Volume: 3: Issue-2: April-June-2014

Copyrights@2014
ISSN: 2278-0246
th
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Received: 24 March-2014
Revised: 25 April-2014
Accepted: 29th April-2014
Coden: IJAPBS www.ijapbs.com Research article
ISOLATION OF CATECHIN FROM STEM BARK OF ALBIZIA LEBBECK
Sonal Desai1, Pratima Tatke*1, S. Y. Gabhe 2
1

Department of Pharmaceutical Chemistry, C. U. Shah College Of Pharmacy, S. N. D. T. Womens

University, Santacruz (W), Mumbai-400 049, India.
2
Department of Pharmaceutical Chemistry, Poona College of Pharmacy, Bharati Vidyapeeth Deemed
University, Erandwane, Pune-411 038, India
Phone: +91-22-2660 9577 Mobile: +919920685857, Email:patatke@gmail.com
ABSTRACT: Albizia lebbeck Benth. is reported to have many significant medicinal properties. This medicinal tree is
well known for its anti-allergic and anti-asthmatic properties. There are number of marketed herbal formulations
containing stem bark extract of Albizia lebbeck which are used for treatment of ailments related to respiratory track.
Catechin is a phytomarker present in stem bark of Albizia lebbeck having antiallergic activity. The present work
discusses a simple, cost effective preparative TLC technique for isolation of catechin from extract of Albizia lebbeck
stem bark. Structure of isolated catechin was confirmed by spectral studies such as IR, NMR and Mass spectroscopy.
Isolated catechin can be used as marker compound for marker-based standardization of extracts and formulations
containing stem bark of Albizia lebbeck.
Key words: Albizia lebbeck, Catechin, Fabaceae, Preparative TLC, Shirish

INTRODUCTION
Albizia lebbeck, commonly known as Shirish, belonging to Fabaceae family is well known medicinal tree used in
Ayurvedic and Unani system of medicine. [1, 2] The tree is big and tall having 17 to 20 meters height. Leaves are
compound, unctuous and hairy while leaflets are wide and 4 to 8 in pairs. Flowers are tender and white in colour.
Legumes are 15 to 30 cm long and 1.5 to 3 cm wide, tough and contain 6 to 10 seeds. Seeds are flat, round and brown
in colour. [3] Stem bark of this plant contains catechin, betulinic acid and its glycosides, albizzia saponins A, B and C,
isomer of leucocyanidin, melacacidin, leuco-anthracyanidin, lebbecacidin, friedelin, -sitosterol, phenolic glycoside,
albizinin and anthraquinone glycosides.[4] Catechin present in stem bark of Albizia lebbeck said to possess antiallergic
activity.[5,6] Catechin is chemically, (2R,3S)-2-(3,4-dihydroxyphenyl)-3,4-dihydro-2H-chromene-3,5,7-triol belongs
to the group of flavan-3-ols (Figure 1).

Figure 1: Structure of catechin

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Many methods have been reported for isolation of catechin from different medicinal plants. These methods utilize
tedious and expensive chromatographic techniques such as column chromatography using Sephadex and supercritical
fluid extraction technique. [7-13] Thus, the present work aims to isolate catechin from stem bark of Albizia lebbeck by
simple, cost effective preparative TLC technique.

