Rhodiola rosea Tincture
DEFINITION
Rhodiola rosea Tincture is prepared as follows.
| Rhodiola rosea | 1 part (g) |
| A mixture of Alcohol and Water (40:60), a sufficient quantity to make | 5 parts (mL) |
Prepare the Tincture as directed for Botanical Extracts 
565
, Tinctures, Maceration Process. It contains NLT 0.06% (w/v) of phenylpropenoid glycosides calculated as the sum of rosarin, rosavin, and rosin; and NLT 0.016% of salidroside.
565, Tinctures, Maceration Process. It contains NLT 0.06% (w/v) of phenylpropenoid glycosides calculated as the sum of rosarin, rosavin, and rosin; and NLT 0.016% of salidroside.IDENTIFICATION
• A. Thin-Layer Chromatography
Standard solution A:
1.0 mg/mL of USP Rosavin RS in methanol
Standard solution B:
50 mg/mL of USP Rhodiola rosea Root and Rhizome Dry Extract RS in methanol. Sonicate for 10 min, centrifuge, and use the supernatant.
Sample solution:
Centrifuge a portion of Tincture, and use the supernatant.
Chromatographic system
Adsorbent:
Chromatographic silica gel with an average particle size of 5 µm (HPTLC plates)
Application volume:
3 µL of Standard solution A, 5 µL of Standard solution B, and 10 µL of Sample solution; as 8-mm bands
Relative humidity:
Condition the plate to a relative humidity of about 33% using a suitable device.
Developing solvent system:
A mixture of ethyl acetate, methanol, water, and formic acid (77:13:10:2)
Developing distance:
6 cm
Derivatization reagent:
Dissolve 1 g of diphenylamine in 40 mL of acetone, add 1 mL of aniline, and mix. Carefully add 7.5 mL of phosphoric acid, and mix.
Analysis
Samples:
Standard solution A, Standard solution B, and Sample solution
Apply the samples as bands to a suitable high performance thin-layer chromatographic plate, and dry in air. Develop the chromatograms in a saturated chamber, remove the plate from the chamber, and dry in air. Derivatize the plate with Derivatization reagent, heat at 120
for 5 min, and examine under visible light.
for 5 min, and examine under visible light.
System suitability:
The chromatogram of Standard solution B exhibits, in the lower half, three gray bands and two brownish bands, one above and the other below the gray bands; the most intense band in the chromatogram is the brownish band with an RF below the gray bands; the most intense gray band is the lower band at an RF corresponding to the band due to rosavin in the chromatogram of Standard solution A; the upper gray band due to rosarin is less intense.
Acceptance criteria:
The chromatogram of the Sample solution exhibits a gray band corresponding to the band due to rosavin in the chromatogram of Standard solution A, and the following bands corresponding to similar bands in the chromatogram of Standard solution B: two additional gray bands and two brownish bands, one above and the other below the gray bands; the most intense band in the chromatogram is the brownish band with an RF below the gray bands; the most intense gray band is the lower band due to rosavin.
• B. HPLC
Analysis:
Proceed as directed in the test for Content of Phenylpropenoid Glycosides and Salidroside.
Acceptance criteria:
The chromatogram of the Sample solution exhibits peaks at the retention times corresponding to the peaks due to salidroside, tyrosol, rosarin, rosavin, rosin, and rosiridin in the chromatogram of Standard solution C.
The ratio of the contents of rosarin, rosavin, and rosin is about 2.5: 6.0: 1.5.
COMPOSITION
• Content of Phenylpropenoid Glycosides and Salidroside
Solution A:
Water
Solution B:
Acetonitrile
Mobile phase:
See Table 1.
