EndofModuleExercise

LaboratoryExercise#2

ExtractionoftheUnknownOrganicComponentsfromtheAqueous
LagundiExtract

Introduction
Lagundi or Vitex negundo has been traditionally used as herbal medicine by Philippine
and Indian folks. It;'s main health benefit is to ease respiratory complaints. Lagundi is
generally used for the treatment of coughs, asthma symptoms, and other respiratory
problems. Commonly known in the Ilocos region as “dangla”, Lagundi has beenclinically
tested to be effective in the treatment of colds, flu, bronchial asthma,chronic bronchitis,
and pharyngitis. Studies have shown that Lagundi can preventthe body's production
ofleukotrienes, which are released during an asthmaattack. Lagundi
containsChrysoplenol D, a substance with anti-histamine andmuscle relaxant properties.

Liquid/liquid extraction is another technique of the chemist used in the

laboratoryforisolatingcompoundsfromamixture.Itinvolvestwophasesasthe
liquid/liquidextractiontitleiscalledandoneisusuallyanorganicphaseandone is in the aqueous
phase. Extraction is process of transferring a solute from one
phasetoanotherphase.Thegoalistomovethingsbetweenthesetwophases.

Liquid-liquid extraction, also known as partitioning, is a separation process consisting of

the transfer of a solute from one solvent to another, the two
solventsbeingimmiscibleorpartiallymisciblewitheachother.Frequently,oneof the solvents
is water or an aqueous mixture and the other is a nonpolar organic liquid. As in all
extraction processes, liquid-liquid extraction comprises a step of mixing (contacting),
followed by a step of phase separation. It is important to consider both steps in the
selection of solvents and modes of operation. Thus, while vigorous mixing is favorable to
the transfer of the extractable from one
solvent to the other, it may also impair the ease of phase separation by forming emulsions.

Equilibrium is reachedwhenthe chemicalpotentialoftheextractablesolute is the same

in the two phases. Practically, this rule leads to the definition of a
“distributioncoefficient,”K,asfollows:

K=C1/C2

where C1 and C2 are the equilibrium concentrations of the solute in the two phases,
respectively. The distribution coefficient is an expression of the relative preference of the
solute for the solvents. In ideal solutions (i.e., where the
chemicalpotentialmaybeassumedtobeproportionaltotheconcentration),the distribution
coefficient at a given temperature is practically constant, that is,
independentoftheconcentration.

ZekiBerk,inFoodProcessEngineeringandTechnology(ThirdEdition),2018

It is very common for a desired compound to be dissolved in a solvent during regular

manipulations in the laboratory. Solvents are used in separatory funnel extractions and
column chromatography, and the solvent must be removed in order to isolate te desired
compound. Solvents are regularly chosen that have lower boiling points than the
compound of interest, so that there is some mechanism for their removal. In theory, a
solution could simply be placed on a heat source to boil away the lower-boiling solvent,
but this approach is not oftenused.

The preferred method for solvent removal in the laboratory is by use of a rotary
evaporator, also known as a "rotovap".A rotaryevaporator is essentially a
reducedpressuredistillation:asolutioninaroundbottomedflaskisplacedinthe water bath of
the apparatus and rotated while the system is partially evacuated (by a water aspirator or
vacuum pump). The reduced pressure in the apparatus causes the solvent to boil at a lower
temperature than normal (see vacuum
distillation),androtatingtheflaskincreasestheliquid'ssurfaceareaandthusthe rate of
evaporation. The solvent vapor condenses when it comes into contact
 with a water condenser and drips into a receiving flask. When the solvent is
removed,theconcentratedcompoundisleftintheflask.Onedifferencebetween distillation and
rotary evaporation is that the distillate is most often retained in
distillationwhiletheresidueisretainedinrotaryevaporation.

Lisa Nichols (Butte Community College). Organic Chemistry Laboratory

Techniques

Whiledichloromethane isn'tmiscible withwater, itis able todissolveawide range of

organic compounds. These properties, combined with its volatility,
makesDCMahighlyeffectivesolventinmanyindustrialprocesses.

Objectives
1. Extracttheunknownorganiccomponentsfrom theAqueousLagundiExtract usingliquid-
liquidextraction
2. DrytheDCMextractusingsodiumsulfateandgravityfiltration
3. EvaporateDCMusingrotaryevaporator
4. Determine the % of the unknown organic components in the sample if the
concentrationoftheis1000ppm.
MaterialsandMethods
Method:
a. Measure 100 mL of Aqueous Lagundi Extract and place it to separatory funnel
b. Add 20 mL dichloromethane to the extract. Facilitate liquid-liquid extraction
c. PlacetheDCMextractinanErlenmeyerflask.
d. Place the aqueous extract in the separatory funnel and repeat the liquid-
liquidextractiontwiceusing15mLofDCM.
e. CombinetheDCMextracts
f. Dry the DCM extract with sodium sulfate and gravity filter into a pre-
 weighed round-bottom flask. Evaporate the solvent using a rotovap and
determinethemassofsolidyouobtain.

