Actaea racemosa (root and rhizome)

From AHPA Botanical Identity References Compendium
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Revision as of 15:20, 5 February 2014

Contents

Nomenclature

Actaea racemosa L. Ranunculaceae

Botanical Voucher Specimen

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Actaea racemosa Tropicos 100105518.jpg
Source: Tropicos.org. Missouri Botanical Garden. 05 Aug 2013 <http://www.tropicos.org/Image/100105518> [1]

Organoleptic Characteristics

Rhizome externally dark brown; internally whitish and mealy or dark brown and waxy; odor slight; taste bitter and acrid.

Roots somewhat cylindrical or obtusely quadrangular, from 1 to 3 mm. in thickness, externally dark brown; internally bark dark brown, wood yellowish. The odor, though not strong, is peculiar and rather disagreeable, and is gradually lost with age.

Source: United States Dispensatory (1918) [2]

Macroscopic Descriptions

"... [A] tall stately plant, having a perennial root, and a simple herbaceous stem, which rises from four to eight feet in height. The leaves are large, and ternately decompound, having oblong-ovate leaflets, incised and toothed at their edges. The flowers are small, white, and disposed in a long, terminal, wand-like raceme, with occasionally one or two shorter racemes near its base. The calyx is white, four-leaved, and deciduous; the petals are minute, and shorter than the stamens; the pistil consists of an oval ovary and sessile stigma. The fruit is an ovoid capsule containing numerous flat seeds.

"Rhizome horizontal, more or less branching, from 2 to 12 cm. in length, and from 1 to 2.5 cm. in thickness; externally dark brown, slightly annulate from circular scars of bud scale-leaves, the upper surface with numerous stout, erect or somewhat curved branches terminated by deep, cup-shaped sears, each of which usually shows a distinct radiate structure; inferior and lateral portions with numerous root-scars and a few short roots; fracture horny; internally whitish and mealy or dark brown and waxy, bark thin, wood distinctly radiate and of about the same thickness as the pith... Roots somewhat cylindrical or obtusely quadrangular, from 1 to 3 mm. in thickness, externally dark brown, longitudinally wrinkled; fracture short; internally bark dark brown, wood yellowish, 4 to 6-rayed."

Source: United States Dispensatory (1918) [3]

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Alkemists Actaea racemosa macro.jpg
Root, cut and sifted
Source: Alkemist Laboratories [4]

Microscopic Characteristics

"Under the microscope, sections of the rhizome ... show a yellowish-brown suberized epidermis, a cortex made up of about 30 layers of starch-bearing parenchyma cells; the fibro-vascular bundles collateral, the xylem consisting of tracheae with bordered pores, and resembling tracheids in that the ends are rather acute; wood-fibers numerous, thin-walled, strongly lignified and with simple, oblique pores, the bundles separated by starch-bearing parenchyma strands from 5 to 30 cells wide; pith cells numerous, resembling those of the cortex.

"... sections of the root ... show a hairy epidermis, which becomes suberized in older roots; the cortex shows about 12 rows of starch-bearing parenchyma cells; endodermis distinct; fibro-vascular bundles 4 to 6, showing in older roots as separate collateral bundles. The powder is light to dark brown; starch grains numerous, single or compound, the individual grains spherical or more or less polygonal, each with a somewhat central cleft, from 0.003 to 0.015 mm. in diameter; fragments showing trachea with bordered pores and lignified wood-fibers; irregular, yellowish-brown fragments of suberized epidermis made up of more or less tabular cells, sometimes elongated and considerably thickened."

