CHM 510 ANALYTICAL SEPARATION METHOD

EXPERIMENT 10 Qualitative Analysis of Ginger by Gas Chromatography Mass

Spectrometry (GC-MS)
Done on
28th March 2014
By
ASHRAF BIN AHMAD

2012706315

ABDUL IZZULISLAM BIN ABDUL HAMID

2013433056

MOHD MAUSA JASNIME BIN NAZRI

2013161343

For
Dr. Tay Chia Chay
Submission date: 4th April 2014

Introduction
In this experiment, we want to perform an extraction of ginger using liquid-liquid extraction
(LLE) technique to analyse and to identify the constituents in the essential oil of ginger by
using gas chromatography (GC). The ginger used is about 8g and was dissolved in a
methanol solution. The chopped up ginger was then refluxed for 30 minutes. Then the
solution was transferred to a separating funnel and extraction is done using dichloromethane.
When the solution formed two layers, the organic layer was supposed to be transferred into a
beaker and dried with anhydrous magnesium sulphate to remove excessive water. Then the
mixture was supposed to be filtered and put into a vial for qualitative analysis using GC-Mass
Spectrometry (GC-MS).
Plant extract have been used by humans for their antifungal, antimicrobial,
insecticidal, cytostatic and therapeutic activities. Many compounds such as antimicrobial,
antiseptic, sedative, and diuretic compounds can be isolated from a single herb. The
therapeutical activities of herbs depend on some vegetal active principle contained in the
plants. It is necessary to take care when a drug is prepared for a complete extraction of the
active principles from plants, otherwise they can be destroyed in the extraction process with
the decrease and even loss of therapeutical activity. Conventional sample preparation
technique based on liquid-liquid extraction (LLE) have been widely used to separate analyte
from sample matrix and to preconcentrate trace compound. This technique consumes a lot of
time and involve a large waste of solvent.
Ginger is widely used as either a food product or an herbal medicine in our life. In this
experiment, we want to determine volatile components in essential oils from ginger by use of
gas chromatography-mass spectrometry (GC-MS). With the chromatograms and related mass
spectra obtained, the chemical components can be qualitatively identified based on the
similarity searches in the MS databases. GC-MS has good ability in separation technique and
assist qualitative and quantitative determination of chemical components separated from
complicated practical systems such as foods, herbal medicines, and environmental samples.
Objective
1. To perform an extraction of ginger by liquid-liquid extraction.
2. To analyze and identify the constituents in the essential oil of ginger by using GC-MS

Reagents
1. Methanol
2. Dichloromethane
3. Anhydrous magnesium sulphate

Sample
Fresh ginger

Apparatus
1. Round-bottom flask
2. 250 mL separating funnel
3. West condenser
4. 13 mm diameter syringe filter (0.45 mm pore size)
5. 50 mL beaker and sample bottle.

Instrument
Gas

chromatograph

(Agilent

Technologies

6890N)

equipped

with

5%

poly(phenylmethylsiloxane) capillary column (30m X 0.25mm) and a 5972 MSD Quadrupole

detector

Procedure
1. Sample preparation
-

3-4g of ginger was weighed and recorded.

Then the ginger was chopped into small pieces and put into a round bottom flask.

Then 25 ml of methanol was added to the flask.

Then the sample was refluxed for about 30 minute.

Then the solution was transferred into a separating funnel and added with 5 ml of
dichloromethane.

Then the organic layer was drained into a beaker and added with anhydrous
magnesium sulphate.

The mixture was then filtered by using filter funnel.

The final product was filtered to remove any particulate matter and put into vial for
analysis by using GC.

2. Instrument set-up.
-

Injection port

: Split (20:1)

Injection port temperature

: 280C

Oven temperature

: initial temperature 70C for 4 min. Then ramp the

temperature at 10C/min up to 140C and 20C/min to
280C

Column flow rate

: 40 mL/min

Detector temperature

: 290C

3. Separation of essential oils in spearmint herb.

-

Inject 2 L of each sample extract. If necessary, temperature ramping conditions may

be altered to optimize the resolution.

4. Identifications of essential oil constituents in each sample extract.

-

Identify the major compounds in the extract by comparing their mass spectra with the
flavor library in the GC-MS database.

Results
No result was obtained from this experiment as the amount of essential oil extracted from the
ginger is too small to be separated and used in any analysis, and no other method was
available at the time to get a better extraction in terms of volume and purity.

