1. Making Sense of Science Megan Tichy, Ph. D
Making Sense of Science Megan Tichy, Ph.D. Lecturer, Texas A&M University

2. Highlights Toxic grains Scientific and non-scientific names
Quantification
Parts per million
Gluten
Chemical structure
Processing
Distillation

3. Scientific names
Classification based on physical, behavioral or genetic diversity
Subject to revisions
Human
Pea
Wheat
Domain
Eukarya
Kingdom
Animalia
Plantae
Phylum
Chordata
Magnoliophyta
Class
Mammalia
Magnoliopsida
Lilliopsida
Order
Primates
Fabales
Cyperales
Family
Hominidae
Fabaceae
Poaceae
Genus
Homo
Pisum
Triticum
Species
H. sapiens
P. sativum
T. aestivum
More specific
Mnemonic device: Kids Prefer Cheese Over Fried Green Spinach

4. The grass family TRIBE Oryzeae - rice SUBFAMILY Festucoideae
KINGDOM – Plantae – plants
PHYLUM – Magnoliophyta – flowering plants
CLASS – Liliopsida – monocotyledons
ORDER – Cyperales – grasses/sedges
TRIBE
Oryzeae - rice
SUBFAMILY
Festucoideae
Aveneae - oats
FAMILY
Triticeae - wheat, rye, barley
Poaceae
Paniceae - millet
Panicoideae
Andropogoneae - sorghum
Tripsaceae - corn
Donald D. Kasarda, Ph.D., Research Chemist (Retired), USDA

5. Toxic grains Modern wheat varieties Wheat Triticum aestivum Durum
Barley
Hordeum
vulgare
Hard Red
Winter
Hard Red
Spring
Hard White
Wheat
Rye
Secale
cereale
Soft Red
Winter
Soft White
Wheat
Images: Home-Grown Cereals Authority (HGCA);

6. Wheat products Atta whole wheat, stone-grinded
Bulgur whole wheat, precooked, dried, cracked, and sifted
Couscous moistened semolina, rolled and shaped,
coated with finely ground wheat four
Farina endosperm milled to fine granular consistency, sifted
Seitan washed dough; water removes starch Semolina inner endosperm of wheat not ground into flour
Atta (India)
Farina
Seitan (Taiwan)
Images: Wikipedia commons

7. Ancient forms of wheat Genetics: Diploid = 14 chromosomes
Tetraploid = 28 chromosomes
Hexaploid = 42 chromosomes
More chromosomes = easier to breed varieties suitable for non-ideal soils/climates
Einkorn Triticum monococcum Diploid
Emmer (farro) Triticum dicoccum Tetraploid
Spelt (dinkel) Triticum spelta Hexaploid
Kamut Triticum turanicum Tetraploid

8. Wheat proteins Protein Soluble in Albumin WATER - neutral pH Globulin
WATER - salt solutions
Prolamin
ETHANOL % alcohol
Glutelin
NONE OF THE ABOVE
dilute acids/bases, detergents,
or reducing agents
Cause of most
wheat allergies
Gluten

9. Gluten proteins Prolamine = proline + glutamine All coded by different
genes (DNA)    
HMW
LMW
“It is highly probable that glutenin proteins are toxic. Attempts to breed wheat with satisfactory baking properties tolerated by coeliac patients will be very difficult.” Eur J Gastroenterol Hepatol May;18(5):

10. Grain proteins Grain Prolamin fraction % total protein Wheat Gliadin69
Rye
Secalin
30-50
Barley
Hordein
46-52
Oats
Avenin 16
Millet
Panicin 40
Corn
Zien 55
Rice
Orzenin 5
Sorghum
Kafirin 52
toxic
safe
still questioned
contaminated? pure?

11. Anatomy of a kernel
Manufacturers remove the germ and bran to make refined bread flour
Endosperm
Storage tissue
starch, gluten
Bran
Rich source of magnesium and iron. ¼ cup contains 6 grams fiber.
Germ
Embryo
vitamin E, magnesium, riboflavin, thiamin, phosphorus, niacin, iron and zinc. Some fat and protein.
Whole-grain: endosperm, bran, and embryo (all three parts of the kernel!) left intact

12. Wheat flour: gluten and starch
H2O
dough
starch (sugars)
75%
gluten (protein)
12% (varies)
dough
yeast
bread
C6H12O6 + 6 O2 → 6 CO2 + 6 H2O + energy
glucose + oxygen → carbon dioxide + water
Image:

13. How much gluten? Flour contains 10-15% protein
The protein is 80% gluten
Q: If 1 slice of bread uses 40 g flour
how much gluten is in it?
A: solve for protein 40 g X 0.13 = 5.2 g protein
solve for gluten g X 0.8 = 4 g gluten
Q: How many mg gluten is in 1/8 teaspoon flour?
A: Conversion for wheat flour 1 tsp = 2.5 g
1/8 tsp all purpose flour = 0.31 g flour
0.31 g X 0.13 X 0.8 = g gluten
1/8 tsp  33 mg gluten
Measuring spoon image:

14. How much is too much?
“Ingestion of contaminating gluten should be kept lower than 50 mg/day in the treatment of CD” – C. Catassi et al. AmJ Clin Nutr 2007;85:160–6. Some people are far more sensitive*
Normal gluten intake by healthy individuals = 13 g gluten per day
Compare – 50 mg (0.05 g) vs. 13 g
Normal intake is 260 times as much as the suggested threshold
Codex Alimentarius (WHO/FAO), has two limits for “gluten-free”:
0.02% gluten content - rendered gluten-free
0.002% gluten content - naturally gluten-free food
A person could ingest 3-6 mg gluten per day by consuming
6-8 ounces of “naturally gluten free” grain food
0.002% = 20 parts per million or ppm
*Reference: A milligram gluten a day keeps the mucosal recovery away: a case report,
F. Biagi, J. Campanella and S. Martucci et al., Nutr Rev 62 (2004), pp. 360–363

15. A part per what? Examples of a part per twelve One egg out of a dozen
One cupcake out of a dozen
One pane from this stained-glass window
A part per hundred (%)
per cent means one part per hundred
X 100 =
A part per million (ppm)
One bushel of wheat (60 lbs) contains 1 million individual kernels

16. A part per million (ppm)
Use % for larger fractions
1 part per = = 1%
Use ppm for smaller fractions
1 part per 1,000,000 = = %
Calculating ppm:
Start with the fraction (part/whole)
Rather than 2 decimal places to the right → %
Move 6 decimal places to the right → ppm
20 ppm = 20 minutes in:
4 days
B) 10 weeks
C) 2 years

17. Quiz answer 20 minutes 1 hour 1 day 20 = 4 days 60 minutes 24 hours
5760 =
= 3,472 ppm
20 minutes
1 hour
1 day
1 year 20 =
2 years
60 minutes
24 hours
365 days =
= 19 ppm

18. Perspective: How much is 20 ppm?
20 seconds in 11.5 days
20 inches in 15.8 miles
20 ounces in 62,500 pounds
20¢ in $10,000
Tiny amounts are biologically significant!
Most fish need  4 ppm dissolved oxygen to survive; Bass and trout seek out waters with 9-12 ppm oxygen

19. Tsp non-dairy GF creamer
Using ppm to calculate quantity gluten
ppm gluten 
2 g
Tsp non-dairy GF creamer
20 g
10 GF crackers
200 g
4 slices GF bread 20
0.04 mg
0.4 mg
4 mg 50
0.1 mg
1 mg
10 mg
100
0.2 mg
2 mg
20 mg
200
40 mg
Reminder: The ingestion of contaminating gluten should be kept lower than 50 mg/day in the treatment of CD

20. Starch vs. Protein Starches Contain carbon, oxygen, hydrogen
Broken down to only one material, glucose
Proteins
Contain carbon, oxygen, hydrogen, nitrogen, often sulfur
Broken down to 22 naturally occurring amino acids
glucose
amino
acid

21. Essential amino acids (from diet)
All amino acids have a
1-letter abbreviation
Mnemonic device:
I Have Received Much Kudos
For Learning These Very Well

22. Protein primary structure
Proteins are polymers of amino acids with directionality
C (carboxy)-terminus → N (amino)-terminus
“Beads on a string”
Stability: Proteins vary widely in their stability to heat
Proline is known to provide stability to proteins

23. Gliadin primary structure (sequence)
MKTFLILALLAIVATTATTAVRVPVPQLQPQNPSQQQPQ
EQVPLVQQQQFLGQQQPFPPQQPYPQPQPFPSQQPYLQL
QPFLQPQLPYSQPQPFRPQQPYPQPQPQYSQPQQPISQQQ
QQQQQQQQQQQQQQQQIIQQILQQQLIPCMDVVLQQHN
IVHGKSQVLQQSTYQLLQELCCQHLWQIPEQSQCQAIHN
VVHAIILHQQQKQQQQPSSQVSFQQPLQQYPLGQGSFRP
SQQNPQAQGSVQPQQLPQFEEIRNLARK
MKTFLILALLAIVATTATTAVRVPVPQLQPQNPSQQQPQ
EQVPLVQQQQFLGQQQPFPPQQPYPQPQPFPSQQPYLQL
QPFLQPQLPYSQPQPFRPQQPYPQPQPQYSQPQQPISQQQ
QQQQQQQQQQQQQQQQIIQQILQQQLIPCMDVVLQQHN
IVHGKSQVLQQSTYQLLQELCCQHLWQIPEQSQCQAIHN
VVHAIILHQQQKQQQQPSSQVSFQQPLQQYPLGQGSFRP
SQQNPQAQGSVQPQQLPQFEEIRNLARK
Q: Why does gliadin survive in the presence of enzymes and heat?
Reference: L. Shan, O. Molberg and I. Parrot et al., Structural basis for gluten intolerance in celiac sprue, Science 297 (2002), pp. 2275–2279.

