Phenolics in
Echinacea
HPLC
Pinnacle II™C18
Hyperforin inSt. John’sWort
HPLC
Pinnacle II™C18
FOODS, FLAVORS,
& FRAGRANCES
27
Dietary Supplements
Phone: 800-356-1688 or 814-353-1300
DietarySupplements
Over the past decade, thenutraceutical industry
has seen rapid growth asmorepeople add flow-
ers, leaves, roots, and fruits of botanicals to their
diets inhope of gaininghealthbenefits.
Herbal products are very complex, often con-
taining hundreds of compounds, and it is not
always clear which compounds are responsible
for the beneficial properties. Marker com-
pounds—phytochemicals that havebeen identi-
fied and areknown tohave some relationship to
the reported health benefit—can be evaluated
qualitatively to identify a rawmaterial or to ver-
ify purity. To determine the concentration or
strength of a material, quantitative analysis is
necessary.
Hyperforin inSt. John’sWort
According to the Institute for Nutraceutical
Advancement (INA), INAMethod112.001—the
HPLC analysis of hyperforin and adhyperforin
in St. John’s wort—the samples are extracted
with methanol in an ultrasonic bath.
Chromatographic separation is performed on a
C18 reversed phase column with acetonitrile
and phosphate buffer as the mobile phase.
Analysis of encapsulated St. John’s wort using a
Pinnacle II™C18
column shows excellent peak
shapes for the active ingredients in this herbal
product.
Phenolics in
Echinacea
Theactivecompounds in
echinacea
are thought to
be caffeic acid derivatives such as caftaric acid,
cichoricacid, chlorogenicacid,andechinacoside.
Conclusion
Gas and liquid chromatography are powerful
tools for the analysis of foods, flavors, and fra-
grances. This technical guide describes columns
and analytical conditions that will help you
achievemoreaccurate identificationandquanti-
tation of target analytes. However, if you have
questions, please don’t hesitate to contact
Restek’s
technical
service
by
e-mail
(
) or by phone (exten-
sion 4), or contact your Restek representative.
Wewill do everythingwe can tohelp you find a
solution.
Peak List:
1. hyperforin
2. adhyperforin
Sample:
St. John’swort capsule
Inj.:
20µL
Conc.:
4.17mg/mL
Solvent:
methanol
Autosampler:
15°C
Column:
Pinnacle II
™
C18
Cat.#:
9214565
Dimensions:
150 x 4.6mm
Particle size:
5µm
Pore size:
110Å
Conditions:
Mobile phase:
0.01N phosphate buffer, pH 2.5:
acetonitrile (15:85, v/v)
Flow:
1.2mL/min.
Temp.:
30°C
Det.:
UV@270nm
LC_0175
LC_0195
Column:
Pinnacle
II
™
C18
Cat.#:
9214565
Dimensions: 150 x 4.6mm
Particle size: 5µm
Pore size: 110Å
Conditions:
Mobile phase: A= 0.1% phosphoric acid
B= acetonitrile
Time
%B
0
10
13
22
14
40
14.5
40
15
10
20
10
Flow:
1.5mL/min.
Temp.:
35°C
Det.:
UV@330nm
Peak List:
1. caftaric acid
2. chlorogenic acid
3. echinacoside
4. cichoric acid
Sample:
Echinacea
capsule
Inj.:
10µL
Conc.:
6.7mg/mL capsule
contents in sample diluent
Solvent:
ethanol:water (70:30, v/v)
Sample temp.: 25°C
References
1. Fennema, O. R. FoodChemistry (1996), 3rd edition.
2. Bensinger, M. “HowHot is that ‘Devil’ Sauce?” in Fiery Foods
Magazine (1997), Sept/Oct.
3. Brandt, Laura. “TheCreation andUse of Vanilla”, Food
Product Design (1996), editorial archives.
4. AOACOfficial Methods of Analysis (2000), 17th edition, AOAC
International.
5. Official Methods andRecommendedPractices (1998), 5th edi-
tion, AmericanOil Chemists’ Society.
6. AACCApprovedMethods (2000), 10th edition, American
Association of Cereal Chemists.
References are not available fromRestek.
for
more
info
Request ApplicationsNote AnalyzingNutraceutical Products by Liquid andGas Chromatography
(cat.# 59364).
