I suggest steam distillation method for extraction of essential oil.

It is so small amount of plant for extraction essential oil. You need more than 100-200 g, better dry material. And you need an installation for extraction by steam water. It called neo-Clevenger, according with British Pharmacopeea. You can find a good method in each Pharmacopeea. Good luck!

You may try Solvent-free microwave extraction coupled with head-space single-drop micro-extraction.

You can try hydrodistillation or SDE

I suggest hydrodistillation, you must work with a little glass-chamber (about 10 cm high/ 2 cm diameter. Obviously you must take in mind that your eventually present EO could be at least in part soluble in aromatic water, so I can suggest you to harvest steam and aromatic waters in CH2Cl2, in order to prevent any lost of you precious sample.

Another option is to use solvent extractionwith a Lickens-Nickerson apparatus. I 've used it myself with aromatic plants when I had small amounts of sample 2-3g. This method is good for qualification purposes and will allow you to find out the essential oil composition or differences in the composition of various samples. However, with the aid of GC-FID and an internal standard you can quantify tour results.

steam distillation would be the best. Use a small (micro Kit) distillation setup (100 mL) round bottom flask and 50 mL collection flask. Then extract the distillate using about 40 mL (10 ml * 4) of CH2Cl2. Removed the solvent using rotary evaporator. Then evaporate the left over solvent in a 5 mL reaction vial using mild heat and mild vacuum. (Still you may need about 10 g of plant material and grind it with water and make a slurry).

You can try clavenger's apparatus. It is especially designed to isolate the essential oil. You can also measure the yield directly on graduated tube attached on apparatus.

Dear colleagues

Thanks for all answer.

I looking for a protocol based on solvent extraction not hydro or steam distillation.

So, if somebody have protocol related to solvent extraction, please send me.

Thank one more time for your aswers

Clevenger apparatus for water vapour distillation is a good method for essential oil extraction

Since you have such a small sample of plant material and want to use solvent extraction, then your option is to use a Lickens-Nickerson apparatus, as already suggested by S. Petropoulos

Lickens-Nickerson

You can check out this paper for a protocol

Or this

The conventional technique and the most used is the steam distillation using Clevenger apparatus and the electric flask heating , but in recent years, heating with microwaves is widely used and the advantages are significant with respect to saving energy and reducing the extraction time, hence reducing artifacts. In addition, if the plant is quite rich in water 'the humidity of the fresh plant exceeding 60%' the extraction of essential oil could be done without adding water. Tis process is called 'Sovent Fee Microwave Extraction SFME ', in this case we minimize the dégradation reactions of many compounds. We never worked with masses of this order. I think the little that can be used is in the range of 80 to 100g.

Lickens-Nickerson apparatus would be the wisest choice in your case :)

Hydrodistillation method

Micro-SDE apparatus designed by Godefroot et al. 1981, New method for quantitative essential oil analysis, Journal of Chromatography A 203 (C) , pp. 325-335 .

The best way is claevenger but if you don't want to use this way must refer to the related article.

I would suggest the conventional clevenger apparatus as the loss will be minimum.

There are four methods of volatile oil isolation

1.Direct steam distillation, 2. Expression, 3. Extraction, 4. Enzymatic hydrolysis

1. Direct steam distillation -3 sub methods according to nature of volatile oil constituents

1a. Water distillation- dried plant ( not degraded at 100°C) e.g. turpentine oil

1b. Water & steam distillation- dried/ fresh drug macerated first for hrs (undergo degradation by direct boiling) e.g. clove oil

1c. Direct steam distillation- fresh drug e.g. peppermint & spearmint oil

2. Expression(best for Rutaceae family fruits)

If No. of compound of vol. oil undergo decomposition by distillation than this technique is used ; 4 sub techniques

2a.Sponge method

2b.Scarification process(Ecuelle)

2c.Rasping process

2d.Mechanical process

3.Extraction

For plant which contain very small amount of volatile oil or oil contents decompose upon steam distillation - 2 sub techniques

3a. Extraction with volatile solvent e.g. hexane benzene, pet. Ether (jasmine species flower)

3b. Extraction with non-volatile solvent e.g. lard tallow & olive oil - sub technique

-Enfleurage method

-Pneumatic method

-Maceration method

4. Enzymatic hydrolysis

Volatile oil found in the form of odorless glycosidal combinations

Above is protocol according to type of chemical constituents one wants to isolate.

