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Presented by :
Kipa Pape
M.Pharma Pharmacognosy &
Phytochemistry
82/MPG/SPS/21
Under the Supervision of:
Prof.(Dr.) Ajay Sharma
HOD (Pharmacognosy &
Phytochemistry)
Co-guide: Dr. Foziyah Zakir
Assistant Prof.
Development and Validation of High-Performance Thin-
layer Chromatographic Method for quantification of
Berberine in Rhizomes of ,an Endangered
Species Collected from Arunachal Pradesh, India
Coptis teeta Wall.
1

LOCATION: Department of
Pharmaceutical Sciences, Dibrugarh
University, Dibrugarh 786004,
Assam, India
PUBLISHER: Oxford Academic
AUTHORS: Ashis Kumar Goswami,
Neelutpal Gogoi, Anshul Shakya, and
Hemanta Kumar Sharma.
Journal of Chromatographic Science,
2019, 1–7 doi: 10.1093/chromsci/bmz009
IMPACT FACTOR: 1.645
2

Introduction
Methods and Materials used
Results
1
2
3
4
CONTENTS :
5 Discussion and Conclusion
Validation of HPTLC method
3

INTRODUCTION
Arunachalee has its traditional methods of
treatment, which have been reported earlier
but their scientific study has not been carried
out in most of the plants to validate the
traditional claims.
One such plant is
Coptis teeta Wall.
Synonym: Mamira, Golden thread root
Biological Source: Obtained from the dried rhizomes of
Common Name: Tiktmoola, Rangkeh ( in Arunachalee)
Family: Ranunculaceae
Chemical Constituents: Berberine, palmitine, epiberberine,
jatrorhizine
uses
SCABIES
ANTI-
INFLAMMATOY
DYSENTERY
ANTIBACTERIAL
SEVERE
HEADACHE
Coptis teeta
Wall.
Coptis teeta Wall.
4

This study was designed to develop
and validate a thin layer
chromatography (TLC)
densitometric-based method using
high - performance thin layer
chromatography for quantification
of berberine.
Lv. and his co-workers reported
that high-performance liquid
chromatography (HPLC) is a
versatile technique in
determination of protoberberine
alkaoids
But Ding and his co-workers
reported that HPLC analysis for
these alkaloids gives poor peak
and resolution and it requires long
time, which leads to column
contamination and damage.
History
Aim and Objectives
5

METHODOLOGY AND MATERIALS USED
The rhizomes of C.teeta were collected from Roying,
Lower Dibang Valley dist. A.P and identified and
authenticated by Dr. A.A Mao ( BSI, ERC, Shillong)
Collection of plant material
Chemicals and standards
Purchased from Merck ( Mumbai, India)
Berberine procured from Sigma-Aldrich
TLC fingerprint and chromatography
TLC plate: Pre-coated with silica gel 60F254
Mobile phase : Butanol: ethyl acetate: formic acid:
water 3.0:5.0:1.0:1.0
Spotter: CAMAG Linomat Vsemi-automatic sample
spotter.
Chamber: CAMAG twin through chamber
Preparation of plant extract
100 mg of powdered rhizomes of C.teeta was
sonicated for 15 min with 1mL of methanol.
After centrifugation, the supernatant was used
as the sample solution.
TLC densitometric quantification of berberine
using HPTLC
C.teeta
Quantittative analysis of the compound was done
in CAMAG Scanner IV by scanning at 351 nm.
The Rf value of methanol extract of rhizomes
of (0.70) is same as that of standard
berberine
The band at 0.70 nm was identified as berberine.
6

Validation
Parameters
Linear range
(ng/band)
90-210 ng/band
Intraday (n=6) :
0.77-1.63
Percentage
recovery: 93.32
(Within 90-110%)
Correlation
coefficient :
0.997386
(0.9-1.1)
LOD : 30ng/band
LOQ: 70ng/band
Interday (n=6) :
1.19-1.88
Repeatability(%CV,
n=10) : 1.0582
Method Validation Parameters for the Quantification
of Berberine in Methanol extract of by TLC
Densitometric Method
C.teeta
7

The validated method
gave compact bands for
berberine at an Rf of
0.70.
RESULTS
The Rf value of berberine
matched with the Rf
value of methanol
extract of rhizomes of
which was found to be
0.70.
C.teeta
1)
2)
3) The berberine
concentration in the
methanol extract of coptis
teeta was found to be
30.97±0.55 mg in 100 mg of
the crude drug.
8

The spectra generated for berberine and methanol extract of rhizomes of
at wavelength range 200-400 nm also matched.
C.teeta
9

CONCLUSION
DISCUSSION
The amount of berberine
found in (30.97%) in
this method was highest in
comparison to earlier studies.
Clear separation was
obtained through this
validated HPTLC method.
Requires urgent attention for
its preservation and
protection.
The method developed was
reproducible and selective for
quantifying berberine in herbal
drug formulation.
The abundance of berberine in
rhizomes of can be used for
further modification to produce
lead molecules with low toxicity
and high potency.
C.teeta
C.teeta
10

References :
Goswami AK, Gogoi N, Shakya A, Sharma HK. Development and Validation of
High-Performance Thin-layer Chromatographic Method for Quantification of
Berberine in Rhizomes of Coptis teeta Wall, an Endangered Species Collected
from Arunachal Pradesh, India. J Chromatogr Sci. 2019 May 1;57(5):411-417. doi:
10.1093/chromsci/bmz009. PMID: 30753457.
Bajpay, Anurag & Nainwal, Rakesh & Singh, Devendra. (2019). Coptis teeta: A
potential endemic and endangered medicinal plant of Eastern Himalayas. 245-
248.
Rathee, D., Rathee, S., Rathee, P., Deep, A., Anandjiwala, S., Rathee, D.; HPTLC
densitometric quantification of stigmasterol and lupeol from Ficus religiosa;
Arabian Journal of Chemistry, (2015); 8: 366–371
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2)
3)
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THIS IS THE END OF MY
PRESENTATION.
WISHING YOU ALL A VERY HAPPY
NEW YEAR IN ADVANCE !
THANK YOU!
12

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HPTLC ANALYSIS OF COPTIS TEETA

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