pH Meter, CALIBRATION (INSTRUMENTS) and Gatot Ciptadi FAKULTAS 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 ModernAnimal 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 Biotech Before 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 Code RED 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 nucleus from egg cell Add somatic cell from adult donor Grow in culture to produce an early embryo (blastocyst) Implant blastocyst in surrogate mother Remove embryonic stem cells from blastocyst and grow in culture Induce stem cells to form specialized cells (therapeutic cloning) Clone of donor is born (reproductive cloning) Donor cell Nucleus from donor cell Animal Cell Culture: Biotechnology In vItro Technology: pH, etc.

Technique and instrument Laminar 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

pHscale: 0-14 Acid: dissociates hydrogen ions into solution pH below 7 Base: dissociates hydroxide ions into solution or accepts hydrogen ions pH above 7 Acidic solution Neutral solution Basic solution Increasingly ACIDIC (Higher concentration of H + ) Increasingly BASIC (Lower concentration 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+H+ OH – H+H+ 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 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 pH pH is defined as the negative decimal logarithm of the hydrogen ion activity a H. pH scale is based on the pH values of a series of standard buffer solutions determined by the potentiometric technique. pH meterpH-indicator glass electrode,reference electrodeautomatic temperature compensator Routine potentiometric measurements of pH are carried out by means of a pH meter equipped by a pH-indicator glass electrode, a reference electrode and an automatic temperature 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 meter adjustment pH meter is a high impedance voltmeter. Direct pH readings are possible after adjustment of a pH meter. Dr. Elena Kardash - Calibration of pH meters, glass electrodes and conductometers at INPL

21 systematic errors Some factors lead to systematic errors in pH measurement results, which can not be compensated by the adjustment procedure: asymmetry potential asymmetry potential of glass electrode, liquid junction potential liquid junction potential of cell (1), clogging ofdiaphragm clogging of glass electrode diaphragm etc. a calibration of the pH meter–electrodes assembly This problem can be solved by a calibration of the pH meter–electrodes assembly. Dr. Elena Kardash - Calibration of pH meters, glass electrodes and conductometers at INPL