Tannins2024 Print

CRUDE DRUGS CONTAINING TANNINS
CHEMICAL TESTS
1. General test tube reactions
Crude drugs: Quercus cortex, Alchemillae herba, Cotini folium, Agrimoniae herba, Camelliae
sinensis folium non fermentatum, Vaccinii macrocarponis fructus
Sample preparation:
 Boil 2 g of the powdered drug with 200 mL of water for 5 minutes (put a funnel onto the
flask)
 Filter the extract through cotton plug.
 Use 5 mL portions of the filtrate for the tests described below (add the reagents drop by
drop).
 Note the results.
Crude drug 1% quinine- R-basic lead R-Fe(III)-chloride
hydrochloride acetate
Quercus cortex
Alchemillae herba
Cotini folium
Agrimoniae herba
Vaccinii macrocarponis fructus
Camelliae sinensis folia
Explanation:
Tannins give precipitate with heavy metals, proteins (leather tanning) and alkaloids.
All phenolic compounds give colorful products with mineral salts (Fe (III) ions). The color of
the complex depends on the number and the position of the phenolic groups. Gallotannins and
ellagitannins (hydrolysable or pyrogallol-type tannins) give bluish-black color and
precipitates, while condensed or non-hydrolysable (=catechin type) tannins give greenish-
brown precipitate. The reaction proceeding in neutral or weekly acidic medium is a
complexation of electron donor OH groups in ortho position and Fe3+ ions. It is a non-specific
reaction.
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2. Determination of tannins in herbal drugs (Ph. Eur.)
Sample: Agrimoniae herba 1.000 g

Quercus cortex 0.700 g
Conduct all the extraction and dilution steps protected from light!
 Add 150 mL of water to the stated amount of the powdered drug (m1) in a 250 mL
Erlenmeyer flask.
 Heat on a water-bath for 30 min.
 Cool under running water and transfer quantitatively to a 250 mL volumetric flask.
 Rinse the Erlenmeyer flask and collect the washings in the volumetric flask,
 Dilute to 250.0 mL with water.
Total polyphenols.
 Dilute 5.0 mL of the filtrate to 25.0 mL with water.
 In a 25.0 mL volumetric flask mix
o 2.0 mL of this solution with
o 1.0 mL of phosphomolybdotungstic reagent R (Folin solution)
o and 10.0 mL of water
 Dilute to 25.0 mL with a 290 g/l solution of sodium carbonate R.
 After 30 min measure the absorbance at 760 nm (AT), using water R as the compensation
liquid.
Polyphenols not adsorbed by hide powder.

 To 10.0 mL of the filtrate, add 0.10 g of hide powder and shake vigorously for 30 min
in ultrasonic bath.
 Filter and dilute 5.0 mL of the filtrate to 25.0 mL with water.
 In a 25.0 mL volumetric flask mix
o 2.0 mL of this solution with
o 1.0 mL of phosphomolybdotungstic reagent R
o and 10.0 mL of water
 Dilute to 25.0 mL with a 290 g/l solution of sodium carbonate R.
 After 30 min measure the absorbance at 760 nm (ANT), using water R as the
compensation liquid.
Standard. NOT TO PREPARE

 Dissolve immediately before use 50.0 mg (m2) of pyrogallol R in water
 dilute to 100.0 mL with the same solvent.
 Dilute 5.0 mL of the solution to 100.0 mL with water.
 Mix 2.0 mL of this solution with 1.0 mL of phosphomolybdotungstic reagent R and 10.0
mL of water R and dilute to 25.0 mL with a 290 g/l solution of sodium carbonate R.
After 30 min measure the absorbance at 760 nm (AP), using water as the compensation
liquid. AP= 0.36
Calculate the percentage content (m/m %) of tannins expressed as pyrogallol!
62.5 × (AT − ANT) × m2

%=
AP × m1
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Explanation:
Tannin content of the crude drug is the fraction of polyphenols adsorbed by
hide powder (protein- tannin complex). Polyphenols are polar molecules
and can be dissolve in water (extraction with H2O). Total polyphenol
content can be determined by the addition of Folin reagent to a portion of
the extract. By the addition of hide powder to the aqueous extract, tannins
are adsorbed (complex formation with proteins), thus after filtering, only polyphenols which are
not tannins will be present in the filtrate. Tannin content can be determined by determining the
amount of these non-tannin-polyphenols and deducting from the total polyphenol content.
Mtotal polyphenol – Mnon-tannin-polyphenol=Mtannin
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3.Procyanidin detection by Bate-Smith reaction

Drugs: Quercus cortex, Agrimoniae herba, Cameliae sinensis folium non fermentatum, Crataegi
folium cum flore, Vaccinii macrocarponis fructus, Vaccinii macrocarponis fructus containing
capsule
Add 25 ml 2 M hydrochloric acid to the content of a capsule or 1 g-of a drug in an Erlenmeyer

with a stopper then boil it on water bath for 30 min using a reflux condenser. Filter the solution
after cooling. Shake 2 ml-of solution with 2 ml n-butanol in a test tube. Write the observations in
your report!
Explanation:
Name of “non-hydrolysable” tannins originate from the fact that the C-C bond of the units can
only be leaved (quantitatively) in strong acidic medium. Cleaving the bond results in a catechol
and a cyanidin unit. The latter shows intensive red color in acidic medium and can be extracted
with n-butanol.
Results into the report
1. General test tube reactions of tannin containing drugs

2. Tannin quantitative determination (%)
3. Procyanidin detection with Bate-Smith reaction

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CRUDE DRUGS CONTAINING TANNINS

1. General test tube reactions

Vaccinii macrocarponis fructus

Camelliae sinensis folia

2. Determination of tannins in herbal drugs (Ph. Eur.)

Sample: Agrimoniae herba 1.000 g

Polyphenols not adsorbed by hide powder.

Standard. NOT TO PREPARE

Calculate the percentage content (m/m %) of tannins expressed as pyrogallol!

62.5 × (AT − ANT) × m2

Mtotal polyphenol – Mnon-tannin-polyphenol=Mtannin

3.Procyanidin detection by Bate-Smith reaction

Add 25 ml 2 M hydrochloric acid to the content of a capsule or 1 g-of a drug in an Erlenmeyer

Results into the report

1. General test tube reactions of tannin containing drugs

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