American Ginseng Capsules
DEFINITION
American Ginseng Capsules contain Powdered American Ginseng Extract. Capsules contain NLT 90.0% and NMT 110.0% of the labeled amount of Extract, calculated as the sum of ginsenosides Rg1, Re, Rb1, Rc, Rb2, and Rd.
IDENTIFICATION
• A. Thin-Layer Chromatographic Identification Test 
201
201
Sample solution
Soft-shell gelatin Capsules:
Transfer a portion of the contents of the Capsules, equivalent to 100 mg of Powdered Extract, to a separatory funnel containing 30 mL of a mixture of hexanes, methanol, and water (20:15:10), dissolve in this mixture, and collect the lower layer. Wash the upper layer with three 15-mL portions of a mixture of methanol and water (15:10), and combine the washings with the lower layer. Evaporate to dryness under vacuum at 45
–50. Dissolve the residue in 5 mL of methanol.
–50. Dissolve the residue in 5 mL of methanol.
Hard-shell gelatin Capsules:
Transfer a portion of the contents of the Capsules, equivalent to 100 mg of Powdered Extract, to a conical flask. Extract at 55 with three 20-mL portions of a mixture of methanol and water (2:8). Evaporate the combined extracts to dryness under vacuum at 45–50. Dissolve the residue in 5 mL of methanol.
with three 20-mL portions of a mixture of methanol and water (2:8). Evaporate the combined extracts to dryness under vacuum at 45–50. Dissolve the residue in 5 mL of methanol.
Standard solution A:
20 mg/mL of USP Powdered American Ginseng Extract RS in methanol
Standard solution B:
20 mg/mL of USP Powdered Asian Ginseng Extract RS in methanol
Application volume:
20 µL
Developing solvent system A:
The lower phase of a mixture of chloroform, methanol, and water (13:7:2)
Developing solvent system B:
The upper phase of a mixture of butyl alcohol, ethyl acetate, and water (4:1:5)
Spray reagent:
Dissolve 0.5 mL of anisaldehyde in 10 mL of glacial acetic acid, add 85 mL of methanol, mix, carefully add 5 mL of sulfuric acid, and mix.
Analysis
Samples:
Sample solution, Standard solution A, and Standard solution B
Develop the chromatograms in a chamber containing Developing solvent system A until the solvent front has moved 10.5 cm from the origin. Remove the plate from the chamber, and allow to dry. Turn the plate 90, and develop in a chamber containing Developing solvent system B until the solvent front has moved 10.5 cm from the origin. Remove the plate from the chamber, and allow to dry. Spray with Spray reagent. Heat the plate at 105–110 for 10 min, and examine. The order, from top to bottom, of ginsenosides on the plates is Rg2 (on left) and Rg1 (on right), Rf, Re, Rd, Rc, Rb2 (on left) and Rb1 (on right), and Ro.
, and develop in a chamber containing Developing solvent system B until the solvent front has moved 10.5 cm from the origin. Remove the plate from the chamber, and allow to dry. Spray with Spray reagent. Heat the plate at 105–110 for 10 min, and examine. The order, from top to bottom, of ginsenosides on the plates is Rg2 (on left) and Rg1 (on right), Rf, Re, Rd, Rc, Rb2 (on left) and Rb1 (on right), and Ro.Ginsenosides Rg2, Rg1, Rf, Re, and Rd are found on the upper half of the plates; the remaining ginsenosides are found on the lower half after chromatographing with Developing solvent system B.
Acceptance criteria:
Standard solution A does not exhibit a spot for ginsenoside Rf. Standard solution B exhibits a spot for ginsenoside Rf. The spots from the Sample solution correspond to those from Standard solution A.
• B.
The retention times of the peaks for ginsenosides Rg1, Re, Rb1, Rb2, Rc2, and Rd in the chromatogram of the Sample solution correspond to those from the Standard solution, as obtained in the test for Content of Ginsenosides. The ratio of the peak response for Rb2 to the peak response for Rb1 is less than 0.4; and the ratio of the peak response for Rg1 to the peak response for Rb1 is less than 0.3. There is no significant peak at the retention time corresponding to that of ginsenoside Rf in the System suitability solution, as obtained in the test for Content of Ginsenosides.
COMPOSITION
• Content of Ginsenosides
Method 1
Diluent:
Water and alcohol (3:2)
Solution A:
Water
Solution B:
Acetonitrile and water (4:1)
Mobile phase:
See the gradient table below.