Since you want to use only 1 or 2 g, in case you have access to an ASE Dionex machine, you could use Accelerated Solvent Extraction with Hexane as a solvent of choice

You can opt for clevenger apparatus or Likens and Nickerson SDE Method at lab scale level

If you want to extract Essential oil by definition, you have to have higher amount of plant material, since you need a hydro distillation or steam distillation apparatus. Alternatively you design a micro extraction apparatus if your plant yields is high> But if you wan to analyze volatiles by other methods, I suggest solvent extraction by Hexane and dissolving the concentrated extract in absolute ethanol filtering and injection the filtrate to GC. or SPME-GC but the results are not the same as you obtain for essential oils.

Steam distillation

Aromatic plants usually do not contain more than a percent (0.05-1%, w/w) of volatile compounds, and this amount (that means maximally ca. 10 mg from the amount of plant material you have available) of an essential oil sample obtained by most of the mentioned techniques would literally get lost in a Clevenger apparatus (for instance), and you would end up with mostly nothing in the receiver portion of the apparatus. Of course if you are in the possession of schizocarps of a prolific Apiaceae species (that are reported to yield up to 10% of essential oil) then hydrodistillation of finely crushed fruits would be the method of choice (since with 100 mg of essential oil you could manage to do almost all analyses with ease).

In a situation like yours I usually decide to make a diethyl ether solvent extract (since I am not very patient I use an ultrasound bath to speed up the extraction). After the separation of plant material by filtration, drying over anhydrous MgSO4 or Na2SO4, most of the ether is removed by a stream of N2 (in order not to lose highly volatile compounds from the sample) and the concentrated (and filtered if any solids formed) sample analyzed GC-MS and GC. Please, note that with this procedure you will not extract only essential oil constituents but also other (non-volatile with water steam) plant metabolites. Although it is not the same thing as essential oils it would give you a good appreciation what to expect from that specific plant next year when you collect a much more substantial amount of the material.

I agree, it is a good alternative.

You can use Soxhlet method for extraction of oil using diethyl ether of petroleum ether as solvent

You can use Headspace Methods.This is our Methods in Damask rose:

The headspace volatiles extraction

Three to five rose branches from each germplasm that had the most flowers were harvested in the early morning and immediately placed inside a vase with clean water, and were then quickly transported to the laboratory for testing. Five flowers from each flower development were selected. Their petals were gently plucked and mixed. Up to 3 g of petals were quickly placed in 20 mL sample vials, internal standard (3-nonanone, 10 μL, 0.8 μg.μL-1) were added, and the vials were then sealed rapidly with septa (PTFE-butyl synthetic rubber). Extraction and concentration of the floral headspace volatiles from the fresh flowers were performed using a Turbo Matrix 40 Trap (Automated headspace sampler with built-in trap, PerkinElmer, USA). The extraction conditions was as follows: The heating temperature was at 80 °C for 30 min, the temperature of the sampling needle and transmission line was 80 °C and trap was maintained for 5 min.

I agree with Niko for small quantity of Raw materials.

In general, when dealing with a small amount of plant material, it is inpossible to proceed by conventional techniques. Personally I opt for the innovative techniques (ultrasound, mico oven, ...). Adiquate a small amount of solvent will be used.

Cleavenger is the best

Go either with Enfleurage method or use Clavenger.

Direct steam distillation esp using glass equipment will lead to least changed plant profile.

Actually, distillation and extraction methods are not a very good choice for you, because your material is insufficient. If your aim is identification of essential oil compounds, I suggest you to use headspace GC/Mass. You can manage to use this devise in Central Laboratory of Food and Drug Administration of the Health Ministry. If you can not manage, I suggest using Clevenger apparatus (model of 5 mL, not model of 25 mL).

