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078G-23
PHENOLIC CONTENTS AND ANTIOXIDANT ACTIVITIES OF
DIFFERENT HERBS (BLACKBERRY LEAF, ELDERBERRY
FLOWER AND DANDELION ROOT) USED IN MIX HERBAL
TEAS MARKETED IN TURKEY D. NILÜFER, S. Eruçar, D.
Boyacioglu, and E. ÇAPANOGLU Department of Food
Engineering, Istanbul Technical University, 34469
Maslak Istanbul, Turkey. e-mail
niluferd_at_itu.edu.tr boyaci_at_itu.edu.tr
capanogl_at_itu.edu.tr IFT Annual Meeting and Food
Expo 2006, June 24-28 2006, Orlando, FL.
RESULTS AND DISCUSSION
MATERIALS AND METHODS
ABSTRACT
TOTAL FLAVONOID CONTENTS
DPPH METHOD
TOTAL PHENOLIC CONTENTS
Infusion Preparation 0.2 g sample 15ml boiled
water (95?C 100?C) Incubation for 3 min
Filtration from Whatman No.4 Freeze drying at
-50?C
Extract Preparation 0.2 g sample 10ml
methanol 80 Centrifuge for 15 min
Supernatant collection (3 times) Filtration
from Whatman No.4
Different extraction solvents (ethanol, methanol
and methanolwater) were used for determination
of total phenolic contents of those plants. For
the determination of total phenolic content Folin
Ciocalteu method was used. Antioxidant activities
were evaluated using DPPH radical scavenging
activity assay, ABTS radical cation
decolorisation assay, super oxide anion
scavenging activity, reductive potential,
antioxidant activity in linoleic acid emulsion
system, hydroxyl radical scavenging and metal
chelating activity methods and RP-HPLC with PDA
detection was used for profiling the phenolic
content. Methanolic extracts gave the highest
phenolic contents for each of the herb, so
further antioxidant assays were carried out in 80
methanol. Average total phenolic contents of
blackberry leaf, elderberry flower, and dandelion
root were found to be 44.17, 30.83 and 7.92 mg
GA/g sample, respectively. Radical scavenging
activity of methanolic extracts of blackberry
leaf at 0.4 mg/ml concentration was determined to
be 93.0, whereas for elderberry flower and
dandelion root at 4 mg/ml concentration
scavenging activities (SA) were 84.43 and
14.41, respectively.
Elderberry flower
Blackberry leaf
Dandelion root
Elderberry flower
Blackberry leaf
Dandelion root
Elderberry flower
Blackberry leaf
Dandelion root

• Blackberry leaf extract was found to have the
  highest phenolic content, DPPH and ABTS radical
  scavenging activities, AOX activity in linoleic
  acid emulsion system and reducing power among
  other samples analyzed. Whereas flavonoid
  contents, metal chelating activities and hydrogen
  peroxide scavenging activities were lower than
  that of elderberry flower. This can be attributed
  to the presence of phenolic acids.
• Dandelion root had significantly lower phenolic,
  flavonoid contents and AOX activities (except for
  metal chelating and hydrogen peroxide scavenging
  activites) than other samples.
• In their infusions all samples had significant
  losses for phenolic contents and AOX activities.

ABTS METHOD
LINOLEIC ACID EMULSION METHOD

• Extracts analyzed for
• Total phenolic content (Folin Ciocalteu method)
• Total flavonoid content (TFC)
• Antioxidant activity analysis
• DPPH radical scavenging activity method,
• ABTS radical cation decolorisation assay method,
  reducing power, metal chelating activity, AOX
  activity in linoleic acid emulsion system, H2O2
  scavenging activity.
• Phenolic profiles by RP- HPLC with PDA detection.
  
• Waters 2695 Seperation module, Waters 2996 PDA
  detector Supelcosil C18, 5 µm, (25 x 4,6 mm)
  Gradient system 3 acetic acid - H2O 3 acetic
  acid, 25 CH3CN, 72 H2O, 278 nm.
• Statistical analysis Differences between the
  means of leaves and infusions for each of the
  sample were evaluated by t-test assuming equal
  variances (Plt0.05). Linear regression variance
  analysis is performed using SPSS software
  (version 14.0) for antioxidant potential
  determination methods versus total phenolic
  contents and total flavonoid contents for all
  samples (Plt0.01).

