Glycyrrhiza glabra (root)

From AHPA Botanical Identity References Compendium
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|caption3=''G. uralensis'' reference standard
 
|caption3=''G. uralensis'' reference standard
 
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| description='''Sample Preparations:''' Extract 1.0 g of coarsely powdered Glycyrrhiza root in 50 mL of water by boiling for about 5 minutes, and filter. Repeat for 4-5 times or until the extract is colorless. Combine the extracts, concentrate to about 100 mL, and cool to room temperature. Before injection, filter through a membrane filter of 0.45-um or finer pore size, discarding the first 5 mL of the filtrate.
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| description=Sample Preparations:
 +
Extract 1.0 g of coarsely powdered Glycyrrhiza root in 50 mL of water by boiling for about 5 minutes, and filter. Repeat for 4-5 times or until the extract is colorless. Combine the extracts, concentrate to about 100 mL, and cool to room temperature. Before injection, filter through a membrane filter of 0.45-um or finer pore size, discarding the first 5 mL of the filtrate.
  
 
'''Column:''' C18, 25-cm x 4.6 mm, 5-um, XXXXX, XXXXX
 
'''Column:''' C18, 25-cm x 4.6 mm, 5-um, XXXXX, XXXXX
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'''Injection volume:''' 20 uL
 
'''Injection volume:''' 20 uL
  
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{|
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}}
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===Table===
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{| border=1
 
| Time (min) || Solution A (%) || Solution B (%)
 
| Time (min) || Solution A (%) || Solution B (%)
 +
|-
 
|0-18 || 95-55 || 5-45
 
|0-18 || 95-55 || 5-45
 +
|-
 
|18-25 || 55-20 || 45-80
 
|18-25 || 55-20 || 45-80
 +
|-
 
|25-28 || 20 || 80
 
|25-28 || 20 || 80
 +
|-
 
|28-35 || 20-55 || 80-45
 
|28-35 || 20-55 || 80-45
 +
|-
 
|35-40 || 55-95 || 45-5
 
|35-40 || 55-95 || 45-5
 +
|-
 
|40-45 || 95 || 5
 
|40-45 || 95 || 5
 
|}
 
|}
 +
=High Performance Thin Layer Chromatographic Identification=
 +
{{HPTLC | source=AHPA Practical, CAMAG HPTLC
 +
            | companyimage=Camag_logo.png
 +
            | companyURL=http://www.camag.com/index.php
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            | mainimage=Camag-blackcohosh-1-d-uv254.png
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            | caption1=Black cohosh HPTLC ID - Developed, UV 254 nm
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            | description=Black cohosh (root) (''Actaea racemosa'')
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            | image2=Camag-blackcohosh-1-d-uv366.png
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            | caption2=Black cohosh HPTLC ID - Developed, UV 366 nm
 +
            | image3=Camag-blackcohosh-1-H2SO4-uv366.png
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            | caption3=Black cohosh HPTLC ID - Sulfuric acid reagent, UV 366 nm
 +
            | image4=Camag-blackcohosh-1-H2SO4-whiteRT.png
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            | caption4=Black cohosh HPTLC ID Method - Sulfuric acid reagent, white RT
 +
            |
 +
            | lanes=Lanes, from left to right (Track, Volume, Sample):
 +
 +
            | notes=
 +
'''Reference Standard Solution:''' 0.1 mg/mL ammonium glycyrrhizate in ethanol and water (7:3). Or 0.1 mg/mL glycyrrhizic acid in ethanol and water (7:3).
 +
 +
'''Reference Sample Preparations:''' Mix 0.5 g of powdered sample with 10 mL of ethanol and water (7:3, v/v), sonicate for 10 minutes, centrifuge or filter the solution, and use the supernatant / filtrate.
 +
 +
'''Stationary Phase:''' HPTLC, Silica gel 60 F254
 +
 +
'''Mobile Phase:''' Ethyl acetate, acetic acid, formic acid, water (15:1:1:2)
 +
 +
'''Development:''' Saturate chamber for 20 minutes; developing distance 70 mm from lower edge of the plate; relative humidity 33%, temperature 25°.
 +
 +
'''Derivatization reagent:''' Methanol sulfuric acid reagent- 180 mL of ice-cooled methanol are mixed with 20 mL of sulfuric acid.
 +
 +
'''Detection:'''
 +
a. Examine under UV light at 254 nm
 +
b. Dip (time 0, speed 5) in Derivatization reagent, heat at 100°C for 10 min, leave to cool, and examine under visible light.
 +
 +
'''Procedure:'''
 +
 +
''Reference Standard Solutions, Stationary Phase, Mobile Phase, Development, Derivatization reagent, and Detection, as described above.''
 +
 +
'''Test Sample Preparation:''' Prepare test sample as described under Reference Sample Preparations and apply 2 uL.
 +
 +
'''Identification:''' Compare Test Sample Preparation chromatogram with chromatograms of Reference Sample Preparations. The Test Sample Preparation chromatogram is similar to that of the Reference Sample Preparations chromatograms. Additional weak zones may be present.
 +
 +
Under UV light, the Test Sample Preparation chromatogram exhibits a quenching zone in the lower-third section of the chromatogram corresponding to the zone due to ammonium glycyrrhizate in the Reference Standard Solution chromatogram. Above it there are four quenching zones corresponding to those marked with red arrows in the Reference Sample Preparations chromatograms.
 +
 +
After derivatization and under visible light, the Test Sample Preparation chromatogram exhibits a brown zone in the lower-third section of the chromatogram corresponding to the zone due to ammonium glycyrrhizate in the Reference Standard Solution chromatogram. Above it there are four yellow zones corresponding to those marked with black arrows in the Reference Sample Preparations chromatograms.
 +
 +
''Note: Images presented in this entry are examples and are not intended to be used as a bases for setting specifications for quality control purposes.''
 +
            | }}
  
