Powdered Chaste Tree Extract
DEFINITION
Powdered Chaste Tree Extract is prepared from Chaste Tree by extraction with hydroalcoholic mixtures or other suitable solvents. It contains NLT 90.0% and NMT 110.0% of the labeled amount of casticin and agnuside, calculated on the dried basis. It may contain suitable added substances.
IDENTIFICATION
• A. Thin-Layer Chromatographic Identification Test
Standard solution:
100 mg of USP Powdered Chaste Tree Extract RS in 1 mL of methanol. Heat in a water bath at 60
for 10 min. Centrifuge, and use the clear supernatant.
for 10 min. Centrifuge, and use the clear supernatant.
Sample solution:
Shake a quantity of Extract, equivalent to about 10 mg of the labeled amount of agnuside, in 10 mL of methanol. Heat in a water bath at 60. Centrifuge or filter before use.
. Centrifuge or filter before use.
Adsorbent:
Chromatographic silica gel with an average particle size of 10–15 µm (TLC plates)
Application volume:
90 µL, Standard solution; 60 µL, Sample solution; in bands that are 2 cm in length
Developing solvent system:
Ethyl acetate, methanol, and water (77:15:8)
Spray reagent:
10 mg/mL of p-dimethylaminobenzaldehyde in 1 N hydrochloric acid
Analysis
Samples:
Standard solution and Sample solution
Develop the chromatograms to a length of NLT 12 cm, and dry the plate in a current of air. Spray the plate with Spray reagent, and heat for 10 min at 120.
.
Acceptance criteria:
The Sample solution shows the following: a blue zone (at an RF value of about 0.21) due to the presence of aucubin and that corresponds in color and RF value to a similar zone for the Standard solution; a blue zone (at an RF value of about 0.44) as a result of the presence of agnuside that corresponds in color and RF value to a similar zone for the Standard solution; and one broad zone, violet in the middle, near the solvent front and that corresponds in color and RF value to a similar zone for the Standard solution. Other colored zones of varying intensities may be observed in the Sample solution.
• B.
In the test for Content of Casticin, the chromatogram of the Sample solution exhibits a peak at the retention time corresponding to casticin.
• C.
In the test for Content of Agnuside, the chromatogram of the Sample solution exhibits a peak at the retention time corresponding to agnuside.
COMPOSITION
• Content of Casticin
Standard solution:
About 0.05 mg/mL of USP Casticin RS in methanol, with sonication. Pass through a cellulose membrane filter of 0.45-µm or finer pore size.
Sample solution:
Transfer a quantity of Extract, equivalent to about 2.5 mg of the labeled content of casticin, into a 50-mL volumetric flask. Add 25 mL of methanol, and sonicate in a bath at 40 for 10 min, shaking to disperse the solid. Cool to room temperature, and dilute with methanol to volume. Centrifuge or pass through a filter of 0.45-µm or finer pore size.
for 10 min, shaking to disperse the solid. Cool to room temperature, and dilute with methanol to volume. Centrifuge or pass through a filter of 0.45-µm or finer pore size.
Solution A:
Methanol
Solution B:
5.88 g/L of phosphoric acid in water
Mobile phase:
See Table 1.
Table 1
| Time (min) |
Solution A (%) |
Solution B (%) |
|---|---|---|
| 0 | 50 | 50 |
| 0 | 50 | 50 |
| 13 | 65 | 35 |
| 18 | 100 | 0 |
| 23 | 50 | 50 |
Chromatographic system
Mode:
LC
Detector:
UV 348 nm
Column:
3.1-mm × 12.5-cm; 5-µm packing L1
Column temperature:
25
Flow rate:
1 mL/min
Injection size:
10 µL
System suitability
Sample:
Standard solution
Suitability requirements
Tailing factor:
NMT 2.0 for the casticin peak
Relative standard deviation:
NMT 2.0% for the casticin peak, in repeated injections
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentage of casticin, Pc, in the portion of Extract taken:
Pc = (rU/rS) × (CS/CU) × 100
| rU | = | = peak response of casticin from the Sample solution |
| rS | = | = peak response of casticin from the Standard solution |
| CS | = | = concentration of USP Casticin RS in the Standard solution (mg/mL) |
| CU | = | = concentration of Extract in the Sample solution (mg/mL) |
Calculate the percentage of the labeled amount of casticin in the portion of Extract taken:
Result = (Pc/L) × 100
| Pc | = | = content of casticin as calculated above (%) |
| L | = | = labeled amount of casticin (%) |
Acceptance criteria:
90.0%–110.0% on the dried basis
• Content of Agnuside
Solvent:
Methanol and water (1:19)
Standard solution:
Dissolve a quantity of USP Agnuside RS in Solvent, with sonication. Dilute with methanol to obtain a concentration of about 0.125 mg/mL. Pass through a cellulose membrane filter of 0.45-µm or finer pore size.
