This document summarizes research conducted on Number Six Lake in Franklin County, Alabama. The research involved several chemical and biochemical analyses: 1. Analysis of iron concentration in the lake water through colorimetric methods. Iron was found to decrease with depth. 2. Classification and analysis of calcium content in mussel shells collected from the lake. Six mussel species were identified and the mean calcium content was found to be 6.89% with a standard deviation of 1.16%. 3. Suggestions for future research include increasing the efficiency of shell grinding, analyzing more samples to correlate calcium content with species, and continuing analysis of iron in the hematite ore found at the lake.

1. A Chemical Profile of
the First Mining Spoils
Lake in Alabama
Number Six Lake
Franklin County, AL
by
Mitchell Luckie
Chelsea Huneycutt
CHEM 385/485
Spring 2015

2. Determination of Iron in Number Six Lake Water
Number Six Lake-Franklin County, Russellville, AL

3. Research Focus
Chemical and Biochemical Analysis
○ Fe in lake water
○ Percentage Fe in ore
○ Number & kinds of
mussels in lake
○ Types of carotenoids in flora and
fauna at lake
○ DNA analysis of beaver colonies
○ Percentage Ca in mussel shells
as a function of mussel species

4. Research Focus
Chemical and Biochemical Analysis
○ Fe in lake water (Murphy et al.)
○ Percentage Fe in ore (Chem 485)
○ Number & kinds of mussels in
lake (Chem 485)
○ Percentage Ca in mussel shells
as a function of mussel species
(Chem 485)
○ Types of carotenoids in flora and
fauna at lake
○ DNA analysis of beaver colonies

5. HISTORY of NUMBER SIX LAKE
Russellville, AL
Franklin County
FACTS ABOUT LAKE:
• Mining spoils lake
• 68 acres
• Was a water reservoir for city until 1995
when Murphys bought it
• Located at first iron mine in the state
• High % Fe reported in ore
• Mine operated from 1918-1942
• Employed more than 100 people
• Furnace on site

6. “Shipment of the First Iron
Produced in Russellville", Mural
at Russellville Post Office, 1938
Project type: Art, Mural
New Deal Agencies: Treasury Section of Fine Arts
Completed: 1938
Artists: Conrad A. Albrizzio

7. ○ A Section of Fine Arts fresco entitled “Shipment of the First
Iron Produced in Russellville” was painted for the Russellville,
Alabama post office in 1938 by Conrad A. Albrizzio.
○ The mural for Russellville turned out to be one of the most
controversial in Alabama. Albrizzio submitted two sketches of
local industry shortly after he was invited to undertake the
commission. The Section office chose the scene of a local
quarry over that of an early iron mine. The Section apparently
made their decision in early July 1937 and by the end of the
month they had received numerous telegrams in protest from
Russellville clubs and business concerns.

8. The local population proposed a slightly different theme of the old Alabama Iron
Works, the first iron furnace in Alabama, built in 1817. Similar letters were also
sent from Alabama to Senator John H. Bankhead, Jr., in Washington. One of the
letters described the general theme and the details the Russellville businessmen
wished to have included: “We know the beehive shape of all charcoal furnaces
erected at the date. We know that the furnace and forge were motivated by water
power through a race that still exists. We know they used a five hundred pound
hammer to shape the pig–we have the hammer. We know the ore was collected
by slave labor and hauled in ox carts to the furnace. We know the pig iron, much
of it, was hauled by ox wagon thirty-five miles to the Tennessee River and
shipped to Liverpool, England, and we have the records where it was sold at one
hundred dollars a ton. The rock wall foundation of the warehouse still stands
along the creek bank.” After a series of letters between Albrizzio, Bankhead, the
citizens of Russellville, and the Section, Albrizzio redesigned to the wishes of
the local population and eventually went to Russellville and painted the mural in
fresco on the walls of the post office.

