The Pharma Innovation Journal 2017; 6(2): 179-182

ISSN: 2277- 7695

TPI 2017; 6(2): 179-182
© 2017 TPI Isolation and standardization of gingerol from ginger
www.thepharmajournal.com
Received: 29-12-2016 rhizome by using TLC, HPLC, and identification tests
Accepted: 30-01-2017

Dr. Gaikwad DD
Department of Pharmaceutics, Dr. Gaikwad DD, Shinde Sachin K, Kawade Ashwini V, Dr. Jadhav SJ
VJSM’s Vishal Institute of and Dr. Gadhave MV
Pharmaceutical Education and
Research, Ale, Pune,
Maharashtra, India Abstract
Many drugs commonly used today are herbal origin. About 25% of prescription drug in the US contains
Shinde Sachin K atleast one active ingredient derived from plant material. Herbal medicine is the oldest form of healthcare
Department of Pharmaceutics, known to mankind. Herbs has been used by all cultures throughout history. Herbal medicine are also in
VJSM’s Vishal Institute of great demand in developed world for primary healthcare because of their efficacy, safety and lesser side
Pharmaceutical Education and effects. The aim of present study is to isolate and standardize gingerol obtained from Zingiber officinale
Research, Ale, Pune, rhizome. Zingiber officinale rhizomes were extracted with ethanol (95%) by simple maceration process.
Maharashtra, India This extract was studied phytochemically and gingerol is isolated by using TLC and HPLC technique.
The isolated compound was found to be oleoresins. Gingerol constituents responsible to reduce effects
Kawade Ashwini V
Department of Pharmaceutics,
such as emesis and nausea.
VJSM’s Vishal Institute of
Pharmaceutical Education and Keywords: Herbal medicine, Zingiber officinale, gingerol, emesis, nausea
Research, Ale, Pune,
Maharashtra, India 1. Introduction
Zingiber officinale belonging to family Zingiberaceae is an ancient Indian medicine used in
Dr. Jadhav SJ
several disorders. Zingiber officinale commonly called as ‘Adrak’ in Hindi and Urdu, ‘Sunthi’
Department of Pharmaceutics,
VJSM’s Vishal Institute of in Marathi. It has various vernacular names such as Ginger, Srngaveram, Adrak, Sunthi.
Pharmaceutical Education and Zingiber officinale is possibly native to India and China. It is now widely grown as a
Research, Ale, Pune, commercial crop in south and Southeast Asia, tropical Africa (especially Sierra Leone and
Maharashtra, India Nigeria), Latin America, the Caribbean (especially Jamaica) and Australia. Ginger has a
distinctive thickened, branched rhizome (underground stem) which sometimes looks
Dr. Gadhave MV
Department of Pharmaceutics, somewhat like a swollen hand. The rhizome has a brown corky outer layer (usually removed
VJSM’s Vishal Institute of before use) and a pale yellow centre with a spicy lemon-like scent. Shoots (pseudostems), up
Pharmaceutical Education and to 1.2 m tall, arise annually from buds on the rhizome. Ginger is a well-known tropical herbs
Research, Ale, Pune, whose root is used in both Traditional Chinese Medicine and Western Herbal Medicine. It has
Maharashtra, India
a long history of medicinal used dating back 2500 years in China and India for conditions such
as headaches, nausea, rheumatism, and colds, has been used since ancient times for a variety of
condition, including fevers, and digestive problems, and as an appetite stimulant.

2. Materials and Methods

2.1 Chemical and Reagents
FTIR Spectrophotometer (Perkin elmers), UV Spectrophotometer (SHIMADZU 1800), HPLC
Waters 1650, all chemicals and solvents used were of A.R. Grade and IPA was obtained from
Alkem Labs. Ltd. Mumbai.

2.2 Plant material

The ginger rhizome were purchased from local market of Pune. The material were cleaned &
dried under shade & then placed in oven at 20-40 0C. The dried rhizomes were weighed and
stored in desicator.