Dear Dr. Jafari,

Thank you so much for your valuable comments.

Regards

Dear friend you can extract with Super critical CO2- good result you will get it.

You could use a volatile solvent like light petroleum ether, using soxhlet at 50 o C for three hours to extract essential oils and fatty content, then you could use GC-MS for determination of the constituents

Pr Mahrouz mostafa UCA FSSM Marrakech

Our Work about free (no combined with sugar or polyphénol,?.?) with out hydrolis there many method shep longer, destructiv, ecoextraction...

Super critic CO2 is the best

with out température just varaiotion diffusion withsolvant volti 5° to 30°C

t with temperature quikly diffusion with solvant volatail like acetone , hexane ether ethylic ether de petrol in soxlhet at TEb 30à 60°c

and steam water at 98°c

You could use a volatile solvent like light petroleum ether, using soxhlet at 50 o C for three hours to extract essential oils and fatty content, then you could use GC-MS for determination of the constituents

I have ever exprereince about the extraction of lipid component from rice bran. You can try this way. Use methanol to extract the lipid (such as tocopherol, tocotrienol and oryzanol) by shaker for 3-4 h., filter, evaporate the filtrate. Then you can obtain the lipid fraction.

Because of low content of essential oil (less than 1 %), in my experience is too dificult to obtain that essential oil fom 1-2 gram plant material. Steam distillation and clavenger apparatus is a good choice if you have enough material. In my opinion you can try to extract the oil of that sample by using solvent hexane /petr ether and then analyzed by gcms.

Use n-hexane solvent and steam distillation

or soxhlet apparatus

Dear Researchers

Thanks for all the replies

Best Regards

Steam-distillation using Clevenger Apparatus is the easy, cheap and hence, best way to extract essential oils from any source.

I think using ultrasound extraction and SFE are the best methods for oil extraction from 1-2 grams plant

2gm sample for the oil are very very little, easy way to determine the volitile componet in the oil, using GCMS with SPME apparatus will help. Direct injection proses from the SPME extraction.

Dear Friends, All of you explain methods that extract essential oils but Mr ramezani need to a methods suitable for small amount of Herb (1-2 g) therefore I think that the best option is using Micro-clevenger or Head Space Analysis .

Dear Ramezani, thank you for your question and Dear Friends thank you for you responds. I have many information concerning extraction from you.

Thank you all

you better use SPME to directly analyse the components using GCMS

Good Luck

If you really have small amount of plant material then I will recommend to use head space GCMS to characterize the volatile components present on the interested plant sample. If you have enough sample then you may used the above described method to isolate volatile components from medicinal plants. Best of luck.

you can find your wa of extraction in a similar study: "Rapid essential oil screening of Rosmarinus officinalis L. by hydrodistillation–headspace solvent microextraction". it is decribed that how you can extract essential oil from a small sample

Hi mr ramezani

i recomended you ,use n-hexane solvent for extraction because you have less than 1-2 gr of plant material.material of plant should be dry.

The analytical technic is good for you.microextraction is very good for your research

You can use SPME method

Good luck

It will be best way get from Soxhlet extraction apparatus..............find yourself the protocol....

Thanks

The steam distillation is among the best methods to extract essenteilles oils.

Many conventional and modern methods are available of which steam distillation and hydrodistillation are popular. True steam distillation uses an outside source of steam which pipes the steam into distillation unit some times at high pressure. In hydrodistillation, the plant materials are submerged in water producing a stew (soup). The steam of which contain the volatile oil. This method is best when we work

with powders (spice powders, ground wood, stem bark or roots, nuts, oleoresins etc, (There are some risks also, overheating, burning etc), Cheap and versatile.

If you have the facility SPME is ok, but if you don't have this then try for DCM in place of hexane.

Simultaneous distillation extraction (SDE) is suitable for the available sample.

Dear Colleagues

Thank you all for answer and help me.

