Making Solutions In chemistry, we use a lot of solutions.

It is important that we know the composition of the solutions that we are using in order to calculate expected yields, maintain proper salt concentrations or pH levels for optimal reaction conditions, etc. Depending on the needs of the experiment, the chemicals involved, industry standards and personal preferences, solutions can be made in a number of different ways. We can make them as a percent solute to solvent or use a mole relationship to determine concentration. We can also create working solutions from stock solutions. Each solution is made slightly differently, and it is important to understand the slight nuances that distinguish one type of solution from another. Below are a short list of some of the types of solutions we may use in a chemistry lab. Relevant Vocabulary Solute: The dissolved substance in a solution Solvent: A substance in a solution that is present in the largest amount Solution: A homogenous mixture consisting of a solvent and one or more solutes. Mole: the amount of a substance that contains as many elementary entities (atoms, molecules, formula units, etc.) as there are atoms in exactly 0.012 kg of Carbon12 (6.02x1023) Molar Solution: the number of moles of solute per liter of solution Stock solution: A concentrated solution that will be diluted to a lower concentration for use in an experiment. They are used to save preparation time, increase accuracy, conserve materials and reduce storage space. Working solution: A solution that is actually used in an experiment. Miscible: mutually soluble in any proportions. General notes on solution making: Do not add water/solvent at the final volume amount - the final volume of the solution will be greater than desired if you do this, as the solute DOES change the volume!! When you are adding the water, make sure that you do not exceed the final volume you are trying to achieve! For many solutions, this can be very close to the final volume, but the more solute you have, the less water you will be able to use, so adjust accordingly. Although beakers/flasks are not volumetric glassware, the markings on the side can tell you if you are too close to your desired volume. Always determine your FINAL volume using a graduated cylinder or volumetric flask. Even when adding liquid to liquid, the final volume may not be what you expect due to the miscibility of some liquids.

A. Percent solution Weight/Volume (w/v) Definition: Example: % solution(w/v) = 10% solution of NaCl = 10 g of salt in 100 mL of solution. 10% NaCl (w/v) = How to make it: 1. Weight 10.000 grams of salt in a tared weigh boat on scale. 2. Transfer NaCl to 250 ml or larger beaker. 3. Dissolve NaCl COMPLETELY in about 50-75 ml of water. 4. Bring final volume up to 100 mL using graduated cylinder or volumetric flask. B. Percent solution Weight/weight (w/w) Definition: Example: % solution(w/w) = 10% solution of NaCl = 10 g of salt in 100 g of solution. 10% NaCl (w/w) = How to make it: 1. Weigh 10.000 grams of salt in a tared weigh boat on scale. 2. Transfer NaCl to 250 ml or larger beaker. 3. Dissolve NaCl COMPLETELY in 90.000 g of water. Note: weight, unlike volume is additive you can measure these independent of each other and mix to create your solution Note: Weighing solutions can be tedious and messy. In order to alleviate this, we can use the density of water (or other solvent), calculate the required amount of solvent by taking the mass of the solution and subtracting the mass of the solute. The remaining mass is the mass of the solvent. (Remember, masses are additive volumes are NOT always additive!). Once you know the mass of the solvent, you can then determine the volume using the density. For pure water, which is the most common solvent, the density of water is 1 g/mL. This way, we can still weight out the dry solute and use a graduated cylinder to measure out the water. How to make it: 1. We want 10.000 grams of salt in a 100 g solution. 2. Weigh out 10.000 grams of salt in a tared weigh boat on scale.

3. Transfer NaCl to 250 ml or larger beaker. 4. The remaining mass is solvent so we need 90 grams of solvent. a. 100 g of solution 10 g NaCl= 90 g of solute. 5. Use the density of the solvent to calculate the volume of the solute required a. 90 g solute * 6. Dissolve NaCl COMPLETELY in 90.000 mL of water. C. Percent solution Volume/Volume (v/v) Definition: Example: % solution(v/v) = 10% solution of Ethanol = 10 mL of ethanol in 100 mL of solution. 10% Ethanol (v/v) = How to make it: 1. Measure 10.0 mL of ethanol using a 10 mL graduated cylinder or 10.0 mL volumetric pipette. 2. Transfer ethanol into 100 mL graduated cylinder or volumetric flask. 3. Bring the final volume to 100 mL/ D. Molar Solution Definition: Example: Molar solution = 1 M solution of NaCl (mm = 58.44 g/mol) 1 M NaCl = How to make it: 1. Calculate the number of grams in 1 mole of NaCl in 1 L of solution. Adjust volume/mols for desired volume. a. Using molar mass, we know that 1 mol of NaCl is 58.44 g of NaCl. b. For 1 L of a 1 M solution of NaCl, we would use 58.44 g of NaCl. For a 2 M solution, we would use 2 times that (116.88 g NaCl). c. To change the volume, we can use a simple ratio expression as below. i. d. If desired volume is 100 mL(0.1 L), we can plug in and solve for X i.