2.3 Isolation of gingerol from ginger

Correspondence
Kawade Ashwini V Dry ginger was crushed to a coarse powder and extracted with95% ethanol by simple
Department of Pharmaceutics, maceration process. Solvent was evaporated by distillation to obtain thick pasty mass. The
VJSM’s Vishal Institute of thick pasty mass was suspended in water. The Ginger resin precipitates in water which was
Pharmaceutical Education and removed by filtration and the residue obtained was dried under vacuum.
Research, Ale, Pune,
Maharashtra, India
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2.4. Standardization of gingerol from ginger 100mg of crude extract was dissolved in methanol and diluted
The gingerol, active constituent of ginger rhizome extract was upto 100ml to get concentration of 1000ppm which is treated
standardized by various methods specified in the compendias. as stock solution. This stock solution was diluted further to
The various tests such as TLC. HPLC, Identification test are get different concentrations. Resultant solutions were scanned
performed to identify the gingerol present in extract. for λmax in the range of 200‐400 nm using UV-
spectrophotometer.
2.4.1 TLC Method
2.4.1.1 Preparation of plates 3.2.2 IR spectrum interpretation
Prepare a suspension of coating substance and spread a Fourier transform infrared (FTIR) was used to identify the
uniform layer of suspension, 0.25 to 0.30 mm thick, on flat characteristic functional groups in the extract. A small
glass plate of 20 cm long. Dried in air and heat at 100 to 1050 quantity (5 mg) of the extract was dispersed in dry potassium
for atleast 1 hr. store the plates protected from moisture, dry bromide (KBr). The mixture was thoroughly mixed in a
the plates at time of use if necessary. mortar and pressed at pressure of 6 bars within 2 min to form
a KBr thin disc. Then the disc was placed in a sample cup of a
2.4.1.2 Mobile phase diffuse reflectance accessory. The sample was scanned from
Hexane: Diethyl ether (30:70) 4000 - 400 cm-1.
Determination of purity:
2.4.1.3 Test solution The purity of extract was determined by HPLC analysis.
Reflux 1 g of the coarsely powdered substance under
examination with 25 ml of methanol for 15 minutes, cool and 3.3 Calibration curve of gingerol
filter. Wash the residue with 10 ml of methanol. Combine all In Methanol Stock solution of ginger extract were pipette out
the filtrates and concentrate to 10 ml. in to series of 10ml volumetric flasks and diluted with
methanol to get final concentration of 20-100mcg/ml. The
2.4.1.4 Reference solution absorbance of resultant solution was measured at 281.40nm.
Reflux 0.5 g of coarsely powdered sunthi RS with 5 ml
methanol for 15 minutes, cool and filter. Apply to the plate 10 4. Result and Discussion
μl of each solution as bands 10 mm by 2 mm. 4.1 Standardization of gingerol from Zingiber officinale
rhizome extract
2.4.2 HPLC Method 4.1.1 TLC
Reflux about 3 g of the coarsely powdered substance under Gingerol is analysed for retention factor. TLC plate showed
examination with 100 ml of methanolon a water–bath for 15 result illustrated in figure 10 of TLC chromatogram. Clear
minutes cool and filter. Reflux the residue further with spot observed from ethanolic extract when visualized by eye,
methanol till the last extract turns colourless, cool and filter. however under UV lamp in long wavelength 365nm the spot
Combine all the filtrates and concentrate to 50.0 ml. colour were fluorescent blue.
Reference solution:
A 0.1 per cent w/v solution of 6-gingerol RS in methanol. Table 1: Rf Values for ethanolic extract of Zingiber officinale by
Chromatographic system TLC
 a stainless steel column 25 cm x 4.6 mm packed with Solvent Front No. of Spot Rf
Solution
octadecylsilane bonded to porous silica (5 μm), Height (cm) spots height(cm) Value
 mobile phase: 55 volumes of acetonitrile and 45 volumes Reference Solution 5.5 1 5.4 0.98
of water, Test Solution 6.2 1 6.0 0.97
 flow rate. 1.3 ml per minute,
 spectrophotometer set at 278 nm, 4.1.2 HPLC
 a 20 μl loop injector. The standard curve for the concentration Vs peak height was
drawn and line of equation was originated. From the line of
2.4.3 UV Method development equation of standard drug y= 0.2228x + 37.658 and based on
For the quick estimation of this extract there is no reported the calculations and findings, it was consequently found that
UV-Visible method, so simple UV spectroscopic method was 100 mg of extract would be consist of 18.276 mg i.e. 18.276%
developed. Calibration curve of rhizome extract was prepared of active content.
in methanol at maximum wavelength of 281.40nm.

3. Pre-formulation study
It needed to ensure the development of stable, effective and
safe dosage form.

3.1 Solubility analysis

Solubility is made by adding solute in small incremental
amount to fixed volume of solvents such as distilled water,
ethanol, chloroform, acetone and ether. Then it is examined
for undissolved particles.

3.2. Confirmation of drug

3.2.1 UV-VIS Spectrophotometric method
Methanol was selected for preparation of calibration curve. HPLC chromatogram of Ginger extract
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Table 2: Standard calibration curve of gingerol by HPLC
Sr. No. Conc. ug/ml of standard solution Peak height %
1 20 40.59
2 40 48.0
3 60 51.69
4 80 55.96
5 100 58.89

Table3 4. Pre-formulation study

Sr. No. Conc. of test solution Peak height % 4.1 Determination of Solubility
1 Unknown 41.73
Table 5: Solubility profile of the ginger extract
4.1.3 UV Method Solvent Solubility behaviour
Extract was found to obey Beer‐Lambert’s law in the Water Insoluble
concentration range of 20‐100 μg/ml with regression Acetone Insoluble
coefficient (r2) values 0.9995. The regression equations were Ethanol Soluble
calculated as y = 0.0097x + 0.0132 for methanol. Methanol Soluble
Chloroform Soluble
Table 4: Different validation parameters of the calibration
4.2 Conformation of Drug
Parameters Results 4.2.1 UV Spectroscopy
Linearity correlation coefficient 0.9995
After studying the UV- spectra of ginger rhizome extract, it
y‐ intercept 0.0132
was found that it shows maximum absorbance at 281.4 nm.
Slope 0.0097
Range 20-100 µg/ml
So absorbance at 281.4 nm was considered as λmax for Ginger
LOD 4.5 µg/ml extract.
LOQ 13.6 µg/ml

UV Spectra of Ginger extract

4.1.2 IR spectrum interpretation characteristic absorption bands in the infrared absorption

The identity of drug was confirmed by comparing IR spectrum of gingerol and a summary of the description of the
spectrum of drug with reported spectrum of Gingerol. The characteristic IR bands for gingerol are described.

