kv¡tha c£r¸a, Punarnav¡di ma¸·£ra, Punarnav¡sava, Sukum¡ra gh¤ta
Therapeutic uses:
Dh¡tu kÀaya (tissuewasting), P¡¸·u (anaemeia), áotha (inflammation)
Rf
1.0
0.5
0.0 Punarnav¡ Hydro-alcoholic Extract is a dried and
powdered extract prepared from Punarnav¡ (appropriately powdered). The extract contains not less than 0.025 per cent of boeravinone B when assayed.
Method of preparation:
Take Punarnav¡ suitably sized (powder or pieces) in an extractor. Add 50 per cent aqueous alcohol, about 3 times the quantity of raw material and heat under a reflux at a temperature between 80- 850 for 3-4 hours. Filter the extract through a filter
(preferably 10 m pore size) to a suitable sized vessel. The marc is extracted three times more, filtering the extract each time into the same vessel. Concentrate the combined filtrate to a syrupy consistency and dry under vacuum (between 400-600 mm of Hg) at a temperature not exceeding 800 till the moisture is below 5 per
cent. Mill the mass and sieve the powder through 500 m mesh to obtain the extract and pack. The yield obtained is about 10 per cent.
Identity, Purity and Strength:
Thin-layer chromatography:
Carry out thin-layer chromatography on a precoated silica gel 60F254 plate (Appendix 3.5)
using boeravinone Bas a reference standard. Test
solution: Extract 2 g of substance by refluxing
with 50 ml of methanol for a period of 15 min. Filter and concentrate the extract to 25 ml.
Standard solution: Dissolve 2 mg of boeravinone B RS in 10 ml of methanol. Procedure: Apply 10
l each of the test and standard solutions as bands at a height of 10 mm from the base of a 10 x 5 cm TLC plate and develop up to 8 cm from the base of the plate using the mobile phase: toluene : ethyl
acetate : formic acid (4.0 : 5.0 : 1.0). Dry the plate
in air and examine under UV 254 nm. The chromatographic profile of the test solution shows a band corresponding to that of the standard solution (Fig. 1).
254 nm
RS T
Fig. 1: Thin-Layer Chromatogram of Punarnav¡ water extract
RS: Boeravinone B, T: Test solution
Quantitative parameters:
Loss on drying: not more than 7.0 per cent
(Appendix 2.1.4); Total ash: not more than 12.0 per cent (Appendix 2.1.5); Acid-insoluble ash: not more than 2.0 per cent (Appendix 2.1.7); pH: 6.0- 8.0 (Appendix 2.1.10); Total soluble solids: not less than 90.0 per cent (Appendix 2.1.11) (Method-I)
Other requirements:
Heavy metals: Complies with the prescribed
limits, (Appendix 3.1); Microbial contamination: Complies with the prescribed limits, (Appendix 3.2); Pesticide residues: Complies with the prescribed limits, (Appendix 3.3); Residual
solvent: Complies with the prescribed limits,
(Appendix 3.8); Aflatoxins: Complies with the prescribed limits, (Appendix 3.4)
Assay:
Carry out the assay by liquid chromatography (Appendix 3.6). Test solution: Take about 5 g, accurately weighed, of the substance being examined and reflux with methanol (25 ml x 3)
on water bath for 15 min each, cool and filter. Combine all the filtrates and transfer to a 100-ml volumetric flask and make up the volume. Filter through 0.42 m membrane. Standard solution: Take about 2.5 mg, accurately weighed,
boeravinone B RS in a 100-ml volumetric flask
and dissolve in about 50 ml of methanol and make up the volume with methanol. Filter through 0.42
m membrane. Chromatographic system: High performance liquid chromatography. Column and
stationary phase: C18 (250 mm x 4.6 mm, 5 m).
Mobile phase: Filtered and degassed gradient
mixture of water and acetonitrile in the following proportions:-
Time
(min) (per cent) Water Acetonitrile (per cent) 0.01 80 20 5 55 45 15 20 80 20 20 80 23 55 45 25 80 20 30 80 20
Injection volume: 20 l. Flow rate: 1.5 ml per min. Detection: UV 280 nm. Procedure: Inject 20
l of the standard solution and record the chromatogram. Inject 20 l of the test solution, record the chromatogram and measure the response for the analyte peak. Calculate the content of boeravinone B in the substance being examined from the peak response of analyte. The test is not valid unless the relative standard deviation for replicate injections is not more than 2.0 per cent.