MATERIALS AND METHODS

Reagents and standards
All the solvents and purified water used for isolation were of AR grade from S. D. Fine chemicals, Mumbai, India.
Standard of catechin was procured from Sigma-Aldrich.
Instruments
To visualise TLC plates, CAMAG TLC visualizer was used. IR spectrum of isolated compounds was carried out using
FTIR-4100 spectrophotometer. NMR spectrum was recorded using Nuclear Magnetic Resonance spectrophotometer,
Bruker Advance 300 MHz having programme Zg30. The probe used was 5 mm BBO probe. Data were analysed by
Xwin NMR version 1.3. Mass spectrophotometer used was Bruker Daltanics, Model Microtof Q. Ionization technique
used was Electron spray ionization technique. All the data were analysed by Hystar software version 3.4.
Plant materials
Stem bark of Albizia lebbeck was collected from Valsad district of Gujarat in February 2012 and authenticated at
Botanical Survey of India (BSI), Pune under voucher specimen number ALGSOD8. The barks were dried in oven
with air circulation at 35 0C, for seven days, powdered and sieved.
Preparation of methanol extract
Methanol extract of bark was prepared by macerating 10 g of powdered bark with 100 ml of methanol for 24 h and
then filtered. The process was repeated thrice to ensure complete extraction and all the filtrates were combined. The
combined methanol extract was dried under reduced pressure using rotary evaporator and kept in an amber colored
container till used.
TLC studies of methanol extract
The following chromatographic conditions were optimized for TLC/HPTLC studies:
Stationary phase: Silica gel GF254 (E. Merck)
Mobile phase: Ethyl acetate: toluene: formic acid (5:5:1, v/v/v)
Sample: Methanol extract of bark of Albizia lebbeck and standard of catechin
Derivatising reagent: 5 % ferric chloride solution, vanillin sulfuric acid (VSR)
Procedure: Bark extract and standard of catechin were dissolved in minimum amount of methanol and spotted in the
form of bands on precoated silica plates. The plates were developed in the mobile phase, ethyl acetate: toluene: formic
acid (5:5:1, v/v/v). After development the plates were dried and observed under short (254 nm) and long (366 nm) UV
light. The plates were derivatised with 5 % ferric chloride solution and VSR.
Isolation of catechin
Catechin was isolated from the methanol extract of bark of Albizia lebbeck by preparative thin layer chromatography
using same chromatographic conditions as used in TLC studies.
Procedure: Bark extract was dissolved in minimum amount of methanol and spotted in the form of bands on silica
plates. The plates were developed in the mobile phase. After development the plates were dried and band at Rf 0.33
was scrapped and dissolved in methanol, filtered through 0.2 PTFE filter and evaporated to dryness to get catechin
residue. Catechin residue thus obtained was further purified by recrystallization using methanol. Isolated catechin was
confirmed with comparison with standard catechin by TLC and HPLC studies. Structure of isolated catechin was
established by spectral studies like IR, NMR and Mass spectroscopy.

RESULTS AND DISCUSSION

TLC studies showed presence of well isolated band of catechin in the methanol extract at Rf of 0.33 (Figure 2).

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At 254 nm
E=Extract S=Standard

At 366 nm

Sprayed with FeCl3

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Sprayed with VSR

Figure 2: TLC images of methanol extract and standard of catechin

It produced brown and pink colour on derivatization with 5 % FeCl3 and VSR, respectively. Isolated catechin was
confirmed by comparison with standard by TLC (Rf 0.33) and HPLC (RT 1.79 min) studies. Purity of isolated catechin
was found to be 97.12 % by HPLC. Structure of isolated compound was further confirmed
by IR, NMR and Mass spectroscopy (Figures 3, 4 and 5).

Figure 3: IR spectrum of isolated catechin

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Figure 4: NMR spectrum of isolated catechin

Figure 5: Mass spectrum of isolated catechin

Catechin: Light brown powder, max (MeOH): 279 nm, IR (KBr): 3301.54 cm-1(exchangeable protons from alcohol),
1285 cm-1 and 1242 cm-1 (C-O-C stetching),
1
H NMR (CD3OD) (ppm): 6.837 (d, 1H), 6.708 (dd, 1H), 5.92 (s, 1H), 5.85 (s, 1H), 4.552 (d, 1H), 3.941 (m, 1H),
2.817 (dd, 1H), 2.491(dd, 1H),
Mass analysis (m/z): 290.2 (M+), 289.4 (M+-H).

CONCLUSION
Catechin was successfully isolated from methanol extract of stem bark of Albizia lebbeck by a simple and cost
effective preparative TLC method. Structure of catechin was confirmed by chemical and spectral studies. The isolation
method can be employed to isolate catechin which can be used as a phytomarker for marker-based standardization of
extracts and formulations containing stem bark of Albizia lebbeck.

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Conflict of Interest
The authors confirm that this article content has no conflict of interest.

ACKNOWLEDGEMENTS
The authors would like thank National Medicinal Plants Board (NMPB), Dept. of AYUSH, New Delhi, Government of
India for funding the project.

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