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2014 Cell 9 Presentation

Rebecca!

Kirby!

Ashley!

Ian!David!

Boshra!

Thomas!

Lauren!

Jeannelle!

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Howdy!

My name isArchibald and we

have an analysis that

we need help with. I

heard that this

Chem 315 class isreally smart. Can

you help us?

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I heard that some

VeggieTales mighthave some special

powers called

FLAVONOIDS Can

you guys teach us

about them and tellus which of us has a

lot of them?

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OkSo What are Flavonoids?

Over 6000 different compounds Secondary plant metabolites

Latin word flavus means yellow

Albert Szent-Gyorgyi in 1938 First named vitamin P

Classes of flavonoids: Quercetin Flavones

etc.

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Why Quantify

Flavonoids?

Reduce risk of cancer

Reduce cardiovascular diseases

Posses anti-oxidants Free radical scavenging activity

Horticultural approaches towardhealth promotion

Optimizing growth and storageconditions

Cooking advices to increase

flavonoid intake

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(SHHHHH. Dont tell them)

YOU HAVE TO EAT

THE VEGGIETALES

TO GET THE

FLAVONOIDS.

But there is ONE LITTLE Problem:

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Objective of Analysis

So whyshould I

volunteer forthis study?

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LiquidNitrogen is

COLD!!!!

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Fundamental Principles

Retention time, Partition coefficient,Gradient Elution, Isocratic Elution,

Chromatogram

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More Fundamentals

Hydrophobic Interactions:

Hydrophobic regions

(Need to know structure ofmolecule)

Advantages: Takes advantage of

hydrophobicity characteristicsof protein or molecule

Large volume of sample canbe loaded

High Ionic strength samplecan be loaded

sample eluted with low salt

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Fundamentals of Elution

Gradient Elution:

Salt concentration in mobile phase changesIsocratic Elution:

Salt concentration in mobile phase remains

constant

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Reverse Phase High Performance

Liquid Chromatography

non-polar stationary phase and an aqueous, polarmobile phase

Retention time is longer for non-polar molecules

Flavonoids have varying levels of polarity, so thisallows separation and identification

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What is the Measurable

Quantity?

Retention Time

To identify specific

flavonoid glycoside Compare to reference

standards

Peak Area

Determine the amountof quercetin

Detective

Larry is going

to help!

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Lets Measure It!

Standards are used Mass spectroscopy is used

to confirm identification

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Calculate Results/ Interpret Data

Finding theArea

Find the Percent

Composition

Matching the

Peaks

Take MassSpectrometry to

verify the

fractions identity.

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So what did we learn about

Bob?

He contains 5 differentflavonoids! (4-26 mg/100 g)

Chalconaringenin (35-

71%) Quercetin 3-rutinoside

Another form of

quercetin kaempferol 3-rutinoside

A form of phloretin

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Accuracy and Precision

Variation in results could be due to:

Light regimes under which the plants weregrown

UV-B irradiation can induce accumulation offlavonoids

Varietal differences in crops Conditions it was kept under during

transport from grower to supermarket

Data presented is in line with previous

studies that utilized similar tissues Accurate measurement of flavonoids in

foods is relatively cheap and not especiallytime consuming and thereby offers method

for product quality assurance

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Th k Ch f

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Thank you Chem 315 for

teaching us the concepts of

chromatography and thespecial powers of Flavonoids!

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The Flavonoids of Tomatoes, RuneSlimestad, Torgils Fossen, and Michel J.

Verheul,Journal of Agricultural and Food

Chemistry2008 56(7), 2436-2441

Quantitative Analysis of the FlavonoidContent of Commercial Tomatoes,

Onions, Lettuce, and Celery, Alan Crozier,

Michael E. J. Lean, Morag S. McDonald,

and Christine Black,J. Agric. Food Chem,1997, 45, 590-595

Sources