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“Standardization of volatile oils”
A
PROJECT REPORT
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“STANDARDIZATION OF VOLATILE OILS”
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FOR ELECTIVE SUBJECT
SUBMITTED TO THE HEMCHANDRACHARYA NORTH GUJARAT
UNIVERSITY, PATAN
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2004-2005
IN
PARTIAL FULFILLMENT
OF THE REQUIREMENT FOR THE
DEGREE OF BACHELOR OF PHARMACY
SUBMITTED BY
MITUL. M. SHAH
DEPARTMENT OF PHARMACHEMISTRY
SHREE S.K PATEL COLLEGE OF
PHARMACEUTICAL EDUCATION AND RESEARCH
GANPAT VIDYANAGAR,
KHERVA-GUJARAT
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This is to certify that the project work for elective subject entitled
“Standardization of volatile oils” represents the bonafide work of
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Ms. Mitul .M .Shah carried out under my guidance & supervision at
the department of Pharmaceutical Chemistry, Shree S. K. Patel college
of Pharmaceutical Education & Research, Ganpat Vidyanagar, during
the academic year 2004-2005. She has collected the literature very
sincerely & methodologically. This work is up to my satisfaction.
GUIDE
Prof.Dimple Sankhala
(M.Pharm)
Department of pharmachemistry,
Shree S.K.Patel College of
Pharmaceutical Education and research.
Ganpat Vidyanagar, Kherva.
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Dr. N.J.Patel
(M.Pharm, PhD)
I/C Principal
Shree S.K.Patel College of
Pharmaceutical Education and research.
Ganpat Vidyanagar, Kherva.
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Date:
Place: Ganpat vidyanagar, Kherva.
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This work has become successful by the blessing of god, my parents, & my family.
This project report has been prepared to give a brief introduction of
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“Standardization of volatile oil “which was undertaken for the fulfillment of degree
course in pharmaceutical science. This project is completed as per the direction of
syllabus
Numerous people have been instrumental in enabling me to give a concrete shape
to my thesis constraints of time & space preclude the mention of all of them here.
However I must mention the names of a few people who have made a catalytic impact
on the development of this thesis.
With profound pleasure I express my deepest gratitude to my esteemed guide
“Ms Dimple Sankhala & Mr. Falgun M Mehta” as professor in department of
pharmachemistry, who fulfilled very limited acknowledgement of mine & also for his
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erudite guidance, timely suggestion, continuous, encouragement & critical remarks for
the entire span of my work. I also want to express my gratitude to professor
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K.I.Maulvy sir & P. U. Patel sir for helping me to complete my project work.
I owe a special word of thanks to professor N.J.Patel (Principal of Shree
S.K.Patel college of Pharmaceutical education & research), & Shree Dr. M.M.Patel
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(Chancellor of Hemchandracharya north Gujarat University), for their precious gift of
knowledge & who has been a constant source of inspiration to steer throughout my
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pharmacy curriculum.
I also thank to Mr. P.I Patel, Mr. Mahadev bhai, & Mr.Mukesh bhai, for
helping me in maximum utilization of the library. I am also thankful to staff members,
who helped me lot to complete this project work.
I also want to express my gratitude to my friends & my classmates for being a
great support to me in completing my thesis. I greatly want to thanks to Nimishadidi,
Snehal, Kinjal, Jaldhai, Sweta, Sweeti, Darshana, Neha, Sonal, Hardi, Ashish, Ankit
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&Deepan.
Finally I want to express my deep gratitude to my revered & loving MOM, DAD
& my caring & loving sisters ANKITA & DHARA, for their moral support, constant
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encouragement & patience essentially needed to complete my entire graduation.
MITUL.M. SHAH
INDEX
 Production & uses of Volatile oils.
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 Evaluation of volatile oils.
1-12
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1. Introduction
 Instrumentation for natural product analysis.
 Essential oil analysis: Principle Criteria.
2. Official methods for determination of essential oils
13-24
 Lemon grass oil.
 Eucalyptus oil.
 Cinnamon oil.
 Chenopodium oil.
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 Nutmeg oil.
 Clove oil.
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 Determination of free alcohols in volatile oil.
 Determination of aldehydes in volatile oil.
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 Test parameters of various pharmacopoeias.
 USP method for volatile oil determination.
3. Reported methods for determination of essential oils
 Lemon grass oil.
25-36
 Eucalyptus oil.
 Cinnamon oil.
 Lavender oil.
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 Sandal wood oil.
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 Clove oil.
4. Different reported spectra of essential oils
37-51
 Chromatograph of cinnamon oil.
 Chromatograms & mass spectras of oils.
 Gas chromatograph of nutmeg oil.
 Specifications for oils.
 1, 8 – cineole reference standard.
 Different spectra of eugenol.
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5. Abstracts
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6. Bibliography
52-54
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1. INTRODUCTION
Volatile or essential oils, as their name implies, are volatile in steam. They
differ entirely in both chemical & physical properties from fixed oils. They
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are secreted in oil cells, in secretion ducts or cavities or in glandular hairs.
Production & uses of volatile oils
Large quantities of volatile oil are produced annually; for 1987, for example it
is estimated that the total world production, in metric tons, was for lemon oil
3000, for eucalyptus oil 2500, for clove leaf oil 2000, for peppermint
6000.Although the production of major oils is highly organized, a no. of
developing countries has volatile oil rich flora fully utilized or cultivated &
the United Nations. Industrial development organization has taken steps to
inform on the setting up of rural based small scale essential oil industries.
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Volatile oils are used for their therapeutic action,
For flavouring (oil of lemon),
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In perfumery (oil of rose),
As starting material for synthesis of other compounds (oil of turpentine),
For therapeutic purposes (eucalyptus oil as inhalation, orally peppermint oil),
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Gargles & mouthwashes (thymol),
Those oils with a high phenol content, example are clove and thyme .They
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have antiseptic properties.
Standardization of volatile oils
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Evaluation:
The evaluation of the crude drugs entering the trade is obviously of
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considerable importance. This operation involves the identification of the
material and the determination of its quality, purity and, if adulterated, the
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nature of the adulterant.
Preliminary Examination: In the case of whole drugs the microscopical
and organoleptic tests are usually sufficient to enable the drug to be
identified.
Foreign Matter: The difficulty of obtaining vegetables drugs in an entirely
pure condition is fully recognized, and pharmacopoeias contain statements as
to the percentage of other of the plant or of other organic matter which may
be permitted.
Microbial Contamination: The official references specify limits of
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microbial contamination of crude drugs. Common tests performed are for the
Total Viable Count (or Total Plate Count) and for organisms such as
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Escherichia coli, Salmonella, Staphylococcus, Yeast and Mould.
Moisture Content: Not only is the purchase of drugs (eg., aloes, gelatin,
gums) which contain excess water, uneconomical, but also in conjunction
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with a suitable temperature moisture will lead to the activation of enzymes
and, given suitable conditions, to the proliferation of living organisms.
Loss on Drying: This method is commonly used for crude drugs to
determine the amount of moisture and volatile oils in the crude drug.
Standardization of volatile oils
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Extractive values: The determination of water-soluble or ethanol-soluble
extractive is used as a means of evaluating drugs the constituents of which are
not readily estimated by other means.
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Ash Values: When vegetable drugs are incinerated, they leave an inorganic
ash which in the case of some drugs (e.g.. rhubarb) varies within fairly wide
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limits and is therefore of lesser value for purposes of evaluation.
