Antrodia camphorata Fruiting Body Powder

Antrodia camphorata Fruiting Body Powder

Proposed For Development Version 0.1

Antrodia camphorata Fruiting Body Powder

 


DEFINITION

The article consists of the dried fruiting bodies of Antrodia camphorata (M. Zang & C.H. Su) Sheng H. Wu, Ryvarden & T.T. Chang (Family Polyporaceae) reduced to a powder or very fine powder. It is also commonly known as Antrodia cinnamomea T.T. Chang & W.N. Chou. It contains NLT 5% 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.

 

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. Botanical Characteristics

Macroscopic: Red brown powder.

 

• B. 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.

 

• C. 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.

• D. HPLC

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

Acceptance criteria: The Sample solution chromatogram exhibits peaks at the retention times corresponding to antroquinonol 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: Transfer 0.5 g of Antrodia camphorata Fruiting Body Powder to a 50-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.

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 Powder taken:

 

Result = (rU/rS) × CS x (V/W) × 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

V = volume of the Sample solution (mL)

W = weight of Antrodia camphorata Fruiting Body Powder taken to prepare the Sample solution (μg)

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.]

Add the percentages of antcin A, antcin B, antcin C, antcin H, antcin K, dehydrosulfurenic acid, and dehydroeburicoic acid.

Acceptance criteria: NLT 5% on the dried basis

• Content of Antroquinonol

Solution A: 0.3% Acetic acid in water

Solution B: Methanol

Mobile phase: See Table 3.

Table 3

Time

(min)

Solution A

(%)

Solution B

(%)

0 95 5
10 20 80
20 10 90
35 10 90
40 95 5

 

Standard solution A: 500 μg/mL of USP Antroquinonol 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: Transfer about 1 g of Antrodia camphorata Fruiting Body Powder to an Erlenmeyer flask, add 10.0 mL of methanol, and weigh the flask and contents. Sonicate for 30 min and adjust to initial weight with methanol. Before injection, pass through a PTFE filter having a 0.2-μm pore size and discard the first portion of the filtrate.

Chromatographic system

(See Chromatography <621> System Suitability.)

Mode: HPLC

Detector: UV 254 nm

Column: 4.6-mm × 25-cm; 5-μm packing L1 (similar to Hypersil C-18)

Flow rate: 1.0 mL/min

Injection volume: 20 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.

Tailing factor: 0.9–1.3 for the antroquinonol peak, Standard solution A

Relative standard deviation: NMT 2% determined from the antroquinonol 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 peak corresponding to antroquinonol.

Separately calculate the percentage of antroquinonol in the portion of Antrodia camphorata Fruiting Body Powder taken:

 

Result = (rU/rS) × CS × (V/W) × 100

 

rU = peak response of the analyte from the Sample solution

rS = peak responses of antroquinonol from Standard solution A

CS = concentration of antroquinonol in Standard solution A

V = volume of the Sample solution (mL)

W = weight of Antrodia camphorata Fruiting Body Powder taken to prepare the Sample solution (mg)

[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 antroquinonol.

Acceptance criteria: NLT 0.4% 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

Articles of Botanical Origin, Alcohol-Soluble Extractives, Method 1 <561>: NLT 20%

Articles of Botanical Origin, Water-Soluble Extractives, Method 2 <561>: NLT 8.0%

Loss on Drying <731>

Sample: 1.0 g Antrodia camphorata Fruiting Body Powder

Analysis: Dry at 105° for 4 h.

Acceptance criteria: NMT 75%

Articles of Botanical Origin, Total Ash <561>: NMT 4%

 

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 Antroquinonol RS

USP Dexamethasone RS

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1 comment

Jent-turn L.posted Friday, March 13, 2015 - 3:06am
Hi, This is Taiwan Leader Biotech. Corp.. We are focus on the research, manufacture, and sell of Antrodia cinnamomea. According to the Herbal Medicines Compendium in USP web site (https://hmc.usp.org), we would like to suggest the item of “Antrodia camphorata Fruiting Body Powder” as below: 1. In the previous study published in J. Agric. Food Chem. 2011 59:7626-35. The authors established 13 representative metabolites from the ethanol extract of A. cinnamomea fruiting body. A comparison of the metabolite profiles of different ethanol extracts from A. cinnamomea strains showed silmilar metabolites when the strains were grown on the original host wood (Cinnamomum kanehirai) and harvested after the same culture time period (9 months). Furthermore, the amounts of typical ergostane-type triterpenoids in A. cinnamomea increased with culture age. Culture substrates also influenced metabolite synthesis; with the same culture age, A. cinnamomea grown on the original host wood produced a richer array of metabolites than A. cinnamomea cultured on other wood species. Therefore, analysis of a fixed group of compounds including triterpenoids, benzolics, and polyacetylenes constitutes a suitable, reliable system to evaluate the quality of ethanol extract from A. cinnamomea fruiting bodies. 2. We would like to suggest the method described in above reference that may provide a better platform for analysis of “Antrodia camphorata Fruiting Body Powder”.

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