Rhodiola rosea
DEFINITION
Rhodiola rosea consists of the dried roots and rhizomes of Rhodiola rosea L. (Fam. Crassulaceae). It contains NLT 0.3% of phenylpropenoid glycosides calculated as the sum of rosarin, rosavin, and rosin; and NLT 0.08% salidroside; both calculated on the dried basis.
IDENTIFICATION
• A.
Rhodiola rosea meets the requirements under Specific Tests, Botanic Characteristics.
• B. 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:
Sonicate for 10 min about 0.5 g of Rhodiola rosea, finely powdered, in 5 mL of methanol, centrifuge, 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 each of Standard solution B and 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.
• C. 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 content of phenylpropenoid glycosides: rosarin, rosavin, and rosin, to the content of salidroside is about 3:1.
COMPOSITION
Change to read:
• 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 |
Solvent:
75% methanol in water
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:
Transfer about 5.0 g of Rhodiola rosea, finely powdered and accurately weighed, to a 25-mL flask. Add 7 mL of Solvent, sonicate for 15 min, and filter into a 10-mL volumetric flask. Wash the residue on the filter paper twice, using 1 mL of Solvent each; add the washings to the volumetric flask; adjust the volume using Solvent; and mix. Before injection, pass through a membrane filter of 0.45-µm or finer pore size, discarding the first few mL of the filtrate.
Chromatographic system
(See Chromatography 
621
, System Suitability.)
621, System Suitability.)
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 Rhodiola rosea taken:
Result = (rU/rS) × CS × (V/W) × 100
| rU | = | = peak area of the relevant analyte in the Sample solution |
| rS | = | = peak area of rosavin in Standard solution A |
| CS | = | = concentration of rosavin in Standard solution A (mg/mL) |
| V | = | = volume of Sample solution (mL) |
| W | = | = weight of Rhodiola rosea taken to prepare the Sample solution (mg) |
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 Rhodiola rosea taken:
Result = (rU/rS) × CS × (V/W) × 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 (mg/mL) |
| V | = | = volume of the Sample solution (mL) |
| W | = | = weight of Rhodiola rosea taken to prepare the Sample solution (g) |
Acceptance criteria:
NLT 0.3% of phenylpropenoid glycosides and NLT 0.08% salidroside; both calculated on the dried basis.
CONTAMINANTS
• Elemental Impurities—Procedures 233
233
Acceptance criteria
Arsenic:
NMT 2.0 µg/g
Cadmium:
NMT 1.0 µg/g
Lead:
NMT 5.0 µg/g
Mercury:
NMT 1.0 µg/g
• Articles of Botanical Origin, General Method for Pesticide Residues Analysis 561:
Meets the requirements
561:
Meets the requirements
• Microbial Enumeration Tests 2021:
The total aerobic bacterial count does not exceed 105 cfu/g, the total combined molds and yeasts count does not exceed 103 cfu/g, and the bile-tolerant Gram-negative bacteria count does not exceed 103 cfu/g.
2021:
The total aerobic bacterial count does not exceed 105 cfu/g, the total combined molds and yeasts count does not exceed 103 cfu/g, and the bile-tolerant Gram-negative bacteria count does not exceed 103 cfu/g.
• Absence of Specified Microorganisms 2022:
Meets the requirements of the tests for absence of Salmonella species and Escherichia coli
2022:
Meets the requirements of the tests for absence of Salmonella species and Escherichia coli
• Articles of Botanical Origin, Test for Aflatoxins 561:
Meets the requirements
561:
Meets the requirements
SPECIFIC TESTS
• Botanic Characteristics
Macroscopic:
Underground parts consist of numerous rhizomes united at their base into a long taproot. Both the rhizome and root exhibit secondary growth. Pharmacopeial article consists of dry pieces of rhizomes and roots of various shapes. Pieces of rhizomes are thick, wrinkly, with remains of stems and scales, and pieces of roots branching off the rhizome. The surface of the rhizome and the roots is shiny, grayish-brown; after peeling off the cork, a golden-yellow layer is revealed; fracture, pinkish-brown or light brown.
Microscopic
Transverse section of rhizome:
Cork, narrow to broad, depending on the sample; cork cells may be dark brown, greenish, or nearly colorless; cortex, large, loosely arranged parenchymatous cells, with slightly thickened walls; secondary phloem, parenchymatous, no sclereides; small narrow vascular bundles occur in a ring surrounding a broad parenchymatous pith; pith includes scattered vascular bundles; parenchyma of the rhizome is filled with starch granules, simple, round or oval-shaped, 5–20 µm in diameter; hilum, if present, appears as a small dot.
Transverse section of root:
Cork, narrow to broad, depending on the sample; cortex, large, parenchymatous cells, may contain orange-brown tannin ducts; tannin ducts are also found embedded in the cork of old roots; secondary phloem, parenchymatous, no sclereides; small narrow vascular bundles occur in a ring surrounding a narrow parenchymatous pith; starch granules, simple, round or oval-shaped, 5–20 µm in diameter; hilum, if present, appears as a small dot.
• Loss on Drying 731
731
Sample:
1.0 g of finely powdered Rhodiola rosea
Analysis:
Dry at 105 for 2 h.
for 2 h.
Acceptance criteria:
NMT 12%
• Articles of Botanical Origin, Total Ash 561
561
Sample:
2 g of finely powdered Rhodiola rosea
Acceptance criteria:
NMT 12%
• Articles of Botanical Origin, Acid-Insoluble Ash 561
561
Sample:
2–4 g of finely powdered Rhodiola rosea
Acceptance criteria:
NMT 3%
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in well-closed containers, protected from light and moisture, and store at room temperature.
• Labeling:
The label states the Latin binomial and, following the official name, the part of the plant contained in the article.
• 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 |
| 2021 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison (301) 816-8339 |
(GCM2010) General Chapters - Microbiology |
| 2022 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison (301) 816-8339 |
(GCM2010) General Chapters - Microbiology |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP38–NF33 Page 6193
Pharmacopeial Forum: Volume No. 39(3)