Picrorhiza kurrooa Root and Rhizome Dry Extract

Picrorhiza kurrooa Root and Rhizome Dry Extract

Proposed For Comment Version 0.2

Picrorhiza kurrooa Root and Rhizome Dry Extract


DEFINITION

The article is prepared from Picrorhiza kurrooa Royle (Family Plantaginaceae) root and rhizome by extraction of 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 iridoid glycosides picroside I and picroside II, calculated on the anhydrous basis.

 

CONSTITUENTS OF INTEREST

Iridoid glycosides: Picroside I and picroside II

 

IDENTIFICATION

• A. Thin-Layer Chromatography

Standard solution A: 0.5 mg/mL of USP Picroside I RS in methanol

Standard solution B: 80 mg/mL of USP Picrorhiza kurrooa Root and Rhizome Dry Extract RS in methanol

Sample solution: 80 mg/mL of Picrorhiza kurrooa Root and Rhizome Dry Extract in methanol. Sonicate for 10 min, centrifuge, and use supernatant.

Chromatographic system

(See Chromatography <621>.)

Adsorbent: Use a suitable chromatographic material with an average particle size of 5 µm (HPTLC plates).

Application volume: 10 µL each of Standard solution A, Standard solution B, and Sample solution, as 10-mm bands

Relative humidity: Condition the plate to a relative humidity of about 33% using a suitable device.

Developing solvent system: Ethyl acetate, methanol, and water (82: 10: 0.8)

Developing distance: 6 cm

Derivatization reagent: Anisaldehyde-sulfuric acid reagent

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 3 min at 100°, and examine under visible light.

System suitability: Under visible light, Standard solution B exhibits, in the lower half, about seven dark brown bands with the most intense two bands close to the upper section of the lower half; the band with higher RF corresponds to the band due to picroside I in Standard solution A.

Acceptance criteria: Under visible light, the Sample solution exhibits a strong dark brown band in the middle corresponding to the band due to picroside I in Standard solution A. The Sample solution exhibits, in the lower half, about seven dark brown bands, with the second strongest band having lower RF than picroside I, corresponding to the band due to picroside II in Standard solution A.

• B. HPLC

Analysis: Proceed as directed in the Assay for Content of Iridoid Glycosides.

Acceptance criteria: The Sample solution exhibits peaks at the retention times corresponding to the peaks due to picroside I and picroside II of Standard solution B.

 

ASSAY

• Content of Iridoid Glycosides

Solution A: Dissolve 0.136 g of anhydrous potassium dihydrogen orthophosphate (KH2PO4) in 900 mL of water and add 0.5 mL of orthophosphoric acid. Dilute with water to 1000 mL.

Solution B: Acetonitrile

Mobile phase: See Table 1.

Table 1

Time (min)

Solution A

(%)

Solution B

(%)

0

85

15

7

80

20

15

70

30

20

20

80

22

20

80

25

85

15

30

85

15

 

Standard solution A: 0.015 mg/mL of USP Picroside I RS in methanol

Standard solution B: 0.6 mg/mL USP Picrorhiza kurrooa Root and Rhizome Dry Extract RS in methanol, sonicate if necessary. Before injection, pass through a membrane filter of 0.45-μm or finer pore size. Discard the first few mL of the filtrate.

Sample solution: Picrorhiza kurrooa Root and Rhizome Dry Extract in methanol at a concentration equivalent to 0.015 mg/mL of picroside I according to the label claim, sonicate if necessary. Before injection, pass through a membrane filter of 0.45-μm or finer pore size. Discard the first few mL of the filtrate.

Chromatographic system

(See Chromatography <621>, System Suitability.)

Detector: UV 263 nm

Column: 4.6-mm × 25-cm; 5-µm packing L1 (similar to Luna 5-µ C18 (2)-100A)

Flow rate: 1.5 mL/min

Injection volume: 20 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 Picrorhiza kurrooa Root and Rhizome Dry Extract RS being used.

Resolution: NLT 2.0 between picroside I and picroside II

Tailing factor: NMT 1.5

Relative standard deviation: NMT 2.5%

Analysis

Samples: Standard solution A, Standard solution B, and Sample solution

Using the chromatograms of Standard solution A, Standard solution B, and the reference chromatogram provided with the lot of USP Picrorhiza kurrooa Root and Rhizome Dry Extract RS being used, identify the retention times of the peaks corresponding to picroside I and picroside II. The approximate relative retention times of the peaks for picroside I and picroside II are 1.00 and 0.77, respectively.

Calculate the percentage of picroside I and picroside II:

 

Result = (rU/rS) × (CS/CU )× 100

 

rU   = response from the Sample solution

rS   = response from Standard solution A

CS  = concentration of Standard solution A (mg/mL)

CU  = concentration of Picrorhiza kurrooa Root and Rhizome Dry Extract in the Sample solution (mg/mL)

 

Calculate the percentage of the labeled amount of iridoid glycosides as the sum of picroside I and picroside II:

 

Result = (P/L) × 100

 

P  = content of iridoid glycosides, as determined above (%)

L  = labeled amount of iridoid glycosides

Acceptance criteria: 90.0%–110.0% of the labeled amount of iridoid glycosides on the anhydrous basis

 

CONTAMINANTS

 Elemental Impurities—Procedures <233>

Acceptance criteria

Arsenic: NMT 2.0 µg/g

Cadmium: NMT 0.5 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

Water Determination, Method III <921>: NMT 7.0%

 

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(s) 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 Picrorhiza kurrooa Root and Rhizome Dry Extract RS

USP Picroside I RS

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Other Versions

Proposed For Development Version 0.1