ii. Therefore, x = 5.844 g NaCl in 0.1 L solution 2. Weight 5.844 of NaCl in a tared weigh boat on scale. 3. Transfer NaCl to an appropriate sized beaker for your final volume. 4. Dissolve NaCl COMPLETELY in about 50-75 ml of water. 5. Bring final volume up to 100 mL using graduated cylinder or volumetric flask. Dilution from stock solutions Many solutions used in a chemistry laboratory often use the same component(s). It is often easier to make a single stock solution of higher concentration and then dilute this stock solution to the desired working concentration to save you from making many different solutions of the same thing but at different concentrations. Example: We have a 1 M stock solution of NaCl We want a 0.1 M working solution of NaCl How to make it: 1. Calculate the number of required volume of stock solution to achieve the concentration and volume you want of your working solution. a. To do this, use M1V1 = M2V2, where M1 = concentration of stock solution, M2 = concentration of desired working solution and V2 = volume of desired working solution. i. (1M NaCl)*V1 = (0.1M NaCl)* (1 L solution) ii. Solving for V1, V1 = 0.1L or 100 mL of 1M NaCl solution 2. Measure out calculated amount of NaCl solution using a graduated cylinder or other appropriate volumetric glassware 3. Bring final volume up to 1 L using graduated cylinder or volumetric flask. E. Serial Dilutions When you are diluting a stock solution substantially to obtain a working solution, or in cases where you want a number of different concentrations, such as in the generation of a standard curve, we use a process of step-wise dilutions. This is referred to as serial dilutions. This is done in the same manner described above, only as you step-down from one dilution to the next, you have to do a new set of calculations. Often, these dilutions are done in a logarithmic manner, such that your steps are factors of 10. (ie// your stock concentration is 1M, and your steps are 0.1M 0.01M, and 0.001M.) To achieve this, you simply take the same amount of the first solution and bring it up to the same volume in the second solution.

Example:

We have a 1 M stock solution of NaCl We want 0.1 M, a 0.01M and a 0.001M working solutions of NaCl

How to make it: a. To do this, use M1V1 = M2V2, where M1 = concentration of stock solution, M2 = concentration of desired working solution and V2 = volume of desired working solution. i. (1M NaCl)*V1 = (0.1M NaCl)* (1 L solution) ii. Solving for V1, V1 = 0.1L or 100 mL of 1M NaCl solution b. REPEAT, but start with M1 as the concentration of the solution you just made (so 0.1M NaCl) i. (0.1M NaCl)*V1 = (0.01M NaCl)* (1 L solution) ii. Solving for V1, V1 = 0.1L or 100 mL of 1M NaCl solution c. REPEAT again and again until you have achieved the desired concentration you ar(s)e looking for

PRE-LAB QUESTIONS TYPE these answers on a separate sheet of paper you may handwrite calculations. Please include relevant calculations in your lab notebook as well!!!! Due at the beginning of Lab

1. Describe how you would make 50.0 mL of a 30.0% (w/w) solution of sucrose. (density of 30.0% sucrose = 1.127 g/mL). Show all calculations. Please note: 50 g of solution does not equal 50 mL of solution! 2. Describe how you would make 250 mL of a 0.1 M Na2HPO4 (disodium phosphate, MM =141.96). Include the calculations for the preparation of 250 mL of a 0.10 M NaH2PO4 (monosodium phosphate, MM = 119.98), but you do not have to describe both, as the protocol will be the same. 3. PLEASE NOTE: THIS IS SLIGHTLY DIFFERENT THAN IN THE PROTOCOL YOU DO NOT NEED TO ENTER THIS TABLE IN YOUR NOTEBOOK, BUT RATHER THE TABLE FROM THE SECTION BELOW THIS IS ONLY FOR A PRELAB EXERCISE! We want to create a standard curve using multiple concentrations. To do this, we will start with of our stock solution and then dilute this solution to create the next desired concentration, and then use this new solution to create the next solution and so on (so use solution 1 as the starting solution for solution 2, and solution 2 as the starting solution for solution 3). The stock solution of dye is 25 mM. Fill in the graph below with the appropriate calculations. Note that the first solution is the stock solution, undiluted and the last solution contains ONLY water (so the concentration =0). This last solution is called the blank. Please show your work. (You may handwrite the calculations, but you must show your work.)
Test Tube # 1 2 3 4 8 Starting Concentration 25 mM Volume of Starting Solution (mL) 8 mL 3 mL 3 mL 3 mL 0 mL Volume of water (mL) 0 mL Final Volume 8 mL 5 mL 5 mL 5 mL 5 mL Final Concentration