Fig. 2: HPLC chromatogram of Punarnav¡ hydro-alcoholic extract with
Boeravinone B as RS
Additional requirements:
Storage: Store in well closed container protected
from heat, light, moisture and against attack by insects and rodents.
Labelling: The label states the official name,
followed by the Latin binominal name and the part of the plant contained in the article.
API reference standard:
API Boeravinone B RS
Rf
1.0
0.5
0.0 Punarnav¡ Water Extract is a dried and powdered
extract prepared from Punarnav¡. The extract contains not less than 0.0025 per cent of
boeravinone B when assayed.
Method of preparation:
Take Punarnav¡ suitably sized (powder or pieces) in an extractor. Add water, about 3 times the quantity of raw material and heat at a temperature between 80–850 for 3-4 hours. Filter the extract
through a filter (preferably10 m pore size) to a suitable sized vessel. The marc is extracted three times more, filtering the extract each time into the same vessel. Concentrate the combined filtrate to a syrupy consistency and dry under vacuum (between 400-600 mm of Hg) at a temperature not exceeding 800 till the moisture is below 5 per cent.
Mill the mass and sieve the powder through 500
m mesh and pack. The yield obtained is about 8 per cent.
Identity, Purity and Strength:
Thin-layer chromatography:
Carry out thin-layer chromatography on a precoated silica gel 60F254 plate (Appendix 3.5)
using boeravinone Bas a reference standard. Test
solution: Extract 2 g of substance by refluxing
with 50 ml of methanol for a period of 15 min. Filter and concentrate the extract to 25 ml.
Standard solution: Dissolve 2 mg of boeravinone B RS in 10 ml of methanol. Procedure: Apply 10
l each of the test and standard solutions as bands at a height of 10 mm from the base of a 10 x 5 cm TLC plate and develop up to 8 cm from the base of the plate using the mobile phase : toluene :
ethyl acetate : formic acid (4.0 : 5.0 : 1.0). Dry the
plate in air and examine under UV 254 nm. The chromatographic profile of the test solution shows a band corresponding to that of the standard solution (Fig. 1).
254 nm
RS T
Fig. 1: Thin-Layer Chromatogram of Punarnav¡ water extract
RS: Boeravinone B, T: Test solution
Quantitative parameters:
Loss on drying: not more than 7.0 per cent
(Appendix 2.1.4); Total ash: not more than 12.0 per cent (Appendix 2.1.5); Acid-insoluble ash: not more than 2.0 per cent (Appendix 2.1.7); pH: 6.0-8.0 (Appendix 2.1.10); Total soluble solids: not less than 90.0 per cent (Appendix 2.1.11) (Method-II)
Other requirements:
Heavy metals: Complies with the prescribed limits,
(Appendix 3.1); Microbial contamination: Complies with the prescribed limits, (Appendix 3.2); Pesticide residues: Complies with the prescribed limits, (Appendix 3.3); Aflatoxins: Complies with the prescribed limits, (Appendix 3.4)
Assay:
Carry out the assay by liquid chromatography (Appendix 3.6). Test solution: Take about 5 g, accurately weighed, of the substance being examined and reflux with methanol (25 ml x 3) on water bath for 15 min each, cool and filter. Combine all the filtrates and transfer to a 100-ml
volumetric flask and make up the volume. Filter through 0.42 m membrane. Standard solution: Take about 2.5 mg, accurately weighed,
boeravinone B RS in a 100 ml volumetric flask
and dissolve in about 50 ml of methanol and make up the volume with methanol. Filter through 0.42 m membrane. Chromatographic system: High performance liquid chromatography.
Column and stationary phase: C18 (250 mm x 4.6
mm, 5 m). Mobile phase: Filtered and degassed gradient mixture of water and acetonitrile in the following proportions:
Time
(min) (per cent) Water Acetonitrile (per cent) 0.01 80 20 5 55 45 15 20 80 20 20 80 23 55 45 25 80 20 30 80 20
Injection volume: 20 l. Flow rate: 1.5 ml per min. Detection: UV, 280 nm. Procedure: Inject 20 l of the standard solution and record the chromatogram. Inject 20 l of the test solution, record the chromatogram and measure the response for the analyte peak. Calculate the content of boeravinone B in the substance being examined from the peak response of analyte. The test is not valid unless the relative standard deviation for replicate injections is not more than 2.0 per cent.
Fig. 2: HPLC chromatogram of Punarnav¡ water extract with Boeravinone B as RS
Additional requirements:
Storage: Store in well closed container protected
from heat, light, moisture and against attack by insects and rodents.
Labelling: The label states the official name,
followed by the Latin binominal name and the part of the plant contained in the article.