COMPARING THE CALCIUM

CONTENT OF VARIOUS TYPES

OF MILK
By: Rebecca Allen
PROBLEM
 Purpose: To determine how raw goat’s milk, pasteurized
goat’s milk, raw cow’s milk, UHT 2% cow’s milk, 2%
organic cow’s milk, 2% cow’s milk, whole cow’s milk,
and skim cow’s milk compare in terms of calcium
content by means of an EDTA titration.
 Importance: Calcium is an essential mineral for a healthy
body, and it is important for consumers to know which of
the wide variety of types of milk on the market provides
the greatest calcium content.
BACKGROUND INFORMATION
 Multiple studies have shown a greater content of calcium
in raw milks as compared to pasteurized milk varieties.
Also, studies have shown a greater content of calcium in
goat milk varieties as compared to cow’s milk varieties.
 Important Vocabulary:
 Pasteurization
 UHT
 EDTA Titration
 OrganicMilk
 Raw Milk
BACKGROUND INFORMATION
 Calcium is an essential mineral for…
 Healthy bones and teeth
 Regulation of metabolic processes
 Lowering risk of colon cancer, heart disease, osteoporosis,
and obesity
 Prevention of preeclampsia, rickets, and strokes
 Regulation of blood pressure
 Improved premenstrual moods
 Proper blood clotting
HYPOTHESIS
 If raw goat’s milk, pasteurized goat’s milk, raw cow’s
milk, UHT 2% cow’s milk, 2% organic cow’s milk, 2%
cow’s milk, whole cow’s milk, and skim cow’s milk are
tested for calcium content, then raw goat’s milk will
contain the highest quantity of calcium.
CONTROLS AND EXPERIMENTAL
VARIABLES
 Independent Variable: Type of Milk
 Dependent Variable: Calcium Content
 Controls:
 EDTA titration process used for all milk samples.
 Milk products were tested within the expiration date.
 Milk products were of the same brand when possible.
 Each milk variety had its own set of labeled beakers and pipettes to
eliminate contamination.
 Each trial was compared to the color of the control sample respectively.
 Extraneous Variables:
 Different brands of milk
 Different shelf life lengths
 Added vitamins (can alter % digestion of calcium)
 Retrieved from sources at different times
 Different dairy farm locations and animal diets
MATERIALS
 800 mL NH₃ + NH₄Cl Buffer
 1600 mL H₂O

 Eriochrome Black T Solution

 15 mL of each milk variety

 EDTA Solution

 48 Labeled Beakers

 8 Pipettes and rubber bulb

 2 Burettes and 2 Burette Clamps

 Magnetic Stirring Objects

PROCEDURE
 Label each of the 48 beakers with its appropriate title.
 Prepare necessary solutions (Eriochrome Black T, EDTA, and
Buffer to maintain pH > 7.0).
 Rinse burettes with EDTA solution to remove prior residue.
 Assemble burettes and magnetic stirrers.
 Pipette 3 mL of milk into the corresponding labeled beaker (X Milk
– Trial 1).
 Add 40 mL of H₂O.
 Add 20 mL of Buffer.
 Add 10 drops Eriochrome Black T solution.
 Magnetically stir.
 Test pH – Color change only occurs at pH > 7.0
 Titrate the mixture with the EDTA solution in the burette until a
distinct color change from purple/magenta to blue.
 Repeat this process for 5 trials of each of the 8 varieties of milk.
CALCULATIONS
 EDTA⁻⁴ + Ca2+  CaEDTA⁻2
 1:1 mole ratio of EDTA : Ca

 Molarity of EDTA = 0.01 mol/L

 Atomic Weight of Ca = 40.08 g

 Sample Calculation:
Type of Milk Trial Number Initial EDTA Burette Reading Final EDTA Burette Reading Volume EDTA (mL)
(mL) (mL)