5 mL

SOLUTION MAKING EXERCISES A. Making a percent solution (w/w) a. Prepare 50.0 mL of a 30.0% solution of sucrose. (density of 30.0% sucrose = 1.127 g/mL). Please note: 50 g of solution does not equal 50 mL of solution! Show all calculations in your notebook. b. Determine the density of your solution. i. Weight of the beaker ___________________________ ii. Weight of the beaker + solution ___________________________ iii. Weight of the solution ___________________________ iv. Volume of the solution ___________________________ v. Temperature of your solution ___________________________ vi. Calculate the density of the solution ___________________________ vii. Show your calculations for density in your notebook. viii. The reported density for a 30% sucrose solution is 1.127 g/mL. What is the % difference between your value and the reported value? (% difference is the experimental value divided by the actual value and multiplied by 100). Show calculation in your notebook. B. Make a molar solution of a weak acid. a. Prepare 250 mL of a 0.1 M solution of 0.10 M Na2HPO4 (disodium phosphate, MM =141.96) b. Prepare 250 mL of a 0.1 M solution of 0.10 M NaH2 PO4 (monosodium phosphate, MM = 119.98) c. Prepare a phosphate buffer by mixing 8.34 mL of Na2HPO4 and 21.66 mL of NaH2PO4. d. Determine the pH of the buffer solution with the help of your TA with a pH meter i. pH of the prepared buffer solution ___________________ e. The calculated pH of this buffer should be 6.4. What is the % difference between your solution and the actual value? i. Divide experimental value by the expected value and multiply by 100. Note: Please label your solutions with the chemical, concentration, date and your initials and place in the cabinet. We will be using these buffers in a later experiment.

C. Serial dilutions from a concentrated stock solution and Beers Law a. Read attached documents explaining Beers Law. b. To create a standard curve using multiple concentrations, we will use a serial dilution protocol to create multiple concentrations from one stock solution. To do this, we will start with of our stock solution and then dilute this solution to create the next desired concentration, and then use this new solution to create the next solution and so on (so use solution 1 as the starting solution for solution 2, and solution 2 as the starting solution for solution 3). The stock solution of dye varies year to year and can be found on the bottle you are using in class. Fill in the graph below with the appropriate calculations, and use this table to determine the amount and concentration of each solution you will be making to generate your standard curve. Note that the first solution is the stock solution undiluted, and the starting concentration for each test tube is the final concentration of the solution above it. Also note, that that the last solution contains ONLY water (so the concentration=0). This last solution is called the blank. Please show your work in your notebook.
Test Tube # 1 2 3 4 5 6 7 8 9 Starting Conc Volume Starting Solution (mL) Volume of water (mL) 0 mL Final Volume 15 mL 15 mL 15 mL 15 mL 15 mL 15 mL 15 mL 6 mL 6 mL Final Conc Absorbance

15 mL 9 mL 9 mL 9 mL 9 mL 9 mL 9 mL 0 mL UNKNOWN 6 mL

c.

d. e. f.

g. h.

0 0 Calculated from graph Once you are ready, prepare the above solutions by carefully pipetting the calculated amount of water into each test tube. Assume that since these are both aqueous solutions, the volumes are additive, so you do NOT need to worry about bringing up to volume. Carefully pipette the appropriate volume of stock solution into the first test tube. Use parafilm to cover the test tube and carefully and thoroughly mix the solution until it is homogenous. Repeat c and d with each solution until all solutions are made. You will know you did a good job if you see a nice gradient of color with your solutions AND all of the volumes are exactly the same. Obtain ~5 mLs of an unknown solution from your TA. Be sure to write down the number/letter associated with your unknown! Once all of your solutions are ready, you need to read them in the spectrophotometer to determine the absorbance of each sample.

