QUALITATIVE PHYTOCHEMICAL TESTS

TEST FOR CARBOHYDRATES

CHEMICAL TEST HOT EXTRACT COLD EXTRACT

1. MOLISH’S TEST (GENERAL TEST)

2. FEHLING’S TEST

3. BENEDICT’S TEST

4. BARFOED’S TEST

5. BIAL’S ORCINOL TEST

6. ANILINE ACETATE TEST

MIX EQUAL AMOUNT OF T.S. AND HCL +

7. HEAT +ADD A CRYSTAL OF
PHOLOROGLUCINOL

8. COBALT CHLORIDE TEST

9. SELWINOFF’S TEST

10. TOLLEN’S PHLOROGLUCINOL TEST

T.S. DOES NOT GIVE RESPONSE TO

11.
FEHLING’S AND BENEDICT’S TEST
HYDROLYSE TEST SOLUTION AND
12. PERFORM FEHLING’S AND BENEDICT’S
TEST

13. IODINE TEST

14. TANNIC ACID TEST

HYDROLYSE T.S. USING DILUTE HCL AND
15.
PERFORM FEHLING’S OR BENEDICT’S TEST
IN POWDERED DRUG ADD FEW DROPS OF
16.
RUTHENIUM RED REAGENT
IN POWDERED DRUG ADD WATER OR
17.
AQUEOUS KOH

TESTS FOR PROTEINS

CHEMICAL TEST HOT EXTRACT COLD EXTRACT

1. BIURET TEST (GENERAL TEST)

2. MILLION’S TEST

3. XANTHOPROTEIN

MIXED 5ML T.S. WITH 2ML 40% NaOH AND 2

4.
DROPS 10% LEAD ACETATE SOLUTION, BOIL
TESTS OF AMINO ACIDS

CHEMICAL TEST HOT EXTRACT COLD EXTRACT

1. NINHYDRIN TEST (GENERAL TEST)

2. MILLION’S TEST

3 ML T.S. + FEW DROPS GLYCOXALIC ACID

3.
AND CONC. H2SO4
5 ML T.S. + FEW DROPS OF 40% NaOH AND
4.
10% LEAD ACETATE SOLUTION +BOIL

TESTS FOR FATS AND OILS

CHEMICAL TEST HOT EXTRACT COLD EXTRACT

PLACE A THIN SECTION OF DRUG ON GLASS
SLIDE + DROPS OF SUDAN RED III REAGENT
1. +WASH WITH 50%ALCOHOL +MOUNT IN
GLYCERIN AND OBSERVE UNDER
MICROSCOPE
TO THIN SECTION OF DRUG ADD A DROP OF
2. 1% OSMIC ACID + OBSERVE UNDER
MICROSCOPE
SAPONIFICATION TEST
3. I.
II.

TESTS FOR VOLATILE OILS

1. ODOUR
2. STAIN TEST

TESTS FOR STEROID

1. SALKOWSKI REACTION
2. LIEBERMANN – BURCHARD REACTION
3. LIEBERMANN’S REACTION

TESTS FOR FLAVONOIDS

1. SHINODA TEST
SMALL QUANTITY OF RESIDUE +LEAD
2.
ACETATE SOLUTION
ADDITION OF INCREASING AMOUNT OF
3.
SODIUM HYDROXIDE TO THE RESIDUE

TESTS FOR GLYCOSIDES

1. BALJET’S TEST
2. LEGAL’S TEST
3. KELLER-KILLIANI TEST
 4. LIEBERMANN’S TEST
5. FOAM TEST
6. HEAMOLYTIC TEST
7. BRONTAGER’S TEST
8. MODIFIED BRONTAGER’S TEST
9. SODIUM PICRATE TEST
EXTRACT + 3 % AQ. MERCUROUS NITRATE
10.
SOLUTION
DIP A PIECE OF FILTER PAPER IN GUAIACUM
RESIN AND MOST IT WITH DILUTE COPPER
11.
SULPHATE SOLUTION. EXPOSE IT TO
FRESHLY CUT SURFACE OF DRUG
12. ODOUR
13. MAKE ALCOHOLIC EXTRACT ALKALINE
TAKE MOISTENED DRY POWDER IN TEST
TUBE. COVER TEST TUBE WITH FILTER
14. PAPER SOAKED IN DILUTE NaOH. KEEP IN
WATER BATH. AFTER SOMTIME EXPOSE
FILTER PAPER TO U.V. LIGHT

TESTS FOR ALKALOIDS

1. DRAGENDORFF’S TEST
2. MAYER’S TEST
3. HAGERS’S TEST
4. WAGNER’S TEST
5. MUREXIDE TEST

TESTS FOR TANNINS AND PHENOLIC COMPOUNDS

1. 5% FeCl3 SOLUTION
2. LEAD ACETATE SOLUTION
3. GELATIN SOLUTION
4. BROMINE WATER
5. ACETIC ACID SOLUTION
6. POTASSIUM DICHROMATE
7. DILUTE IODINE SOLUTION
8. DILUTE HNO3
ONE DROP NH4OH, EXCESS 10% AgNO3
9. SOLUTION. HEAT FOR 20 MIN. IN BOILING
WATER BATH