Raw Goat 1 0.45 12.55 12.10

2 13.81 28.20 14.39
3 31.70 41.47 9.77
4 16.51 28.23 11.72
5 28.23 40.13 11.90
Pasteurized Goat 1 8.16 16.35 8.19
2 19.98 37.74 17.76
3 1.30 14.70 13.40
4 14.70 27.79 13.09
5 27.79 40.64 12.85
Raw Cow 1 17.60 27.97 10.37
2 18.40 31.52 13.12
3 27.79 39.51 11.54
4 39.51 50.00 10.49
5 31.52 40.90 9.39
UHT 2% Cow 1 12.61 23.60 10.99
2 0.92 13.40 12.48
3 23.60 38.79 15.14
4 13.40 26.91 13.51
5 26.91 40.94 14.03
2% Cow 1 3.52 18.01 14.49
2 0.83 16.40 15.57
3 18.01 32.10 14.09
4 30.35 44.40 14.05
5 17.00 31.69 14.69
Whole Cow 1 1.00 16.05 15.05
2 18.35 33.90 15.55
3 16.05 28.94 12.89
4 33.90 46.27 12.37
5 28.94 42.60 13.66
Skim Cow 1 2.99 15.00 12.01
2 2.75 14.50 11.75
3 15.00 25.70 10.70
4 14.45 25.65 11.15
5 25.65 36.11 10.46
Organic Cow 1 25.50 40.38 14.88
2 36.11 49.89 13.78
3 5.97 19.78 13.81
4 5.48 20.70 15.22
5 7.18 20.30 13.12
 Type of Milk Average Amount of Standard Deviation
Calcium (mg)
Raw Goat’s Milk 8.385 1.64169

Pasteurized Goat’s Milk 7.455 3.38997

Raw Cow’s Milk 4.401 1.41717

UHT 2% Cow’s Milk 5.303 1.57691

2% Cow’s Milk 5.844 0.6167

Whole Cow’s Milk 5.571 1.36597

Skim Cow’s Milk 4.493 0.6629

Organic Cow’s Milk 5.675 0.86465

STATISTICAL ANALYSIS
 One Way Analysis of Variance (ANOVA) –
Compares the means of each group

 Fisher F-Value was calculated for the data: 21.453

 Fisher F-Value corresponds to P-Value of significance.
 P < 0.0000001 = Less than a million to one chance that
results were not significant.
 In summation, each of the 8 groups were significantly
different from each other, in terms of calcium content.
ANALYSIS
 Possible explanations for the results:
 Pasteurization process and UHT processing.
 Partial removal / full removal of fat content in 2% cow’s milk
and skim cow’s milk
 Calcium content diminishes with freshness
 Feed quality/characteristics
 Lactation cycle stage
 Seasonal changes in calcium content
 Daily fluctuation
ANALYSIS
 Possible errors:
 Milk products were not purchased directly from the
manufacturer in some cases, so superior freshness could not
be insured.
 Milk products were purchased from different sources.
 One cannot be sure of the amount of variability allowed on
either side of the “best by” date, despite all milk products
being well within this estimate.
 One cannot be sure of the lactation cycle stage of the goats
from which the goat milk products were produced.
 One cannot be sure of the specific weather conditions of each
farm from which milk products were produced at the time of
milking.
CONCLUSION
 The milk variety shown to have the highest calcium
content: raw goat’s milk (8.385 mg Ca)
 The milk variety shown to have the lowest calcium
content: raw cow’s milk (4.401 mg Ca)
 The hypothesis is accepted because the data showed raw
goat’s milk to have the highest calcium content of all
varieties of milk tested.
 In conclusion, the consumer may be better off
purchasing raw goat’s milk as compared to other
varieties of milk from a nutritional standpoint.
RECOMMENDATIONS
 Purchase milk varieties from the same manufacturer to
rule out variables dealing with weather conditions and
location.
 Purchase directly from the manufacturer to insure
superior freshness.
 Test milk varieties for calcium content immediately after
production and processing to insure superior freshness.
 Determine lactation cycle stage of goats used for milk
production prior to milking.
EXTENSIONS
 Comparing calcium content of alternative milks, such as
soy, almond, and rice milk.
 Exploring the effects of farm location and climate on
calcium content of milk.
 Exploring the effects of diet on cow and goat milk
varieties.
 Comparing the calcium content of various type of
cheeses, yogurts, or other dairy products.
 Investigating the impact of age and lactation cycle stage
of animals on calcium content of milk.
 Determining which variety milk has the most humanly
digestible calcium content.
RESOURCES
Reykdal, Olafur, and Ken Lee. “Soluble, Dialyzable, and Ionic Calcium in Raw and Processed
Skim Milk, Whole Milk, and Spinach.” Journal of Food Science 56.3 (2006) : 864-866.
Web. 29 Oct. 2009.

Pettifor, John M. “Nutritional Rickets: Deficiency of Vitamin D, Calcium, or Both?” The

American Journal of Clinical Nutrition 80.6 (2004) : n.p. Web. 29 Oct. 2009.

