pH Meter, CALIBRATION

(INSTRUMENTS) and

Gatot CiptadiFAKULTAS PETERNAKAN - UB

Biotech Laboratory(Animal Cell Biotechnology)

Biotechnology• Technology based on biology (molecular)

• Biotechnology is the integration of natural sciences and engineering sciences in order to achieve the application of organisms, cells, parts thereof and molecular analogues for products and services

Traditional vs. Modern Biotechnology

Traditional• Based on microorganisms• Cells are not manipulated• Production of antibiotics

by fermentation

Modern Animal Cell culture ?• Based on microorganisms and cell culture• Cells are manipulated• Production of protein products based on recombinant

DNA and cell culture• Modern biotech refers to Molecular Biology, Molecular

Genetics and Genetic Engineering

Current Applications of BiotechBefore going in-depth with a few areas of biotech research, it is

important to gain a brief understanding of the many applications of biotechnological advancements.

• Microbial

• Agricultural

• Animal

• Forensic

• Environmental

• Aquatic

• Medical

Biotechnology Color CodeRED BIOTECHNOLOGY: Medicine / Pharmaceuticals• Gene therapy• Production of proteins, antibodies, and vaccines• Tissue engineering

WHITE BIOTECHNOLOGY: Chemical industry• Basic chemicals• Biopolymers• Biorefinery

GREEN BIOTECHNOLOGY: Agriculture• Crops improvement (Agriculture)• Natural pesticides• Pest and disease resistance

Desinfection step

Tissue isolation

Incubation&growth

Primary cells

Animal Cell Biotechnology: pH?

Remove nucleusfrom egg cell

Add somatic cellfrom adult donor

Grow in culture to produce anearly embryo (blastocyst)

Implant blastocyst insurrogate mother

Remove embryonic stemcells from blastocyst andgrow in culture

Induce stem cells toform specialized cells(therapeutic cloning)

Clone of donor is born(reproductive cloning)

Donorcell

Nucleus fromdonor cell

Animal Cell Culture: Biotechnology

In vItro Technology: pH, etc.

Technique and instrumentLaminar flow

Why scientists research water pH?

• To determine what can live in a water body, both animals and plants

• To track the mixing of waters in estuaries and along coasts

• To help relate water quality to surrounding soil, geology, and the pH of rain and snow melt

• Cell culture• Live and dead Cell, etc

PH METER :

Draft 2005

Science Content: What is pH?pH measures the acid content of water

pH scale: 0-14

Acid: dissociates hydrogen ions into solutionpH below 7

Base: dissociates hydroxide ions into solution or accepts hydrogen ionspH above 7

Acidic solution

Neutral solution

Basic solution

Incr

easi

ng

ly A

CID

IC(H

igh

er c

on

cen

trat

ion

of

H+)

Incr

easi

ng

ly B

AS

IC(L

ow

er

con

cen

trat

ion

of

H+)

NEUTRAL[H+] = [OH–]

Lemon juice; gastric juice

Grapefruit juice, vinegar, beer, wine, cola

Tomato juice

Urine

PURE WATER

Seawater

Milk of magnesia

Household ammonia

Household bleach

Oven cleaner

Human blood

H+

OH–

H+

OH–

Black coffee, rainwater

Introduction

Draft 2005

Science Content: What affects water pH?• Local geology

• Soil conditions

• Surrounding vegetation

• Mining activities/limbah

• Industry runoff/water pollution

• Other human activities near water body

• pH of rain water and up-wind air pollution sources

• Life-dead cells (cultured)

• Sperms-Oocyte, Somatic cell etc

Photo taken by Carlye Calvin

© University Corporation for Atmospheric Research

Biotech Laboratory

•Most widely used biotech products are recombinant proteins (produced by gene cloning in cell culture)

•Cell culture refers to the technique of growing cells in a lab under controlled conditions; similar to in vitro

•“in vitro” refers to working in a controlled environment outside of a living organism

•Bioreactors (large culturing “containers” where DNA of interest can be mass produced) are also used

Tissue culture Ware

Changing Medium

Your standards should resemble the following colors.

* You did not prepare a solution with pH 8. I added this test tube so you can see the change in color.

pH 2 pH 3 pH 5 pH 7 *pH 8 pH 12

Using indicators to measure pH Experiment

Measuring pH of various solutions using a pH meter Record the pH of the solutions on your report sheet.

The pH meter and pH paper Experiment (dry lab)

The pH meter and pH paper Experiment

Measuring pH of various solutions using a pH paper

The pH paper is orange in color (before use) and changes color depending on the pH being tested. Look at the pH key to determine the pH of the solution ranging from 1-12.

A pH strip has been dipped into the solution and is displayed sitting outside the beaker

II. The pH meter and pH paper Experiment (dry lab)

The pH meter and pH paper Experiment

19

pHpH is defined as the negative decimal logarithm of the hydrogen ion activity aH.

pH scale is based on the pH values of a series of standard buffer solutions determined by the potentiometric technique.

Routine potentiometric measurements of pH are carried out by means of a pH meterpH meter equipped by a pH-indicator pH-indicator glass electrode,glass electrode, a reference electrodereference electrode and an automatic automatic temperature compensatortemperature compensator.

Dr. Elena Kardash - Calibration of pH meters, glass electrodes and conductometers at INPL

20

The following cell is required for the pH measurements:

glass electrode│test solution bridge solution│ref. electrode, ║ (1)

where │ is the electrode-solution interface, and ║ is the liquid junction denoting an interface between the test solution and the salt bridge solution.

pH meterpH meter is a high impedance voltmeter. Direct pH readings are possible after adjustmentadjustment of a pH meter.

Dr. Elena Kardash - Calibration of pH meters, glass electrodes and conductometers at INPL

21

Some factors lead to systematic errorssystematic errors in pH measurement results, which can not be compensated by the adjustment procedure:

asymmetry potentialasymmetry potential of glass electrode,

liquid junction potentialliquid junction potential of cell (1),

clogging ofclogging of glass electrode diaphragmdiaphragm etc.

This problem can be solved by a calibration of the pH a calibration of the pH

meter–electrodes assemblymeter–electrodes assembly.

Dr. Elena Kardash - Calibration of pH meters, glass electrodes and conductometers at INPL