Rhodiola rosea Root and Rhizome Dry Extract

Rhodiola rosea Root and Rhizome Dry Extract

Final Authorized Version 1.0

Rhodiola rosea Root and Rhizome Dry Extract


 

DEFINITION

The extract is prepared from Rhodiola rosea L. roots and rhizomes (Family Crassulaceae) by extraction with hydro-alcoholic mixtures. It may contain suitable added substances as carriers. It contains NLT 90.0% and NMT 110.0% of the labeled amount of the phenylpropenoid glycosides rosarin, rosavin, and rosin calculated as rosavin, and NLT 90.0% and NMT 110.0% of the labeled amount of salidroside, calculated on the dried basis.

 

POTENTIAL CONFOUNDING MATERIALS

Related Rhodiola species including R. kirilowii, R. yunnanensis, R. crenulata, R. sacra, and R. sachalinensis

 

CONSTITUENTS OF INTEREST

Phenylpropenoid glycosides: Rosarin, rosavin, rosin

Phenylethanoids: Salidroside, tyrosol

Monoterpene glycoside: Rosiridin

 

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: 50 mg/mL of Rhodiola rosea Root and Rhizome Dry Extract in methanol. Sonicate for 10 min, centrifuge, and use the supernatant.

Chromatographic system

(See Chromatography <621>, Thin-Layer Chromatography.

Adsorbent: Chromatographic silica gel with an average particle size of 5 µm (HPTLC plates)

Application volume: 3 µL of Standard solution A and 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.

Temperature: 25°

Developing solvent system: 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 HPTLC plate and dry in air. Develop the chromatograms in a saturated chamber, remove the plate from the chamber, and dry. Treat with Derivatization reagent, heat for 5 min at 120°, and examine under visible light.

System suitability: Standard solution A shows a gray band at about one-fourth of the chromatogram. The chromatogram of Standard solution B exhibits, in the lower half, three gray bands, the most intense gray band at an RF corresponding to the band due to rosavin in the chromatogram of Standard solution A; the other two are above the band corresponding to rosavin. The chromatogram of Standard solution B exhibits the most intense band as a brownish band with an RF below rosavin.

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. It exhibits the following bands corresponding to similar bands in the chromatogram of Standard solution B: two additional gray bands and two brownish bands, one above the group of gray bands; the most intense band in the chromatogram is the brownish band with an RF below the rosavin; the most intense gray band is the lower band due to rosavin. 

• B. HPLC

Analysis: Proceed as directed in the Assay 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 B. The ratio of the contents of rosarin, rosavin, and rosin is about 2.5: 6.0: 1.5.

 

ASSAY

• 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: 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.

Standard solution C: 1.0 mg/mL of USP Salidroside RS in methanol

Sample solution: 4.0 mg/mL of Rhodiola rosea Root and Rhizome Dry Extract in methanol. Sonicate to dissolve, if necessary. 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.)

Mode: LC

Detector: UV 205 nm

Column: 3.0-mm × 10-cm; 2.5-µm packing L1 (similar to Luna C18-HST)

Column temperature: 40 ± 1°

Flow rate: 1.0 mL/min

Injection volume: 1 µL

System suitability 

Samples: Standard solution A and Standard solution B

Suitability requirements 

Chromatogram similarity: The chromatogram of Standard solution B 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 rosarin and rosavin peaks, Standard solution B

Tailing factor: NMT 2.0 for the rosavin peak, Standard solution A

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:

 

P1 = (rU/rS) × (CS/CU) × 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)

CU    = concentration of Dry Extract in the Sample solution (mg/mL)

 

Calculate the percentage of phenylpropenoid glycosides as the sum of the percentages of rosarin, rosavin, and rosin.

Calculate the percentage of salidroside:

 

P2 = (rU/rS) × (CS/CU) × 100

 

rU    = peak area of salidroside in the Sample solution

rS    = peak area of salidroside in Standard solution C

CS   = concentration of salidroside in Standard solution C (mg/mL)

CU   = concentration of Dry Extract in the Sample solution (mg/mL)

 

Calculate the percentage of the labeled amount of phenylpropenoid glycosides in the Dry Extract:

 

Result = (P1/L) × 100

 

P1   = content of phenylpropenoid glycosides, as determined above (%)

L      = labeled amount of phenylpropenoid glycosides (%)

 

Calculate the percentage of the labeled amount of salidroside in the Dry Extract:

 

Result = (P2/L) × 100

 

P2    = content of salidroside, as determined above (%)

L      = labeled amount of salidroside (%)

Acceptance criteria

Phenylpropenoid glycosides: 90.0%–110.0% of the labeled amount on the dried basis

Salidroside: 90.0%–110.0% of the labeled amount on the dried basis

 

CONTAMINANTS

Elemental Impurities—Procedures <233>

Acceptance criteria

Arsenic: NMT 2.0 µg/g

Cadmium: NMT 1.0 µg/g

Lead: NMT 5.0 µg/g

Mercury: NMT 0.2 µg/g

Articles of Botanical Origin, General Method for Pesticide Residues Analysis <561>: Meets the requirements

Articles of Botanical Origin, Test for Aflatoxins <561>: Meets the requirements

Microbial Enumeration Tests <61>: The total aerobic bacterial count does not exceed 104 cfu/g and the total combined molds and yeasts count does not exceed 103 cfu/g.

Tests for Specified Microorganisms <62>: Meets the requirements of the tests for the absence of Salmonella species and Escherichia coli

 

SPECIFIC TESTS

Loss on Drying <731>

Sample: 1.0 g of Dry Extract

Analysis: Dry the Sample at 105° for 2 h.

Acceptance criteria: NMT 5.0%

 

ADDITIONAL REQUIREMENTS

Packaging and Storage: Preserve in well-closed containers, protected from light and moisture. Store at controlled room temperature.

Labeling: The label states the Latin binomial and, following the official name, the parts of the plant from which the article was derived. It meets other labeling requirements under Botanical Extracts <565>.

USP Reference Standards <11>

USP Aflatoxins RS

USP Rhodiola rosea Root and Rhizome Dry Extract RS

USP Rosavin RS

USP Salidroside RS

Other Versions

Proposed For Comment Version 0.2