ResultsandDiscussion
Parameters Value Volumeofaqueousextract
100mL ConcentrationofExtract 1000ppm
VolumeofDCMExtract 51mL
Volume of DCM Extract after drying 49mL
usingsodiumsulfate
Mass of a pre-weighed round-bottom flask 122.621g
Mass of round-bottom flask after
Rotaryevaporation 122.652g
Mass of unknown organic
components 0.031g
%oftheunknownorganiccomponents
inthesample
63.27%

Base on the given result

s, the
in our
unknown
parameter
mass in
grams of organic components is 0.031g by subracting the mass of pre- weighed
round bottom flask from the mass of round bottom flask after rotary evaporation.
The percent of the unknown organic components in the sample was obtained by
converting 1000ppm to g/mL and the result
was0.001g/mLthenmultiplyittothevolumeofDCMextractafterdrying
usingsodiumsulfatewhichhas49mLtocancelthemLthenyouwillhave a mass of 0.049
g. To get the percent unknown of the unknown organic components use the mass
of organic components which is 0.031g then
divideitin0.049gandmultiplyto100%havinga63.27%.Obtainingahigh
percentageofunknown.
Lagundi, Vitex negundo, leaves ethanolic extract is the active ingredient of
the formulation. Lagundi leaves were extracted using
exhaustiveextractionthroughSoxhletapparatus.Itwasthenconcentrated into asyrupy
consistency in vacuum using rotary evaporator. The product
obtainedisnowthelagundileavesextract.Theextractwassubjectedtoa protein
stabilization test to determine its anti-inflammatory action. The
resultconcludedthattheextractindeedhasanti-inflammatoryproperty.
Ethanol,alsocalledalcohol,ethylalcoholandgrainalcohol,isaclear, colorless
liquid and the principle ingredient in alcoholic beverages like beer, wine or
brandy. Because it can readily dissolve in water and other organic compounds,
ethanol also is an ingredient in a range of products,
frompersonalcareandbeautyproductstopaintsandvarnishestofuel. Ethanol is a
common ingredient in many cosmetics and beauty products. It acts as anastringent
to helpcleanskin, as apreservativeinlotions and to help ensure that lotion
ingredients do not separate, and it helps hairsprayadheretohair.

Because ethanol is effective in killing microorganisms like bacteria, fungi

and viruses, it is a common ingredient in many hand sanitizers. The
 U.S. Centers for Disease Control and Prevention (CDC) recommends the use of
hand sanitizers in situations where soap and water are not available. Practicing
hand hygiene is also an important part of helping to stopthespreadofCOVID-
19.Usinghandsanitizersoralcoholbasedhand rubs (ABHR) can help to inactivate
SARS-CoV-2, the strain of coronavirus that causes COVID-19. In fuel ,more than
97 percent of U.S. gasoline contains ethanol, typically in a mixture called E10,
made up of 10 percent ethanol and 90 percent gasoline, to oxygenate the fuel and
reduce air pollution. Ethanol has a higher octane number than gasoline, providing
premiumblendingproperties,accordingtotheU.S.DepartmentofEnergy. Minimum
octane number requirements prevent engine knocking and maintaindrivability.

ConclusionsandRecommendation
Upon researching the extracted unknown organic components from the
Aqueous Lagundi Extract using liquid -liquid extraction is ethanol. By final traces
of water are removed by treating the organic solution with a drying agent. A
drying agent is an inorganic salt which readily takes up water to become hydrated.
Calcium chloride, magnesium sulfate, and sodium sulfateare
thethreemostcommonlyusedagents.Thereis no set
ruleastohowmuchdryingagentneedstobeadded.Theamountrequired depends on the
amount of water in the solvent solution which you are drying, and this amount
varies from experiment to experiment. Use as much as it takes to drythe solution.
In most cases, drying is as complete as it will get in 20 minutes.When drying is
complete, you need to remove the dried organic solution from the drying
agent.Gravity filtration is an easy way to remove solid impurities or the
precipitation from an organic
liquid.Theimpurityistrappedinthefilter.Gravityfiltrationcancollectany insoluble
solid. As when the DCM was evaporated by the use of rotovap.
 Having a low percent of unknown organic component in the sample concentrayion
of 1000ppm, thus a single extraction is not reasonable underthesecondition.

References
https://www.sciencedirect.com/topics/agricultural-and-biological-
sciences/liquid-liquid-extraction
https://www.sciencedirect.com/topics/chemistry/rotary-evaporation
https://www.chemicalsafetyfacts.org/ethanol/
J.Chem.Educ.1944,21,9,449
PublicationDate:September1,1944 https://doi.org/10.1021/ed021p449
https://www.sciencedirect.com/science/article/abs/pii/S0095955315321
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