Terminalia chebula Fruit Dry Extract

Terminalia chebula Fruit Dry Extract

Proposed For Development Version 0.1

Terminalia chebula Fruit Dry Extract

 


DEFINITION

The extract is prepared from the dried fruit of Terminalia chebula Retz. (Family Combretaceae) by extraction with hydroalcoholic mixtures. It contains NLT 90% and NMT 110% of the labeled amount of hydrolyzable tannins calculated as the sum of chebulagic acid and chebulinic acid, on the dried basis.

 

POTENTIAL CONFOUNDING MATERIALS

Terminalia citrine

 

CONSTITUENTS OF INTEREST

Hydrolyzable tannins: Chebulagic acid and chebulinic acid

Triterpene: Arjungenin

 

IDENTIFICATION

• A. Thin-Layer Chromatography

Standard solution A: 0.2 mg/mL of USP Chebulagic Acid RS in methanol

Standard solution B: 10 mg/mL of USP Terminalia chebula Fruit Dry Extract RS in methanol. Sonicate for 10 min, centrifuge, and use the supernatant.

Sample solution: Sonicate about 0.1 g of Terminalia chebula Fruit Dry Extract in 100 mL of boiling water for 10 min, centrifuge, pass through a membrane filter of 0.45-μm pore size, and use the supernatant.

Chromatographic system

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

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

Application volume: 15 µL each of Standard solution A, 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, toluene, formic acid, and methanol (6:2:2:1)

Developing distance: 6 cm

Derivatization reagent: Ferric chloride reagent created by dissolving 12 g of ferric chloride in 100 mL of water and filtering.

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. Examine under visible light.

System suitability: Under visible light, Standard solution B exhibits the most intense band in the middle where two dark blue bands appear; the lower band corresponds in RF and color to the chebulagic acid band in Standard solution A, and the upper band is consistent with chebulinic acid. One or two minor blue bands appear close to the origin, and two additional light blue bands appear above chebulinic acid. One blue band appears close to the solvent front.

Acceptance criteria: Under visible light, the Sample solution exhibits the most intense band in the middle where two dark blue bands appear; the lower band corresponds in RF and color to the chebulagic acid band in Standard solution A, and the upper band is consistent with chebulinic acid.

• B. HPLC

Analysis: Proceed as directed in the Assay for Content of Constituents of Hydrolyzable Tannins.

Acceptance criteria: The chromatogram of the Sample solution exhibits peaks at retention times corresponding to the peaks due to chebulagic acid and chebulinic acid in Standard solution B.

 

ASSAY

• Content of Constituents of Hydrolyzable Tannins

Solution A: Dissolve 0.136 g of monobasic potassium phosphate in 900 mL of water, add 0.5 mL of o-phosphoric acid, dilute with water to 1 L, and mix.

Solution B: Acetonitrile

Mobile phase: See Table 1.

Table 1

Time
(min)

Solution A
(%)

Solution B
(%)

0

95

5

18

65

35

25

45

55

28

45

55

35

65

35

40

95

5

45

95

5

 

Standard solution A: 1.0 mg/mL of USP Chebulagic Acid RS in boiling water

Standard solution B: Dissolve 0.1 g of USP Terminalia chebula Fruit Dry Extract RS in 100 mL of boiling water, sonicate, and pass through a membrane filter of 0.45-μm pore size.

Sample solution: Transfer 0.1 g of Terminalia chebula Fruit Dry Extract to a 100-mL volumetric flask. Add 50 mL of boiling water and sonicate for 10 min. Dilute with water to 100 mL, mix well, and pass through a membrane filter of 0.45-μm pore size.

Chromatographic system

(See Chromatography <621>, System Suitability.)

Detector: UV 270 nm

Column: 4.6-mm × 25-cm; L1 (similar to Merck kGaA Purospher Star LP HPLC Column, RP-18)

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 of Standard solution B is similar to the reference chromatogram provided with the lot of USP Terminalia chebula Fruit Dry Extract RS being used.

Resolution: 11.4 between chebulagic acid and chebulinic acid peaks, Standard solution B

Tailing factor: NMT 1.5, Standard solution A

Relative standard deviation: NMT 2.5%, Standard solution A

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 Terminalia chebula Fruit Dry Extract RS being used, identify the retention times of the peaks corresponding to chebulagic acid and chebulinic acid. The approximate relative retention times of the peaks for chebulagic acid and chebulinic acid are 1.0 and 1.15, respectively.

Separately calculate the percentages of chebulagic acid and chebulinic acid in the portion of Terminalia chebula Fruit Dry Extract taken:

 

Result = (rU/rS) × (CS/CU) × 100

 

rU     = peak area of the analyte from the Sample solution

rS     = peak area of the analyte from Standard solution A

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

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

 

Calculate the percentage of the labeled amount of chebulagic acid in the Extract:

 

Result = (P/L) × 100

 

P     = content of chebulagic acid as determined above (%)

L     =  labeled amount of chebulagic acid (%)

Acceptance criteria: 90.0%–110.0% of the labeled amount of chebulagic acid 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

• 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 10cfu/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

• Identification of Arjungenin

Standard solution: 0.2 mg/mL of USP Arjungenin RS in methanol

Sample solution: Use the Sample solution prepared in Identification test A.

Chromatographic system

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

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

Application volume: 15 µL each of Standard solution A, 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, toluene, and glacial acetic acid (55:45:2)

Developing distance: 6 cm

Derivatization reagent: Anisaldehyde-sulfuric acid reagent created by combining 0.5 mL of anisaldehyde mixed with 10 mL of glacial-acetic acid, 85 mL of methanol, and 5 mL of sulfuric acid.

Analysis

Samples: Standard solution 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. Examine under visible light.

Acceptance criteria: Under visible light, the Sample solution exhibits the most intense band in the middle as a dark blue band corresponding in color and RF to the band due to arjungenin in the Standard solution.

• Articles of Botanical Origin, Foreign Organic Matter <561>: NMT 1.0%

• Articles of Botanical Origin, Alcohol-Soluble Extractives, Method 1 <561>: NLT 40.0%

• Articles of Botanical Origin, Water-Soluble Extractives, Method 2 <561>: NLT 60.0%

• Loss on Drying <731>

Sample: 1 g of Terminalia chebula Fruit Dry Extract, finely powdered

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

Acceptance criteria: NMT 8%

• Articles of Botanical Origin, Total Ash <561>: NMT 15%

• Articles of Botanical Origin, Acid-Insoluble Ash <561>: NMT 5%

 

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 the part(s) of the plant contained in the article.

• USP Reference Standards <11>

USP Arjungenin RS

USP Chebulagic Acid RS

USP Terminalia chebula Fruit Dry Extract RS

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Other Versions

Proposed For Comment Version 0.3
Proposed For Comment Version 0.2