Powdered Eleuthero Extract
Powdered Eleuthero Extract is prepared from Eleuthero using hydroalcoholic mixtures. The ratio of the starting crude plant material to Powdered Extract is between 13:1 and 25:1. It contains NLT 0.8% of eleutherosides B and E, calculated on the anhydrous basis. It may contain added substances.USP35
A.USP35Thin-Layer Chromatographic Identification Test 
201
Standard solution A:
1 mg/mL of USP Eleutheroside E RS in methanol
• B. HPLC:
The chromatogram of the Sample solution obtained in the test for Content of Eleutherosides B and E shows a peak at a retention time corresponding to that of eleutheroside B in the chromatogram of Standard solution B and a peak at a retention time corresponding to that of eleutheroside E in the chromatogram of Standard solution A.USP35
Standard solution A:
0.1 mg/mL of USP Eleutheroside E RS in methanol. Transfer 2.0 mL to a 5-mL volumetric flask, and dilute with Solvent to volume.
System suitability
USP35
• Microbial Enumeration Tests 2021:
The total aerobic microbial count does not exceed 104 cfu/g. The total combined yeasts and molds count does not exceed 103 cfu/g.USP35
• Absence of Specified Microorganisms 2022:
Meets the requirements of the tests for absence of Salmonella species and Escherichia coliUSP35
11
USP Eleutheroside B RS
-d-Glucopyranoside, 4-(3-hydroxy-1-propenyl)-2,6-dimethoxyphenyl.
C17H24O9 372.37
USP35
DEFINITION
Change to read:
Powdered Eleuthero Extract is prepared from Eleuthero using hydroalcoholic mixtures. The ratio of the starting crude plant material to Powdered Extract is between 13:1 and 25:1. It contains NLT 0.8% of eleutherosides B and E, calculated on the anhydrous basis. It may contain added substances.USP35IDENTIFICATION
Change to read:
• A.USP35Thin-Layer Chromatographic Identification Test 201
Standard solution A:
1 mg/mL of USP Eleutheroside E RS in methanol
Standard solution B:
1 mg/mL of USP Eleutheroside B RS in methanol
Standard solution C:
0.1 g of USP Powdered Eleuthero Extract RS in 5 mL of aqueous ethanol 50%. Sonicate for 10 min, centrifuge, and use the supernatant.
Sample solution:
0.1 g of Powdered Extract in 5 mL of aqueous ethanol 50%. Sonicate for 10 min, centrifuge, and use the supernatant.
Adsorbent:
Chromatographic silica gel with an average particle size of 5 µm (HPTLC plates)
Application volume:
10 µL, as bands
Developing solvent system:
Chloroform, methanol, and water (35:15:2)
Spray reagent:
Place 18 mL of methanol in a glass flask, and cool in a water–ice–salt bath or in a freezer. To the ice-cold methanol, slowly and carefully add 2 mL of sulfuric acid, and mix well. Allow the mixture to adjust to room temperature.
Analysis
Samples:
Standard solution A, Standard solution B, Standard solution C, and Sample solution
Before the development of the chromatogram, saturate the chamber for 20 min with Developing solvent system. Record the temperature and humidity in the laboratory. If the relative humidity exceeds 50%, condition the plate to about 30% relative humidity, using a suitable device. Develop the plate over a path of 6 cm, dry, and spray the plate with Spray reagent. Heat the plate at 100
for 5 min, and examine under visible light and UV light at 365 nm.
for 5 min, and examine under visible light and UV light at 365 nm.
Acceptance criteria:
Under visible light, the Sample solution exhibits two brown bands due to eleutheroside E and eleutheroside B at RF values of about 0.34 and 0.45, corresponding in color and RF to the bands exhibited by Standard solution A and Standard solution B, respectively. The Sample solution also exhibits two additional brown bands near the application zone, corresponding in color and RF values to the bands exhibited by Standard solution C. Other bands may be observed in the Sample solution and Standard solution C chromatograms. Under UV light, the Sample solution shows a brown band due to eleutheroside E corresponding in color and RF to the band exhibited by Standard solution A.USP35
USP35
Add the following:
• B. HPLC:
The chromatogram of the Sample solution obtained in the test for Content of Eleutherosides B and E shows a peak at a retention time corresponding to that of eleutheroside B in the chromatogram of Standard solution B and a peak at a retention time corresponding to that of eleutheroside E in the chromatogram of Standard solution A.USP35
COMPOSITION
Change to read:
• Content of Eleutherosides B and E
Solvent:
Methanol and water (1:1)
Solution A:
Acetonitrile and water (5:95)
Solution B:
Acetonitrile and water (60:40)
Mobile phase:
See Table 1.