Determination of Volatile Oil: Minimum standards for the percentage of
volatile oil present in a number of drugs are prescribed by many
pharmacopoeias.
Refractive Index: If any impurity is present in the liquid the refractive
index of the liquid will change thus showing impurities. Density is also
responsible for changing it.
Optical Rotation: This is the property of a liquid to rotate the plane of
plane polarized light, if there is a change from the standard value there may
be impurity present in the liquid.
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The application of the knowledge of plant histology of crude drugs should
make possible recognition of plant part sections under a microscope. The
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identity of a powder is not regarded as established until it has been compared
with one of known authenticity. Powdered drugs are often tested based on the
presence or absence of characteristic starch grains, epidermal trichomes and
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calcium oxalate crystals.
Standardization of volatile oils
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METHODS FOR NATURAL PRODUCTS ANALYSIS
Common Instruments employed for analysis of natural products are for
and
authentication
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drugs,
determination
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Identification
of phytochemical constituents, detection of impurities and substandard quality
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of drugs. Listed below are some of the analytical techniques we employ for
Herbal and Pharmaceutical Analysis.
Thin layer chromatography
In Chromatography one essentially needs a supporting medium i.e. a filter or
thin, layer of powder (silica gel) spread on glass plate commonly known as
TLC (Thin Layer Chromatography). A solution/ liquid in which the paper of
TLC plate is dipped, is technically called mobile phase which is normally a
mixture of two to five components, and Water which is used for spreading the
adsorbent layer, is called stationery phase. In case of some techniques one
uses silicon fluid, liquid paraffin or standard oil as a stationary phase instead
of water, and, the irrigating liquid is aqueous solution. This technique is
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called reverse-phase chromatography.
The same principles when applied to a column of supporting medium held in
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a tubular container it is called as 'column chromatography'. Very large
columns are used in manufacturing technique. In case of columns where
specific type of polymer powders (called ion exchange resins) is used, it is
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called 'ion-exchange chromatography'. One of the very common examples
of, ion-exchange chromatography is water purification plants producing
highly purified water called 'de-ionized' or 'de-mineralized' water.
AdsorptionChromatography
Because of a physical property of the supporting medium called
"adsorptivity" various chemical ingredients in the sample gets "adsorbed" on
Standardization of volatile oils
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the particles of the supporting medium. The property of "Adsorptivity" is a
surface phenomenon, which due to electrical charge on the surface layer of
the "Supporting Medium". By designing the composition of the "eluting"
solvent, the substance adsorbed can be "eluted" step-by-step i.e. dissolved
chromatography.
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Muffle furnace
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away one after other and collected separately. This is called "Adsorption"
This is used for ash estimation. This is a small cabinet heavily insulated with
ceramic wall in which the heating elements are embedded.
Polarimetry
When an organic substance has an asymmetric carbon atom, i.e. when a
carbon atom in the structure of the molecule has all the 4 valancies occupied
by different groups, it is called "asymmetric" carbon atom, and the substances
has a capacity to change angle of plane of light [polarized light] passing
through its solution by an angle, which is measured using an instruments
called 'polarimeter', and the measure is of the angle of rotation of the plane of
light, called optical rotation. This instrument, called polarimeter, consists of
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source of light (normally Sodium Vapour lamp) which passes through a
polarizes set of prisms. The light emerging from it is uniplaner. This light
passes through solution of the sample which may turn the plane of the light to
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left (levo) or to right (dextro) side of which analysis is done by analyzer set of
prisms which measures the extract angle in which the plane of the light is
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rotated. This angle of rotation of 'Polarized' light is a quality parameter of
liquids (especially, essential oils), where one or more of its components is
optically active, i.e. has 'asymmetric' carbon atom. It can be used for
quantization of solutes which are "optically active".
Turbidometry (Nephelometry)
The colorimeter can be used for measuring of the sample if red filters are used
i.e. in the range of 600-700 nm. Alternatively, when the light is measured
Standardization of volatile oils
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from all the sides placing the photo tubes at the circumference and the light
comes from the bottom of the sample holder. The light is measured at right
angles to the incident light then it is called the Nephelometry or
Turbidometry. There are a very few applications of these techniques,
technique in Vitamin assays.
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Fluorimeter
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important amongst which is measurement of bacterial concentration using the
Fluorescent substances absorb light of a certain wavelength and the light
emerging out is at two different wavelengths. The intensity of which is
measured at right angles to the incident light. Important applications of this
technique are chemical estimation of Vitamin B1& B2. There are many other
applications of the principles of colorimeters like Infra red spectrometry etc.
Which are basically used in investigations of chemical structures and so not
discussed here.
A few techniques related to these principles which are used in analysis are
Flame Photometer and Atomic Absorption Spectrometer
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In these techniques the sample is introduced into a 'flame' using a 'nebulizer'.
When the inorganic atoms get excited and emit light of specific wavelength, it
is proportional to their concentration. The arrangement to introduce the
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solution of the inorganic substance into a flame replaces the light source as
explained in given earlier.
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I.C.P.-MS
There is an advanced in instrument where in place of the burner an electric
are (Plasma) is used and the colour emitted is measured at different
wavelengths. This instrument is called Inductively Coupled Plasma-Mass
Spectrometry. There are other variations of these techniques, an important
example of which is densitometry where an arrangement is provided to hold a
plate on which samples are separated by techniques like chromatography/
Standardization of volatile oils
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electrophoresis, stained if necessary and the colour of the separate bands or
sport is measured at distance of mm or less. This arrangement replaces the
Densitometer
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sample holder compartment of the colorimeter.
in HPTLC, sample separated by simple techniques of TLC are quantified by
measuring colours. Such densitometers are also used to measure protein
Application of
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fractions separated by techniques of Electrophoresis.
Colorimetry is in estimation of colours. The substance itself may be coloured
a substance without colour can be developed into a coloured derivative. The
example given here is estimation of iron from 'Kasis' which itself is not much
coloured but on converting it into a complex with 'Orthophenathroline' gives
a red colour which is measured in a colorimeter.
Extractors & extraction
when one tries to take out i.e. extract components soluble in a solvent, one is
using principle of "diffusion" of the substance from the samples under
extraction into the solvent. In Ayurved, one uses "water" as solvent. This
perhaps because the water solubles are more "saatmya" to body, as it has
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about 80% water in its composition. The process of "diffusion" depends upon
temperature of solvent, and, adequacy of contact between the material and the
solvent, as well as degree of "saturation" of the solvent, making it necessary
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to continuously provide fresh solvent.
Soxhlet Apparatus
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In the usual extractor the condensed solvent passes through the material being
extracted, and returns to the flask. In Soxhlet Apparatus, the bottom of sample
hold is closed, but it connected to the flask through 'Syphon'. This allows
collection of solvent, and contact with the material being extracted. It gets
drained into the solvent flask, when the solvent gets syphoned out.
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Essential oil extraction
this apparatus collects the condensate in a graduated side tube, from where,
the lower (heavier) aqueous phase in continuously returned to the flask. The
oil coming out is measured, using the graduation. This apparatus is also used
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for 'aquametry' as well as for carrying out esterification, when the water
collected does not return, but is separately collected, while the water carrying
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volatile solvent like Benzene/toluene returns. This modification is called
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'Dean-Stark apparatus'
Standardization of volatile oils
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Essential Oil Analysis : Principle Criteria
There are many criteria which can be used to establish the quality of essential
oils. In order to provide quality standards for the essential oil trade, these
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criteria have now been standardized for individual oils by the International
Standards Organization in Geneva. These standards are known as I.S.O.