9. Fe Analysis of Water at
Number Six Lake
Fe Analysis Method
Sampling
Grid made of lake
Samples taken at 12 different locations around lake
Samples taken from 1-25 feet depth in lake center (M)
Analysis
SCN- added to samples, Absorbance measured at 490
nm for [Fe(SCN)]2+ determination
Determine Fe concentration from calibration curve made
with standard solutions
PREVIOUS WORK
Fe concentration
in lake as function
of depth of water
A B C D E
F G H I J
K L M N O
P Q R S T
U V W X Y

10. Colorimetric Complex Formed
Fe(III) + SCN- ↔ [Fe(SCN)]2+
In sample excess thiocyanate iron(III) thiocyanate
Deep orange-red
Absorbs at 490 nm
Fe Analysis of Lake
Water
Calibration curve for Fe concentration

11. Fe Analysis of Lake Water

12. Percentage of Calcium in Mussel Shells
as a Function of Mussel Species
RESEARCH DESIGN
• Literature search on topic
• Why calcium measurements?
• Sample collection from Number Six Lake
• Classification of mussel shells
• Physical measurements, photographs of mussel shells
• Grinding of shells/sample preparation
• Selection of method for analysis of calcium in shells
• Ca Analysis
• Data mining, statistics, error analysis
• Conclusions

13. Calcium Analysis of
Mussel Shells from
Lake Number Six
Ca Analysis in Mussels from
Number Six Lake
The Collector of Mussels
At Number Six Lake
Mink (genus Mustela), either of two species of the
weasel family (Mustelidea) native to the Northern
Hemisphere. The European mink (Mustela
lutreola) and the American mink (M. vison) are
both valued for their luxurious fur. The American
mink is one of the pillars of the fur industry and is
raised in captivity throughout the world. In the
wild, mink are small, discreet, and most often
nocturnal, and they live in close proximity to
water.

14. Alabama rivers and waterways are home to the largest
and most diverse population of freshwater mussel
species in the nation, roughly 60% of U.S. mussel fauna.
The Mobile River Basin, which drains portions of
Tennessee, Georgia, and Mississippi waterways, also
contains diverse mussel populations. However, many of
these species have been significantly depleted in the last
century due to habitat alteration (river damming,
channelization, siltation), pollution, and invasive species,
and many more are in imminent danger of extinction.
The authors offer encyclopedic entries on each of the 178
mussel species currently identified in Alabama and the
Mobile River Basin—the scientific and common names; a
morphological description as well as color photographs of
the shell appearance; analysis of the soft anatomy;
information about ecology, biology, and conservation
status; and a color distribution map.

15. IDENTIFICATION OF MUSSELS
Identification of umbo
location, shell margins, and
shell characteristics. Used
reference text to classify
mussel shells.

16. SAMPLES OF MUSSEL SHELLS COLLECTED FROM NUMBER SIX LAKE
Six (6) Different Species Represented Here
A- Elliptio nigella B-Epioblasma haysiana C-Alasmidonta mccordi
D- Elliptio mcmichaeli E-Ellipsaria lineolata F-Elliptio crassidens
A
D
F
B
C
E

17. CLASSIFICATION OF MUSSELS USED IN OUR STUDY
(includes picture and scientific name)
A- Elliptio nigella

18. This analysis will be done volumetrically by using a
characteristic reaction of carbonate compounds, namely
their reaction with acids. Calcium carbonate (limestone) is
very insoluble in pure water but will readily dissolve in acid
according to the reaction:
2HCl (aq) + CaCO3(s) ----> Ca2+ (aq) + CO2(g) + H2O + 2Cl- (aq)

19. This reaction cannot be used directly to titrate the
CaCO3 because it is very slow when the reaction is
close to the endpoint. Instead the determination is
achieved by adding an excess of acid to dissolve all
of the CaCO3 and then titrating the remaining H3O+
with NaOH solution to determine the amount of acid
which has not reacted with the calcium carbonate.
The difference between amount of the acid (HCl)
initially added and the amount left over after the
reaction is equal to the amount used by the CaCO3.
The reaction used to determine the leftover acid is
HCl (aq) + NaOH (aq) ----> H2O + Na+(aq) + Cl- (aq)

20. 1. Accurately weigh between 0.450 and 0.550 g of dried shell into each of 3
labeled 125 ml conical flasks. Be certain you record the mass of shell for
each flask in your notebook. Add several drops of ethanol to each flask.
This acts as a wetting agent and helps the HCl dissolve the CaC03.
2. Slowly pipet 10.00 ml of 1.0 M HCl solution into each flask. Swirl the flask
to wet all of the solid. Any excess HCl should be disposed of in the sink by
diluting with water. Heat the solutions in the flasks until they begin to boil
and allow to cool. Rinse the walls of the flask with water from your wash
bottle.
3. Add 3-4 drops of phenolphthalein to each flask. Using a funnel, partly fill a
clean buret with 0.100 M NaOH solution to rinse it. Empty the buret into the
sink. Fill the buret with the NaOH solution. Run some solution out to
remove all bubbles from the tip. Replenish the solution in the buret if
necessary. Read and record the initial volume to +/- 0.01 ml.
4. Titrate one sample to the first persistent pink color. When you are close to
the endpoint the color will fade slowly. Add the remaining NaOH dropwise
until the color remains for at least 30 sec. Read and record the final volume
to + 0.01 ml.
5. Repeat the titration for the other two samples.
6. Calculate the percent calcium carbonate in each sample and the mean
value. Calculate the average deviation from the mean.

