Journal of Liquid Chromatography &

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Volume 30, Issue 15, 2007 Translator disclaimer
Special Issue: Thin Layer
Chromatography

Original Articles
Separation of Selected Flavonoids by use of RPHPLC/NP
HPTLC Coupled Methods
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DOI:
10.1080/10826070701451621
Mirosaw A. Hawrya, Monika WaksmundzkaHajnosa* & Janusz Makara
pages 2253-2265

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Received: 21 Dec 2006
Accepted: 26 Jan 2007
Published online: 28 Jun 2007
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Abstract

Phenolic compounds, such as phenolic acids, flavonoids, flavonoid glycosides, and resveratrol, were
chromatographed on thinlayers of silica in various eluent systems. Systems with the highest
selectivity were chosen. Also, reversedphase HPLC systems were optimized for the separation of
investigated compounds and most the selective system was applied for the separation of two phenolic
extracts Polygonum avicularis andPolygonum hydropiper. Partly separated fractions were
evaporated and dissolved in methanol. Fractions with partly separated phenolic compounds were
spotted onto the silica layer and developed in an appropriate eluent system. After drying, the plate
 was derivatised with the Naturstoff reagent and videoscanned. Several phenolic compounds were
identified, according to their retention coefficients in both systems

Journal of Liquid Chromatography &

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Volume 35, Issue 10, 2012 Translator disclaimer
Special Issue: Thin Layer
Chromatography

Original Articles
COMPARISON OF TLC AND HPLC FINGERPRINTS OF
PHENOLIC ACIDS AND FLAVONOIDS FRACTIONS DERIVED
FROM SELECTED SAGE (SALVIA) SPECIES
DOI:
10.1080/10826076.2012.676463
Mieczyslaw Sajewicza, Dorota Staszeka, Monika Waksmundzka-Hajnosb & Teresa Kowalskaa*
pages 1388-1403

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Accepted author version posted online: 03 May 2012
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Abstract

This study is part of a larger project on the contents and composition of essential oils, phenolic acids,
and flavonoids originating from over twenty species belonging to the Salvia genus. These herbs are
recognized in the traditional European pharmacy and cuisine for their curative and aromatic
properties, and yet they have not attracted sufficient attention from the side of phytochemists. Thus, it
seemed inevitable to focus on the contents of the phenolics in the individual sage species, basically
due to the recognized antioxidant properties of this class of compounds. In this paper, we compared
the results of a spectrophotometric assessment of the overall contents of phenolic acids and
 flavonoids (selectively extracted from twenty three different Salvia species) with those of a more
detailed analysis carried out with use of HPLC/DAD and the TLC-based video imaging. Six species
with the highest contents of phenolic acids and five species with the highest contents of flavonoids,
and additionally Salvia officinalis, were chosen for the chromatographic fingerprinting of the selectively
extracted fractions of the free and bonded phenolic acids, and the flavonoid aglycons and glycosides.
As Salvia officinalis is the only representative of the discussed genus recognized as a medicinal plant
by Polish Pharmacopoeia, our goal was to compare its phenolic contents with those of the other
phenolic-rich sage species. This part of our sage investigation project has methodological importance
and shows how thin-layer chromatographic video imaging can be used in a phytochemical study in
combination with other analytical techniques.

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Volume 34, Issue 10-11, 2011 Translator disclaimer
Special Issue: THIN LAYER
CHROMATOGRAPHY

Original Articles
TLC-MS VERSUS TLC-LC-MS FINGERPRINTS OF HERBAL
DOI:
10.1080/10826076.2011.571131
Mieczysaw Sajewicza,Dorota Staszeka, Maja Natiab, ukasz Wojtala, Monika Waksmundzka-Hajnosc & Teresa
Kowalskaa*
pages 864-887

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Published online: 17 May 2011