−
}}
 
  
−
=High Performance Thin Layer Chromatographic Identification=
 
  
 
=Supplementary Information=
 
=Supplementary Information=

Revision as of 15:57, 28 February 2014

Contents

Nomenclature

Botanical Voucher Specimen

bottomright

Glycyrrhiza glabra - Tropicos.jpg
Source: Tropicos.org. Missouri Botanical Garden. 27 Feb 2014 <http://www.tropicos.org/Image/100253567> [1]

Organoleptic Characteristics

Color: Unpeeled – yellowish or purplish brown to dark brown externally and yellowish internally. Peeled – pale yellow.

Source: Natural Remedies Pvt Ltd [2]
Natural Remedies Logo.png
Aroma/Odor: Characteristic

Flavor/Taste: Sweet, sugary

Source: American Herbal Products Association. March 2013. Organoleptic Analysis of Herbal Ingredients. AHPA: Silver Spring, MD [3]

Macroscopic Characteristics

"...Root nearly cylindrical up to 2 cm in diameter, externally wrinkled with patches of cork. Fracture, coarsely fibrous in bark and splintery in wood."

Source: Natural Remedies Pvt Ltd [4]
Natural Remedies Logo.png
bottomright bottomright

G. glabra - Flora von Deutschland.jpg
Source: Flora von Deutschland, Österreich und der Schweiz- Otto Wilhelm Thomé (1885) [5]

Glycyrrhiza glabra - foliage - eol.jpg
Foliage
Source: Encyclopedia of Life http://eol.org/data_objects/2447928 [6]

Glycyrrhiza glabra - inflorescence - eol.jpg
Inflorescence
Source: Encyclopedia of Life http://eol.org/data_objects/24932881 [7]
bottomright bottomright

G. glabra - Dried Root - Quality Assessment of Selected IndianMedicinal Plants.png
Dried Roots
Source: Natural Remedies Pvt Ltd http://www.naturalremedy.com/ [8]

G. glabra - dried root - EOL 76628 orig.jpg
Dried Roots 2
Source: Encyclopedia of Life http://eol.org/data_objects/19163752 [9]

Microscopic Characteristics

"Rhizome: Transverse section of stolon more or less rounded. Phellem severeal layered with tabular cells; outer layers filled with reddish brown contents, inner colourless. Phellogen indistinct; phelloderm three to five layered, collenchymatous; some of the cells contain calcium oxalate and minute starch grains. Secondary phloem with numerous concentrically arranged bundles of phloem fibres and surrounded by a parenchymatous sheath containing prisms of calcium oxalate. Medullary rays distinct, bi-to multiseriate, parenchymatous, in continuation with those of xylem. The rays are narrower in xylem and wider in phloem region. Xylem consists of vessels, fibres and lignified wood parenchyma. The unpeeled drug shows the presence of polyhedral tubular brownish cork cells. In case of stolons, the pith is present and is parenchymatous. The root is characterized by the presence of tetrarch xylem and absence of pith.

Powder Microscopy: It shows plenty of starch grains, hexagonal crystals vessel elements are with reticulate wall pitting."

Source: Natural Remedies Pvt Ltd [10]
Natural Remedies Logo.png


bottomright bottomright bottomright

Glycyrrhiza Root - NR transverse section.JPG
Transverse section
Source: Natural Remedies Pvt Ltd http://www.naturalremedy.com/ [11]

Glycyrrhiza Powder microscopy-starch granules - NR.jpg
Starch granules and hexagonal crystals in powder
Source: Natural Remedies Pvt Ltd http://www.naturalremedy.com/ [12]

Glycyrrhiza Powder microscopy - vessels - NR.jpg
Vessels with reticulate thickening in powder
Source: Natural Remedies Pvt Ltd http://www.naturalremedy.com/ [13]

High Performance Liquid Chromatographic Identification

Natural Remedies Logo.png
(thumbnail)
G. glabra reference standard

Sample Preparations: Extract 1.0 g of coarsely powdered Glycyrrhiza root in 50 mL of water by boiling for about 5 minutes, and filter. Repeat for 4-5 times or until the extract is colorless. Combine the extracts, concentrate to about 100 mL, and cool to room temperature. Before injection, filter through a membrane filter of 0.45-um or finer pore size, discarding the first 5 mL of the filtrate.

