2011

NPC Natural Product Communications Vol. 6

No. 10
Variability of Total Flavonoid, Polyphenol and Tannin 1417 - 1420

Contents in Some Lythrum salicaria Populations

Tímea Bencsik*, Györgyi Horváth and Nóra Papp

Department of Pharmacognosy, University of Pécs, Pécs 7624, Hungary

timea.bencsik@aok.pte.hu

Received: May 27th, 2011; Accepted: June 27th, 2011

Lythrum salicaria L. can vary in some morphological and phytochemical features according to its habitat. The present study investigates and
compares the total flavonoid, polyphenol and tannin composition of twelve populations of L. salicaria collected from different ecological
habitats in south-west Hungary. We studied the plant heights and soil moistures of the habitats analyzing the potential correlation between the
morphological and environmental factors and chemical compositions. Total flavonoid, polyphenol and tannin contents were determined
according to the valid spectroscopic methods of the European Pharmacopoeia. Total flavonoid content was higher in the populations
collected during the main blooming period in August than at the beginning of flowering in July. Higher values for total polyphenol and tannin
were detected in flowering branch tips in August than in July, and higher ones in leaves and shoots in July than in August. The highest
flavonoid content was measured in the leaves, followed by the flowering branches and shoots as opposed to the total polyphenol and tannin
contents, which were higher in the flowering branch tips than in the other organs.

Keywords: Lythrum salicaria L., total flavonoid and polyphenol composition, phytochemical polymorphism.

Lythrum salicaria L. (purple loosestrife) is a widely
distributed plant in Europe, North America, Asia,
northwest Africa and southeastern Australia. In Hungary it
is a common perennial species found growing in marsh
meadows, fen communities, wetlands, edge of streams,
rivers and lakes [1].
The official drug is the whole flowering plant with branch
tips (Lythri herba – European Pharmacopoeia 6th ed.),
which is used traditionally in the treatment of diarrhea,
chronic intestinal catarrh, hemorrhoids, eczema, varicose
veins and bleeding of the gums [2]. In recent years
research has confirmed the traditional uses of the
decoctions and extracts of Lythri herba for their
astringent, antidiarrheal, hypoglycemic, antioxidant,
antiinflammatory, antinociceptive, antifungal, antibacterial
and calcium antagonistic properties [3-8], which are
attributed to the major group of compounds, tannins and
flavonoids [9-11]. Flavonoids play an important role in the
protection against UV-B, microbial infections and
herbivores. Among the tannins, the presence of
ellagitannins is characteristic of the Lythraceae with a
protective effect against insect herbivores and
microorganisms [13-16].
This comparative work studied the total flavonoid,
polyphenol and tannin composition of some populations of
L. salicaria collected from 12 habitats in south-west
Hungary with various water supplies and total light
exposure.
Moderate correlations were detected between the plant
height and the water content of the soil at the studied
habitats. Selected populations were classified into three
clusters according to these criteria. Our groups, in
accordance with the measured heights of the specimens,
were as follows (Table 1): 100-140 cm (SI, CR), 141-160
cm (SO, KC, SA, AR, CB, KP, LS) and 161-200 cm (LA,
LD, LM). According to the soil moisture content, three
groups were created with high (SI, SA, LA, LD, LS, LM),
medium (SO, KC, AR) and low water contents (CB, KP,
CR). Anthropogenic disturbance can be assumed at the
dikes of the roads in drainage ditches (SO, SI, KP, CR),
next to marsh meadow (AR) and at the edge of Lake
Deseda.
In all populations, higher total flavonoid contents were
measured in the samples collected in the main blooming
period in August than at the beginning of flowering in
July. The highest flavonoid content was measured in the
leaves, followed by the flowering branch tips and shoots.
The detected values ranged from 0.04 to 2.72%, including
0.28-2.72% in leaves, 0.10-0.67% in the flowering branch
tips and 0.04-0.24% in shoots (Figure 1). Among the
populations, the highest flavonoid content was recorded for
the population of Lake Almamellék in July and Lake
Desesa in August.
The polyphenol and tannin contents showed variance
among the studied plant organs and populations. In the
flowering branch tips the highest total polyphenol and
1418 Natural Product Communications Vol. 6 (10) 2011 Bencsik et al.

