MODX311 LABORATORY | 2nd Semester

WEEK 2: LABORATORY PH METER

INTRODUCTION The most common buffer solutions to conduct a 3-point

calibration are neutral (pH 7.0), acidic (pH 4.0), and alkaline
● pH is a measure of hydrogen ion concentration in a solution. (pH 10.0).
● pOH is a measure of hydroxide ion concentration. ● A two point calibration will also provide you with high
● pH is the more commonly used measure. However, either accuracy results. The buffer solutions used for 2-point
scale can describe acidity or alkalinity. calibration are 7.0 and 4.0. We lean towards 3-point
● When a solution contains more hydrogen ions than calibration as they provide a higher accuracy over a full pH
hydroxide ions, it is acidic. range.
○ An acidic solution has a pH of < 7 at 25ºC. 6.999 acidic na
● When a solution contains more hydroxide ions than
hydrogen ions, it is basic.
pag Two-Point
○ A basic solution has a pH of > 7 at 25ºC. 7.0001 Calib (7.0 & 4.0) Three-point Calib (4.0, 7.0, 10.0)

0-8 lang yung accuracy pag ito ginamit may 0-14 accuracy
Steps 1: Mid-Point Calibration
Components of pH Meter

● The mid-point calibration must be done first, as it clears the

● pH meter Inside nung probe doon nakalagay yung other calibration points.
● pH electrode electrode. Inside the electrode mag-
papass yung voltage and minemeasure ● Enable continuous readings.
● pH probe yung voltage na narereceive nung another
electrode kaya nalalaman yung pH value ● Rinse the pH probe well with demineralize, deionized (DO), or
reverse osmosis (RO) water.
The two electrodes inside the pH probe measures the hydrogen-ion
● Remove the top of the pH 7.0 calibration pouch or solution
activity in a solution, such as water. The exchange of ions generated a
bottle.
voltage, which is then measured and converted by the pH meter into a
● Place the pH probe inside the pouch/bottle, and let it sit in
readable pH value.
the solution until the readings stabilize (this usually take 1-2
minutes).
● Once the pH readings have stabilized, issue the mid-point
calibration commandL cal, mid, 7

Step 2: Low-Point Calibration

e.g, Domex ● Rinse the pH probe before calibrating the low-point.

● Open the calibration pouch or bottle of the pH 4.0 calibration
● Determining the pH provides the user with a number ranging solution.
from 0 to 14, where 7.0 is neutral, anything above 7.0 is ● Insert the pH probe inside the acidic buffer solution.
alkaline (or basic), and a pH value above 7.0 is acidic. ● Wait for the pH readings to stabilize.
● Measuring pH is essential in a wide range of applications, ● After the pH reading has stabilized, issue the low-point
but it is more critical in industrial applications such as calibration command: cal, low, 4
environmental monitoring, quality control in food and
beverages, water pre-treatment systems, and medical and Step 3: High-Point Calibration
laboratory applications. In MolBio we use ph Meter para
imeasure yung ph ng agarose

● Again, rinse the pH probe.

CALIBRATION OF A PH METER ● Open the pouch or bottle of the pH 10.0 calibration solution.
● Place the pH probe inside the pouch or bottle.
● Calibrating the pH meter correctly ensures the electrode ● Issue the low-point calibration command (cal, high, 10) once
inside the pH probe has not drifted off or is damaged. the readings have stabilized.
● To calibrate a pH meter correctly, you will need at least two
yung mga calib sol. are standard sol. - highest purity / grade of chemicals
different buffer solutions. The two main buffer solutions are Note: so one time used lang sila since onec na maopen it is considered na contaminated na agad
the pH4 and pH7 solutions.

● As opened calibration solutions are no longer considered

How to Calibrate pH Meter Correctly accurate after 20 minutes, dispose of the used and unused
solution.
● If it is the first time using your pH meter, the 3-point ● Once a buffer solution has been opened, we can assume that
calibration is performed in the following order of buffer they are contaminated and therefore, are no longer suitable
solutions: to calibrate pH meters.
a. Mid-point pH (neutral) ● In addition to general cleaning your pH meter, ensure you
b. Low-point pH (acidic) have removed salt deposits by submerging the pH electrode
c. High-point pH (alkaline) in 0.1M HCl for 5 minutes followed by 0.1M NaOH for

JEANINE YSABELLE B. PAZ 4

 another 5 minutes. Finish off by rinsing the electrode with
distilled water.
● Wash the sensor bulb in methanol or a mild detergent to
remove oil and grease buildup.
● You also want to prevent the reference junction from
clogging. Place the pH electrode into a heated diluted KCl preferably 3M of KCl

solution for 10 minutes and allow it to cool before use or

storing.

PH IN LIVING SYSTEMS

INDICATOR COLOR AT TRANSITION COLOR AT

LOWER PH PH RANGE HIGHER PH

Thymol Blue Red 1.2 - 2.8 Yellow

(First
Transition)

Methyl Red Red 4.4 - 6.2 Yellow

Bromothymol Yellow 6.0 - 7.6 Blue

Blue (Second
Transition)

Thymol Blue Yellow 8.0 - 9.6 Blue

Phenolphthalein Colorless 8.3 - 10.0 Fuchsia

PH IN LIVING SYSTEMS

● In the case of blood, pH is 7.365 (slightly basic) and it is

kept between 7.35 to 7.45 to be able to undergo all
physiological reactions.
● The pH of gastric fluids is highly acidic in order to digest
proteins. The acidity is due to its composition: high
concentration of hydrochloric acid (HCl) and potassium
chloride (KCl) and sodium chloride (NaCl). Cells in the
stomach produce mucus to form a protective barrier that
prevents damages in the organ.

Fluid pH

Gastric Acid 1.0

Urine 6.0

Cerebrospinal Fluid (CSF) 7.5

Blood 7.35 - 7.45

Pancreas Secretions 8.1

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