| Time (min) |
Solution A (%) |
Solution B (%) |
|---|---|---|
| 0 | 76 | 24 |
| 12 | 76 | 24 |
| 28 | 65 | 35 |
| 51.5 | 56.5 | 43.5 |
| 52.5 | 0 | 100 |
| 64.5 | 76 | 24 |
| 77 | 76 | 24 |
Standard solution:
A solution of USP Powdered American Ginseng Extract RS in Diluent containing the equivalent of 0.2 mg/mL of ginsenoside Rb1
Sample solution (soft-gelatin Capsules):
Open NLT 20 Capsules, transfer the contents to a suitable container, and mix to homogenize. Transfer a portion, expected to contain an amount of Extract equivalent to 12 mg of ginsenosides, to a suitable flask with a stopper. Add 5.0 mL of tetrahydrofuran, and sonicate for 5 min. Add 25.0 mL of a mixture of methanol and water (4:6), and shake for 50 min in an automatic shaker. Transfer 15.0 mL of the obtained emulsion to a centrifuge tube with a stopper, add 800 mg of sodium chloride, shake for 30 s, and centrifuge to obtain a clear upper phase. Dilute 1.0 mL of the upper phase with 4 mL of water in a suitable tube, and transfer the solution to a column containing 360 mg of packing L2 that has been previously treated with 3.0 mL of methanol followed by 8.0 mL of water. [Note—Elute slowly, not faster than 1 drop/s, in all elution steps. Do not use vacuum. ] Rinse the tube with 5 mL of water, transfer to the column taking the precaution of slow elution, and discard the eluate. Repeat the elution with 5 mL of a mixture of methanol and water (4:6), and discard the eluate. Elute the ginsenosides with 5.0 mL of methanol. Evaporate the solution under a stream of nitrogen at 40 (50 min), and dissolve the residue with 1.0 mL of a solution of acetonitrile and water (1:4).
(50 min), and dissolve the residue with 1.0 mL of a solution of acetonitrile and water (1:4).
System suitability solution:
24 mg/mL of USP Powdered Asian Ginseng Extract RS in Diluent. Filter.
Chromatographic system
Mode:
LC
Detector:
UV 203 nm
Column
Guard column:
4.6-mm × 2.0-cm; packing L1
Analytical column:
4.6-mm × 15-cm; 3-µm packing L1
Column temperature:
25
Flow rate:
1.5 mL/min
Injection size:
10 µL
System suitability
Sample:
System suitability solution (inject 20 µL)
Suitability requirements
Chromatogram similarity:
The System suitability solution chromatogram is similar to the Reference Chromatogram provided with the lot of USP Powdered Asian Ginseng Extract RS being used.
Relative standard deviation:
NMT 2.0%, determined for the sum of the peak areas for the six major ginsenosides, in repeated injections
Analysis
Samples:
Standard solution and Sample solution
Identify ginsenosides Rg1, Re, Rb1, Rc, Rb2, and Rd in the Standard solution and the Sample solution by comparing the chromatograms with the Reference Chromatogram provided with USP Powdered American Ginseng Extract RS being used, and measure the peak responses.
Calculate the quantity, in mg, of each relevant ginsenoside (Rg1, Re, Rb1, Rc, Rb2, and Rd) in the portion of Capsule contents taken:
Result = 0.3 × (rU/rS) × CS × P
| rU | = | = peak area for each relevant ginsenoside from the Sample solution |
| rS | = | = peak area for each relevant ginsenoside from the Standard solution |
| CS | = | = concentration of USP Powdered American Ginseng Extract RS in the Standard solution (mg/mL) |
| P | = | = labeled amount, in percentage, of each relevant ginsenoside in the USP Powdered American Ginseng Extract RS lot being used |
Calculate the content of total ginsenosides, T, in mg, by adding the amounts of individual ginsenoside.
Calculate the percentage of Powdered Extract with respect to the label claim:
Result = T × (AWT/W) × (100/LE) × (100/L)
| T | = | = content of total ginsenosides in the portion of Capsule contents taken (mg) |
| AWT | = | = average weight of Capsule contents (mg/Capsule) |
| W | = | = weight of the portion of Capsule contents taken (mg) |
| LE | = | = content of total ginsenosides, mg, in 100 mg of the Extract used to prepare the Capsules |
| L | = | = amount of Extract per Capsule according to label claim (mg/Capsule) |
Method 2
Diluent, Solution A, Solution B, Mobile phase, System suitability solution, Chromatographic system, and Suitability requirements:
Proceed as directed under Method 1.
Solvent A:
Upper phase of a mixture consisting of hexane, methanol, and water (4:3:2)
Solvent B:
Lower phase of a mixture consisting of hexane, methanol, and water (4:3:2)
Standard solution:
A solution of USP Powdered American Ginseng Extract RS in Diluent containing the equivalent of 1 mg/mL of ginsenoside Rb1
Sample solution A (for soft-gelatin Capsules):
Open NLT 20 Capsules and transfer the contents to a suitable container. Mix to homogenize and transfer a portion, expected to contain an amount of Extract equivalent to 15 mg of total ginsenosides, to a 50-mL flask. Add 10.0 mL of Solvent A, and sonicate for 3–5 min at 25–30. Transfer the solution to a 125-mL separatory funnel. To the residue add 10 mL of Solvent B, and sonicate for 3–5 min at 25–30. Transfer the solution to the same separatory funnel. Repeat the above procedure twice (the total volume will be about 60 mL). Shake, and then allow the phases to separate. Collect the combined lower phase in a round-bottom flask, and wash the combined upper phase twice with 10 mL of Solvent B. Evaporate the combined lower phase to dryness under vacuum at 45–50. Transfer the residue to a 10-mL volumetric flask using small volumes of methanol, and dilute with methanol to volume.
