Antrodia camphorata Fruiting Body Dry Extract

Antrodia camphorata Fruiting Body Dry Extract

Proposed For Development Version 0.1

Antrodia camphorata Fruiting Body Dry Extract

 


DEFINITION

The article is prepared from the dried fruiting bodies of Antrodia camphorata (M. Zang & C.H. Su) Sheng H. Wu, Ryvarden & T.T. Chang (Family Polyporaceae), also commonly known as Antrodia cinnamomea T.T. Chang & W.N. Chou, by ethanol extraction. The ratio of crude material to dry extract is 10:1. It contains NLT 90% and NMT 110% of the labeled amount of triterpenoic acids, calculated as the sum of antcin A, antcin B, antcin C, antcin H, antcin K, dehydrosulfurenic acid, and dehydroeburicoic acid on the dried basis. It may contain suitable added substances as carriers.

 

POTENTIAL CONFOUNDING MATERIALS

Ganoderma camphoratum

 

CONSTITUENTS OF INTEREST

Triterpenoids: Antcin A, antcin B, antcin C, antcin H; antcin K, dehydrosulfurenic acid, and dehydroeburicoic acid

Methoxy-methylenedioxy aromatics: 1,4-Dimethoxy-2,3-methylenedioxy-5-methylbenzene

Polyacetylenes: Antrocamphin A and antrocamphin B

 

IDENTIFICATION

• A. Thin-Layer Chrom​atography

 

CALL FOR SUBMISSION OF VALIDATED INFORMATION

 

Additional information including validation data will be required to complete the development of the Identification. For requirements, please see under “Identification” and related sections of the guidelines document “Monographs in the Herbal Medicines Compendium” at http:// hmc.usp.org/about/general-noticesguidelines.

 

Interested parties are encouraged to submit their proposals to complete the monograph.

 

• B. HPLC

Analysis: Proceed as directed in the Assay for Content of Triterpenoids

Acceptance criteria: The Sample solution chromatogram exhibits peaks at the retention times corresponding to antcin A, antcin B, antcin C, antcin H, antcin K, 1,4-dimethoxy-2,3-methylenedioxy -5-methylbenzene , dehydrosulfurenic acid, dehydroeburicoic acid in the chromatogram of Standard solution B.

 

ASSAY

• Content of Triterpenoids

Solution A: 0.1% Formic acid in water

Solution B: Acetonitrile

Mobile phase: See Table 1.

 

Table 1

Time

(min)

Solution A

(%)

Solution B

(%)

0.0 70 30
7.5 50 50
10.8 40 60
13.8 35 65
16.8 32 68
18.5 25 75
30.6 15 85
35.6 0 100
35.7 70 30
38.0 70 30

 

Standard solution A: 100 μg/mL of USP Dexamethasone RS in methanol

Standard solution B: Transfer 10 mg of USP Antrodia camphorata Fruiting Body Dry Extract RS to a 10-mL volumetric flask, dilute with methanol, and sonicate. Before injection, pass through a PTFE filter having a 0.2-μm pore size and discard the first portion of the filtrate.

Sample solution: Use 100 mg of Antrodia camphorata Fruiting Body Dry Extract and proceed as directed for Standard solution B.

Chromatographic system

(See Chromatography <621> System Suitability.)

Mode: UHPLC

Detector: UV 248 nm

Column: 2.1-mm × 10-cm; 1.8-μm packing L1 (similar to ACQUITY UPLC HSS T3, Zorbax SB C-18, and Eclipse Plus C-18)

Column temperature: 40 ± 1°

Flow rate: 0.3 mL/min

Injection volume: 5 μL

System suitability

Samples: Standard solution A and Standard solution B

Suitability requirements

Chromatographic similarity: The chromatogram of Standard solution B is similar to the reference chromatogram provided with the lot of USP Antrodia camphorata Fruiting Body Dry Extract RS being used.

Resolution: NLT 1.0 between dexamethasone and antcin K peaks, Standard solution B

Tailing factor: NMT 2.0 for the dexamethasone peak, Standard solution A

Relative standard deviation: NMT 2.0%, determined from the dexamethasone peak in repeated injections, 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 Antrodia camphorata Fruiting Body Dry Extract RS being used, identify the retention times of the peaks corresponding to antcin A, antcin B, antcin C, antcin H, antcin K, dehydrosulfurenic acid, and dehydroeburicoic acid. The approximate relative retention times of the different peaks are provided in Table 2.

 

Table 2

Analyte

Approximate
Relative

Retention Times

Factor
Dexamethasone 1.00 1.00
(R,S)-Antcin K 1.29 0.34
1,4-Dimethoxy-2,3-methylenedioxy-5-methylbenzene 1.69 NA
(R,S)-Antcin C 2.38 0.21
(R,S)-Antcin H 2.50 0.15
Dehydrosulfurenic acid 2.84 0.33
(R,S)-Antcin B 3.07 0.21
(R,S)-Antcin A 3.76 0.24
Dehydroeburicoic acid 5.87 0.47

 

Separately calculate the percentage of each of the triterpenes in the portion of Antrodia camphorata Fruiting Body taken:

 

Result = (rU/rS) × (CS/CU) × F × 100

 

rU = peak response of the analyte from the Sample solution

rS = peak responses of dexamethasone from Standard solution A

CS = concentration of dexamethasone in Standard solution A (μg/mL)

CU = concentration of Antrodia camphorata Fruiting Body Dry Extract taken in the Sample solution (μg/mL)

F = conversion factors for the analytes (See Table 2.)

[Note—Incorporate a factor in the calculation to account for the aliquot taken compared to the volume of the Sample solution.]

Calculate the content of Triterpenoids as the sum of the percentage of the different triterpenoids.

Separately calculate the percentage of the labeled amount of triterpenoids in the extract.

 

Result = (P/L) × 100

 

P = content of triterpenoids as determined above (%)

= labeled amount of triterpenoids (%)

Acceptance criteria: 90.0%-110.0% of the labeled amount of triterpenoids 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 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

Articles of Botanical Origin, Test for Aflatoxins <561>: Meets the requirements

 

SPECIFIC TESTS

Loss on Drying <731>

Sample: 1.0 g Antrodia camphorata Fruiting Body Dry Extract

Analysis: Dry at 105° for 4 h.

Acceptance criteria: NMT 2.0%

 

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

USP Reference Standards <11>

USP Aflatoxins RS

USP Antrodia camphorata Fruiting Body Dry Extract RS

USP Dexamethasone RS

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Other Versions

Proposed For Comment Version 0.2