Rhodiola rosea Root and Rhizome Tincture

Rhodiola rosea Root and Rhizome

Proposed For Comment Version 0.2

Rhodiola rosea Root and Rhizome Tincture


 

DEFINITION

Rhodiola rosea Tincture is prepared as follows.

 

Rhodiola rosea Root and Rhizome

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 the phenylpropenoid glycosides rosarin, rosavin and rosin calculated as rosavin, and NLT 0.01% (w/v) of salidroside.

 

Constituents of Interest

Phenylpropenoid glycosides: Rosarin, rosavin, and rosin

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 Powdered 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

(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 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: 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. 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 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 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: 0.1 mg/mL of USP Rosavin RS in methanol

Standard solution B: 4.0 mg/mL of USP Rhodiola rosea Powdered 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: 0.1 mg/mL of USP Salidroside RS in methanol

Sample solution: Before injecting the Tincture, pass through a membrane filter of 0.45-µm or finer pore size, discarding the first few mL of the filtrate. [Note—The sample can be weighed and converted to volume using the density of the Tincture.]

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 from Standard solution B is similar to the reference chromatogram provided with the lot of USP Rhodiola rosea Powdered Extract RS being used.

Resolution: NLT 1.5 between the rosarin and rosavin peaks, Standard solution B

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 Powdered 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 the phenylpropenoid glycosides rosarin, rosavin, and rosin as rosavin:

 

Result = (rU/rS) × CS × 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 (g/mL)

 

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

Calculate the percentage of salidroside:

 

Result = (rU/rS) × CS × 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 (g/mL)

 

Acceptance criteria

Phenylpropenoid glycosides: NLT 0.06% (w/v)

Salidroside: NLT 0.01% (w/v)

 

OTHER COMPONENTS

Alcohol Determination, Method I <611>: 90.0%–110.0% of the labeled amount of alcohol (C2H5OH)

 

CONTAMINANTS

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

 

ADDITIONAL REQUIREMENTS

Packaging and Storage: Preserve in tight, light-resistant containers. Store at room temperature.

Labeling: The label states the Latin binomial and the parts of the plant from which the article was prepared. Label it to indicate the content of phenylpropenoid glycosides as the sum of rosavin, rosarin, and rosin as rosavin, the content of salidroside, the alcohol content, and the ratio of the starting plant material to Tincture.

USP Reference Standards <11>

USP Rhodiola rosea Powdered Extract RS

USP Rosavin RS

USP Salidroside RS

 

The commenting period for this monograph has expired.

Other Versions

Final Authorized Version 1.0
Final Authorized Version 1.0
Proposed For Comment Version 0.2