Table 1
| Time (min) |
Solution A (%) |
Solution B (%) |
|---|---|---|
| 0 | 97 | 3 |
| 5 | 97 | 3 |
| 30 | 60 | 40 |
| 31 | 5 | 95 |
| 45 | 5 | 95 |
| 45.1 | 97 | 3 |
| 60 | 97 | 3 |
Standard solution A:
0.1 mg/mL of USP Eleutheroside E RS in methanol. Transfer 2.0 mL to a 5-mL volumetric flask, and dilute with Solvent to volume.
Standard solution B:
0.1 mg/mL of USP Eleutheroside B RS in methanol. Transfer 2.0 mL to a 5-mL volumetric flask, and dilute with Solvent to volume.
Standard solution C:
5.0 mg/mL of USP Powdered Eleuthero Extract RS in Solvent. Sonicate for 30 min, cool to room temperature, and decant. Before injection, pass through a nylon filter of 0.45-µm or finer pore size, discarding the first few mL of the filtrate.USP35
USP35
Sample solution:
Transfer 500 mg of Powdered Extract to a 100-mL volumetric flask, add 80 mL of Solvent, and sonicate for 30 min. Cool to room temperature, dilute with Solvent to volume, and mix. Before injection, pass through a nylon filter of 0.45-µm or finer pore size, discarding the first few mL of the filtrate.USP35
Before injection, pass through a nylon filter of 0.45-µm or finer pore size, discarding the first few mL of the filtrate.USP35
Chromatographic system
Mode:
LC
Detector:
UV 220 nm
Column:
4.0-mm × 25-cm; 5-µm packing L1
Flow rate:
1 mL/min
Injection size:
10 µL
System suitability
Samples:
Standard solution B and Standard solution C
Suitability requirements
Chromatogram similarity:
The chromatogram from Standard solution C is similar to the reference chromatogram provided with the lot of USP Powdered Eleuthero Extract RS being used.
Relative standard deviation:
NMT 2.0% determined from the eleutheroside B peak in repeated injections, Standard solution B
USP35
Analysis
Samples:
Standard solution A, Standard solution B, Standard solution C, and Sample solution
USP35
Samples:
Standard solution A, Standard solution B, Standard solution C, and Sample solution
Identify the eleutheroside B and eleutheroside E peaks in the Sample solution by comparison with the chromatograms of Standard solution B and Standard solution A, respectively, and measure the peak responses.
Separately calculate the percentages of eleutheroside B and eleutheroside E in the portion of Powdered Extract taken:
Result = (rU/rS) × (CS/CU) × 100
| rU | = | = peak response of the relevant analyte from the Sample solution |
| rS | = | = peak response of eleutheroside E or eleutheroside B from Standard solution A or Standard solution B, respectively |
| CS | = | = concentration of eleutheroside E or eleutheroside B in Standard solution A or Standard solution B, respectively (mg/mL) |
| CU | = | = concentration of Powdered Extract in the Sample solution (mg/mL) |
USP35
Acceptance criteria:
NLT 0.8% on the anhydrous basis
CONTAMINANTS
• Heavy Metals, Method II 231:
NMT 20 ppm
231:
NMT 20 ppm
Add the following:
• Microbial Enumeration Tests 2021:
The total aerobic microbial count does not exceed 104 cfu/g. The total combined yeasts and molds count does not exceed 103 cfu/g.USP35
Add the following:
• Absence of Specified Microorganisms 2022:
Meets the requirements of the tests for absence of Salmonella species and Escherichia coliUSP35
SPECIFIC TESTS
• Water Determination, Method Ia 921:
NMT 5.0%
921:
NMT 5.0%
• Articles of Botanical Origin, Total Ash 561:
NMT 10.0%
561:
NMT 10.0%
• Alcohol Determination, Method II 611:
NMT 0.5%
611:
NMT 0.5%
• Other Requirements:
It meets the requirements for Residual Solvents and Pesticide Residues in Botanical Extracts 565.
565.
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight, light-resistant containers.
• 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 eleutherosides, the extracting solvent used for preparation, and the ratio of the starting crude plant material to Powdered Extract. It meets the requirements for Labeling in Botanical Extracts 565.
565.
Change to read:
• USP Reference Standards 11
USP Eleutheroside B RS
-d-Glucopyranoside, 4-(3-hydroxy-1-propenyl)-2,6-dimethoxyphenyl.
C17H24O9 372.37
USP Eleutheroside E RS
-d-Glucopyranoside, (tetrahydro-1H,3H-furo(3,4-c)furan-1,4-diyl)bis(2,6-dimethoxy-4,1-phenylene)bis-.
C34H46O18 742.70
-d-Glucopyranoside, (tetrahydro-1H,3H-furo(3,4-c)furan-1,4-diyl)bis(2,6-dimethoxy-4,1-phenylene)bis-.
C34H46O18 742.70
USP35
USP Powdered Eleuthero Extract RS
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 |
| 2022 |
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 1287
Pharmacopeial Forum: Volume No. 36(6) Page 1592