TESTS FOR ENZYMES

TO FEW THICK SECTIONS OF DRUG + ADD

1. CATECHOL SOLUTION. REPEAT TEST WITH
OTHER BOILED SECTIONS.
TO DRUG MATERIAL, ADD 1% ALC.
2. SOLUTION OF GUACIUM RESIN AND
WATER
 TO FEW SECTIONS OF DRUG + ADD
CATECHOL SOL. AFTER 2 MINUTES
3.
REMOVE CATECHOL + ADD H2O2
REPEAT TEST WITH BOILED SECTIONS
4. TO THICK DECTION OF DRUG + ADD H2O2
TO THICK SECTIONS OF DRUG ADD 5 ML
COLD WATER. AFTER 5 MIN ADD FEW
5. DROPS TRIPHENYL TETRAOLIUM
CHLORIDE SOLUTION
REPEAT TEST WITH BOILED SECTIONS
TAKE THICK SECTIONS OF DRUG ADD
COLD WATER AND FEW DROPS
6.
METHYLENE BLUE SOLUTION
REPEAT TEST WITH BOILED SECTIONS

TEST FOR VITAMINS

DISSOLVE A QUANTITY EQ. TO 10 – 15 UNITS

1. IN 1 ML OF CHLOROFORM AND 5 ML OF
ANTIMONY TRICHLORIDE SOLUTION
DILUTE 1 ML OF 2% w/v SOLUTION WITH 5 ML
OF WATER AND ADD 1 DROP OF FRESHLY
PREPARED 5% w/v SOLUTION OF SODIUM
2.
NITROPRUSSIDE AND 2 ML OF DILUTE
SODIUM HYDROXIDE SOLUTION. ADD 0.6 ML
OF HCl ACID DROP WISE AND STIR
ADD 2 ML OF 2% w/v SOLUTION TO FEW ML OF
3. 2, 6-DICHLOROPHENOLINDOPHENOL
SOLUTION
TO 2 ML OF A 2% w/v SOLUTION ADD 2 ML OF
WATER, 0.1g OF SODIUM BICARBONATE AND
4.
ABOUT 20 mg OF FERROUS SULPHATE, SHAKE
AND ALLOW TO STAND
DISSOLVE A QUANTITY EQ. TO ABOUT 1000
UNITS OF VITAMIN D ACTIVITY IN
5.
CHLOROFORM AND ADD 10 ML OF ANTIMINY
TRICHLORIDE SOLUTION

TESTS FOR INORGANIC ELEMENTS

TO 10 ML OF FOLTERATE, ADD 1 DROP

1. DILUTE NH4OH AND SATURATED
AMMONIUM OXALATE SOLUTION
MIX TEST SOLUTION WITH SOLUTION OF
2.
AMMONIUM CARBONATE
FILTER AND SEPARATE WHITE CALCIUM
OXALATE PPT OBTAINED ABOBE. HEAT AND
3. COOL THE FILTERATE WITH SOLUTION OF
SODIUM PHOSPHATE IN DILUTE
AMMONIUM SOLUTION
 TO 2 ML TEST SOLUTION ADD LITTLE
4. URANYL MAGNESIUM ACETATE REAGENT.
SHAKE WELL AND KEEP FOR FEW MINUTES.
5. FLAME TEST
TO 10 ,; ASH EXYRACT ADD 2 ML OF
6.
PATASSIUM PYROANTHLLOLLATE
TO 2-3 ML TEST SOLUTION, ADD FEW DROPS
7.
SODIUM COBALT NITRITE SOLUTION
8. FLAME TEST
TO 5 ML TEST SOLUTION ADD FEW DROPS
9.
2% POTASSIUM FERROCYANIDE
TO 5 ML TEST SOLUTION, ADD FEW DROPS
10.
5% AMMONIUM THIOCYANATE
TO 5 ML FILTRATE, ADD FEW DROPS 5%BaCl2
11.
SOLUTION
TO 5 ML TEST SOLUTION PREPARED IN
12. HNO3, ADD FEW DROPS AMMONIUM
MOLYBDATE SOLUTION. HEAT 10 MIN. COOL
TO 3 ML TEST SOLUTION PREPARED IN
13. HNO3, ADD FEW DROPS 10% AgNO3
SOLUTION
TO ABOUT 5 TO 7 ML FILTERATE, ADD 3 TO 5
14.
ML LEAD ACETATE SOLUTION
15. MIX WITH MERCURIC CHLORIDE SOLUTION
WARM WITH THE TEST SOLUTION WITH
16.
SULPHURIC ACID AND COPPER