INDEX

S.No. Experiment Page no.

1 To perform the preliminary phytochemical
screening of alkaloids
2 To perform the preliminary phytochemical
screening of saponins
3 To perform the preliminary phytochemical
screening of steroids
4 To perform the preliminary phytochemical
screening of tannins
5 To perform the preliminary phytochemical
screening of anthraquinones
6 To perform the preliminary phytochemical
screening of flavanoids
7 To extract volatile oil from cinnamon bark by
using Clevenger apparatus
8 To determine the aldehyde content of given
extract of crude drug
9 To prepare herbal cream
10 To study the different methods of extraction
11 To isolate the caffine from tea leaves extract
12 To prepare tea extract and determine its phenol
13 content
To extract piperene from the given sample of
14 black
To pepper the moisture content of given
determine
15 sample
15
 Experiment No. 1

Aim: To perform the preliminary phytochemical screening of alkaloids.

Requirements: Vinca Leaves, Dragondroff’s reagent, Wagner’s Reagent,

Mayer’s Reagent, Hager’s reagent, titration flask, funnel, beaker, petri
plates, droppers.

Procedure:

Test solution:- The test solution was prepared by dissolving the

extract in dilute HCl acid, the solution was filtered. The filtrate was
subjected to the following tests for the detection of the presence of
alkaloids.
Sr. Test Procedure Observation
No
.
1 Dragondroff’s In 3ml of filterate, few drops Formation of orange brown
Test of dragondroff’s reagent were coloured precipitate shows
added. presence of alkaloids
2 Mayer’s Test Few drops of Mayer’s reagent Formation of cream coloured
were added in 3 ml of test precipitate shows presence of
solution. alkaloids
3 Hager’s test Small quantity of Hager’s Formation of yellow coloured
reagent was added in filtrate. precipitate shows presence of
alkaloids
4 Wagner’s test Few drops of Wagner’s Formation of reddish brown
reagent were added in 3ml coloured precipitate shows
filtrate. presence of alkaloids
 Experiment No. 2

Aim: To perform the phytochemical screening of saponins.

Requirements: liquarice,H2O,GAA,concentration,H2OSO4.

Procedure:

Preparation of test solution: It was prepared by dissolving extract in

water and making it aqueous extract.

Sr.No Test Procedure Inference

1 Foam test Formation of foam Saponin may be
present.

2 Libermann Colour change red to violet Saponin may be

Burchard test present.
 Experiment No. 03

Aim: To perform the preliminary phytochemical screening of steroids.

Requirements: Chloroform,concentrated sulphuric acid,acetic anhydride.

Procedure:

Preparation of test solution: The test solution was prepared by dissolving

the extract in Chloroform.

S.No. Test Procedure Observation

1 Salkwoski test A few drops of concentrated sulphuric acid The formation of red
were added to the test solution and allowed to colour in lower layer
stand for some time. indicates the
presence of steroids.
2 Libermann Burchard Some drops of acetic anhydride were added to The formation pf
test test solution;the content were boiled and brown ring at the
cooled then concentration sulphuric acid was junction of two layer
added from side of the tube. and the upper layer
turns green
indicating the
presence of steroids.
 Experiment :-4

Aim:- To perform the preliminary phytochemical screening of tannins.

Requirements: Ferric chloride solution, Gelatin, NaCl, Lead acetate,

conc.HCl.

Procedure:

Test solution: The test solution was prepared by dissolving

the extract in water and alcohol.

Sr.No Test Procedure Observation

1 Ferric Chloride To 1ml extract, ferric chloride Dark blue or
test solution was added greenish black
colour
2 Gelatin test A few ml of 1% gelatine sol. WHitw precipitates
containing 10% NaCl were added
to test solution
3 Lead acetate A few ml of 10% Lead acetate Voluminous white
test was added to test solution ppt.
 Experiment No. 5

Aim: To perform the preliminary phytochemical screening of anthraquinones

Requirements: Sennc, H2O, GAA,H2SO4,Ammonia,Fecl2 and sodium

pictate and ethanol.

Procedure:

Preparation of test solution: It was prepared by dissolving extract in

water and alcohol.