Dear sadrollah

If you have money and logistic and you look for quality you must use CO2 supercritic. But we can use many processus of extraing essen..Oil.with few quality

With low temperature :gradient temperature 4°C to 25°C witn hexanme or ether petrol but other substances are extraing.

with hot temperatture : Hydrodistillation, Dean stark, ....

soxlhet with derivation

others...many

Pr mahrouz mostafa

University cadi Ayyad chemistry

Director of ERIDDECV Green chemistry

It will be best way get from Soxhlet extraction apparatus..............find yourself the protocol....Solvent may be use: acetone, chloroform, hexane, ether, diethyl ether, DCM, DMSO, Glycerinn etc.

It is good to use organic solvent(hexane,pet.ether....etc.)to extract the volatile oil.It is best to use supercritical fluid extraction(SFE) if you have it in which CO2 is used when liquified under pressure(as organic solvent),when the liquid depressurized,CO2 returns to its gaseous state and only pure product remains.

The best technique for the extraction of essential oil is super critical carbon dioxide extraction as the oil is devoid of extractants, resin, waxes and other phytochemicals that are not part of the constituents of essential oils.

2014-04-02 Dear Friend,

To avoid any destruction of the oil two careful methods are recommended.

Supercritical extraction using CO2 and

low temperature extraction with suitable solvent protected by Nitrogen gas.

Excluding heating of the solvent any polymerisation and condensation of the material is avoided. You exclude risk for oxidation by using Nitrogen gas atmosphere.

The extraction can be performed at temps. of minus degrees.

Frank A, Wennerholm M. The Equilibrium Extractor and its use in the study of rancidity.

“In Focus”. 1991;14:4-5. (The Tecator journal of technology for chemical analysis).

Dear Colleagues

Thank you so much for valuable comments.

Best Regards

Sadrollah Ramezani

Hi Several methods available

See my papers in google

Mr Ramezani

This is depend of your purpose.....if you want make some test, you will need of the oil....I like very much of distillation with apparatus Clevenger....it is more cheap and easy method for obtain the volatile oil.....but some compounds have interaction with the water. Look for in the discussion about the methods for extraction and works of essential oil, here in Researchgate....maybe you have some ideas for your study.

Good Luck

Rozilaine

Unfortunately, the most of colleagues don't pay attention to my question, because i have problem with low amount of plant materials. When the plant material is enough, there are different methods that can be used. Anyway, thank you all for comment.

S. Ramezani

Dear Friend, Supercritical extraction can be used for very small amount of material to be extrated . Investigate the possibilities of that technique. Adrian F.

Well, with such a small amount of raw materials may be the best method to use distillation oven, which is not yet widely used. The camera Soksleta, you move tyklo extraction, and oil do not isolate. You can also perform the standard-distillation with water vapor or with the addition of m-xylene, but for that you need a micro cameras, even smaller than for the distillation of essential oil of nasiom. It seems to me that the best would be the use of distillation oven.

Yours.

Using CO2 to change the oil. This is not the best way to destylacjioleju. It is relatively expensive and is not recommended for understanding the qualitative and quantitative composition of essential oil.

Hi Agneszka, Would you kindly mention References of useful articles, please.

Adrian

The best method for essential oil extraction is by steam distillation

You can use solid phase microextraction (SPME) method.

In this method you need just 2 grammes of plant.

Steam distillation is best methods

Hello Sadrollah

Have you used my prpusition? and wath of results?

I have propused many protocols :

witn organic solvant : EP, EE, haxane with gradient température

steamdistillation

soxlhet modifed (ecoprootocoll)with water

DIC classic

DIC with water satured with NaCl and décantation

microweave

CO2 supercritic

.....

I think Soxhlet extraction with pure methanol is a good method for sage. A colleague & I tried it few years ago for a science fair (not for research) and it worked well.

Dear Friend

You are able to use Stahl distillation for your small amount sample.

Dear Sadrollah Ramezani

Hydrodistillation followed by extraction of hydrolate with chloroform or ethyl ether.

Dear Sadrollah;

For small amount of plant material the best method for essential oil extraction in this medicinal plant you can using :

steam distillation

soxlhet with ethyl ether

or optimised : crud material in nirogène and extrraed with few ether de petrol or hexane and s....