Standards. Most are produced by individual countries which produce oils and
these are ultimately agreed by the International Authority. The information
required falls into three broad categories:
SENSORY EVALUATION

APPEARANCE

COLOUR

ODOUR
PHYSICAL CHARACTERISTICS
SPECIFIC GRAVITY/ RELATIVE DENSITY

REFRACTIVE INDEX AT 20 %C

OPTICAL ROTATION AT 20% C

FREEZING POINT

FLASH POINT
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
CHEMICAL CHARACTERISTICS

ACID VALUE

ESTER VALUE

ESTER VALUE AFTER ACETYLATION

CONTENT OF CARBONYL COMPOUNDS

CHROMATOGRAPHIC PROFILE. (GC TRACE)
Standardization of volatile oils
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These criteria are not applicable for all oils. Eg. The acid value is not
appropriate for oils that contain lactones. Furthermore, some of these methods
may have to be modified according to the oil being tested. For example, the
ester value after acetylation in pyridine provides a measure of the free alcohol
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content of essential oils containing two types of free alcohol - primary and
secondary. This method however, is not appropriate for oils which contain
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appreciable amounts of tertiary alcohols or other compounds that are subject
to acetylation (phenols, lactones, aldehydes or enolysable ketones). In this
case formic acid is used instead of pyridine.
The Optical Rotation of an essential oil refers to the angle through which the
plane of polarized light is rotated by a layer 10cm. thick of this essential oil at
a specified temperature usually 20%C.
The Refractive Index of an essential oil is the ratio of the sine of the angle of
incidence to the sine of the angle of refraction, when a ray of light of defined
wavelength passes from air into the essential oil kept at a constant
temperature: 20%C for oils that are liquid at this temperature, or up to 30%C
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for oils that have a higher melting point.
GAS CHROMATOGRAPHY - ANALYSIS
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Gas Chromatography (sometimes called Gas Liquid Chromatography) is used
to provide a chromatographic profile of the constituents of essential oils. The
technique is based upon the partitioning of compounds between a stationary
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phase such as silicone and an inert gas phase such as nitrogen. Partitioning of
compounds refers to the ratio amount of the compound dissolved in the
stationary phase, and the amount of the same compound that remains in the
gaseous phase. The stationary phase in bonded on to the interior surface of a
narrow silica glass or steel tube or column which is placed inside a
temperature controlled oven. The temperature of the column, the nature of the
Standardization of volatile oils
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stationary phase, the type of carrier gas and the flow rate of the carrier gas,
are all factors which have an appreciable effect on the separation process.
In order to achieve a separation, the oil sample is injected into the column
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which is heated so that the compounds of interest vaporise. Different volatile
chemical compounds will differ in the ease with which they dissolve in the
stationary phase at a particular temperature, and it is this fact that is used as
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the basis for separation. Eventually an equilibrium is achieved between the
amount of each sample compound dissolved in the stationary phase, and the
amount still present in the gaseous phase. The sample compounds in the
gaseous phase will then pass down and out of the column. .
At this point it is possible to selectively elute off the different sample
compounds in order of their boiling points if the temperature of the column is
gradually raised. The greater the solubility and/or polarity of each compound,
the higher the temperature required to boil them off. Thus the greater the
differences in the solubility and/or polarity of each sample component in the
stationary phase, the better the separation achievable between the compounds
as they can more easily be separately 'boiled off’. Obviously if all the
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compounds have a very similar polarity and similar solubilities in the
stationary phase, they will boil off at similar temperatures and so will not be
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well resolved.
As the oven temperature rises, the individual chemical components of the oil
are thus released from the liquid phase into the gaseous phase and passed out
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of the column. They then pass through a detector chamber which provides
quantitative data on the individual compounds. This data is plotted as a trace
with a peak for each compound, the height of which is related to the
concentration of the compound.
By running standard compounds through the column under identical
conditions and comparing these with the compounds separated from the oil
Standardization of volatile oils
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sample, it is usually possible to work out the composition of the major peaks.
Alternatively, the compounds are passed on to a mass spectrometer which
usually provides an unequivocal identification of each compound.
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The Specific Gravity (mass density) at 20%C of an essential oil is the mass of
a given volume of the oil at 20% The relative density of an essential oil at
20% or 4% is the ratio of the density of the oil at 20%C to that of distilled
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water at 20%C or 4%C. This quantity is expressed as d20/20 or d20/4.
Other parameters may be used which relate to specific oils. There is an I.S.O.
Standard for the total carotenoid content of oil of sweet orange for example.
Carotenoids produce the orange pigment in oranges and carrots. Some oils
may be tested for the quantity of residue (% mass) after evaporation of the
volatile component under standard conditions.
Analysis: Principal Criteria Essential Oil Analysis: Principal Criteria
essential Oil Analysis: Principal Criteria Essential Oil Analysis:
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Principal Criteria
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2. Official methods for determination of essential
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Lemon grass oil
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oils
Lemon grass oil is the oil which is distilled from cymbopogon flexuous. It
contains not less than 75.0%w/w aldehydes, calculated as CITRAL, C10H18O.
Description:
Reddish yellow to brown mobile oil. Odour that of lemon oil.
Solubility:
Almost entirely soluble in 3 part of alcohol [70%] the solubility gradually
decreasing on storage wt. per ml. at 250 0.892 to 0.9099 optical rotation -30
to +10 refractive index at 250 , 1.4808 to 1.4868.
Assay:
Carry out the determination of aldehydes in volatile oil lemon grass oil.
(Refer page 21)
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Each ml. of 0.5 N KOH in alcohol [60%] is equivalent to 0.07672g of citral.
Storage:
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Preserve lemon grass oil in a well closed container protected from light &
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store in a cool place category insect repellent.
Standardization of volatile oils
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Hindi: Neeli Gond ka tel
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Eucalyptus oil
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Eucalyptus oil is the oil distilled from the fresh leaves of eucalyptus globules
labill.from other species of eucalyptus & rectified. It contains not less than
60.0%w/w of cineole, C10H18O.
Solubility: Soluble in one volume of alcohol (80%).
Weight per ml: At 250, 0.901 to 0.920 gm
Optical rotation: -50 to +100
Refractive index: at 250 , 1.457to 1.469
Heave metals: Shake 10ml with 10ml of water & 1 drop of HCL & pass
H2S through the mix. Until it is saturated no darkening in colour is produced
in the oil or water.
Phellandrone: Mix. 1ml. with 2ml. of glacial acetic acid & 5ml. of light
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petroleum (B.P 600 to 800) add 2ml. of a saturated solution of sodium nitrite &
shake the mixture gently no crystalline precipitate forms in the upper layer
Aldehydes: Carryout the method described under lemon oil for the
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determination of aldehydes in volatile oil is using 10ml. of eucalyptus oil with
4ml. of hydroxylamine hydrochloride reagent in alcohol (60%) & 5ml. of
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benzene; not more than 2ml. of 0.500 KOH in alcohol (60%) is required.
Assay: Carry out the method for the determination of cineole.
