Rhodiola rosea Root and Rhizome

Rhodiola rosea Root and Rhizome

Final Authorized Version 1.0

Rhodiola rosea Root and Rhizome


 

DEFINITION

The article consists of the dried roots and rhizomes of Rhodiola rosea L. (Family Crassulaceae). It contains NLT 0.3% of the phenylpropenoid glycosides rosarin, rosavin, and rosin calculated as rosavin, and NLT 0.08% of salidroside, calculated on the dried basis.

 

SYNONYMS

Rhodiola arctica Boriss.

Rhodiola borealis Boriss.

Rhodiola elongate (Ledeb.) Fisch. & C.A.Mey

Rhodiola iremelica Boriss.

Rhodiola maxima Nakai

Rhodiola minor Mill.

Rhodiola odorata Lam.

Rhodiola odorata Salisb.

Rhodiola roanensis (Britton) Britton

Rhodiola sachalinensis Boriss.

Rhodiola scopolii Simonk.

Sedum altaicum G.Don

Sedum arcticum (Boriss.) Rønning

Sedum elongatum Ledeb.

Sedum rhodiola DC.

Sedum rhodiola Vill.

Sedum roanense Britton

Sedum roseum (L.) Scop.

Sedum rosea (L.) Scop.

Sedum sachalinense (Boriss.) Vorosch.

Sedum scopolii Simonk.

Tetradium odoratum (Lam.) Dulac

Tolmachevia krivochizhinii (Sipliv.) Á.Löve & D. Löve

 

POTENTIAL CONFOUNDING MATERIALS

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

 

SELECTED COMMON NAMES

Chinese: 红景天

Danish: Almindelig rosenrod

English: Rhodiola, arctic rose, golden root, King’s Crown, roseroot, rosewort, snowdown rose, arctic root, Siberian golden root, Siberian rhodiola rosea

French: Rhodiole, racine d'or, orpin rose, racine arctique

German: Rosenwurz

Japanese: イワベンケ (iwa-benkei)

Pinyin: Hong jing tian

Spanish: Raíz dorada siberiana, raíz del ártico

Swedish: Rosenrot

 

CONSTITUENTS OF INTEREST

Phenylpropenoid glycosides: Rosarin, rosavin, rosin

Phenylethanoids: Salidroside, tyrosol

Monoterpene glycoside: Rosiridin

 

IDENTIFICATION

• A. Botanical Characteristics

Macroscopic: 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, uneven, 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 but no sclereids; secondary phloem; small narrow vascular bundles occur in a ring surrounding a broad parenchymatous pith; pith includes scattered vascular bundles xylem surrounded by phloem; 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; starch granules, simple, round or oval-shaped, 5–20 µm in diameter; hilum, if present, appears as a small dot.

• 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 about 0.5 g of Rhodiola rosea Root and Rhizome, finely powdered, in 5 mL of methanol 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.

• C. 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 content of phenylpropenoid glycosides (rosarin, rosavin, and rosin) to the content of salidroside is about 3:1.

 

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

 

 

Solvent: 75% Methanol in water

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: Transfer about 5.0 g of Rhodiola rosea Root and Rhizome, finely powdered and accurately weighed, to a 25-mL flask. Add 7 mL of Solvent, sonicate at 37° 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 time. Add the washings to the volumetric flask, dilute with 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.)

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:

  

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)

     = volume of the Sample solution (mL)

W     = weight of Rhodiola rosea Root and Rhizome 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:

 

Result = (rU/rS) × CS × (V/W) × 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)

     = volume of the Sample solution (mL)

W     = weight of Rhodiola rosea Root and Rhizome taken to prepare the Sample solution (mg)

Acceptance criteria

Phenylpropenoid glycosides: NLT 0.3% on the dried basis 

Salidroside: NLT 0.08% 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 105 cfu/g, the total combined molds and yeasts count does not exceed 103 cfu/g, and the bile-tolerant Gram-negative bacteria 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

Articles of Botanical Origin, Foreign Organic Matter <561>: NMT 2.0%

Loss on Drying <731>

Sample: 1.0 g of Rhodiola rosea Root and Rhizome, finely powdered

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

Acceptance criteria: NMT 12.0%

Articles of Botanical Origin, Total Ash <561>

Analysis: 2.0 g of Rhodiola rosea Root and Rhizome, finely powdered

Acceptance criteria: NMT 12.0%

Articles of Botanical Origin, Acid-Insoluble Ash <561>

Analysis: 2.0 g of Rhodiola rosea Root and Rhizome, finely powdered

Acceptance criteria: NMT 3.0%

 

ADDITIONAL REQUIREMENTS

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

Labeling: The label states the Latin binomial and the parts of the plant contained in the article.

USP Reference Standards <11>

USP Aflatoxins RS

USP Rhodiola rosea Root and Rhizome Dry Extract RS

USP Rosavin RS

USP Salidroside RS

Other Versions

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