S.No. Test Procedure Observation

1 Borntrager test To 3ml of extract dilute H2SO4 was added. Ammonical layer turns
It was boiled and filtered to the cold extract pink.
chloroform was added, after showing
organic solvent get separated ammonia was
added.
2 Modified The extract was boiled with alc. KOH, Ammonical layers turns
Borntrager test filter added dil. HCL to filterate and extract more red.
with other added ammonia and shaken.
 Experiment No. 6

Aim: To perform the preliminary phytochemical screening of flavanoids

Requirements:Onion,ethanol,HCL,mgturning,leadacetate
solution,NAOH.

Procedure:

Preparation of test solution: To small amount of extract equal amount of

2mg HCL was added and heater for about 30-40 min. at 100 degree

celcius.

S.No. Test Procedure Observation

1 Shinoda test 5ml of (95% v/v) ethanol was added Pink colour was observed.
to the test extract and few drops of
concentration HCL 0.5g mgturning
were added
2 Lead acetate test To small quantity of extract lead Yellow colour ppt were
acetate solution was added. formed.
3 Sodium hydroxide NAOH were added in large amount Yellow colour was formed
test to the extract. which decolorized after
addition of dilute acid.
 Experiment No. 7

Aim: To extract volatile oil from cinnamon bark by using clevenger apparatus.

Requirements: NaSo4, H2O and cinnamon bark.

Procedure:

. The apparatus was setted.

. Cinnamon bark (5g) was weighed and added to RBF(50ml).

. Adequate amount of H2O was added to RBF and fitted with assembly

and glycerine was applied on the joints.

. The temp. was setted at 100 degree celcius and after boiling it was

reduced to 60 degree to 65 degree celcius and distillation was allowed to

seen for 2 hrs.

. The volume of volatile oil collected was noted from graduated tube and

then it was collected in glass collecting tube.

. To the volatile oil NaSo4 was added to make it clear and free of any

water.

. The tube was wrapped with carbon paper and labelled.

 Experiment No. 8

AIM : To determine the aldehyde content of given extract of crude

drugs.

Requirement : Stoppened tube, hydroxylamine,hcl reagent in alcohol

60 percent,0.5N icon in alcohol,methyl orange,lemon oil.

Procedure :
• About 10g of cinnamon oil was weighed accurately and get put in
the stoppened tube.

• Hydroxylamine hydrochloride reagent in alcohol and few drop of

solution was titrated with 0.5N icon in alcohol until red colour
change to permanent yellow.

• Then aldehyde contact was calculated.

• 1ml of 0.5N icon in alcohol was equivalent to 0.0767 of cilrus.

 Experiment No. 9

Aim- To prepare the herbal cream.

Requirement- Liquid paraffin, beeswax,methyl paraben.

Procedure- preparation of extracts

1. Neem extract

• Neem leaves were collected and washed with distilled water and
dried in hot air oven.
• After proper drying leaves were powdered.
• Powdered was taken with (10ml) dimethyl sulfoxide and added to
the volumetric flask.
• Heated the mixture at 100 degree on water bath.
• The volumetric flask was shaken on mechanical shakes.
• Again solution was heated and concentration of 20ml and then
filtered using muslin cloth.
• The extract of water leaves obtained and used in the preparation.
•
2.Tulsi leaves extract-

• Tulsi leaves were collected and washed with distilled water and
dried in hot air oven.
• Then after proper drying,leaves were powdered.
• Then 1g of tulsi powder 10ml dimethyl sulfoxide was taken in
volumetric flask and then shaken on mechanical shakes.
• Then the solution was heated on water bath at 80-100 degree
celcius and then concentration upto 5ml and filtered using muslin
cloth.
• Extract was obtained and used in preparation.
 Experiment No. 10

Aim- To study the different methods of extraction.

Requirement- Crude drug, solvent.

Procedure-

• Firstly convert plant material into dry powdered form.

• Now this powdered is placed in a thimble made up of filter paper

then it placed in ride the soxhlet appratus.

• This appratus is filtered to be RBF which contain solvent and also

with a reflex conderer with the help of adapter.

• The solvent in the RBF is boiled gently vapours passes through

the side tube, then it condessed through condenser and falls into
thimble containing the material and slowly fill the soxhlet.

• When the solvent reaches the top of the attached tube it Diphous
over into flask.

• It takes about 8-10 for complete extraction.

• After extraction remove the porus com which contain residue and
repeat process for more.