Storage: Preserve eucalyptus oil in a well closed container protected from
light & store in a cool place.
Category: Counter irritant mild expectorant.
Dose: 0.06to 0.2ml.
Standardization of volatile oils
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Cinnamon oil
Cinnamon oil is the oil distilled from cinnamon. It contains not less than
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55.0% w/w &not cinnamic aldehyde C9H80.
Description: A yellow liquid when freshly distilled gradually becoming
reddish brown with age.
Solubility: Soluble, at 1550
in 3 volumes of alcohol (70%) the solution
showing not more than a slight opalescence.
Wt. per ml: At 200, 0.94 to 1.034g.
Refractive index: At 250, 1.545 to 1.575.
Optical rotation: 00 to -20
Heavy metals: complies with the test for heavy metal;
Cinnamon leaf oil; cassia oil- dissolve one drop in 5ml. of alcohol (9%) &
add one drop of test solution of FeCl3, a slight green , but not a blue or a
deep brown colour may be produced.
Carry
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Assay:
out the assay by the method for the determination of
aldehydes in volatile oil in cinnamon oil. (Refer page 21)
Preserve cinnamon oil in a well closed container protected from
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Storage:
light & store in cool place.
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Category: Carminative.
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Dose: 0.06 to 0.2ml.
Standardization of volatile oils
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Chenopodium oil
Chenopodium oil is mixed oil obtained by steam distillation from the fresh
flowering & fruiting plants excluding roots of various species of C.
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ambrosioides. It contains not less than 65.0% w/w of ascaridoles C10H16O2.
Description: A colorless or light yellow liquid.
Solubility: Soluble at 200, in 10 vol. of alcohol (70%).
Identification:
Heat 1ml. to incipient ebullition in a test tube with a
fragment of unglazed porcelain, remove the flame & cool; adecpyolden
yellow liquid is produced. (This test should be carried out very cautiously as
the oil is liable to explode)
Wt. per ml: At 200, 0.955 to 0.975g.
Refractive index: At 200, 1.474 to 1.480.
Optical rotation: -30 to -80
Assay: Weigh accurately about 25gm. & dissolve in sufficient acetic acid
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(90%) to produce 50ml. & place the solution in a burette. In a stoppered tube
of about 60ml. capacity, place 3ml. of an 83.0% w/w solution of KI. 5ml. of
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HCL & 10ml. of glacial acetic acid, immerse the tube in a freezing mixture.
Until the temperature is reduced to -30 then add 5ml. of the CH3COOH.
Solution of the oil mixing it with the reagent as quickly as possible & making
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due allowance for the draining of the burette set aside in a place below 100 for
five minutes & without diluting, titrate the liberated I2 with 0.1N sodium
thiosulphate. Repeat the experiment with the same quantities of the same
reagents in the same manner omitting the oil, but diluting the solution with
20ml. of H2O before titrating the liberated I2. The difference between two
titrations represents the iodine liberated by ascaridole.
Standardization of volatile oils
16
Storage: Preserve the oil in a well closed container, protected from light &
store in a cool place.
Dose: 0.2 to 1ml.
bn
et
.a
c.
Nutmeg oil
in
Category: Anthelmintic
Nutmeg oil is the volatile oil distilled with steam from the dried kernels of the
ripe seeds of myristica fragrance.
Description: A colourless or pale yellow liquid.
Solubility: Soluble in 3 volume of alcohol.
Wt. per ml: At 250, 0.880 to 0.910g for East Indian oil 0.854 to 0.880 for
West Indian oil.
Optical rotation: + 80 to + 300 for East Indian oil & 250 to + 450 for West
Indian oil.
Refractive index: At 200, 1.4740 to 1.4880 for East Indian oil. 1.4690 to
fli
1.4760 for West Indian oil.
Non volatile matter: when evaporated rapidly in flat bottomed nickel dish
9cm in diameter &1.5cm in depth on a boiling water bath, 3ml leaves not
in
more than 60 mg of the residue for East Indian oil & 50mg for West Indian
oil.
gn
u.
Storage: Preserve in well closed container, protected from light & store in
cool place.
Category: flavouring agent.
Dose: 0.06 to 0.2 ml.
Standardization of volatile oils
17
in
Clove oil
Clove oil is the oil distilled from clove. It contains not less than 85.0% v/v &
bn
et
.a
c.
not more than 90% v/v of eugenol C10H12O2.
Description:
A colourless or a pale yellow liquid when freshly distilled,
becoming darker & thicker by ageing or exposure to air; odour &taste those
of clove.
Solubility: soluble in two volumes of alcohol (70%)
Wt. per ml: At 250, 1.038 to 1.060 g.
Refractive index: At 250, 1.5300 to 1.5310
Optical rotation: 00 to -1.50
Phenol- Shake 1 ml with 20 ml of hot water; the water shows not more than
scarcely perceptible acid reaction with blue litmus paper. Cool the mixture,
pass the water layer through a wetted filter, &treat the clear filtrate with one
drop of test solution of FeCl3.The mixture has only transient grayish green
fli
colour, but not a blue or violet colour.
Assay: The volume of the unabsorbed oil measures not less than 1.0ml. &
in
not more than 1.5ml, indicating the presence of not less than 85% v/v not
more than 90% v/v of eugenol .Determination of free alcohols in volatile oil.
Carry out the method described for the determination of esters & calculate the
gn
u.
ester value of oil.
Storage: Preserve clove oil in a well closed container protected from air &
light & storage in a cool place.
Category: Dental obtundent.
Dose: 0.06 to 0.2ml.
Standardization of volatile oils
18
DETERMINATION OF FREE ALOCOHOLS
bn
et
.a
c.
in
IN VOLATILE OILS
1) Acetylate the oil by the following method; - mix 10ml, of the oil, 20ml, of
acetic anhydride and 2g. of freshly fused anhydrous sodium acetate in a long
necked, round-bottomed 200ml, flask, attached to an air-cooled reflux
condenser. Support the flask on a sheet of asbestos in which a hole about 4 cm,
in diameter has been cut and heat it with a small naked flame, not more than
25mm, in height , which does not impinge on the bottom of the flask . Boil
gently for two hours, allow the flask to cool, and add 50ml of water; then with
reflux condenser in position. Heat on a boiling water-bath for fifteen minutes
with frequent and through shaking. Cool, and transfer the contents of the flask
to a separator, and reject the lower layer. Wash the acetylated oil successively
with 1) 50ml, of brine; 2) 50ml brine, containing in solution 1g. of anhydrous
fli
sodium carbonate; 3) 50ml, of brine. At each washing shake vigorously, and
allow separation to take place completely, before rejecting the lower layer. The
aqueous layer from the final washing is alkaline to solution of phenolphthalein
in
finally, shake gently with20ml, of water, and remove the water layer as
completely as possible. Pour the acetylated oil into a small dish, and 3g. Of
gn
u.
powdered anhydrous sodium sulphate; stir frequently during fifteen minutes or
until a drop of the oil produces no cloudiness when added to 10 drops of carbon
disulphide in a dry tube. Filter the oil through a dry filter paper in a covered
funnel.
2) Determine the ester value of the acetylated oil, using about 2g. Accurately
weighed, by the method for the determination of esters; and calculate the ester
value of the acetylated oil from the formula given.
