagamit siya usually para mabuhay itong mga in vitro
studies na ‘to because dagdagan mo man siya ng acid,
ng base, ng pagkain, hindi mababago yung pH condition,
na sinet mo doon. For example, enzymes–salivary
amylase, laway natin, tatanggalin mo yung laway sa
bunganga mo, gagamitin mo siya to test kung gaano
katagal mag breakdown ng starch. Of course, yung pH
condition sa bunganga natin is 6.5 to 7. Para tumagal
yung salivary amylase mo, hindi siya mamatay, kailangan
mo ng buffer na the same doon sa condition ng mouth
mo which is also around 6.5 to 7 para magamit mo siya
for a week or for a day kasi anything that is beyond or
above 6.5 to 7, mamamatay yung salivary amylase natin.
Another example, normal breast cells. Ilalabas mo siya
sa isang organism (human), its favorite condition is
Figure 1.4 Example of pH paper with multiple indicators around 6.5 to 6.9 pH para tumagal yung normal breast
and a color chart. cells mo kasi kailangan mo for thesis, kailangan mo ng
buffer same condition doon sa organism na ang pH is
BUFFER SOLUTION also 6.5 to 6.9. Kasi anything beyond or below,
● A buffer is an aqueous solution containing a weak mamamatay yung normal breasts cells mo. Buffer
acid and its conjugate base or a weak base and its solution is very important especially in in vitro studies. It
conjugate acid. A buffer’s pH changes very little is highly resistant to pH.
when a small amount of strong acid or base is added
to it. It is used to prevent any change in the pH of a
solution, regardless of solute. Buffer solutions are
used as a means of keeping pH at a nearly constant
value in a wide variety of chemical applications. For
example, blood in the human body is a buffer
solution.

Buffer solution is a combination of a weak acid and its

conjugate base. One is an acid and one is a conjugate
base so therefore it makes your solution not prone to
removal and addition of hydrogen. It is very resistant to
pH change. Isang acid plus a conjugate base na
pinagsama natin will become what we call a buffer
Figure 1.5 Buffer Solution
solution. Since one is an acid and the other one is not a
base, it is a conjugate base, therefore, it is not prone to
addition or removal of hydrogen on its structure therefore HENDERSON-HASSELBALCH EQUATION
it is very resistant to any solution that will be added to our ● Henderson–Hasselbalch equation can be used to
buffer solution. So lagyan mo man ng strong acid yung estimate the pH of a buffer solution containing given
namixed na buffer solution, hindi magchachange yung concentrations of an acid and its conjugate base (or
pH niya. For example, 7 yung pH niyan, dagdagan natin a base and its conjugate acid). The numerical value
ng acid, hindi magchachange yung pH. Magdagdag man of the acid dissociation constant of the acid must
ng base solution na may pH of 10, hindi pa rin also be known.
magbabago yung buffer pH natin. Why do we need to
have a buffer solution? This is important especially if we

COURSE CODE TRANSCRIBER/S: Gilbuena 3

 pH and Buffers TRANS # 3

How do we test/how do we check the pH of a certain

buffer? Kailangan lang natin is yung pKa value or yung
tinatawag nating acid dissociation. Logarithm of 10 of the
conjugate base and the weak acid.

Figure 1.6 Henderson-Hasselbalch Equation

Example:
What is the pH of a solution consisting of 0.75 M
HC2H3O2 and 0.50 M NaC2H3O2? Ka of HC2H3O2 is
−5
1. 8 × 10 .
𝑏𝑎𝑠𝑒
𝑝𝐻 = 𝑝𝐾𝑎 + 𝑙𝑜𝑔( 𝑎𝑐𝑖𝑑 )
𝑝𝐾𝑎 =− 𝑙𝑜𝑔(𝐾𝑎)
−5
𝑝𝐾𝑎 =− 𝑙𝑜𝑔(1. 8 × 10 ) Figure 1.7 pH Scale
𝑝𝐾𝑎 = 4. 745
● What is a solution called that has more OH-ions? H+
Look for pH using Henderson-Hasselbalch equation ions? Does a solution at pH 4 have more or less
𝑏𝑎𝑠𝑒
𝑝𝐻 = 𝑝𝐾𝑎 + 𝑙𝑜𝑔( 𝑎𝑐𝑖𝑑 ) hydrogen ions than the same amount of another
0.50 solution at pH 8?
𝑝𝐻 = 4. 745 + 𝑙𝑜𝑔( 0.75 ) ○ A solution that has a pH 4 has more hydrogen
𝑝𝐻 = 4. 57 ions and is considered as an acidic solution than
0.75 M HC2H3O2 is an acid because it has extra a solution with pH 8.
hydrogen, it has 4 hydrogen. 0.50 M NaC2H3O2 has
only 3 hydrogen. Kung sino yung mas madaming REFERENCES
hydrogen, that is your acid. Kung sino yung mas kaunti
na tanggalan ng isang hydrogen, that is considered as
● Professor’s PPT
your conjugate base.
● Recorded Video Lecture
Ka is the dissociation constant value for acid. pKa means
gaano katapang yung acid, gaano kastrong yung acid.
Normally, the lower the pKa, the stronger the acid. Ka
value is computed sa product concentration divided by
reactant concentration.
𝑝𝑟𝑜𝑑𝑢𝑐𝑡
𝐾𝑎 = 𝑟𝑒𝑎𝑐𝑡𝑎𝑛𝑡 𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛
For example,

+ −
[𝐻3𝑂 ][𝑁𝑂2 ]
𝐾𝑎 = [𝐻𝑁𝑂2][𝐻2𝑂]

POST-LAB
● What do acidic solutions have high concentrations
of? On the pH scale, where can you find the strong
acids and the strong bases?
○ Acidic solutions have high concentrations of H+
ions. The higher the concentration of hydrogen
ions (measured in mol/l or moles per litre) the
stronger the acidity of the solution

COURSE CODE TRANSCRIBER/S: Gilbuena 4