24. Protein folding
Secondary structure: Regularly repeating local structures stabilized by hydrogen bonds. Ex: alpha helix, beta sheet
Tertiary structure: Overall shape or “fold”; the spatial relationship of the secondary structures relative to one another
Quaternary structure: Results from interaction of more than one protein molecule (subunit) which functions as part of the larger assembly or protein complex.

25. Protein function requires folding
amino acids
binding site
5 common protein functions:
Structure: keratin
Transport: hemoglobin
Storage: myoglobin
Hormones: insulin
Catalysis: lactase
Just a few “beads” from the string ≠ gluten

26. MSG ≠ gluten The average person consumes 10 - 20 g bound glutamate/day
g MSG/day
L versus D
In higher mammals, all proteins
contain “L” amino acids.
During hydrolysis “L” → “D”
The “D” form is tasteless

27. Other small molecules ≠ gluten
Not all symptoms are related to celiac disease
The structure of caffeine
milligrams daily appears to be safe
Starbucks’ Tall Coffee = 260 milligrams
The structure of calcium oxalate
Oxalate binds with calcium in your urinary tract
Calcium oxalate can form crystals which eventually grow
into kidney stones

28. Hydrolyzed wheat starch
Hydrolysis = hydro (water) + lysis (cleavage/breakage)
A chemical reaction in water that
breaks down wheat starch
North America – starch products usually derived from corn
Any of the following can be made from wheat starch:
Maltodextrin, glucose syrup, dextrose
Poly-ols, sugar alcohols (sorbitol, xylitol, mannitol etc…)
Lactic acid, citric acid, acetic acid (vinegar), ascorbic acid
How do we test for gluten?
ELISA – Enzyme Linked Immunosorbent Assay
ELISA detection limit between 1-2 ppm, with <0.5 ppm reported*
*Sousa et al. Am J Clin Nut, Vol. 87, No. 2, , February 2008

29. Alcoholic fermentation
Safe liquors
Wine/brandy - natural sugars present in fruits
Rum - cane sugar or molasses
Whiskey - amylase-treated grain
Vodka - amylase-treated grain or potatoes
Gin - grain infused with juniper berry
A vessel allows carbon dioxide to escape,
but prevents outside air from coming in
Without oxygen present:
C6H12O6 → 2 C2H5OH + 2 CO2
glucose → ethanol + carbon dioxide

30. Distillation: separation based on volatility
Volatility - how readily substances become gases
Stronger intermolecular forces decrease volatility
It takes more energy to break these molecules apart
Volatile
Non-volatile
Also not volatile:
large peptides (proteins)
starches, any polymer
Not solely based on size
of the molecule

31. Volatility ≠ size 3 C’s 1 O 2 C’s 1 O Increasing boiling point 4 C’s
Acetone
Molecular Weight: g/mole
Boiling Point: 56.5 C
3 C’s
1 O
Ethanol
Molecular Weight: g/mole
Boiling Point: 64.7 C
2 C’s
1 O
Increasing boiling point
Ethyl acetate
Molecular Weight: 88 g/mole
Boiling Point: 77 C
4 C’s
1 O
Butanol
Molecular Weight: g/mole
Boiling point: C
4 C’s
1 O

32. Simple distillation procedure
Ethanol gas floats up
circulates through
condensing tubes
Volatile portion
enters gas phase
Tubes are cooled
gas → liquid
Liquid heated
in boiling chamber
Peptides/amino acids
do not evaporate
The separated liquid (distillate)
drips out into a new chamber

33. Conclusions There are MANY other names for wheat
Spelt is NOT an alternative for wheat!
A ppm is not a quantity
20 ppm can be many different quantities, depends on how
much you ate!
Gluten is a highly stable protein fraction
Several different fragments of gliadin and glutenin are toxic
Small molecules do not mimic gluten, but can have negative
health effects of their own
Distillation is separation based on volatility
Distilled products are gluten-free as long as gluten was not
added post-distillation

34. THANK YOU! Brazos Valley Gluten Intolerance Group
Denise Fries, Taylor Jensen Julie Hoyle (secretary), Jen
Sara Boswell Kim Melissari (treasurer), Lauren
Monica Michalka
GIG mentors
Cynthia Kupper, RD
Betty Barfield
Texas A&M University
Dr. Ganesa Gopalakrishnan
Family
Shane (husband), Nathan (son)
Karen Jones & Family
McLean family – Rochester, NY
Santiago family – Bryan, TX