Standardization of volatile oils
19
3) The percentage of free alcohols is then obtained from the following
formula:(b –a) y
0.42*(1335-b)
Where, a=ester value of oil,
bn
et
.a
c.
b=ester value of acetylated oil,
in
Percentage of free alcohols =
y=molecular weight of the alcohol
gn
u.
in
fli
Value of borneol, 154.3; menthol, 156.3
Standardization of volatile oils
20
bn
et
.a
c.
VOLATILE OIL
in
DETERMINATION OF ALDEHYDES IN
1) LEMON OIL
Weigh accurately about 10 gm of the oil into a stoppered tube
approximately 25mm in diameter and 150mm in length; add7ml of
hydroxylamine hydrochloride reagent in alcohol (60%) and one drop of
solution of methyl orange; shake & neutralize the liberated acid with 0.5N
potassium hydroxide in alcohol (60%) until the red colour changes to yellow;
continue shaking & neutralizing, until the full yellow colour of the indicator
is permanent in lower layer, after shaking vigorously for two minutes &
allowing separation to take place. The reaction is completed in about 15
minutes.
fli
Each ml of 0.5N KOH in alcohol (60%) is equivalent to 0.07672 gm of citral.
This procedure gives an approximate determination of the citral in the oil.
Carryout a second determination in exactly the same manner, using, as the
in
colour standard for the end point, the titrated liquid of the first determination
with the addition of 0.5ml of 0.5N potassium hydroxide in alcohol(60%).
gn
u.
Calculate the accurate value from the second determination.
NOTE. -
The volume of the hydroxylamine hydrochloride reagent in
alcohol (60%) used is varied according to the citral content of the oil, and
must exceed by 1 or 2 ml the volume of 0.5N potassium hydroxide in alcohol
(60%) required.
Standardization of volatile oils
21
2) CINNAMON OIL
Carry out the process described for lemon oil using about 1gm,
in
accurately weighed, of the cinnamon oil with 5ml of benzene, and from 10 to
12mlof hydroxylamine hydroxide reagent in alcohol (60%), according to the
bn
et
.a
c.
aldehyde content of the oil. The volume of hydroxylamine hydrochloride
reagent in alcohol (60%) used must exceed by 1 to 2ml the volume of 0.5N
potassium hydroxide in alcohol (60%) required.
Each ml of 0.5N KOH in alcohol (60%) is equivalent to 0.06661 gm of
cinnamic aldehyde
3) LEMON GRASS OIL
Carry out the process described for lemon oil, using about 1g,
accurately weighed, of the lemon grass oil, with 5mlof benzene and 10
to15ml of hydroxylamine hydrochloride reagent in alcohol (60%), according
to the aldehyde content of the oil. The volume of hydroxylamine
fli
hydrochloride reagent in alcohol (60%) used must exceed by 1 to 2 ml the
volume of 0.5N potassium hydroxide in alcohol (60%) required.
Reagents
gn
u.
in
Each ml of0.5N KOH in alcohol (60%) is equivalent to 0.07672g of citral.
0.5N potassium hydroxide in alcohol (60 %)
A solution containing in 1,000ml 28.05g of potassium hydroxide prepared
with alcohol (60%), and standardized by means of 0.5N hydrochloric acid by
running the alkali into the acid, until the full yellow colour of the indicator, a
0.2%W/V solution of methyl orange in alcohol (60%), is obtained.
Standardization of volatile oils
22
B
SR.
PARAMETER
H
P
2
3
4
5
6
7
Macroscopic Description
1991
J.S.H.M. 1993
A.P.I
W.H.O.
Sensory evaluation (Touch,
+
-
Organoleptic
+
+
-
+
+
Quantitative Microscopy
+
-
+
+
-
+
+
+
+
+
+
-
Purity tests
+
+
+
-
And in Ether
+
+
-
-
-
-
+
+
+
Identification
+
-
bn
et
.a
c.
1
B.H.C.
in
Below is a compilation of the test parameters recommended by
various Pharmacopoeias for Crude Drugs /Herbs.
(Morphology)
Microscopy of the section
Ash (Total, Acid insol.,
Water soluble)
Moisture
Foreign matter Extractives
in water
Methanol extractives
Finger printing of water
extractives
Odour, Test)
Specific active or marker
component (Alkaloid,
Glycoside, Tannin,
fli
8
Phenolics)
Fixed oil/ essential oil
resinoid (their related
in
9
+
-
+
+
+
-
-
-
-
+
tests)
Heavy metals Pb, Cd
gn
u.
10
11
Toxic Metals As, Hg
SPECIAL TESTS
-
-
-
-
+
-
-
-
-
+
Organic Chlorine/
12
Phosphorus (Pesticides
residue)
13
Swelling factor
-
-
-
-
+
14
Foaming index
-
-
-
-
+
Standardization of volatile oils
23
16
Microbiology
Radio activity (as per
Vienna Conventions)
+
-
-
-
+
-
-
-
-
+
Abbreviations
British Herbal Pharmacopoeia
BHC
bn
et
.a
c.
BHP
in
15
British Herbal Compendium
JSHM Japanese Standard for Herbal Medicines
API
Ayurvedic Pharmacopoeia of India
WHO
World Health Organization, Guidelines
USP method for volatile oil determination
Set up a one liter round bottom flask, (short necked) on a heating mantle
fli
over a magnetic stirrer. Insert an egg shaped stirring bar magnet in the flask,
& attach a cold finger condenser & an appropriate volatile trap of the type
in
illustrated.
Coarsely comminute a sufficient quantity of the drug to yield from one
to three ml of volatile oil. Small seeds, fruits, or broken leaves of herbs
gn
u.
ordinarily do not need comminution. Very fine powders are to be avoided. If
this is not possible, it may be necessary to mix them with purified sawdust or
purified sand. Place a suitable quantity of the drug, accurately weighed, in the
flask & fill it half with water. Attach the condenser & the proper separator.
Boil the contents of the flask, using a suitable amount of heat to
maintain gentle boiling for two hours, until the volatile oil has been
completely separated from the drug & no longer collects in the graduated tube
of the separator.
Standardization of volatile oils
24
LEMON GRASS OIL:
Botanical Name: Cymbopogon citratus (DC.) Stapf
bn
et
.a
c.
Family: Poaceae
in
3. Reported methods for determination of essential oils
Vernacular Name: Lemon grass, Lomi sar (Amharic)
General: Cymbopogon (Poaceae) represents an important genus of about 120
species and several varieties. Cymbopogon species are well known as a
source of commercially valuable compounds like geraniol, geranyl acetate,
citral (neral and geranial), citronellal, piperitone, eugenol, etc., which are
either used as such in
perfumery and allied industries, or as starting
materials for the synthesis of other products commonly used in perfumery
(Shahi and Tava, 1993).
Previous Work:
Lemongrass oil is the commercial name given for the volatile oil obtained
from Cymbopogon citratus (DC.) Stapf. In contrast to the present study (see
fli
below), geraniol was reported to be the major constituent of lemongrass oil
growing in Ethiopia (Abegaz et al., 1983). It is well known that lemongrass
in
oil is the most important source of citral, which is the starting material for the
preparation of ionone. The oil is also used in low cost perfumes, for soap and
other laundry products.
gn
u.
Present Work:
For this study, the plant material was collected from Wondo Genet, Essential
Oil Research Center (EORC) farm site. Citral A (47.7%), citral B (33.2%)
and myrcene (10.7%) were found as the major constituents.