Figure 1: Total flavonoid content of the studied L. salicaria populations. Abbreviations see Table 1.

Figure 2: Total polyphenol content of the studied L. salicaria populations. Abbreviations see Table 1.

Figure 3: Total tannin content (polyphenols not adsorbed by hide powder) of the studied L. salicaria populations. Abbreviations see Table 1.

tannin values were detected, followed by the leaves and
shoots. Summarizing the data, these components represent
higher quantity in the flowering branch tips collected in
August than in July as opposed to the values for the leaves
and shoots, which were higher in July than in August. The
measured total polyphenol values ranged from 1.68 to
27.28%, including 8.27-27.28% in flowering branch tips,
5.26-23.30% in leaves, and 1.68-9.86% in shoots (Figure
2). Total tannin values varied between 0.97 and 21.88%,
with 6.60-21.88% in the flowering branch tips, 3.94-0.87%
in leaves and 0.97-8.37% in shoots (Figure 3). The highest
tannin content was detected in the population of
Szentlőrinc in July and Cserénfa in August.
 Polyphenol composition of Lythrum salicaria Natural Product Communications Vol. 6 (10) 2011 1419

Table 1: Plant heights and categories of soil moisture of the selected flavonoid and low polyphenol/tannin contents were
habitats, with abbreviations.
detected in leaf (LA, LD, SI), flower (LA, LD) and shoot
Plant
Habitats Latitude Longitude Abbr. height
Soil (SI). A positive relationship could also be observed
moisture between total flavonoid and polyphenol/tannin contents in
(cm)
1 Szentlőrinc 46°2'39.85" 18°0'18.48" SO 140 medium some samples. In the following cases the three investigated
2 Kacsóta 46°2'19.26" 17°56'38.79" KC 155 medium
3 Szigetvár 46°3'20.6" 17°48'40.78" SI 100 high secondary metabolites of the plants were detected in high
4 Szentlászló 46°8'33.9" 17°49'49.99" SA 145 high quantity in leaf (KP) and flower (SI), and in low quantity
5 Almamellék 46°10'2.99" 17°53'30.22" AR 150 medium in leaf (LD) and shoot (CB, LD, SI).
6 Lake Almamellék 46°10'8.23" 17°53'57.26" LA 200 high
7 Csebény 46°11'2.69" 17°55'20.84" CB 140 low
8 Kaposvár 46°20'20.57" 17°47'11.09" KP 140 low Our results prove the variability of total flavonoid and
9 Lake Deseda 46°26'29.38" 17°47'19.51" LD 165 high polyphenol/tannin contents in L. salicaria populations
10 Cserénfa 46°18'12.8" 17°52'52.06" CR 100 low
11 Lake Sikonda 46°10'53.26" 18°12'59.89" LS 160 high originating from different habitats. The amount of
12 Lake Malomvölgy 46°1'6.54" 18°12'0.2" LM 170 high flavonoids can be influenced also by the presence of heavy
metals in the soil, so we plan to study the composition of
The content of flavon C-glycosides (e.g. vitexin, the soil samples collected from selected habitats. Further
isovitexin, orientin, isoorientin) in Lythri herba is well- studies aim at investigating the annual effect on the
documented [17]. In the European Pharmacopoeia (6th), phytochemical composition.
there are two methods for measuring flavonoids, the one
(selected method in the Experimental section) is Experimental
recommended for the measurement of O-glycosides, the Chemicals: Hexamethylene-tetramine, aluminum chloride
second method (60% ethanol as solvent, final measurement and anhydrous sodium sulfate were purchased from
of boric acid complexes at 405 nm) is recommended for Spektrum 3D Ltd. (Debrecen, Hungary), hydrogen
the study of C-glycosides. According to our studies, higher chloride, acetone, ethyl acetate and methanol from Molar
total flavonoid contents were detected in the samples with Chemicals Ltd. (Budapest, Hungary), Folin-Ciocalteu's
