# Describe how pH and H3O+ concentration are related and explain why diluting an acid raises the pH, but diluting a base lowers the pH.

BIO 1121 Unit 2 Assignment
Accurately measuring the volume of liquids, weighing chemicals, and adjusting the pH of solutions are routine procedures in a working laboratory environment.  This assignment is designed to provide you with an overview of the general skills and knowledge you would need to perform such tasks.
Before completing this assignment you should ensure you have read chapter 2 of your text book – particularly the section entitled pH, Buffers, Acids, and Bases.  Answers should be concise and well written.  Make sure you correctly explain your thought process and provide all the necessary information.
The questions are available on the following worksheet:
Question 1:  The pH of a solution describes its acidity or alkalinity: Describe how pH and H3O+ concentration are related and explain why diluting an acid raises the pH, but diluting a base lowers the pH.
Phosphate Buffered Saline (PBS) is a commonly used buffer for experiments in biology, because its pH and ion concentrations are similar to those in mammalian organisms.  It works in a similar fashion to the blood plasma buffer mentioned in the textbook, but using dihydrogen phosphate ions and hydrogen phosphate ions for buffering.
aq)
Question 2: Based on the equation below, which ion plays the role of hydrogen-ion donor (acid) and which ion plays the role of hydrogen-ion acceptor (base) in PBS?
H2PO4-(aq)  ⇆  H+(aq) + HPO42-(
The composition of PBS is 0.137M NaCl,  0.012M Phosphate,  0.0027M KCl,  pH 7.4.  Below is the protocol to make 1 liter of 10x concentrate PBS.
Combine the following:
• 80g NaCl
• 2g KCl
• 14.4g Na2HPO4 (dibasic anhydrous)
• 2.4g KH2PO4 (monobasic anhydrous)
• 800mL distilled H2O
1. Adjust pH to 7.4 with HCl
3. Autoclave for 20 minutes on liquid cycle. Store at room temperature.
Question 3:  Which ions are being produced by this process, assuming that each of the chemical compounds dissociate into their constituent parts once they are dissolved in water?
Preparation of the correct buffer is key to any good biological experiment and it is important that you understand how to calculate the mass of each chemical required to make that buffer and what the resulting concentration of those constituents will be in moles per liter.
Your text book explains that moles are just a way to express the amount of a substance, such that one mole is equal to 6.02 x 1023 particles of that substance.  These particles can be can be atoms, molecules, ions etc, so 1 mole of water is equal to 6.02 x 1023 water molecules, or 1 mole of Na+ is equal to 6.02 x 1023  Na+ ions.  Since different chemicals have different molecular weights (based on the number of protons and neutrons each atom contains) 1 mole or 6.02 x 1023 atoms of oxygen (O) will have a mass of 16g whereas 1 mole or 6.02 x 1023 atoms of sodium (Na) will have a mass of 23g
You can read more about the definition of moles here if you wish https://www.britannica.com/science/mole-chemistry
Although you may sometimes see it written as g/liter, the concentration of solutions is more often described in term of molarity since it better defines the chemical properties of a solution because it is proportional to the number of molecules or ions in solution, irrespective of molecular mass of its constituents.  However, it is not possible to measure moles on a laboratory balance, so in the first instance chemicals are measured by mass (milligrams, grams, kilograms etc) and the number of moles is calculated using the known molecular mass (often called molecular weight and abbreviated to M.W.) of the chemical.  As indicated earlier, the molecular mass of a chemical is based on the number of protons and neutrons that is contained in each atom (eg NaCl is made up of one molecule of Na, M.W. = 22.99g and one molecule of Cl, M.W. = 35.45g, so the M.W. of NaCl is 58.44g).  These values can be found in the periodic table however the molecular mass of chemicals is generally provided by any vendors of the products and so can also be found on various suppliers’ websites.
When the concentrations of solutions are as described as ‘molar’, this refers to number of moles per liter eg a 3 molar solution of NaCl will contain 3 moles of NaCl in 1 litre of water.  As indicated above, the M.W. of NaCl is 58.44g, so in 58.44g there are 6.02 x 1023 NaCl molecules ie 1 mole.  So for 3 moles of NaCl you would need to dissolve 175.32g in 1 liter of water (175.32/58.44 =3) whereas If you only dissolved 29.22g of NaCl in 1 liter of water this would result in a 0.5 molar solution (29.22/58.44= 0.5)
Question 4:  Using periodic table found in your textbook, calculate (to 2 decimal places) the molecular mass for each of the compounds used to make PBS.
Create the following table and fill it in with the mass of each component required to make 1 liter of 10 x PBS (the recipe for 10x PBS is below question 2) and their final molar concentration in the buffer calculated as described above.
Compound formula Molecular mass (in g/mol) Mass of compound per liter of 10x PBS (in g) Molar concentration (in mol/l)
NaCl
KCl
Na2HPO4
KH2PO4
Question 5:  As previously stated, the concentration of NaCl, KCl and Phosphate in working strength 1 x PBS is 0.137M NaCl, 0.012M Phosphate, 0.0027M KCl,  pH 7.4   How do they compare to the concentrations you calculated for 10x PBS?
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