Plant Source: Essential Oil Research Center (EORC), Addis Ababa, Ethiopia
Serial Number: HP GC, 0495
Standardization of volatile oils
25
Components Identified: 1) myrcene (10.7%); 2) citral B (33.2%); 3)
fli
bn
et
.a
c.
GC Chromatogram:
in
geraniol (2.4%); 4) citral A (47.7%) [All the peaks confirmed by GC-MS].
gn
u.
in
Structure of Major Compounds:
Standardization of volatile oils
26
Botanical Name: Eucalyptus globulus Labill.
Family: Myrtaceae
Vernacular Name: Blue Gum Eucalyptus, Netch Bahrzaf
in
Eucalyptus oil:
bn
et
.a
c.
General: Eucalyptus trees with over 700 species are native to Australia and
are widely grown in many parts of the world for production of timber, fuel
wood, and essential oil as a source of nectar in honey production. In 1895 a
French Forester, Mondon-Vidaillet, brought seeds of several Eucalyptus
species to Ethiopia upon the request of Emperor Menelik II and found two
species to be particularly suitable, Eucalyptus globulus Labill. growing on the
highland areas and Eucalyptus camaldulensis Dehnh. For the lowland areas
(Zrira et. al, 1996).
PreviousWork:
The essential oil of Eucalyptus globulus Labill. contains 1, 8-cineole as its
major constituent. Monoterpene hydrocarbons (e.g. alpha-pinene) and
oxygenated monoterpenes (e.g. 4-terpineol) are also present (Dagne et al.,
fli
2000).
Present Work:
in
Same as above.
Plant Source: Essential Oil Research Center (EORC), Addis Ababa, Ethiopia
Serial Number: HP GC, 0422
u.
Components Identified: 1) alpha-pinene (19.0%); 2) 1, 8-cineole (71.0%);
3) alpha-terpineol (1.3%); 4) alpha-terpinyl acetate (2.5%); 5) caryophyllene
gn
oxide (1.3%) [All peaks confirmed by GC-MS].
Standardization of volatile oils
27
bn
et
.a
c.
in
GC Chromatogram:
u.
in
fli
Structure of Major Compounds:
gn
Other Data: Optical rotation: -15 (neat); Yield: 1.1% (w/w).
Standardization of volatile oils
28
Botanical Name: Cinnamomum zeylanicum Blume
Family: Lauraceae
bn
et
.a
c.
Vernacular Name: Cinnamon, Kerefa (Amharic)
in
Cinnamon oil:
General: Cinnamomum cinnamon comprises 250 species and is widely
distributed throughout the tropical countries, and used to flavour food, baked
goods, beverages and pharmaceutical preparations
Previous Work:
Cinnamaldehyde and eugenol were reported to be the main constituents in the
oil and extract of Cinnamomum zeylanicum Blume (Tateo and Chizzini,
1989).
Present Work:
As in the literature the analysis here also shows, cinnamaldehyde, eugenol
and eugenyl acetate to be the main constituents of cinnamon oil.
Plant Source: Addis Ababa Market, Ethiopia
fli
Serial Number: HP GC, 0529
Components Identified: 1) cinnamaldehyde (96.7%); 2) eugenol (0.5%); 3)
gn
u.
in
eugenyl acetate (2.2%)
Standardization of volatile oils
29
bn
et
.a
c.
in
GC Chromatogram:
u.
in
fli
Structure of Major Compounds:
gn
Other Data: Yield: 2% (W/W)
Standardization of volatile oils
30
Clove oil:
Family: Myrtaceae
Vernacular Name: Clove, Kerunfud (Amharic)
in
Botanical Name: Eugenia caryophyllus (Spreng.) Bullock & S. G. Harrison
bn
et
.a
c.
General: The biggest clove producing country is Tanzania, which produces
80% of the total world production. Clove oil is mainly used as flavouring
agent and also in perfumery and medicines. Medicinal oil has a phenol
content of about 82 to 90 per cent. The oils which are relatively low phenol
content are mainly used in pharmacy while the strong oils are used in
manufacture of Vanillin (Chopra and Kapur, 1982)
Previous Work:
The steam distillate of clove oil contains phenols, sesquiterpenes and small
quantity of esters, ketones and alcohols (Chopra and Kapur, 1982).
Present Work:
In this study, clove buds originating from Pemba were hydro distilled and
analyzed. Eugenol and eugenyl acetate were found to be the major
fli
constituents.
Plant Source: Tanzania
in
Serial Number: HP GC, 0526
gn
u.
Components Identified: 1) eugenol (75.3%); 4) eugenyl acetate (4.4%)
Standardization of volatile oils
31
bn
et
.a
c.
in
GC Chromatogram:
u.
in
fli
Structure of Major Compounds:
gn
Other Data: Optical rotation: -13 (1g/100mL CHCl3).
Standardization of volatile oils
32
Botanical Name: Lavandula latifolia Medik.
Family: Lamiaceae
bn
et
.a
c.
Vernacular Name: Spike Lavender (from Ethiopia)
in
Lavender oil:
General: Lavender oil is obtained by steam distillation from the fresh
flowering tops of the dwarf shrub Lavandula officinalis Chaix, growing in
Mediterranean area, Europe, North America and North Africa. It has a fresh
sweet, floral-herbaceous odour and is used extensively in perfumes, colognes,
toiletry articles, etc. (Formacek and Kubeczka, 1982). Lavandin oil is
obtained from Lavandula hybrid Rev., a hybrid between Lavandula latifolia
Medik. (Spike lavender) and Lavandula officinalis Chaix (true lavender).
Spike lavender oil is obtained from Lavandula latifolia Medik. (Masada,
1976)
Previous Work:
Spike lavender contains linalool, camphor and 1,8-cineole as major
components (Masada, 1976).
fli
Present Work:
Spike lavender from Addis Ababa was found to contain 1, 8-cineole, linalool,
in
camphor and linalyl acetate. The oil was initially named to be Lavender oil
but from its constituents though not botanically identified it agree with the
name given.
gn
u.
Plant Source: Asni Gallery, Addis Ababa, Ethiopia
Serial Number: HP GC, 0657
Components Identified: 1) alpha-pinene (1.4%); 2) camphene (0.5%); 3)
beta-pinene (3.7%); 4) 1, 8-cineole (39.4%); 5) linalool (3.4%); 6) camphor
(32.3%); 7) linalyl acetate (4.2%) (All peaks confirmed by GC-MS; peaks 4
and 6 also confirmed by C-13 NMR).
Standardization of volatile oils
33
in
fli
bn
et
.a
c.
in
GC Chromatogram:
gn
u.
Structure of Major Compounds:
Standardization of volatile oils
34
Botanical Name: Santalum album L.
Family: Santalaceae
bn
et
.a
c.
Vernacular Name: Sandal wood
in
Sandalwood oil:
General: Sandalwood oil (East Indian oil) is derived from the evergreen tree,
Santalum album, growing in India and Malaysia. Sandalwood oil is used in
perfumery, because of its characteristic woody oriental note and its fixative
properties (Masada, 1976).
Previous Work:
The most important chemical components of sandalwood oil are alpha-and
beta-isomers of santalol (Masada, 1976).
Present Work:
The major constituent of the oil is santalol (90%).
