Mangifera indica Bark Dry Extract

Mangifera indica Bark Dry Extract

Proposed For Comment Version 0.2

Mangifera indica Bark Dry Extract

 


 

DEFINITION

The article consists of the dried bark of Mangifera indica L. (Family Anacardiaceae) by extraction with methanol. The ratio of starting crude plant material to Dry Extract is between 12:1 and 10:1. It contains NLT 90.0% and NMT 110.0% of the labeled amount of mangiferin, calculated on the dried basis.

 

POTENTIAL CONFOUNDING MATERIALS

None known

 

CONSTITUENTS OF INTEREST

Xanthon: Mangiferin and isomangiferin

Triterpenes: Cycloart-24-en-3β,26-diol

Phenolic acid: Protocatechuic acid

 

IDENTIFICATION

• A. Thin-Layer Chromatography

Standard solution A: 0.15 mg/mL of USP Mangiferin RS in methanol

Standard solution B: 5 mg/mL of USP Mangifera indica Bark Dry Extract RS in methanol. Sonicate for 10 min, centrifuge, and use the supernatant.

Sample solution: Sonicate about 50 mg of Mangifera indica Bark Dry Extract in 10 mL of Solvent for 10 min, centrifuge, 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: 4 µL each of Standard solution A and Standard solution B, and 2 µL of Sample solution, as 8-mm bands

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

Developing solvent system: Ethyl acetate, formic acid, and water (80:10:10)

Developing distance: 7 cm

Temperature: 25“

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. Examine under UV 254 and 366 nm.

System suitability: Under UV 254 nm, the chromatogram of Standard solution B exhibits a black band similar in RF to mangiferin in the chromatogram of Standard solution A. One weak band appears right above the origin and two weak bands appear above the mangiferin band. Under UV 366 nm, the chromatogram of Standard solution B exhibits a weak blue band similar in color and RF to mangiferin in the chromatogram of Standard solution A. About six additional bands appear in the chromatogram with increasing RF: a pale white/yellow band near the origin, a bright orange band slightly below the mangiferin band, a pale orange band above the mangiferin band, two blue bands near RF of about 0.75, and a blue band near the solvent front.

Acceptance criteria: Under UV 254 nm, the chromatogram of Sample solution exhibits a band corresponding in color and RF to the mangiferin band in the chromatogram of Standard solution A. One weak band appears right above the origin. Under UV 366 nm, the chromatogram of Sample solution exhibits a weak blue band similar in color and RF to mangiferin in the chromatogram of Standard solution A. About six additional bands appear in the chromatogram with increasing RF: a pale white/yellow band near the origin, a bright orange band slightly below the mangiferin band, a pale orange band above the mangiferin band, two blue bands near RF of about 0.75, and a pale blue band near the solvent front.

• B. HPLC

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

Acceptance criteria: The chromatogram of the Sample solution exhibits a peak at a retention time corresponding to the peak due to mangiferin in Standard solution B.

 

ASSAY

• Content of Mangiferin

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
(%)

0.01

85

15

10

80

20

15

80

20

20

85

15

25

85

15

Solvent: 70% Methanol in water

Standard solution A: 0.1 mg/mL of USP Mangiferin RS in Solvent

Standard solution B: Accurately weigh USP Mangifera indica Bark Dry Extract RS, equivalent to 10 mg of mangiferin, into a 100-mL volumetric flask, add 50 mL of Solvent, and dissolve in a boiling water bath for 15 min with sonication. Cool the solution and dilute with Solvent to volume. Mix well and pass through a membrane filter of 0.45-μm pore size.

Sample solution: Accurately weigh Mangifera indica Bark Dry Extract, equivalent to 10 mg of mangiferin, into a 100-mL volumetric flask, add 50 mL of Solvent, and dissolve in a boiling water bath for 15 min with sonication. Cool the solution and dilute with Solvent to volume. Mix well and pass through a membrane filter of 0.45-μm pore size.

Chromatographic system

(See Chromatography <621>, System Suitability.)

Detector: UV 254 nm

Column: 4.6-mm × 25-cm; 5-µm packing 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 Mangifera indica Bark Dry Extract RS being used.

Tailing factor: NMT 1.5, Standard solution A

Relative standard deviation: NMT 2.0%, 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 Mangifera indica Bark Dry Extract RS being used, identify the retention time of the peak corresponding to mangiferin.

Calculate the percentage of mangiferin in the portion of Mangifera indica Bark Dry Extract taken:

 

Result = (rU/rS) × (CS/CU) × 100

 

rU   = peak area of mangiferin from the Sample solution

rS   = peak area of mangiferin from Standard solution A

C = concentration of USP Mangiferin RS in Standard solution A (mg/mL)

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

 

Calculate the percentage of the labeled amount of mangiferin in the portion of Dry Extract taken:

 

Result = (P/L) × 100

 

P   = content of mangiferin as determined above (%)

L   = labeled amount of magiferin (%)

Acceptance criteria: 90.0%–110.0% 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 <2021>: The total aerobic bacterial count does not exceed 104 cfu/g, the total combined molds and yeasts count does not exceed 102 cfu/g, and the bile-tolerant Gram-negative bacteria does not exceed 102 cfu/g.

• Absence of Specified Microorganisms <2022>: Meets the requirements of the tests for the absence of Salmonella species, Escherichia coli, and Staphylococcus aureus

 

SPECIFIC TESTS

• Loss on Drying <731>

Sample: 1 g of Mangifera indica Bark Dry Extract

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

Acceptance criteria: NMT 10%

• Articles of Botanical Origin, Total Ash <561>: NMT 5%

• Articles of Botanical Origin, Acid-Insoluble Ash <561>: NMT 1.5%

• Other Requirements: It meets the requirements of the Residual Solvents test in Botanical Extracts <565>.

 

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. It meets other labeling requirements in Botanical Extracts <565>.

• USP Reference Standards <11>

USP Mangifera indica Bark Dry Extract RS

USP Mangiferin RS

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Proposed For Development Version 0.1