Source: United States Dispensatory (1918) [5]


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Alkemists Actaea racemosa brown suberized cells.png
Brown suberized cells (400X, Acidified chloral hydrate solution)
Source: Elan M. Sudberg, Alkemist Laboratories [6]

High Performance Thin Layer Chromatographic Identification

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Black cohosh HPTLC ID - Developed, UV 254 nm

Black cohosh (root) (Actaea racemosa)

Lane Assignments Lanes, from left to right (Track, Volume, Sample):

  1. 2 μL Isoferulic acid
  2. 2 μL Norcimifugin
  3. 2 μL Actein
  4. 2 μL 23-epi-26-Deoxyactein
  5. 2 μL Cimifugin
  6. 2 μL C. racemosa
  7. 2 μL C. racemosa
  8. 2 μL C. foetida
  9. 2 μL C. heracleifolia
  10. 2 μL C. dahurica
  11. 2 μL C. americana 

Reference Sample(s) Reference: Dissolve 1mg of actein, 23-epi-26-deoxyactein, isoferulic acid, cimifugin and/or norcimifugin individually in 10 mL of methanol. 

Stationary Phase Stationary phase, i.e. Silica gel 60, F254 

Mobile Phase Toluene, ethyl formate, formic acid 50:30:20 (v/v/v) 

Sample Preparation Method Sample: Mix 0.5 g of powdered sample with 10 mL of an ethanolwater mixture (1:1) and sonicate for 10 minutes, then centrifuge or filter the solutions and use the supernatants/filtrates as test solutions.

Sulfuric Acid Reagent: Preparation: 20 mL of sulfuric acid are mixed with 180 mL of ice-cooled methanol 

Detection Method UV/Vis, Sulfuric Acid Reagent Use: Dip (time 0, speed 5), heat at 100°C for 5 min 

Other Notes Compare result under UV 254 nm and white RT with reference images in Image Comparison Viewer. The fingerprint of the test solution is similar to that of the corresponding botanical reference sample. Additional weak zones may be present.

C. racemosa Identification: Under UV 254 nm the chromatogram obtained with the test solution does not show a dark zone at the position of Cimifugin in the reference solution or just above it. Under white RT a characteristic fingerprint is detected. Below Cimifugin there are two violet zones. Just above the application position two brown zones are detected (blue arrow). There is no brown zone at the position of Cimifugin, but there might be a faint violet zone. There is a brown zone at the position of actein as well as two brown zone just below. Just above the actein zone is a very thin brown zone (green arrow).

Further methodology available here, in Identification of black cohosh by HPTLC.

Source: AHPA Practical, CAMAG HPTLC [7]


Supplementary Information

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Black cohosh (root) (Actaea racemosa, syn Cimicifuga racemosa) 

General Characteristics AHPA recommends in its Known Adulterants list that appropriate steps be taken to assure that this raw material is free of the noted adulterant. Contact AHPA for additional information regarding relevant analytical methods or follow this link for more information. 

Reported Adulterants Chinese cimicifuga (root) (Actaea spp.). Also known as sheng ma or Rhizoma Cimicifugae; consists of Actaea cimicifuga, syn. Cimicifuga foetida; Actaea dahurica, syn. C. dahurica; A. heracleifolia, syn. C. heracleifolia; and possibly other Asian species of Actaea.

Source: AHPA Known Adulterants [8]


AHPA Practical: Detecting Adulteration of Black cohosh root/rhizome

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Reproduced from [1].

Introduction

The American Herbal Products Association is providing here analytical tools and methods to identify adulteration of raw materials and dietary ingredients labeled as black cohosh (Actaea racemosa syn. Cimicifuga racemosa) root/rhizome, or extracts thereof. This information is provided for industry in order to deal appropriately with reports of adulteration of this ingredient and make wise purchasing decisions.

Background

The economic adulteration of black cohosh root and rhizome with other species is well established. AHPA’s guidance policy on Known Adulterants identifies Chinese cimicifuga root/rhizome, also known as sheng ma or Rhizoma Cimicifugae as known black cohosh adulterants. This material commonly consists of Actaea cimicifuga, syn. Cimicifuga foetida; Actaea dahurica, syn. C. dahurica; A. heracleifolia, syn. C. heracleifolia; and possibly other Asian species of Actaea. The citation below provides a review of Rhizoma Cimicifugae for those wanting additional information.