Plant Source: South Africa
Serial Number: HP GC, 0480
fli
Components Identified: 1: beta-ionone (2.5%) 2: santalol (95.5%) (Peaks
gn
u.
in
confirmed by GC-MS and NMR).
Standardization of volatile oils
35
bn
et
.a
c.
in
GC Chromatogram:
gn
u.
in
fli
Structure of Major Compounds:
Standardization of volatile oils
36
4. Different reported spectra of essential oils
bn
et
.a
c.
in
Chromatogram of cinnamon bark oil.
Chromatogram of Cinnamon Bark Oil
in
fli
Column Length 10 m, column I.D. 1 mm, packing Partisil Silica Gel, 20
m, mobile phase 3% v/v ethyl acetate in n-heptane, flow rate 38 l/
min., sample volume 0.5 l, efficiency 160,000 theoretical plates
gn
u.
The column was 10 m long, 1 mm I.D. packed with Partisil Silica Gel 20
m particle diameter. At the optimum flow rate (i.e., 10 l/min.) the
column gave a quarter of a million theoretical plates. However, the
chromatogram shown in figure was obtained at a flow rate of 38 l/min.
and, thus, as it was operated well above its optimum velocity; the
column only gave an efficiency of 160,000 theoretical plates.
Standardization of volatile oils
37
As the chromatographic data was acquired and processed by a computer
portions of the chromatogram could be expanded and these are shown as
inserts in the figure. It is seen that the apparently confused peaks at the
start of the chromatogram are, in fact, well resolved into individual
in
small peaks. It is also seen that the late small peak has retained its
symmetry and is almost perfectly Gaussian in shape. Those familiar with
bn
et
.a
c.
cinnamon bark oil separated on GC capillary columns may wonder at
the relatively few peaks that appear on the chromatogram. It should be
pointed out that a UV detector was employed to monitor the separation
and, thus, only, those substances that adsorb in the UV would be
disclosed. As the majority of the substances in essential oils are UV
transparent, only a limited number of the components will be detected.
The example is given to illustrate the wide range of solutes that can be
separated and that, providing adequate efficiency is available together
with suitable apparatus, multi component mixtures can be separated by
gn
u.
in
fli
LC as well as GC.
Standardization of volatile oils
38
gn
u.
in
fli
bn
et
.a
c.
in
Chromatograms & mass spectra.
FIGURE 1.
Standardization of volatile oils
39
in
(A) CHROMATOGRAM OF AN EXTRACT OF BLUE GUM EUCALYPTUS (EUCALYPTUS
GLOBULUS) FOR EXPERIMENTAL CONDITIONS DESCRIBED IN THE TEXT; THE INSET IS
THE STRUCTURE OF CINEOLE (EUCALYPTOL).
(B) MASS SPECTRUM OF THE CHROMATOGRAPHIC FRACTION THAT ELUTED AFTER 7
MINUTES AS SHOWN IN A.
(C) MASS SPECTRUM OF PURE CINEOLE OBTAINED FROM THE HP LIBRARY.
bn
et
.a
c.
Blue Gum Eucalyptus (Eucalyptus Globulus Var. Globulus)
A representative chromatogram of an extract of fresh eucalyptus leaves is
shown in Figure 1A. The largest peak, which eluted after 7 min, was
identified as cineole, also known as eucalyptol (1, 3, 3-trimethyl-2-oxabicyclo
[2.2.2] octane; MW = 154.24), and its structure is shown in the upper right
hand corner of this figure. This compound has therapeutic activity as an
inhalation expectorant .The mass spectrum of this chromatographic peak and
a mass spectrum of pure cineole from the HP library are shown in Figures 1B
and 1C, respectively. Other minor compounds identified from the mass
gn
u.
in
fli
spectra include pinene (insecticide), camphene, and caryophyllene (perfume).
Standardization of volatile oils
40
in
bn
et
.a
c.
fli
in
u.
gn
FIGURE 2.
Standardization of volatile oils
41
Saltwort (Chenopodium Abrosioides)
in
(A) CHROMATOGRAM OF AN EXTRACT OF SALTWORT (CHENOPODIUM
ABROSIOIDES) FOREXPERIMENTAL CONDITIONS DESCRIBED IN THE TEXT; THE
INSET IS THE STRUCTURE OF ASCARIDOLE.
(B) MASS SPECTRUM OF THE CHROMATOGRAPHIC FRACTION THAT ELUTED AFTER
12 MINUTES, AS SHOWN BY THE ASTERISK IN A.
(C) MASS SPECTRUM OF PURE ASCARIDOLE OBTAINED FROM THE HP LIBRARY.
shown
bn
et
.a
c.
A representative chromatogram of an extract of Chenopodium abrosioides is
in
Figure
2A.
Ascaridole
(1-methyl-4-(methyl
ethyl)-2,3-
dioxabicyclo[2.2.2]oct-5-ene; MW = 168.23), which eluted after about 12
min, has previously been reported to constitute up to 60–80% of the essential
oil of saltwort . Ascaridole is an anthelmintic (antiparasitic) that has been
used in the treatment of roundworm. Although detected, it did not appear as
the major constituent of our extract probably due to the manner in which the
extract was prepared. The experimentally determined mass spectrum of the
peak that eluted after 12 minutes in Figure 2A and the mass spectrum of pure
ascaridole as extracted from the HP library are depicted in Figures 2B and
gn
u.
in
fli
2C, respectively.
Standardization of volatile oils
42
in
bn
et
.a
c.
fli
in
u.
gn
FIGURE 3.
Standardization of volatile oils
43
in
(A) CHROMATOGRAM OF AN EXTRACT OF SPEARMINT (MENTHA SPICATA) EXTRACT
FOR EXPERIMENTAL CONDITIONS DESCRIBED IN THE TEXT; THE INSET IS THE
STRUCTURE OF MENTHOL.
(B) MASS SPECTRUM OF THE CHROMATOGRAPHIC FRACTION THAT ELUTED AFTER
8 MINUTES, AS SHOWN IN A.
(C) MASS SPECTRUM OF PURE MENTHOL OBTAINED FROM THE HP LIBRARY .
bn
et
.a
c.
Spearmint (Mentha Spicata)
Both a fresh spearmint extract and an extract prepared from pure peppermint
tea purchased at a nutritional food store yielded similar chromatograms. A
representative chromatogram of Mentha spicata extract is shown in Figure
3A. The largest peak, which eluted after approximately 8 min, was identified
as menthol by mass spectrometry. Other minor compounds identified in
spearmint
include
methyl
acetate
(perfume),
menthone
(perfume),
caryophyllene, cineole, and pulegone (antifungal). The mass spectrum of the
largest chromatographic peak, and a mass spectrum of pure menthol (5-
methyl-2-(1-methylethyl) cyclohexanol; MW = 156.26) obtained from the HP
library are shown in Figures 3B and 3C, respectively. Menthol has been used
fli
externally as a mild local anesthetic and antiseptic, and it has been used
gn
u.
in
internally as a carminative and gastric sedative.
Standardization of volatile oils
44
bn
et
.a
c.
in
Chromatograph of nutmeg oil
The volatile oil from the seed contains:
fli
1. (2902) -pinene
2. sabinene
in
3. (2903) -pinene
4. myrcene
5. (2633) limonene
gn
u.
6. -terpinene
7. (2248) terpinen-4-ol
among the major ingredients. The numbers in brackets are the FEMA
codes (Flavor and Extract Manufacturers' Association of the USA).