21. CALCULATIONS
1. Calculate the number of moles of HCl added to each shell sample. This is
given by the expression:
moles HCl = (0.01000 l HCl)*(1.00 moles HCl/liter) = 1.00x10-2 moles HCl
2. Calculate the moles of HCl left in each sample after the reaction with CaC03.
moles HCl left = (vol. of NaOH in liters)*(concentration of NaOH in
moles/liter)*(1 mole HCl/1 mole NaOH)
3. For each sample determine the number of moles of HCl that has reacted
with CaCO3 by taking the difference between the moles of HCl added and the
moles of HCl remaining after the reaction is complete.
4. The moles of CaCO3 in each sample is calculated by: moles CaCO3 =
(moles HCl)*(1 mole CaCO3 / 2 moles HCl)
5. Calculate the percent CaCO3 in each sample by using % CaCO3 = [(moles
CaCO3) *(100.09 g CaCO3/mole CaCO3)*(100)] / grams of sample
6. Calculate the mean value and the average deviation from the mean.

22. RESULTS-Sample Calculations
1) Moles HCl added = .010 L x 0.100 M HCl
=.001 moles HCl
2) Moles NaOH delivered = ____ L x 0.1011M NaOH = moles NaOH
Sample #1: 0.000157 moles NaOH
Sample #2: 0.000415 moles NaOH
Sample #3: 0.000374 moles NaOH
3) 1 - 2 = moles HCl reacted with shell
Sample #1: 0.000843 moles
Sample #2: 0.000585 moles
Sample #3: 0.000626 moles
4) Moles HCl react x (1 mole CaCO3/2 moles HCl) x 100.015g CaCO3/1 mole CaCO3
Sample #1: 0.04216 g CaCO3
Sample #2: 0.02925 g CaCO3
Sample #3: 0.03130 g CaCO3
5) % CaCO3= (g CaCO3/ g shell) x 100
Sample #1: 8.53 %
Sample #2: 5.96 %
Sample #3: 6.19 %
Mean: 6.89 Variance Standard Deviation: 1.35
Standard Deviation: 1.16

23. d
DATA ANALYSIS
% Ca in our samples
○ Overall:
○ % Ca
○ Mean Sample #1: 8.53%
○ Mean Sample #2: 5.96%
○ Mean Sample #3: 6.19%
○ Mean % Ca of all samples(9)= 6.89%
○ Std. Dev.= 1.16
○ Variance= 1.35
Calculations done by hand.

24. CONCLUSIONS
○ Classification of
mussels found in
Number Six Lake (list
names of those found)
○ Ca concentration
(mean) of each species
of mussel found
Ca Analysis in Mussels
from Number Six Lake
We have used reference materials to
classify six species of mussels found in
Number Six Lake in Franklin County,
AL in the Tennessee River /Bear Creek
Watershed. These are:
Elliptio nigella
Epioblasma haysiana
Alasmidonta mccordi
Elliptio mcmichaeli
Ellipsaria lineolata
Elliptio crassidens
For Elliptio nigella :
○ Mean % Ca of all samples(9)=
6.89%
○ Std. Dev.= 1.16
○ Variance= 1.35
Other groups will report % Ca in
their species of shells today.
These are the first known
measurements of % Ca in
mussels found in Alabama
waterways.

25. SUGGESTIONS FOR FUTURE
RESEARCH
1. Increase grinding efficiency of shells to
make powder prior to analysis. This will
increase surface area and help make
analysis more reproducible.
2. Analyze more samples and correlate %
Ca with species-specific mussels.
3. Analyze Ca content of lake water.
4. Continue with determination of %Fe in
hematatite (iron ore) found on site at
Number Six Lake.

26. YOUR QUESTIONS???
Ca Analysis in Mussels for
Number Six Lake