Li JX, Yu ZY. Cimicifugae rhizoma: from origins, bioactive constituents to clinical outcomes. Curr Med Chem. 2006;13(24):2927-51.

The black cohosh monograph in the Natural Health Ingredients Database of the Canadian Natural Health Products Directorate at Health Canada, available online here or as a pdf file here, references instances where adverse events associated with ingestion of products labeled as containing black cohosh (Actaea racemosa syn. Cimicifuga racemosa) were found to contain plant species that were not black cohosh upon laboratory analysis. The Canadian Adverse Reaction Newsletter (2010 Jan;20(1):1-2) provided specifics on this situation as covered in a January 14, 2010 AHPA Update.

Analytical tools

There are several analytical tools available for the authentication of genuine black cohosh and its differentiation from other species. Two papers detailing the use of high performance thin-layer chromatography (HPTLC) are cited below. The first focuses on detection of black cohosh adulteration by other North American Actaea species while the second deals more specifically with differentiation of black cohosh from Chinese cimicifuga. The full methodology for the HPTLC analysis of the second citation can be accessed via this: Identification of black cohosh by HPTLC.

The third citation below employed phytochemical “fingerprinting” using high-performance liquid chromatography with a photo-diode array detector (HPLC-PDA) and liquid chromatography with mass spectrometry detection (LC-MS) to authenticate black cohosh and differentiate it from 14 other Actaea species.

Verbitski SM, Gourdin GT, Ikenouye LM, McChesney JD, Hildreth J. Detection of Actaea racemosa adulteration by thin-layer chromatography and combined thin-layer chromatography-bioluminescence. J AOAC Int. 2008 Mar-Apr;91(2):268-75.

Ankli A, Reich E, Steiner M. Rapid high-performance thin-layer chromatographic method for detection of 5% adulteration of black cohosh with Cimicifuga foetida, C. heracleifolia, C. dahurica, or C. americana. J AOAC Int. 2008 Nov-Dec;91(6):1257-64.

Jiang B, Ma C, Motley T, Kronenberg F, Kennelly EJ. Phytochemical fingerprinting to thwart black cohosh adulteration: a 15 Actaea species analysis. Phytochem Anal. 2011 Feb 19. doi: 10.1002/pca.1285. [Epub ahead of print] Supporting information is available here.

The black cohosh monograph in the Natural Health Ingredients Database of the Canadian Natural Health Products Directorate at Health Canada, available online here or as a pdf file here, references instances where adverse events associated with ingestion of products labeled as containing black cohosh (Actaea racemosa syn. Cimicifuga racemosa) were found to contain plant species that were not black cohosh upon laboratory analysis. The Canadian Adverse Reaction Newsletter (2010 Jan;20(1):1-2) provided specifics on this situation as covered in a January 14, 2010 AHPA Update.

The American Herbal Pharmacopoeia and the USP Dietary Supplements Compendium also contain useful information for the differentiation of black cohosh and other species representing common black cohosh adulterants. Authenticated black cohosh reference materials are available from both organizations as well as AHPA member companies Alkemists Laboratories, Botanical Liaisons, and Chromadex. DNA testing can be used to verify authentic species, and rule out the presence of common or other unexpected DNA-containing adulterants or contaminants. Work in this area is currently being conducted by AHPA member AuthenTechnologies.

Sources

  1. Tropicos.org. Missouri Botanical Garden. 05 Aug 2013 <http://www.tropicos.org/Image/100105518>
  2. United States Dispensatory (1918)
  3. United States Dispensatory (1918)
  4. Alkemist Laboratories
  5. United States Dispensatory (1918)
  6. Elan M. Sudberg, Alkemist Laboratories
  7. AHPA Practical, CAMAG HPTLC http://www.camag.com/index.php
  8. AHPA Known Adulterants http://www.ahpa.org/
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