A recent study on nutmeg oil from St Catherine, Jamaica and other West
Standardization of volatile oils
45
Indian nutmeg oils revealed significant differences that could be used to
distinguish between them.
Specification
EUCALYPTUS OIL GLOBULOUS
1.459 at
30°C
Cineole content by O. Cresol method
61.8%
Cineole content by GLC
66%
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Refractive Index
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Specifications for oils
Specification
CUMIN OIL
Light Yellow to Brown
Odour
Disagreeable
Sp. Gravity at
30°C
0.8788
Ref. Ind. At
30°C
1.4848
Optical Rotation
+ 4.5
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Colour
Specification
PEPPERMINT
Colour
Yellow
Odour
Strong Penetrating Peppermint
Odour and pungent taste
Sp. Gravity
0.896 to 0.908
Ref. Index
1.460 to 1.470
Optical Rotation
-18° to -30°
Standardization of volatile oils
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Yellow Liquid
Odour
Spicy Odour
Sp. Gravity at
15°C
0.8959 to 0.9168
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Colour
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Specification
BASIL OIL
Ref. Ind. At 20°C
1.477 to 1.488
Specification
LEMON GRASS
OIL
Brownish yellow coloured
oily liquid having pleasant
odour.
Specific Gravity
0.8889 at 30°C
Optical Rotation
0°
Refractive Index
1.4835 at 30?c
Citral content by GC
76%
Solubility in 70%
Ethyl Alcohol
2 vol.
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Description
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Specification
THYME OIL
Colour
Red Liquid
Odour
Pleasant Odour and pungent
persistent taste
Sp. Gravity at 27°C
0.9494
Ref. Ind. At 27°C
1.5096
Optical Rotation
-3.0
Standardization of volatile oils
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Spectra of Eugenol
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IR SPECTRA
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Eugenol - Proton NMR Spectrum
Standardization of volatile oils
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in
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Eugenol - Carbon 13 NMR Spectrum
Standardization of volatile oils
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in
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Eugenol - Mass Spectrum
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1).
Volatile
oil
constituents
of
the
essential
oil
of
Santolina
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Chamaecyparissus linn. from the southern hills of India.
in
5. Abstracts
Garg, S.N.; Gupta, D.; Mehta, V.K; Sushil Kumar.
The composition of the essential oil of the aerial parts of the Santolina
Chamaecyparissus plants collected from the campus of CIMAP field
Station.Kodaikanal was analysed by GC &GC/MS. The oil yield obtained
upon hydrodistillation was 1.1% (V/W) on fresh weight basis. The analysis of
the oil led to the identification of 25 constituents totaling 97.4 % of the oil.
Through MAPA 2002-01-0490
2). Investigations of the content & the composition of essential oil found
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in leaves & gallenic preparation from sage.
Ludwiczuk, A; Wolski, T.; mardarowicz, M.
in
The investigations were done by GC/MS method.
Through MAPA 2002-01-05-09
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3).Composition of the essential oil of microglosa pyrrhopappa.
Mwangi, J.W; Thoithi, G.N.; Juliani, H.R; zygaldo, J.A.
The essential oil of the microglossa pyrrhapappa var. Analysis of the oil is
done by GC & GC/MS reveals.
Through MAPA 2002-01-0524
Standardization of volatile oils
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4). Chemical analysis, anti microbial activity, & the essential oils from
in
some wild herbs in Egypt.
A1-Gaby, A.M; Allam, RF. (Dept of biochemistry, Faculty of agriculture)
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The composition & anti microbial activity of the essential oils in aerial parts
of Achillae fragrantissima, Artemisia Judaica. collected from southern Sinai
were determined. The volatile oil yield of A.fragrantissima was 0.81% V/W
& consisted of 18 components with Thujone (57.5%) & santolina alcohol
(31.4%) as the major constituents.A.Judaica yielded 1.57% V/W volatile oil
with camphor, 37.3% & piperitone (27.4%) as the major constituents of 20
components. The volatile oil of P.tortuous (0.56% V/W) consisted of 32
components with camphene (31%) as the major constituent, although the
alcoholic components constituted more than 42% of the oil.
Through MAPA 2000-04-2381
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5). Essential oils of annual Siderites species growing in Turkey.
Kirimer, N.; Tabanca, N.; Ozec, T.; Tumen G.; Baser, K.H.C (medicinal &
in
aromatic plant & Drug Research Centre)
Water distilled essential oils of 5 annual siderites species Viz., S. Lanata,
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S.Curvidens, S.Montana ssp. Montana, S.montana ssp. remota & S.Romana
ssp. Romana. Collected from different regions of Turkey analysed by GC/MS
have been tabulated & compared with main components of the essential oils
of perennial siderites species from Turkey.
Through MAPA 2000-04-2446
Standardization of volatile oils
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6). The essential oil of Thymus macedonicus. Subsp. Macedonicus (Deg.et
urum) Ronn. From Macedonia
Faculty of Pharmacy.
in
Kulevanova, S.; Ristic. M.; Stafilov, T.cC institute of pharmacognosy,
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The composition of the essential oils Macedonian Thymus macedonicus ssp.
Macedonicus varied in eight different populations of the taxa.
Through MAPA 2000-04-2452
7). Antibacterial & antifungal studies of essential oil of strobilanthus
Lx iocephala
The essential oil was studied for various physicochemical properties. It has
shown density (0.9009), specific gravity (1.732), and refractive index (1.484).
TLC of essential oil on silica gel G in pure benzene revealed four major & six
minor spots, when spread with 1% vanillin H2SO4.GC-MS study of the
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essential oil indicated total 12 components.
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Through Indian drugs.
Standardization of volatile oils
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6. Bibliography
Delhi; 1966
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 Pharmacopoeia of India, Second edition, Manager of publications,
 Indian Herbal Pharmacopoeia, Indian drug manufacturer’s
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association; edition 1998
 C.K.Kokate, A.P.Purohit, & S.B.Gokhale; Pharmacognosy,
nineteenth edition, Nirali Prakashan;2002
 W.C.Evans, Trease & Evans; Pharmacognosy, Fifteenth
Edition; W.B.Saunders:2002
 The United status pharmacopoeia, XXIII.Rockville, MD: The
United Status Pharmacopoeia convention, Inc; 1990
 S.K.Bhattacharjee, Handbook of medicinal plants, second edition,
pointer publisher;1999
 The complete technology book on herbal perfumes & cosmetics,
National institute of industrial research.
 Masada,Y. Analysis of Essential Oils by Gas Chromatography
and Mass Spectrometry, John Wiley & Sons, New York (1976)
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 http://194.94.42.12/license_materials00897/papers/0003003/33smi8
97.pdf-
in
 http://www.ics.trieste.it/essentialoils/essentialoils.aspx?ID_23
 http://www.samispices.com/prod.list1.html
 http://www.ics.trieste.it/essentialoils/essentialoils.aspx?ID_14
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 http://Ift.confex.com/Ift/2001/techprogram/paper_7642.html
 http://www.Fgb.com.au/oldwebsite/products/cinolestandard.html
 http://laboratorytalk.com/books/chem/chrom/rs_10/rs_10_54.html
 http://www.butterburandsage.com/docs/viewdoitemsasp?doctypeid
Standardization of volatile oils
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