• Extract obtained in RBF

 Experiment No. 11

Aim- To isolate the caffine from tea leaves extract.

Requirement- Ccl4, linse powder,chloroform,H2SO4.

Procedure-

• Tea powder (50g) was mixed lime (15g) by slight moistening with
water to basify the Alkaloid and lime being a base liberated the
alkaloid present in it.

• This mixture was kept for imbibation for proper penetration of

solvent into dry and then it was packed in soxhlet aprratus using
ccl4 as the solvent for extraction.

• The extraction was carried out on water bath for appropriate time.

• The organic solution was filtered after extraction was

completed.The filterate was concentrated under vaccum.

• Now to the concentration residue acidulated water that is 2 percent

H2SO4(50ml) was added.

• Now the solution was basified to ph 7.8 and was taken in seprating
funnel and was extracted with 20ml of chcl2 for 3-4 times.

• The Chcl3 layer were mixed together and were concentrated till
the crystallisation point comes.

• The crystals of caffine were seprated by filteration and mother

liquor was again concentrated and kept for crystallisation.
 Experiment No. 12

Aim- To prepare the tea extract and determine its phenol content.
Requirement- Tea extract, folin reagent,methanol,sod.
Bicarbonate,distilled water.

Procedure-

• Determination of phenolic content in the plant extract. The

concentration of phenolics in plant extract was determined using
spectrophotometric method.

• Methanolic solution of the tea extract in the concentration of

1mg/ml was used in the analysis.

• The reaction mixture was prepared by mixing 0.5ml of methanolic

solution of extract 2.5ml of 10 percent folin ciocal tea reagent
dissolved in water and 2.5ml of 75 percent sodium bicarbonate
blank was concomitantly prepared containing 0.5ml of methanol,
2.5ml of 10 percent.

• Folin ciocal tea reagent dissolved in water and 2.5ml of 75 percent

of sodium bicarbonate.The sample were thereafter incubated in a
thermostat at 45 degree celcius for 45min.

• The absorbance was determined using spectrophotometer at 1mac-

765nm. The sample was prepared in triplicate for each analysis
and then mean value of absorbance was obtained.

• The same procedure was repeated for the standard solution of

gallic acid and the calibration line was constructed.

• Based on the measured absorbance the concentration of phenolin

was read (mg/ml) from the calibration line then the content of
phenolic in extract was expressed in terms of gallic acid
equivalent.
 Experiment No. 13

Aim- To extract piperine from the given sample of black pepper.

Requirement- Ethanol,koH

Procedure-

• Clean and dry soxhlet appratus was taken and thimble was
prepared by using five filter paper.

• 10g of powdered pepper was taken in the thimble.

• Extraction was done by using 250ml of ethanol in soxhlet appratus

for 3hrs on water bath.

• Now the content but in RBF and were vaccum on water bath at
temp. not exceeding 60 degree celcius.

• 20ml of the 10 percent alc. KoH solution and solution was

prepared by taking 10gm of koh in C2H5OH making volume to
100ml and was added to the conc. With constant strining this was
then washed and filtered white warm.

• The filterate was then set a side for few hrs.

• The yellow needle like crystals of piperine was there.

• Mother liquor was then drandoff and crystals were washed with
alc. Koh solution and impurities attached to crystals were
removed.

• The washing were then finally mixed with mother liquor and set
aside for crystallisation.

• The final crystals were weighed and percent yield was calculated.
 Experiment No. 14

Aim- To determine the moisture content of given sample.

Requirement- Vinca

Procedure- Determination of moisture content

1. Accurately 2gm of crude drug was weighed and get put in percolin
dish.

2. Drug was kept in hot air oven for 2hrs. after it get cooled.

3. Crude drug was weighed percent of moisture content get

calculated.
 Experiment No. 15

Aim- To prepare herbal lotion.

Requirement- Beeswax,coconut oil,Aloevera jel,borax,distilled

water.
Procedure-

• Borax was dissolved in rose water on water bath at 70 degree

celcius.

• Wax was melted with mineral oil on water bath is another beaker.

• Temp. was keeped at about 70 degree celcius.

• Borax solution was pound into the motter wax at same temp. with
constant stirring.

• Whole mass was continuously stirred when the temp. drop to

about 45 degree celcius.

• Formulation was poured in wide mouth container and get labelled