INTRODUCTION
Elderberry flower
Blackberry leaf
Dandelion root
Elderberry flower
Blackberry leaf
Dandelion root
H2O2 SCAVENGING ACTIVITY
METAL CHELATING ACTIVITY

• Blackberry leaf, elderberry flower and dandelion
  root materials are oftenly used as ingredients of
  herbal teas in the market for introducing new
  flavors and well-being effects. Herbal infusions
  contribute to the major source of phenolic
  compounds in our diet.
• Several studies have been carried out for the
  presence and activity of antioxidants in herbs
  but mostly, they are focused on organic solvent
  extracts rather than infusions.
• No single assay is available to accurately
  reflect AOX power of plants as the phenolic
  compound profile and AOX activity and mechanisms
  varies among plants.
• In this study the objective is to the phenolic
  contents and the antioxidant activity potentials
  of those plants used as herbal tea.
• Also different methods for assessing AOX activity
  were compared and effectiveness of these methods
  were evaluated.

REDUCING POWER
PHENOLIC PROFILES BY HPLC
Elderberry flower
Blackberry leaf
Dandelion root
Rutin
Elderberry flower
Elderberry flower
Blackberry leaf
Dandelion root
Elderberry flower infusion
Blackberry leaf infusion
Dandelion root infusion
Chlorogenic acid
Elderberry flower
Blackberry leaf
Dandelion root
REGRESSIONS OF METHODS
Caffeic acid
Luteolin
(-)-Epicatechin

• When the relations between the methods are
  evaluated except for hydrogen peroxide
  scavenging activity method all methods showed
  statistically significant regressions with total
  phenolic contents.
• For regressions with total flavonoid contents
  only insignificant differences were similarly for
  hydrogen peroxide scavenging activity method and
  metal chelating activity.
• Highest regressions were found for DPPH, ABTS
  radical scavenging and ABTS Trolox Equivalents
  versus total phenolic contents.

Elderberry flower Chlorogenic acid, caffeic
acid, (-)- epicatechin, rutin and luteolin are
identified. In infusions 50 decrease in rutin,
38 decrease in chlorogenic acid, 44 decrease
in (-)-epicatechin were determined. Dandelion
root Gallic acid, chlorogenic acid, coumaric
acid and rutin are identified. In infusions
except for gallic acid all other components were
determined. Generally there was decreasing trend
for all phenolic compounds of infusions except
for rutin which had an increasing trend (50
increase). Blackberry leaf Chlorogenic acid,
caffeic acid, coumaric acid, ferulic acid, gallic
acid and rutin phenolics were determined. In
infusions losses were 88 for coumaric acid and
76 for chlorogenic acid.

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R E F E R E N C E S
Dandellion root
Rutin
Coumaric acid
Gallic acid
Chlorogenic acid
CONCLUSIONS

• When evaluating antioxidant activity single
  method data is not sufficient. All data from
  various methods should be evaluated to understand
  the action of each mechanism.
• Infusion process reduce phenolic contents and
  antioxidant activities to some extent but do not
  degrade all phenolic content and antioxidant
  activity.
• These results can also provide useful data for
  future studies for optimization of the methods
  for preparing herbal tea infusions.
• Blackberry leaf, elderberry flower and dandelion
  root with their phenolic contents and antioxidant
  activities are promising natural nutraceutical
  ingredients of herbal tea preparations.

Coumaric acid
Blackberryleaf
Chlorogenic acid
Ferulic acid
Rutin
Gallic acid
Caffeic acid
ACKNOWLEDGEMENT The authors gratefully
acknowledge the financial support from the EU
6th Framework STREP Project FLORA (Flavonoids And
Related Phenolics For Healthy Living Using Orally
Recommended Antioxidants).