the previous method, in spite of their characteristic phenol reagent and hide powder from Sigma Aldrich
flavonoid C-glycoside content, and, therefore, it was used (Budapest, Hungary).
in our analysis.
Plant materials: Whole herbs of 12 populations were
According to the European Pharmacopoea (6th), Lythri collected from different ecological habitats in south-west
herba contains not less than 5.0% of tannins, expressed as Hungary in summer 2010: drainage ditches (Szentlőrinc,
pyrogallol, and calculated with reference to the dried drug. Szigetvár, Kaposvár, Cserénfa), marsh meadows (Csebény,
This criterion was met in the specimens of all selected Kacsóta, Almamellék), humid meadow (Szentlászló), and
populations. edge of lakes (Lake of Almamellék, Deseda, Malomvölgy
and Sikonda) (Table 1). Specimens were identified by Dr
In conclusion, higher total flavonoid content was measured Nóra Papp botanist, and voucher specimens have been
in plants living in humid habitats compared with areas with deposited in the Department of Pharmacognosy at the
low water content, except for the leaves and shoots University of Pécs (Lythrum salicaria 10SO, L. s. 10KC,
collected from CR. The plants living in humid habitats L. s. 10SI, L. s. 10SA, L. s. 10AR, L. s. 10LA, L. s. 10CB,
presented higher total polyphenol and tannin values in L. s. 10KP, L. s. 10LD, L. s. 10CR, L. s. 10LS, L. s.
August than in July as opposed to the plants growing in 10LM).
soil with medium water content in July.
Extraction: Herbs were dried at room temperature for 3
The height of plants can be influenced by flavonoids weeks, ground and stored in the dark. Total flavonoid,
through inhibition of auxin oxidase [13]. In our polyphenol and tannin contents were determined according
investigations there was no correlation between the to the valid spectroscopic methods of the European
flavonoid content and plant height. Pharmacopoeia, 6th ed. [12]. A Metertech SP-8001 UV-
VIS spectrophotometer (Spektrum 3D Ltd., Debrecen,
Flavonoids might play an important role as phytoalexins
Hungary) was used for all absorbance measurements.
[13]. Our samples living far from the roads (e.g. CB, LM,
SI, LD) contained lower quantities of flavonoids compared
Determination of total flavonoids: According to the Ph.
with anthropogenic disturbed habitats (e.g. KP, CR, LA),
Eur. 6, the extracts were prepared from 0.500 g of
but correlation between total polyphenol/tannin and
powdered drugs, and absorbance of the aluminum complex
anthropogenic disturbance could not be observed.
was quantified at 425 nm after 30 mins. Total flavonoid
content was determined as hyperoside equivalent.
An inverse relationship could be detected between total
flavonoid and polyphenol/tannin content in the case of
Determination of polyphenols and tannins: In accordance
some samples. In some cases, the total flavonoid contents
with Ph. Eur. 6, the extracts were prepared from 0.750 g of
were low, but total polyphenol/tannin values were high in
powdered drugs and mixed with a determined quantity of
leaf (CB), flower (SA, CB) and shoot (LM), and high
1420 Natural Product Communications Vol. 6 (10) 2011 Bencsik et al.

Folin-Ciocalteu reagent and 290 g/L sodium carbonate
solution. After 30 min the absorbance was measured at 760
nm used distilled water as compensation fluid. The
percentage content of tannins was expressed as pyrogallol
equivalent.
Acknowledgments – The authors are thankful to Ágnes
Farkas for correction of the manuscript and Gyöngyi
Dörmer for technical assistance.

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