Ethanol Lab Preparation
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Arizona State University *
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367
Subject
Chemistry
Date
Dec 6, 2023
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Lab 8 Preparation
Fill in the sample table for the standard curve samples. The unknown ethanol samples and
controls are also given in this document. We will also go over the unknown alcohol samples and
the control samples (other than no ethanol) in class before starting the experiment.
You are bring your only samples of this lab. Please follow the rules for samples provided in the
lab.
Write down AND taking a picture of the alcohol content (in %) of the container.
Notes on Standard curve samples:
Calculation all sample to have a final volume (V
2
) of 250
l
Well concentration of buffer is consistent thus volume added is consistent
o
0.15 M or 150 mM stock concentration (of sodium pyrophosphate in buffer
stock, there are more components but it is the sodium pyrophosphate that we
are paying attention to)
o
Desired sample concentration is 75 mM
Well concentration of NAD+ is consistent thus volume added is consistent
o
Stock concentration: 3 mM
o
Desired well concentration: 1 mM
Amount of enzyme is consistent thus volume added is consistent
o
Stock concentration: 5000 units/ml
o
Desired concentration: 25-50 units/ml
Choose an enzyme concentration in this range
The well concentration of ethanol is DIFFERENT thus the volume of each sample will be
different
o
Stock concentration of ethanol: 2 mM or 2000
M
o
9 concentrations in the range of 0 to 200
M is desired (see table below)
You make adjust the concentrations in your standard curve as long as 200
M remains the top concentration used.
Water to make up the total volume once all other components are accounted for.
Sample table:
Well
concentration
of Ethanol
(
M)
Volume of
buffer (0.15
M)
Volume of
NAD+ (3
mM)
Volume of
Enzyme
(5000
units/ml)
Volume of
Ethanol
Standard (2
mM)
Volume of
water
Total
volume (
l)
200
250
175
250
125
250
100
250
75
250
50
250
10
250
0
250
Remember that 200
M is 0.2 mM
If any ethanol volume is less than 2
l plan to make a dilution (10x, 25x, or 50x) of the stock
solution to raise the volume in the well above 2
l
Background samples:
The 0
μ
M ethanol concentration can be used for the background for the standard curve
since the known ethanol solution is diluted in water.
For the unknown samples background, I suggest replacing the enzyme volume with
water and keeping all other components of you. This way there will be no reaction that
occurs in your sample. Allowing you to consider the buffer, NAD+, any absorbance from
your unknown ethanol sample (that isn’t the NADH product of the reaction), and any
absorbance that could come from the microplate.
o
Note: this background would not consider any absorbance of the enzyme.
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PART 3- LABORATORY EQUIPMENT
EQUIPMENT NAME
FUNCTION
To hold test tubes.
Attaches to a retort stand to support glassware being heated.
To pour liquid from a large container into a small container.
To provide a flame during an experiment.
ones
To light the Bunsen burner.
To attach test tubes to a retort stand.
To clean substances out of a test tube.
To hold a hot crucible or flask.
To hold a hot test tube.
To place on the lab bench to prevent burning or to use with ring clamp.
To transfer small amounts of liquid.
To grow bacteria or contain small experiments.
To hold liquids.
To measure liquids.
To contain experiments (a stopper can be used to close the flask).
To hold a hot beaker.
To perform an experiment with a different substance in each well.
To hold chemicals and perform experiments in.
To scoop and transfer small amounts of substances,
To stir during experiments that involve mixing or dissolving.
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1
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7-10 subparts
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In this treatment level, part of suspended solids and suspended organic
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Secondary
Preliminary
Tertiary
O None of the above
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Multiple Choice
Each of the numbered items or incomplete statements is followed by answers or by completions of the statement.
Select the one lettered answer or completion that is best in each case. Write your answer before each number.
42. Primary Standard reagent used in standardization of 0.1N sodium hydroxide
a.
Sulfamic acid
C. sodium carbonate
KHP
b. sodium bicarbonate
d.
43.An analyst who determines the strength, potency and percentage purity of a drug or pharmaceutical product
is performing:
a. Special method
C.
d.
Pharmaceutical assaying
Identification test
b. Pharmacopoeial test
44.Chemical analysis which determines the amount or percentage content of a certain component in a sample:
a. Qualitative
C.
d.
Proximate
Ultimate
Quantitative
ܩ
b.
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Procedure
1. To 5 ml of permanent hard water, add 2 ml of sodium bicarbonate, NaHCO3, solution.
Filter. To the filtrate, add soap solution. Write the reaction involved.
2. To 5 ml of permanent hard water, add 1 ml of soap solution. Compare the results
obtained from procedures 1 and 2. Compare the amount of bubbles and precipitate
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3. Boil 5 ml of temporary hard water and add 1 mL soap solution.
4. To 5 ml of temporary hard water, add 1 ml soap solution. Compare the results obtained
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Question
1. What is the precipitate formed from the reaction of permanent hard water with
sodium carbonate? Write the balanced chemical reaction involved.
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Which of the following is a valid conclusion to draw at this point?
Select one:
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Pb2+ is definitely present.
Ag+ could be present, or Hg22+ could be present, or BOTH could be present.
Ag+ is definitely present
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Which of the following is a valid conclusion to draw at this point?
Pb2+ is definitely present.
Hg22+ is definitely present.
Ag+ could be present, or Hg22+ could be present, or BOTH could be present.
Ag+ is definitely present
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4. Drag the test tube containing the test strip over to the color chart to determine the pH
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PART ONE: Preparation of FECNS* Solution
Prepare the two solutions in table 1 by accurately measuring the required volumes of distilled
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soon after adding the Fe" solution from a buret.
TABLE 1
Fe
5 ml
5S mL
Solution
Distilled Water
CNS
2 mL
Total Volume
3 ml
2 ml
10 ml
3 mL
10 ml
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PRELIMINARY Exercise, formulate a hypothesis as to what would happen under each of the
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1. 1ml of 0.0020 M CNS' solution is added to the unused portion of
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2. One drop of 0.20 M CNS solution is added to the unused portion of
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