Mahrouz

l

I used steam hydro distillation method to extract the essential oil from fresh ginger rhizome, and it worked very good with me. I could extract 100 ml in 30 kg rhizome. TQ

The use of hypercritical carbon dioxide extraction is a fairly new way to extract essential oils from botanical material and although a bit on the expensive side, does yield good quality oils.

Carbon dioxide becomes hypercritical at 33 degrees Celsius, which is a state in which it is not really gas or liquid, but has qualities of both, and is an excellent solvent to use in the extraction of essential oils since the low temperature required and the fact that the process is near to instantaneous.

The carbon dioxide is furthermore inert and therefore does not chemically interact with the essence that is being extracted. To remove the carbon dioxide solvent, you simply need to remove the pressure under which it is kept.

This process has to take place in a closed chamber for the hypercritical pressure required for carbon dioxide is 200 atmospheres - that is 200 times the pressure of normal atmosphere.

To achieve this type of pressure some heavy-duty stainless steel equipment is required, and this is where high capital investment is required for this extraction method.

Thankful to all.

Best Wishes

Essential oils are extracted best way in n-hexane of pet-ether

Although steam distillation is the most well-known technique for extracting essential oil from plants, there are several other methods that are used to remove and concentrate the aromatic constituents from plant materials. The attached file may provide you detailed information on this aspect.

Hydrodistillation best method 

for small amount of plant material (fresh and dry) less than 1-2 gm, the best method for essential oil extraction is  in- tub extraction  method  (ITEX- method)  dynamic headspace. i used this method and it got very good results..

Supercritical CO2 extraction is best method.

There are so many techniques are available for the extraction of natural product (Phytochemicals) like

1. Classical solvent extraction procedures

2. Ultrasound-assisted extraction

3. Microwave-assisted extraction

4. Extraction with ionic liquids

5. Accelerated (pressurized) solvent extraction

6. Supercritical fluid extraction

7. Extraction on solid phases

8. Distillation methods

All these techniques have their own disadvantages. you can prefer Supercritical fluid extraction (SFE), Microwave distillation and Headspace trapping techniques . for details you can go through the link given below....

http://agritech.tnau.ac.in/horticulture/extraction_methods_natural_essential_oil.pdf

If we don't have Supercritical CO2 Extractor, classical distillation as Hydro distillation, steam distillation still can be used to obtaine the essential oil.

essential oils are in general non-polar, thus required non-polar solvents, such as CO2, hexane, pet ether and others...

Supercrirical carbodioxide is one of the best menthod for extraction of essential oils

Supercrirical carbodioxide is one of the best menthod for extraction of essential oils

Why not simultaneous extraction-distillations with Likens-Nickerson apparatus and later concentration with N2 flow or rotavapor at room temperature? I have work with this method usin no more than 2-3 g of Mint or Artemisia absinthium leaves.

Thank you Juan-Antonio Llorens-Molina for your offer.

This apparatus is not available for me.

Could you send me some real photo with dimensions?

Best Regards

Ramezani

you have to extract using light petroleum ether using soxhlet apparatus and then to evaporate the light petroleum ether to get the crude extract containing essential oils.

you could obtain the volatile oils through hydrodistillation

supercritical fluid extraction using CO2 will afford to you all the constituents in the plant

The best protocol is hydrodistillation, a simple and easy to maintain system. The fresh and intact leaves are mixed with distilled water inside a glass baloom, with heating during three hours. The fresh leaves are necessary to assure obtaining total oil yield. The essential oil is collected in a separatory device or extracted with chloroform or diethyl ether from the distilled water when the concentration is low.The composition of essential oils from medicinal plants in general changes due to many factors such as climate, rain period, time of the day (circadian variation) or time of the year (seasonal variation) , chemotypes etc. Then the most analysis of essential oils of medicinal plants are punctual.

By using the Equilibrium Extractor with any kind of solvent, any

temperature (no heating), even in Coldroom and protecting gas (Nitrogen, Argon).

See : Adrian Frank, The Equilibrium Extractor 1991