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Abstract

In the previous paper from this series, we proposed mass spectrometric fingerprinting of a complex
and volatile botanical sample upon an example of the essential oil derived fromSalvia lavandulifolia. In
that paper, we compared two variants of fractionation of such a mixture. A simpler one-dimensional
variant consisted of the low-temperature thin-layer chromatographic fractionation coupled with mass
spectrometric fingerprinting of each separated fraction (1D LT TLC-MS). A more sophisticated variant
was the two-dimensional liquid chromatographic system composed of the low-temperature thin-layer
chromatography, high-performance liquid chromatography, and mass spectrometric detection (2D LT
TLC-LC-MS). In this study, we present an analogous approach to the non-volatile botanical mixtures
upon an example of the pharmacologically important phenolic acids and flavonoids selectively
extracted from Salvia lavandulifolia. With these non-volatile fractions, the thin-layer chromatographic
separations were carried out at ambient temperature (21 0.5C). Once again, we compared two
variants of fractionation. A simpler one-dimensional variant consisted of the thin-layer
chromatographic mode coupled with mass spectrometric fingerprinting of each separated fraction (1D
TLC-MS). A more sophisticated variant was the two-dimensional liquid chromatographic system
composed of the thin-layer chromatography and high-performance liquid chromatography, with mass
spectrometric detection (2D TLC-LC-MS). As expected, the two-dimensional mode proved better
performing than the one-dimensional mode (1D TLC-MS). It was concluded that thin-layer
chromatography directly or indirectly coupled with mass spectrometric detection can prove very useful
in the analysis of the phenolic acid and flavonoid fraction selectively extracted from botanical material.

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Volume 37, Issue 11, 2014

Original Articles

STUDY OF EFFECT OF EXTRACTION CONDITIONS ON THE

BIOCHEMICAL COMPOSITION AND ANTIOXIDANT ACTIVITY
OF ARTEMISIA ABSINTHIUM BY HPLC AND TLC
DOI:

10.1080/10826076.2013.803200
 H. Ghafooria*, R. Sariria &M. R. Naghavib

pages 1558-1567

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Accepted author version posted online: 28 Aug 2013

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The purpose of the present study was to investigate the effect of extraction temperature and solvent
type on the biochemical compounds and antioxidant capacity of Artemisia absinthium. Alternations on
biochemical composition and antioxidant potential of extracts were studied due to various extraction
conditions, i.e, temperatures (3060 C), solvent types (methanol, ethanol, and acetonitrile), and
solvent concentration (25100%). Total phenolic content (6591033 mg gallic acid equivalents
(GAE)/100 g dry weight (DW)) and total flavonoid content (259392 mg catechin equivalents
(CE)/100 g DW) were examined in all samples. Radical scavenging capacities were determined using
the 2,2-diphenyl-2-picrylhydrazyl (DPPH) (5179%), ferric reducing antioxidant power (FRAP) (151
287 M Trolox equivalents (TE)/100 g DW), and 2,2-azino-di-[3-ethylbenzthiazoline sulphonate]
(ABTS) (1733.7 mM TE/g FM) methods in triplicate. Plant extracts obtained by 75% methanol at
45C showed the highest antioxidant capacity, whereas extracts obtained using 75% methanol at
60C had the highest levels of total phenolics. RP-HPLC (detection at 205 and 258 nm) and thin-layer
chromatography (TLC) of analyses did not detect artemisinin in extracts of leaves. On the other hand,
high-performance liquid chromatography (HPLC) results showed that extraction solvents had
significant effects on anabsinthin amount. This was supported by the result that showed highest
anabsinthin extraction was obtained by 75% methanol (16.58 g/g DW), but lowest by 25% methanol
(9.45 g/g DW).

Encyclopedia of Chromatography, Third Edition

 Flavonoids: CCC Separation
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DOI: 10.1081/E-ECHR3-120041127
Authors: L. M. Yuana
Published: 23 Sep 2009

Abstract
Various countercurrent chromatography (CCC) methods employ two-phase solvent systems for
separation of flavonoids. CHCl3 : CH3OH : H2O (4 : x : 2, for which x is between 2.5 and 4 for different
samples) may be used for their prefractionation. Two-phase solvent systems can be divided into four
types: CHCl3 : CH3OH : H2O, CHCl3 : CH3OH : n-BuOH :H2O, EtOAc : PrOH : H2O or BuOH : HOAc :
H2O, and n-C6H14 : EtOAc : CH3OH : H2O, respectively. CCC fractionation permits the use of either
stepwise elution or gradient elution, and analytical-scale CCC can also be carried out for separation of
flavonoids.