Ott, Christopher. “The Surprising Benefits of Calcium.” Natural Health Jan.- Feb. (2002) : 55.
Web. 28 Oct. 2009.

Krebs, Nancy F. “Optimizing Bone Health and Calcium Intakes of Infants, Children, and
Adolescents.” Pediatrics (2006) : n.p. Web. 29 Oct. 2009.

“Building Strong Bones: Calcium Information for Health Care Providers.” U.S. Department Of
Health and Human Services – National Institute of Child Health and Human Development
(2006) : 1-4. Web. 28 Oct. 2009.

Baron, JA, M. Beach, J.S. Mandel, et al. “Calcium Supplements for the Prevention of
Colorectal Adenomas.” New England Journal of Medicine 340.2 (1999) : 101-107. Web. 27
Oct. 2009.
RESOURCES
Bucher, HC, G.H. Guyatt, R.J. Cook, et al. “Effect of Calcium Supplementation on Pre-
Induced Hypertension and Preeclampsia.” Journal of the American Medical Association
275 (1996) : 1113-1117. Web. 29 Oct. 2009

“Calcium 101: How It Does a Body Good.” The Cleveland Clinic Foundation. 2008. Web. 28
Oct. 2009.

University of Granada. "Goats' Milk Is More Beneficial To Health Than Cows' Milk, Study
Suggests." Science Daily 31 July 2007. 31 January 2010 <http://www.sciencedaily.com­
/releases/2007/07/070730100229.htm>.

"Standard Deviation Calculator." Easy Calculation.com. HIOX India, n.d. Web. 21 Feb. 2010.
<http//:easycalculation.com/statistics/standard-deviation.php>.

Tóth, Á., M. Borbély, and Z. Gyori. "Examination of protein and calcium content of consumer
milks and breakfast drinks." CABI Abstract Database. Tejgazdaság, n.d. Web. 21 Feb.
2010. <http://www.cababstractsplus.org/abstracts/Abstract.aspx?AcNo=2005301745>.

"Raw Milk Vs. Pasteurized Milk." A Campaign for Real (Raw) Milk. Farm-to-Consumer Legal
Defense Fund, n.d. Web. 21 Feb. 2010. <http://www.realmilk.com/rawvpasteur.html>.
RESOURCES
Gillis, Erin . "The Effect of Heat Treatment on the Nutritional Value of Milk." Faculty Articles. California
State University - Los Angeles, 1 Sept. 2005. Web. 19 Feb. 2010.
<www.calstatela.edu/faculty/hsingh2/articles/milk.research.pdf>.

"Goat Dairy Foods." Dairy Research & Information Center. UC Davis - DRINC Program, n.d. Web. 26 Feb.
2010. <http://drinc.ucdavis.edu/goat1.htm>.

Kluiber, RW. "EDTA Titration for Ca+2 in milk.." Milk: Calcium and Nutrition. Rutgers University, 18 Nov.
1998. Web. 20 Feb. 2010. <http://genchem.rutgers.edu/Milk.html>.

Nolte, Molly. "Info on Raw Goat Milk & Pasteurization." Fias Co Farm. N.p., 18 Jan. 2010. Web. 26 Feb.
2010. <http://fiascofarm.com/dairy/rawmilk.htm>.

Shelton, Herbert. "Destructive Effects Of Pasteurization."ChestofBooks.com. N.p., n.d. Web. 20 Feb. 2010.
<http://chestofbooks.com/health/natural-cure/The-Hygienic-System-Orthotrophy/Destructive-Effects-Of-
Pasteurization.html>.

"The Michigan Fresh Unprocessed Whole Milk Workgroup: Benefits and Values." Michigan Food & Farming
Systems - MIFFS. N.p., 23 Oct. 2009. Web. 26 Feb. 2010.
<http://www.miffs.org/MIfuwmilk/benefitsvalues.htm#6>.

"Varieties of Milk." The Dairy Council. N.p., n.d. Web. 20 Feb. 2010. <http://www.milk.co.uk/page.aspx?
intPageID=43>.Top of Form
ACKNOWLEDGEMENTS
 Special thanks to Mrs. Joan Bechtel for all of her
assistance in the lab during this experiment. It would not
have been possible without her expertise and guidance.
 Thank you to my parents for their continued support of
everything I endeavor to achieve.
 This project would not have been possible without the
help of PJAS and CASEF sponsors Mr. Jason Sibbach
and Mr. Jason Ambler.
 Thank you to all the professionals who make PJAS and
CASEF possible for students to learn and compete in
these competitions.