Table 6: Interpretation of Ginger Extract

Sr. No. Functional Group Theoretical Peaks (cm-1) Practical Peaks (cm-1)
1 C-H(stretch) 2700-3300 2927.5, 2852.5
2 C-O (stretch) 900-1300 1017.5
3 N-H (stretch) 3100-3500 3387.5
4 O-H (stretch) 3000-3700 3677.5
5 C=O (stretch) 1600-1900 1652.5
6 C=C (stretch) 1475-1610 1517.5

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63

60

55

50

45

40
1727.5cm-1
35
1652.5cm-1
%T

30
1517.5cm-1
2852.5cm-1
25
3677.5cm-1 1387.5cm-1
20 2927.5cm-1
3387.5cm-1
15

10

5
1017.5cm-1
0
-1
4000 3500 3000 2500 2000 1500 1000 450
cm-1
Name Description
gingerol Sample 017 By vishal Date Thursday, May 03 2012

FTIR spectra of Zingiber officinale extract

4.1.3 Determination of Purity 6. References

The purity of Ginger extract was done by using HPLC 1. Pharmacologicals. The Indian Phamaceutical Industry,
analysis method depending on retention time (Rt Value). Pure ICRA.2010.
gingerol shows the Retention time at 1.0 min. The analytical 2. Chaiyakunapruk N, Kitikannakorn N, Nathisuwan S.et al.
graph of HPLC scanning shows presence of various peaks at The efficacy of ginger for the prevention of postoperative
different retention time. Out of that the peak of gingerol nausea and vomiting: A meta-analysis. Am J Obstet
appears at 0.98. Depending upon that it was found that the Gynecol. 2006; (194):95-99.
extract containing gingerol is 98 % pure. 3. Max Wichtl. et al., Herbal Drugs And
Phytopharmaceuticals., Bisset.NG. Stuttgart: Medpharm
4.1.4 Standard Calibration Curve Scientific Publisher. 1994, 33-35.
4.1.4.1 Standard Calibration Curve of Gingerol in 4. Morgan K. Medicine of the Gods: Basic Principles of
methanol Ayurvedic Medicine. 2002.
[http://www.compulink.co.uk/mandrake/ayurveda.htm]
5. Schulz V, Hänsel R, Tyler VE. Rational Phytotherapy. A
Physician’s Guide to Herbal Medicine, 4th Ed., Berlin,
Springer-Verlag. 2001.
6. Sandberg F, Corrigan D. Natural Remedies. Their
Origins and Uses, New York, Taylor & Francis. 2001.
7. Surh Y. Molecular mechanisms of chemopreventive
effects of selected dietary and medicinal phenolic
substances. Muta. Res. 1999; 428:305-327.
8. Der Marderosian A, Beutlerr JA. The review Of Natural
Products St. Louis, Mo. Wolters Kluwer. 2006.
9. Ali BH, Blunden G, Tanira MO, Nemmar A. Some
λmax value for Ginger extract was found to be 281.4 nm from UV
spectra. phytochemical, pharmacological and toxicological
properties of ginger (Zingiber officinale Roscoe): a
Table7: Hence, this wavelength was selected for preparation of review of recent research. Food Chem Toxicol. 2008;
calibration curve for ginger extract. Calibration curve data of Ginger 46(2):409-20.
extract in methanol solvent 10. Nadkarnis KM. Indian Materia Medica, first edition,
popular Prakashan. 1308-1312.
Medium Equation R2
11. Wallis TE. Textbook of Pharmacognosy, 5th edition 2005,
Methanol y= 0.0097x + 0.0132 0.9995
CBS publishers. 389-91
12. Indian pharmacopoeia, Published by Indian
5. Conclusion
Pharmacopoeial Commission and Govt. of India. 2010.
However, further spectral analysis of isolated compound such
13. The Ayurvedic Pharmacopoeia of India, Published by
as 13C-NMR and mass spectroscopy could not be taken. As far
Govt of India, Ministry of Welfare, Dept. of Indian
as our knowledge goes this bioactive compound is novel.
System of Medicine and Homeopathy, New Delhi. First
Zingiber officinale provides deeper insights into indigenous
edition. 2006.
method of application and effectiveness of plant derivatives in
treating different ailments. Therefore, the structural
ilucidation, nomenclature, validation and pharmacological
screening of isolated gingerol will require for proving their
clinical reliability, safety and efficacy.

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