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How To Make Simple Solutions and Dilutions

This document provides instructions for making simple dilutions, serial dilutions, solutions of specific concentrations using the C1V1=C2V2 method, molar solutions, percent solutions, and conversions between units. It explains how to calculate volumes and masses of reagents needed to make solutions of desired concentrations from stock reagents through dilution or direct preparation. Key methods covered include dilution factor, C1V1=C2V2, use of moles and molarity, percent solutions, and conversions between units.

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Amra ahmed
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99 views

How To Make Simple Solutions and Dilutions

This document provides instructions for making simple dilutions, serial dilutions, solutions of specific concentrations using the C1V1=C2V2 method, molar solutions, percent solutions, and conversions between units. It explains how to calculate volumes and masses of reagents needed to make solutions of desired concentrations from stock reagents through dilution or direct preparation. Key methods covered include dilution factor, C1V1=C2V2, use of moles and molarity, percent solutions, and conversions between units.

Uploaded by

Amra ahmed
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
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How to Make Simple Solutions

and Dilutions
Taken from: Resource Materials for the Biology Core
Courses-Bates College (there may be errors!!)
WHY???
• You want to subject your DNA to a .5M solution
of EDTA. HUM!!!!!!!!
• How would you do that??????
• You want to subject your cells to a .54% KCl
hypotonic solution. HUM!!!!!!!!
• How would you do that?
• You want to dilute your buffer 5 fold. HUM!!!!!
• How would you do that?
1. Simple Dilution
(Dilution Factor Method)

A simple dilution is one in which a unit volume of a liquid material of interest is


combined with an appropriate volume of a solvent liquid to achieve the desired
concentration. The dilution factor is the total number of unit volumes in which your
material will be dissolved.

For example, a 1:5 dilution entails combining 1 unit volume of diluent (the material to
be diluted) + 4 unit volumes of the solvent medium (hence, 1 + 4 = 5 = dilution
factor).

Example: Frozen orange juice concentrate is usually diluted in 4 additional cans of


cold water (the dilution solvent) giving a dilution factor of 5 (1 : 5). So, in a simple
dilution, add one less unit volume of solvent than the desired dilution factor value.
2. Serial Dilution
• A serial dilution is simply a series of simple dilutions which amplifies the dilution factor
quickly beginning with a small initial quantity of material (i.e., bacterial culture, a
chemical, orange juice, etc.).
• Three step 1:100 serial dilution of a bacterial culture. The initial step combines 1 unit
volume culture (10 µL) with 99 unit volumes of broth (990 µL) = 1:100 dilution. In the
next step, one unit volume of the 1:100 dilution is combined with 99 unit volumes of
broth now yielding a total dilution of 1:100x100 = 1:10,000 dilution. Repeated again
(the third step) the total dilution would be 1:100x10,000 = 1:1,000,000 total dilution.
The concentration of bacteria is now one million times less than in the original sample.
3. Making fixed volumes of specific
concentrations from liquid reagents:
C1V1=C2V2 Method
• Suppose you have 3 mLof a stock solution of 100 mg/mL
ampicillin (= C1) and you want to make 200 µL(= V2)of solution
having 25 mg/ mL (= C2). You need to know what volume (V1)
of the stock to use as part of the 200 µL total volume needed.

• V1 = the volume of stock you will start with. This is your


unknown.

C1 = 100 mg/ mL in the stock solution

V2 = total volume needed at the new concentration =


200 ul = 0.2 mL

C2 = the new concentration = 25 mg/ mL


V1C1=V2C2 Method
cont.
• V1 = (V2 x C2) / C1
• V1 = (0.2 mL x 25 mg/mL) / 100 mg/mL
• and after cancelling the units,
• V1 = 0.05 mL, or 50 µL

• So, you would take 0.05 mL = 50 µL of stock solution


and fill to 200 uL with the solvent to get the 200 uL of the
25 mg/ mL solution needed (remember that the amount
of solvent used is based upon the final volume needed.
4. Moles and Molar solutions
(unit = M = moles/L)
• Sometimes it may be more efficient to use molarity
when calculating concentrations.

• A mole is “one gram molecular weight” of an element or


compound, and comprised of exactly 6.023 x 10^23
atoms or molecules (this is called Avogadro's number).
– Sodium Chloride = 58.44 g/mole

• The mass attributed to one mole of any element or


compound is called its atomic weight (element) or
molecular weight, or formula weight for compounds.
The number of moles of a given dry reagent can be
calculated as:
– # of moles = weight (g)/ molecular weight (g)
Moles and Molar solutions cont.
• Molarity is the unit used to describe the number of moles of a
chemical or compound in one liter (L) of solution. For example, a 1.0
Molar (1.0 M) solution is equivalent to one formula weight (FW =
g/mole) of chemical dissolved in 1 liter (1.0 L) of solvent (for example
water). Formula (or molecular) weight is always given on the label of
a chemical bottle.

• Example 1: To prepare a liter of a simple molar solution from a


dry reagent

• Multiply the formula weight (or MW) by the desired molarity to


determine how many grams of reagent to use:

– Chemical FW = 194.3 g/mole; to make 0.15 M solution use:

194.3 g/mole * 0.15 moles/L = 29.145 g/L


Moles and Molar solutions cont.
• Example 2: To prepare a specific volume of a specific molar
solution from a dry reagent
• A chemical has a FW of 180 g/mole and you need 25 mL (0.025 L)
of 0.15 M (M = moles/L) solution. How many grams of the chemical
must be used to make this solution?

180.0 g/mole * 0.15 moles/L * 0.025L = 0.675 g

Procedure:
1. Mass 0.675 grams of chemical and fill to 15mL
in appropriate solvent in appropriate vessel.
2. Agitate until in solution (solubility considered)
3. Fill to final volume of 25 mL
5. Percent Solutions (= parts per
hundred)
• Many reagents are mixed as percent concentrations. When working with a
dry chemical it is mixed as dry mass (g) per volume where #g/100 mL =
percent concentration.
– A 10% solution is equal to 10 g solute dissolved in 50mL solvent,
agitated, and filled (finally) to 100 mL with solvent.

• Example 1: If you want to make 3 % NaCl you would dissolve 3.0 g NaCl
and fill to 50 mL with distilled water, agitate, fill to 100 mL with distilled
water. SOLUBILITY?

• When using liquid reagents the percent concentration is based upon


volume per volume, i.e., # ml/100 mL.

• Example 2: If you want to make 70% ethanol you would measure 70mL of
100% ethanol and fill to 50mL with distilled water, agitate and fill to 100mL.
• To convert from % solution to molarity, multiply the percent solution value by
10 to get grams/L, then divide by the formula weight.

• Molarity = (% solution) * 10
FW
• Example 3: Convert a 6.5 % solution of a chemical with FW = 325.6 to molarity 
• [(6.5 g/100 ml) * 10] / 325.6 g/L = 0.1996 M

• To convert from molarity to percent solution, multiply the molarity by the FW


and divide by 10:
 
• % solution = molarity * FW
10
• Example 4: Convert a 0.0045 M solution of a chemical having FW 178.7 to
percent solution:
•  
• [0.0045 moles/L * 178.7 g/mole] / 10 = 0.08 % solution
ppm?

This means: One part per million parts examined.

So if the concentration of arsenic is measured to


be 1 ppm. That means there is one gram of
arsenic for every million grams of water.

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