Discussion
A few modifications were made to this experiment that differed from the manual. First of all,
ginger was used instead of spearmint herbs because unlike spearmint herbs, ginger is widely
available and rather cheap. This substitution does not require any change to the methodology
used. However, a large portion of ginger is needed to ensure that extraction is possible. This
is where the second modification came to use. The initial mass of sample stated in the manual
is three to four grams. This experiment was run with 7.89g, almost twice the number stated in
the manual, in hope that extraction could be more effective. But that was not the case, as
shown in the pictures inserted.
The extracting layer, which is dichloromethane, and the extracted layer, which is
methanol, are both organic solvents. Being organic solvents, they are both miscible to each
other. Hence, a clear and separated line between these two solvents does not exist. The line
seen in the pictures shown illustrates an alteration in color in the two layers because the oil
has been extracted by the extracting solvent, but no line between the two solvents is visible.
When the extracted layer is to be drained, the essential oil may slip back into the extracted
layer due to two reasons. One is as mentioned above, which is miscibility between the two

solvents. Another reason which may greatly affect the extraction is the polarity of these two
solvents. Although both are polar solvents, with the predicted dipole moments of 1.882
Debye and 1.807 Debye for methanol and dichloromethane respectively, methanol has a
higher polarity index of 5.1 compared to dichloromethanes polarity index of 3.1. This shows
that methanol may likely retain or reverse-extract the essential oils back into its body of fluid.

Extracting solvent (Dichloromethane +

Extracted solvent

One solution to this problem is to increase the mass of sample used. The higher the
mass of ginger used, the greater the volume of essential oil extracted. However, this is only a
stop-gap solution which may not prove handy when other types of samples are used, and can
only help so much in volume, not purity, as the polarity of the solvents still affects the

extraction. Another solution is to change the solvent used, either using a more polar solvent
during extraction or using a solvent that is immiscible to the initial solvent. In fact, the initial
solvent may be the one changed. Using water as the initial solvent may not be a good choice
as it has a higher boiling point than methanol. Having a higher boiling point means that water
needs more heat to be boiled, and more heat may lead towards the deterioration of the
sample.
The best solution is to modify the extraction process. Rather than using liquid-liquid
extraction, it is possible to get the crude essential oil using steam distillation or a rotary
evaporator (Rotavap). Since setting up a steam distillation set is not possible in the lab, the
choices had been narrowed down to only one, which is using the rotavap. Rotavaps are very
useful and efficient in collecting small amounts of product from a solvent-containing mixture.
Another good point of rotavaps is that it does not require the use of an extracting solvent,
which is more eco-friendly compared to LLE and less tedious. However, due to a series of
unforeseen misfortune, the rotavap available at the time this experiment was held has suction
problems which will prolong operation time. Prolonged operation times are not good as it
leads to excessive and wasteful energy use and may break the machine itself, as the cooler of
the rotavap available cannot handle such long periods of usage. It is at this point that the
experiment was halted as extraction using the current sample is near to impossible. Even if
extraction is done, the purity of the sample is very questionable. Therefore, it is better to not
do any extraction given the situation.
If given a rerun with a rotavap in good condition, the experiment may yield better
results. After collecting the essential oil, a minimum amount of anhydrous magnesium
sulphate must be used in order to absorb excessive moisture from the oil. After that, the
essential oil must be filtered, transferred into a sample vial using a syringe equipped with a
0.45mm pore filter, and lastly, analyzed using the GC-MS.
Although the experiment was halted midway, we can still predict the expected
compounds that may be identified by the GC-MS. These compounds can be predicted
through extensive search of results of analysis done on ginger by other people. Among the
chemical compounds usually found in the essential oil of ginger is a-pinene, camphene,
b-pinene, 1,8-cineole, linalool, borneol, y-terpineol, nerol, neral, geraniol, geranial, geranyl
acetate, b-bisabolene and zingiberene.

Conclusion
The experiment did not meet its objective; therefore the experiment is a failure. However,
problems with the method used for this experiment has been identified, and further
improvements can be implemented in order to get better results in the future.

Reference
http://www.essentialoils.co.za/essential-oils/ginger.htm
http://umpir.ump.edu.my/461/1/KHAIRU_AIZAM_BIN_IBRAHIM_2049.pdf
http://infohost.nmt.edu/~jaltig/SteamDistill.pdf
http://agritech.tnau.ac.in/horticulture/extraction_methods_natural_essential_oil.pdf
http://macro.lsu.edu/howto/solvents/Polarity%20index.htm
https://au.answers.yahoo.com/question/index?qid=20091003063830AAFSL4l