–30. Transfer the solution to a 125-mL separatory funnel. To the residue add 10 mL of Solvent B, and sonicate for 3–5 min at 25–30. Transfer the solution to the same separatory funnel. Repeat the above procedure twice (the total volume will be about 60 mL). Shake, and then allow the phases to separate. Collect the combined lower phase in a round-bottom flask, and wash the combined upper phase twice with 10 mL of Solvent B. Evaporate the combined lower phase to dryness under vacuum at 45–50. Transfer the residue to a 10-mL volumetric flask using small volumes of methanol, and dilute with methanol to volume.
Sample solution B (for hard-gelatin Capsules):
Weigh the contents of NLT 20 Capsules, and composite the contents. Transfer a portion of the composite, expected to contain an amount of Extract equivalent to 15 mg of total ginsenosides, to a conical flask. Add 15 mL of methanol, and shake to mix. Sonicate the mixture at 25–30 for 30 min. Cool, pass through filter paper, and return the residue to the conical flask. Add another 15 mL of methanol, sonicate the mixture at 25–30 for 30 min, and filter. Wash the residue with three 15-mL portions of methanol. Evaporate the combined extracts and washing to dryness under vacuum at 45–50. Transfer the residue to a 10-mL volumetric flask using small volumes of methanol, and dilute with methanol to volume.
–30 for 30 min. Cool, pass through filter paper, and return the residue to the conical flask. Add another 15 mL of methanol, sonicate the mixture at 25–30 for 30 min, and filter. Wash the residue with three 15-mL portions of methanol. Evaporate the combined extracts and washing to dryness under vacuum at 45–50. Transfer the residue to a 10-mL volumetric flask using small volumes of methanol, and dilute with methanol to volume.
Analysis
Samples:
Standard solution and Sample solution
Identify ginsenosides Rg1, Re, Rb1, Rc, Rb2, and Rd in the Standard solution and the Sample solution by comparing the chromatograms with the Reference Chromatogram provided with USP Powdered American Ginseng Extract RS, and measure the peak responses.
Calculate the quantity, in mg, of each relevant ginsenoside (Rg1, Re, Rb1, Rc, Rb2, and Rd) in the portion of Capsule contents taken:
Result = 0.1 × (rU/rS) × CS × P
| rU | = | = peak area for each relevant ginsenoside from the Sample solution |
| rS | = | = peak area for each relevant ginsenoside from the Standard solution |
| CS | = | = concentration of USP Powdered American Ginseng Extract RS in the Standard solution (mg/mL) |
| P | = | = labeled amount, in percentage, of each relevant ginsenoside in the USP Powdered American Ginseng Extract RS lot being used |
Calculate the content of total ginsenosides, T, in mg, by adding the amounts of individual ginsenoside.
Calculate the percentage of Powdered Extract with respect to the label claim:
Result = T × (AWT/W) × (100/LE) × (100/L)
| T | = | = content of total ginsenosides in the portion of Capsule contents taken (mg) |
| AWT | = | = average weight of Capsule contents (mg/Capsule) |
| W | = | = weight of the portion of Capsule contents taken (mg) |
| LE | = | = content of total ginsenosides, mg, in 100 mg of the Extract used to prepare the Capsules |
| L | = | = amount of Extract per Capsule according to label claim (mg/Capsule) |
Acceptance criteria:
90.0%–110.0% of Extract, calculated as the sum of ginsenosides Rg1, Re, Rb1, Rc, Rb2, and Rd
PERFORMANCE TESTS
• Disintegration and Dissolution of Dietary Supplements 2040:
Meet the requirements for Disintegration
2040:
Meet the requirements for Disintegration
• Weight Variation of Dietary Supplements 2091:
Meet the requirements
2091:
Meet the requirements
CONTAMINANTS
• Microbial Enumeration Tests 2021:
The total aerobic microbial count does not exceed 104 cfu/g. The total combined molds and yeasts count does not exceed 103 cfu/g.
2021:
The total aerobic microbial count does not exceed 104 cfu/g. The total combined molds and yeasts count does not exceed 103 cfu/g.
• Absence of Specified Microorganisms 2022:
Meet the requirements of the tests for absence of Salmonella species and Escherichia coli.
2022:
Meet the requirements of the tests for absence of Salmonella species and Escherichia coli.
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight containers, protected from light. Store at controlled room temperature.
• Labeling:
The label states the Latin binomial and, following the official name, the article from which the Capsules were prepared. The label also indicates the amount of Extract, in mg/Capsule. Label the Capsules to indicate the percentage of ginsenosides in the Extract contained in the Capsules. For soft-gelatin Capsules, state the method for Content of Ginsenosides with which the product complies only if Method 1 is not used.
• USP Reference Standards 11
11
USP Powdered American Ginseng Extract RS
USP Powdered Asian Ginseng 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 1179
Pharmacopeial Forum: Volume No. 36(1) Page 157