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Determination of The Punicalagins Found in Pomegranate by High Performance Liquid Chromatography

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47 views2 pages

Determination of The Punicalagins Found in Pomegranate by High Performance Liquid Chromatography

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mazahir raza
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© © All Rights Reserved
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Determination of the Punicalagins Found in Pomegranate

by High Performance Liquid Chromatography CAN


106
Steve Baugh, Jessica Revell, and Kim Eastman
Chromadex, Irvine, CA 92618

Introduction Solution Preparation


Antioxidants found in fruits and vegetables or the juices of fruits Mobile Phase: 1 % Formic Acid in Milli-Q water
and vegetables are known to provide our bodies with many health The solution was prepared by adding 10 mL of Formic Acid to 1 L of
benefits. Pomegranate is one such fruit that contains high levels of Milli-Q water and mixing well.
antioxidants; studies have shown that pomegranate contains more
antioxidants than green tea, cranberries, and even red wine. The Stock Standard Preparation
antioxidants in pomegranates include polyphenols, such as tannins The stock standard was prepared by weighing 5.42 mg of
and anthocyanins. The most abundant polyphenols in pomegranate punicalagins A & B into a 10 mL volumetric flask. 5 mL of Milli-Q
juice are the hydrolyzable tannins called punicalagins that have free- water were added and sonicated. The solution was then allowed
radical scavenging properties in laboratory experiments. to re-equilibrate to ambient temperature, brought to volume with
Milli-Q water, and mixed well.
For more details on pomegranate antioxidant properties click the
following link: Instrument Parameters
http://pomegranateinformation.com/pomegranate/antioxidant- Instrument: Dionex HPLC System
properties/ Detection: UV-vis
Mobile Phase A: 1% Formic Acid in Milli-Q Water
Summary
Mobile Phase B: Acetonitrile
Samples
Lot # CDXA # Gradient Program:
Punicalagins A & B Chromadex 16983-1218 CDXA-119.113 Time (min) %A %B
Results 0 95 5
See chromatograms in Data section: 18 85 15
20 35 65
Analytical Method 25 95 5
Laboratory Supplies 30 95 5
Analytical Balance
Ultrasonication Bath Column: Dionex Acclaim® Polar Advantage PA II
Assorted Volumetric glassware 150 × 3.0 mm, 3 µm particle
Syringes and Syringe Filters Flow Rate: 1.0 mL/min
HPLC/GC glass vials and caps Temperature: 30 °C
Injection Volume: 10 µL
Solvents and Reagents
UV Detection: 260 nm
Formic Acid
Milli-Q® Water
Acetonitrile (CH3CN)

Passion. Power. Productivity. This is a Customer submitted application note published as is. No ISO data available for included figures
Data Structure
Figures
OH OH
OH
2 OH O O OH
Peaks: 1. Punicalagin B OH
2. Punicalagin A
OH

O OH

O O
OH O
O
mAU 1 O OH
O O

O OH
O

O OH
0 5 10 15 20 25 30 OH
Minutes OH OH

OH
Figure 1. Punicalagins mixed standard injection.
Figure 2. Punicalagin.

References
1. ChromaDex Analytics Laboratory Notebook 73, pages 114–115.
2. ChromaDex Analytics Laboratory Notebook 119, page 113.
3. ChromaDex SOP “Routine Laboratory Calculations.”
4. Analytical Method: 99.1-CD-1.0-000216.

Milli-Q is a registered trademark of Millipore.


Acclaim is a registered trademark of Dionex Corporation.

Passion. Power. Productivity.


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