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Volume 21, Issue 18, 1998
 Original Articles

Simultaneous Determination of Flavonoids, Phenolic Acids,

and Coumarins in Seven Medicinal Species by HPLC/DIODE-
Array Detector
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DOI:

10.1080/10826079808003444

P. B. Andradea, R. M. Seabraa, P. Valentoa & F. Areiasa

pages 2813-2820

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Received: 30 Nov 1997

Accepted: 20 Feb 1998
Published online: 22 Aug 2006
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Abstract
A simple and accurate reversed phase HPLC procedure is proposed for the determination of 19
phenolic compounds (flavonoids, phenolic acids, and coumarins) in seven medicinal species. The
sample preparation involved extraction, alkaline and acid hydrolysis, and purification through a Bond-
Elut C18 column. The chromatographic separation was achieved using a reversed phase column
Spherisorb ODS2 (5m; 25.0 0.46 cm). Gradient elution was carried out using water-formic acid
(19:1) (A) and methanol (B). A diode-array detector monitored the effluent and chromatograms were
recorded at 280, 320, and 350 nm. The optimised methodology seems to be useful for the
phytochemical analysis of plant extracts. A close correlation between the phenolic compound patterns
and the botanical origin of plants was found.

Natural Product Research: Formerly Select Language

 Natural Product Letters
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Volume 25, Issue 5, 2011

Review Article

Flavonoids: chemical properties and analytical

methodologies of identification and quantitation in foods and
plants
DOI:

10.1080/14786419.2010.482054

Eleonora Corradinia,Patrizia Fogliaa, Piero Giansantia, Riccardo Gubbiottia, Roberto Samperia & Aldo Lagana*

pages 469-495

Publishing models and article dates explained

Received: 17 Dec 2009

Accepted: 23 Mar 2010
Published online: 07 Mar 2011
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Flavonoids have been recognised as one of the largest and most widespread groups of plant
secondary metabolites, with marked antioxidant properties. The general name flavonoid refers to a
class of more than 6500 molecules based upon a 15-carbon skeleton. In this paper a general
overview of flavonoids, their classification, structures and analytical methods for their determination is
presented.
 Journal of Liquid Chromatography & Select Language
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Volume 37, Issue 8, 2014

Original Articles

ISOLATION, IDENTIFICATION, AND SIMULTANEOUS

QUANTIFICATION OF FIVE MAJOR FLAVONOIDS
IN EPIMEDIUM ELATUM BY HIGH PERFORMANCE LIQUID
CHROMATOGRAPHY
DOI:

10.1080/10826076.2013.765465

Shahnawaz N. Sofia,Shakeel-u-Rehmana, Parvaiz H. Qazia, Shabir H. Lonea, Haroon M. Bhata & Khursheed A. Bhata*

pages 1104-1113

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Accepted author version posted online: 01 Aug 2013

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Abstract

A simple and reliable HPLC-UV-DAD method was developed for rapid resolution, identification, and
quantitation of prenylated flavonoids in Epimedium elatum, a plant endemic to Kashmir Himalayas.
Five major prenylated flavonoids namely epimedoside A, epimedin A, epimedin B, epimedin C, and
icariin were isolated using repeated column chromatography and simultaneously quantified using
HPLC in both underground and aerial parts of E. elatum. All calibration curves showed good linearity
(r2 > 0.998) within test ranges and recoveries were 96.8 to 101.2%. The optimized method was
successfully applied for the analysis of five major flavonoids in 18 samples of E. elatum collected from
three different ecogeographical zones in three different harvesting seasons. The contents of five
investigated compounds were greatly variant ranging from 8.73% to 10.96% in aerial parts and 2.52%
to 5.06% in underground parts. The concentration of epimedin C was found to be the highest (6.42%)
among the five quantified compounds in one of the accessions. The method established in this paper
is simple and reliable and could be used for the quality control of E. elatum.

https://www.researchgate.net/publication/245405120_Determination_of_Flavonoids_in_Tea_and_R
ooibos_Extracts_by_TLC_and_HPLC

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%2F10.1007%252FBF02490451&usg=AFQjCNGGEjBx867SvqPTdE_SQPlgVC_HEA&bvm=bv.11015184
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