Analytical Profile of Coleus Forskohlii | Forskolin (Coleonol)

1. ANALYTICAL PROFILE OF HERBAL DRUG
Coleus forskohlii
Presented By –
Mr. Swapnil Therkar
M. Pharm 1st Year
(Pharmacognosy)
Guided By –
Miss. Nikita Takale
(M. Pharm)
Government College of Pharmacy, Amravati

2. 1. Introduction
2. Uses
3. Physico-chemical Properties
4. Analytical Profile
5. References
Contents
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3. INTRODUCTION
Synonyms: Plectranthus barbatus, Coleus barbatus.
1. Sanskrit : Mayani, Makandi
2. Hindi : Patharchur, Pashanabhedi
3. Marathi : Mainmul, Garamar
Biological Source: It consists of extract of roots of
plant Coleus forskohlii belonging to family Lamiaceae
Not less than 0.4 % of forskolin.
Chemical Constituents: Forskolin (Coleonol),
Methylene-quinone, Barbatusin, Colenal B & C,
Deoxycolenol.
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5. Uses 1. As antihypertensive
2. In asthma
3. As anticoagulant
4. In glaucoma
5. Help to loose body weight
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6. Physico-chemical Properties
 Color: White crystalline powder
 Molecular formula : C22H34O7
 Molecular weight: 410.5 g/ mol
 Solubility: It is non-polar in nature.
Soluble in acetone, chloroform, benzene and sparingly
soluble in Pet. ether and water
 Melting Point: 232-235 °C
 Boiling Point: approximately 519-520 °C at a
pressure of 760 mmHg
Forskolin (Coleonol)
Forskolin is a Labdane Diterpenoid obtained from extract
of roots of plant Coleus forskohlii
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7. 1. TLC
2. HPTLC
3.HPLC
4. UV-Spectroscopy
5. IR-Spectroscopy
6. NMR-Spectroscopy
7. MASS-Spectrometry
Analytical
Profile
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8. TLC (Thin Layer Chromatography)
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TLC Method-
 Stationary phase- Aluminium plate pre-coated with Silica gel
60F-254
 Mobile system- benzene: methanol (9:1 v/v)
 Reference solution- 0.1 % w/v solution of pure forskolin in
methanol
 Sample- 500 mg of the extract + 10 ml methanol and filter
 Sample volume- 10uL of band
 Detectors-
I. UV Detector at 210 nm
II. Densitometric analysis- at 545 nm, after spraying with
anisaldehyde-sulfuric acid visualization reagent (dark violet or
purple)
 Rf value- 0.25±0.02

9. HPTLC (High Pressure Thin Layer
Chromatography)
 Stationary phase- Pre-coated Silica gel 60F-254 HPTLC plate.
 Sample- 50mg sample + 25ml Chloroform volume make
up (100 mL) with Chloroform
Mobile Phase St. Phase Detection reagent Rf Value
Benzene: Ethyl
Acetate (80:20
v/v).
HPTLC Plate Anisaldehyde-sulfuric
acid /560nm
0.45
Toluene: Methanol
(18:1.5 v/v)
Silica Gel 60
F254
Anisaldehyde-sulfuric
acid/545 nm
0.27± 0.02
Benzene: Ethyl
Acetate(75:25 v/v)
Perforated
Plate
200 nm 0.49± 0.01
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Fig: HPTLC Fingerprinting of forskolin at (a) 366nm
(b) Visible Light

10. HPLC (High Pressure Liquid
Chromatography)
HPLC Method- The samples was extracted with acetonitrile by sonication.
 Stationary Column – RP C18 Column.
 Solvent system- Water-Acetonitrile gradient elution (50:50 v/v)
 Sample- 250mg sample + 25ml acetonitrile volume make up
(100 mL) with acetonitrile
 Flow rate- at a flow rate of 1.0 ml /min
 Temperature- constant at 25-30 °
 Injection volume- 20uL
 Detector and wavelength- Photodiode array detector at 210 nm.
 RT- approximately 6.9 min
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11. UV Spectroscopy
 Forskolin is not very much UV sensitive. Its UV-absorption maximum is around 200 nm.
 Concentrated Benzene extract of C. forskohlii is used for UV Spectroscopy.
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Fig: UV- Spectrum of Pure Forskolin
isolated from C. forskohlii Root
Fig: UV- Spectrum of Marketed

12. IR Spectroscopy
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The FTIR spectra have been recorded in KBr pellet, using a: Fourier
Transmission Infrared Spectra by: SHIMADZU.
Wavenumber Functional
Group
3394 cm-1 Alcoholic -OH
2957 cm-1
1735 cm-1 C=O
1268 cm-1 C-O-C
1162 cm-1 C-O
Table: IR Functional Groups

13. NMR Spectroscopy
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Carbon δ value In CDCl₃ Multiplicity Type of Carbon
C11 a 205.60 s C=O
C21 b 169.77 s Acetyl C-O
C14 c 146.30 d olefinic carbons
C15 d 110.69 t olefinic carbons
C9 e 82.65 s heterosubstituted
alkyl carbons
C1 i 74.35 d --||--
C12 K 48.75 t alkyl carbons
C3 n 36.09 t --||--
4α-CH₃ P 32.97 q methyl groups
13-CH₃ q 31.52 q methyl groups
Carbon NMR  Proton NMR
Hydrogen δ value In CDCl₃ Multiplicity
9α-OH 6.2 s
H-2β 2.16 d
H-7α 5.46 d
H-12α 3.1 7 d
H-12β 2.38 d

14. MASS Spectroscopy
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 Stationary phase- RP C18 column
 Sample- Benzene extract of C. forskohlii is used followed by
concentration Process.
 Mobile Phase- acetonitrile: water (65:35 v/v).
 The flow rate was modified to 0.5 mL/min
 Retention time increased to about 12.3 min in the LC.
 In LC/MS method positive ESI mode, the spectrum of forskolin
showed signals at m/z 428.2 [M+NH4] + and 411.1 [M+H] +.
Fig- LC/MS of Forskolin
Fig- MASS Spectra of Forskolin

15. REFERENCES
1. Kokate C.K., Purohit AP, Gokhale SB. (2008) “Textbook of Pharmacognosy” Pune: Nirali
Prakashan, Page No:- 14.98-14.99
2. Kumar H. , Spandana M. (2013) “Method of Extraction and Determination of Forskolin from
Coleus forskolii of Nepal; Journal of Pharmacognosy and Phytochemistry; 37-39
3. Thimmegowda S., HussainF.H.M. , Patil R. Et.al.(October 2020) “Antioxidant, antimicrobial and
cytotoxic activity of Curkolin (Curcuma longa and Coleus forskohlii formulation)”; Article in
Indian Journal of Traditional Knowledge.
4. Mohamed Saleem A. (2013)“Methods of Isolation and Analysis of Forskolin from Coleus
forskohlii” Springer-Verlag Berlin Heidelberg
5. Kegler H. and Fehlhaber H.W. (1991) “NMR Investigations of Forskolin Complete Assignment of
Proton and Carbon NMR Spectra and Conformational Analysis” MAGNETIC RESONANCE IN
CHEMISTRY, VOL. 29, 993-998
6. Forskolin Coleus forskohlii, = 98 HPLC, powder 66575-29-9 (sigmaaldrich.com)
7. Forskolin – Wikipedia
8. Forskolin | C22H34O7 | ChemSpider
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16. THANK YOU