6 0

i. Please remember, you MUST use the cuvettes with an F on them, indicating that they have been calibrated and specially made such that there are no blemishes in the glass to impede the light traveling through it. ii. Transfer approximately 5 mL of your sample #8 (blank) to a clean cuvette. You can simply pour carefully or use a transfer pipette to transfer the contents from your test tube to the cuvette! iii. Set the wavelength of the machine to 520 nm. iv. With NOTHING in the spec, set the transmittance to 0 using the left dial on the front panel of the machine. v. Clean off the cuvette with a ChemWipe and place the cuvette in the spec. Close the lid. vi. Set the transmittance to 100 using the right dial on the front panel of the machine. vii. Remove the cuvette and dump out the water into a waste beaker. Carefully tap the cuvette onto a paper towel in your hand to remove as much of the water as possible. viii. Transfer ~5 mL of sample #1 into the clean cuvette. (again, you can just pour your sample in carefully or use a transfer pipette) ix. Change the settings of the machine (for digital machines) to absorbance. x. Clean off the cuvette with a ChemWipe and place the cuvette in the spec. Close the lid. xi. Record the absorbance of your sample and write it down in your notebook. xii. Dump out your sample into a waste beaker and rinse well with water. You do NOT need to wash between samples, but do rinse well between samples. xiii. Repeat viii-xi for each of the standard samples and your unknown sample. i. When you are done, empty your waste beaker into sink. Nothing you are working with is toxic, and can go down the sink. j. Once you have all of your data, you will need to generate a standard curve using the data from samples 1-7. See How to make a scientific graph in Excel 2010 at the bottom of this document for how to make this graph. k. Calculate the concentration of your unknown sample using the line of best fit from your graph. Show your calculations.

POST-LAB QUESTIONS (TYPE these answers due at the beginning of Lab) 1. When making a molar solution, why is it important to mix the solute with only part of the solvent before bringing a solution up to a given volume, rather than simply adding the whole volume of solvent initially.

2. When mixing two liquids of given volumes, why are the volumes not always additive? Explain.

How to make a scientific graph in Excel 2010:

1. Enter your data into the spreadsheet with column A as the controlled variable (what you can control) and Column B as the observed or dependent variable (what you measure).

2. Highlight the data by dragging your cursor over the entire data set. 3. Select the Insert Tab and then click Scatter (highlighted)

4.

Choose Scatter with Only Markers (again highlighted). Excel will automatically plot the data for you, but this is not a proper scientific graph.

5. Click on the border of the graph, and then the Layout tab under Chart Tools (in green). Border and appropriate tabs have been highlighted.

6. Select the Axis Titles button to add titles to each axis.

For Horizontal Axis:

For Vertical Axis:

7.

Double click on the chart where it reads Axis Title and input your desired title (here I replaced it with Absorbance (AU) for the vertical axis and Concentration (uM) for the

horizontal axis).

8. Were not done yet, you still need a title. Pick something that is short and descriptive. The button for this is right next to the axis title button (Chart title). 9. We also need to add a trendline, and make sure to show the equation for the trendline and the r value for the data on the chart. To add a trendline, right click on a data point and click Add Trendline from the context menu.

10. From the window that pops up, make sure to select the radio button for linear and check the boxes for display equation on chart and Display R-squared value on chart. This will help both you and your TA.

11. So now your chart will have the equation displayed for the line drawn, this line is called the line of best fit for your data, and should be used for all subsequent calculations with your unknowns. So can drag the equation around the chart so you can read the equation (excel often puts it in the middle of a gridline) and you can delete the legend (click on it and press the delete key), unless you need it. You can delete the legend by clicking on it and hitting delete. 12. You can resize the chart itself by clicking on a corner of the graph itself (where the grid lines are) and dragging. You can also change the size of the actual graph by clicking on an outside corner and dragging. 13. Your graph should look something like this.

Standard Curve for Dye Absorbance

0.18 0.16 0.14 0.12 0.1 0.08 0.06 0.04 0.02 0 0 1 y = 0.024x + 8E-17 R = 1 Absorbance (AU)

Concentration (uM)

14. Now you can see I have a title, axis titles, a line of best fit, and an equation of the line. 15. Congratulations! Youve finished your plot. Be sure to save the workbook and print the graph for your TA! When you print it off, you need to print off the whole screen - make sure that you click in a cell OUTSIDE of the graph otherwise you will just be printing the graph. Do check on Print Preview that you have the graph and data arranged in a way that will print everything on one page. Move the graph or change the page to landscape if necessary.