Mangifera indica Bark Powder

Mangifera indica Bark Powder

Proposed For Comment Version 0.2

Mangifera indica Bark Powder

 


DEFINITION

The article consists of the dried bark of Mangifera indica L. (Family Anacardiaceae) reduced to a fine or very fine powder. It contains NLT 2.5% 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. Botanical Characteristics

Macroscopic: Powder of bark is brown in color and shows fragments of cork cells in surface and sectional view.

Microscopic: Tannin tubes associated with fiber and stone cells; starch embedded in parenchymatous cells and scattered as such throughout; prismatic crystals of calcium oxalate; fragment of bark in tangential longitudinal section shows medullary ray cells; fibers associated with stone cells and calcium oxalate crystals; fragments of bark in radial longitudinal section shows medullary ray cells crossing over fibers and associated stone cells and calcium oxalate crystals; fragment of bark shows medullary ray cells, fibers, calcium oxalate crystals, and cells containing resin.

• B. 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 0.2 g of Mangifera indica Bark Powder in 10 mL of methanol 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 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 with increasing RF: a pale white/yellow band near the origin, a pale orange band slightly below the mangiferin band, a pale orange band above mangiferin band, two blue bands near RF of about 0.75, and a pale blue band near the solvent front.

• C. HPLC

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

Acceptance criteria: The chromatogram of the Sample solution exhibits peaks at retention times corresponding to the peaks 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: Transfer 0.2 g of Mangifera indica Bark Powder, accurately weighed, into a 50-mL volumetric flask. Add 40 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 Powder taken:

 

Result = (rU/rS) × CS × (V/W) × 100

 

rU     = peak area of mangiferin in the Sample solution

rS     = peak area of mangiferin in Standard solution A

CS    = concentration of USP Mangiferin RS from Standard solution A (mg/mL)

     = volume of the Sample solution (mL)

    = weight of Mangifera indica Bark Powder taken to prepare the Sample solution (mg)

Acceptance criteria: NLT 2.5% 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

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

• Articles of Botanical Origin, Alcohol-Soluble Extractives, Method 1 <561>: NLT 20.0%

• Articles of Botanical Origin, Water-Soluble Extractives, Method 2 <561>: NLT 14.0%

• Loss on Drying <731>

Sample: 1 g of Mangifera indica Bark Powder

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

Acceptance criteria: NMT 8%

• Articles of Botanical Origin, Total Ash <561>: NMT 9%

• Articles of Botanical Origin, Acid-Insoluble Ash <561>: NMT 2%

 

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 Mangifera indica Bark Dry Extract RS

USP Mangiferin RS 

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Proposed For Development Version 0.1