Sample solution:
Transfer an amount of Extract, equivalent to about 6.25 mg of the labeled content of agnuside, into a 50-mL volumetric flask. Add 25 mL of Solvent, and sonicate in a bath at 40 for 10 min, shaking to disperse the solid. Cool to room temperature, and dilute with Solvent to volume. Centrifuge or pass through a filter of 0.45-µm or finer pore size.
for 10 min, shaking to disperse the solid. Cool to room temperature, and dilute with Solvent to volume. Centrifuge or pass through a filter of 0.45-µm or finer pore size.
Solution A:
Acetonitrile
Solution B:
5.88 g/L of phosphoric acid in water
Mobile phase:
See Table 2.
Table 2
| Time (min) |
Solution A (%) |
Solution B (%) |
|---|---|---|
| 0 | 7 | 93 |
| 0.6 | 10 | 90 |
| 5 | 10 | 90 |
| 7 | 14 | 86 |
| 13 | 15 | 85 |
| 13.1 | 100 | 0 |
| 18 | 100 | 0 |
| 18.1 | 7 | 93 |
| 23 | 7 | 93 |
Chromatographic system
Mode:
LC
Detector:
UV 258 nm
Column:
3.1-mm × 12.5-cm; 5-µm packing L1
Column temperature:
25
Flow rate:
1.3 mL/min
Injection size:
10 µL
System suitability
Sample:
Standard solution
Suitability requirements
Tailing factor:
NMT 2.0 for the agnuside peak
Relative standard deviation:
NMT 2.0% for the agnuside peak, in repeated injections
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentage of agnuside, Pa, in the portion of Extract taken:
Pa = (rU/rS) × (CS/CU) × 100
| rU | = | = peak response of agnuside from the Sample solution |
| rS | = | = peak response of agnuside from the Standard solution |
| CS | = | = concentration of USP Agnuside RS in the Standard solution (mg/mL) |
| CU | = | = concentration of Extract in the Sample solution (mg/mL) |
Calculate the percentage of the labeled amount of agnuside in the portion of Extract taken:
Result = (Pa/L) × 100
| Pa | = | = content of agnuside calculated above (%) |
| L | = | = labeled amount of agnuside (%) |
Acceptance criteria:
90.0%–110.0% on the dried basis
CONTAMINANTS
• Heavy Metals, Method II 
231
:
NMT 20 ppm
231:
NMT 20 ppm
• Microbial Enumeration Tests 2021:
The total bacterial count does not exceed 104 cfu/g. The total combined molds and yeasts count does not exceed 1000 cfu/g. It meets the requirements of the tests for absence of Salmonella species and Escherichia coli.
2021:
The total bacterial count does not exceed 104 cfu/g. The total combined molds and yeasts count does not exceed 1000 cfu/g. It meets the requirements of the tests for absence of Salmonella species and Escherichia coli.
• Other Requirements:
It meets the requirements for Botanical Extracts 565, Residual Solvents and Pesticide Residues.
565, Residual Solvents and Pesticide Residues.
SPECIFIC TESTS
• Loss on Drying 731:
NMT 6.0%
731:
NMT 6.0%
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight containers, and store in a cool place, protected from light.
• Labeling:
The label states the Latin binomial and, following the official name, the part of the plant from which the article was prepared. The label also indicates the content of casticin and agnuside, the extracting solvent or solvent mixture used for preparation, the ratio of the starting crude plant material to Extract, the percentage of native extract, and the name and quantity of any added substances. It meets the requirements for Botanical Extracts 565, Labeling.
565, Labeling.
Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Maged H. Sharaf, Ph.D.
Principal Scientific Liaison 1-301-816-8318 |
(DS2010) Monographs - Dietary Supplements |
| 2021 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison 1-301-816-8339 |
(GCM2010) General Chapters - Microbiology |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP35–NF30 Page 1243
Pharmacopeial Forum: Volume No. 29(4) Page 1269