Table 1
| Time (min) |
Solution A (%) |
Solution B (%) |
|---|---|---|
| 0 | 94 | 6 |
| 6 | 83 | 17 |
| 7 | 80.3 | 19.7 |
| 9 | 80.3 | 19.7 |
| 10 | 0 | 100 |
| 12 | 94 | 6 |
| 17 | 94 | 6 |
Standard solution A:
1.0 mg/mL of USP Rosavin RS in methanol
Standard solution B:
0.3 mg/mL of USP Salidroside RS in methanol
Standard solution C:
4.0 mg/mL of USP Rhodiola rosea Root and Rhizome Dry Extract RS in methanol. Sonicate to dissolve, if necessary. Before injection, pass through a membrane filter of 0.45-µm or finer pore size.
Sample solution:
Before injecting the Tincture, pass through a membrane filter of 0.45-µm or finer pore size. [Note—The sample can be weighed and converted to volume using the density of the Tincture. ]
Chromatographic system
Mode:
LC
Detector:
UV 205 nm
Column:
3.0-mm × 10-cm; 2.5-µm packing L1
Column temperature:
40 ± 1
Flow rate:
1.0 mL/min
Injection volume:
1 µL
System suitability
Samples:
Standard solution A and Standard solution C
Suitability requirements
Chromatogram similarity:
The chromatogram obtained from Standard solution C is similar to the reference chromatogram provided with the lot of USP Rhodiola rosea Root and Rhizome Dry Extract RS being used.
Resolution:
NLT 1.5 between the rosarin and rosavin peaks, Standard solution C
Relative standard deviation:
NMT 2% determined from the rosavin peak in repeated injections, Standard solution A
Analysis
Samples:
Standard solution A, Standard solution B, Standard solution C, and Sample solution
Using the chromatograms of Standard solution A, Standard solution B, Standard solution C, and the reference chromatogram provided with the lot of USP Rhodiola rosea Root and Rhizome Dry Extract RS being used, identify the retention time of the peaks corresponding to salidroside, tyrosol, rosarin, rosavin, rosin, and rosiridin in the Sample solution.
Separately calculate the percentages of rosarin, rosavin, and rosin as rosavin in the portion of Tincture taken:
Result = (rU/rS) × CS × 100
| rU | = | = peak area of the relevant analyte from the Sample solution |
| rS | = | = peak area of rosavin from Standard solution A |
| CS | = | = concentration of rosavin in Standard solution A (g/mL) |
Calculate the percentage of phenylpropenoid glycosides as the sum of the percentages of rosarin, rosavin, and rosin.
Calculate the percentage of salidroside in the portion of Tincture taken:
Result = (rU/rS) × CS × 100
| rU | = | = peak area of salidroside from the Sample solution |
| rS | = | = peak area of salidroside from Standard solution B |
| CS | = | = concentration of salidroside in Standard solution B (g/mL) |
Acceptance criteria:
NLT 0.06% (w/v) of phenylpropenoid glycosides; and NLT 0.016% of salidroside
OTHER COMPONENTS
• Alcohol Determination, Method I 611:
NLT 90.0% and NMT 110.0% of the labeled amount of ethanol (C2H5OH)
611:
NLT 90.0% and NMT 110.0% of the labeled amount of ethanol (C2H5OH)
CONTAMINANTS
• Articles of Botanical Origin, General Method for Pesticide Residues Analysis 561:
Meets the requirements
561:
Meets the requirements
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight, light-resistant containers, and store at room temperature.
• Labeling:
The label states the official name of the article, the Latin binomial, and the part of the plant from which the article was prepared. Label it to indicate the content of phenylpropenoid glycosides and salidroside, the solvent mixture used for extraction, and the ratio of the starting crude plant material to Tincture.
• USP Reference Standards 11
11
USP Rhodiola rosea Root and Rhizome Dry Extract RS
USP Rosavin RS 
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USP Salidroside RS
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Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Anton Bzhelyansky, M.S.
Scientific Liaison (301) 230-6303 |
(DS2010) Monographs - Dietary Supplements and Herbal Medicines |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP38–NF33 Page 6195
Pharmacopeial Forum: Volume No. 39(3)