Column: C18, 25-cm x 4.6 mm, 5-um, XXXXX, XXXXX

Mobile Phase: 0.14 g of anhydrous potassium dihydrogen phosphate in 900 mL of water, add 0.5 mL phosphoric acid, mix, complete to volume with water, and mix (Solution A); and acetonitrile (Solution B)

Elution: gradient program, see Table below

Flow rate: 1.5 mL/min

Detection: UV, 254 nm

Injection volume: 20 uL

  • (thumbnail)

    G. glabra reference standard

  • (thumbnail)

    G. inflata reference standard

  • (thumbnail)

    G. uralensis reference standard

Source: Natural Remedies Pvt Ltd [14]

Table

Time (min) Solution A (%) Solution B (%)
0-18 95-55 5-45
18-25 55-20 45-80
25-28 20 80
28-35 20-55 80-45
35-40 55-95 45-5
40-45 95 5

High Performance Thin Layer Chromatographic Identification

Camag logo.png
(thumbnail)
Black cohosh HPTLC ID - Developed, UV 254 nm

Black cohosh (root) (Actaea racemosa)

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

Other Notes Reference Standard Solution: 0.1 mg/mL ammonium glycyrrhizate in ethanol and water (7:3). Or 0.1 mg/mL glycyrrhizic acid in ethanol and water (7:3).

Reference Sample Preparations: Mix 0.5 g of powdered sample with 10 mL of ethanol and water (7:3, v/v), sonicate for 10 minutes, centrifuge or filter the solution, and use the supernatant / filtrate.

Stationary Phase: HPTLC, Silica gel 60 F254

Mobile Phase: Ethyl acetate, acetic acid, formic acid, water (15:1:1:2)

Development: Saturate chamber for 20 minutes; developing distance 70 mm from lower edge of the plate; relative humidity 33%, temperature 25°.

Derivatization reagent: Methanol sulfuric acid reagent- 180 mL of ice-cooled methanol are mixed with 20 mL of sulfuric acid.

Detection: a. Examine under UV light at 254 nm b. Dip (time 0, speed 5) in Derivatization reagent, heat at 100°C for 10 min, leave to cool, and examine under visible light.

Procedure:

Reference Standard Solutions, Stationary Phase, Mobile Phase, Development, Derivatization reagent, and Detection, as described above.

Test Sample Preparation: Prepare test sample as described under Reference Sample Preparations and apply 2 uL.

Identification: Compare Test Sample Preparation chromatogram with chromatograms of Reference Sample Preparations. The Test Sample Preparation chromatogram is similar to that of the Reference Sample Preparations chromatograms. Additional weak zones may be present.

Under UV light, the Test Sample Preparation chromatogram exhibits a quenching zone in the lower-third section of the chromatogram corresponding to the zone due to ammonium glycyrrhizate in the Reference Standard Solution chromatogram. Above it there are four quenching zones corresponding to those marked with red arrows in the Reference Sample Preparations chromatograms.

After derivatization and under visible light, the Test Sample Preparation chromatogram exhibits a brown zone in the lower-third section of the chromatogram corresponding to the zone due to ammonium glycyrrhizate in the Reference Standard Solution chromatogram. Above it there are four yellow zones corresponding to those marked with black arrows in the Reference Sample Preparations chromatograms.

Note: Images presented in this entry are examples and are not intended to be used as a bases for setting specifications for quality control purposes.

  • (thumbnail)

    Black cohosh HPTLC ID - Developed, UV 254 nm

  • (thumbnail)

    Black cohosh HPTLC ID - Developed, UV 366 nm

  • (thumbnail)

    Black cohosh HPTLC ID - Sulfuric acid reagent, UV 366 nm

  • (thumbnail)

    Black cohosh HPTLC ID Method - Sulfuric acid reagent, white RT

Source: AHPA Practical, CAMAG HPTLC [15]


Supplementary Information

Sources

  1. ↑ Tropicos.org. Missouri Botanical Garden. 27 Feb 2014 <http://www.tropicos.org/Image/100253567>
  2. ↑ Natural Remedies Pvt Ltd http://www.naturalremedy.com/
  3. ↑ American Herbal Products Association. March 2013. Organoleptic Analysis of Herbal Ingredients. AHPA: Silver Spring, MD http://www.ahpa.org/
  4. ↑ Natural Remedies Pvt Ltd http://www.naturalremedy.com/
  5. ↑ Flora von Deutschland, Österreich und der Schweiz- Otto Wilhelm Thomé (1885)
  6. ↑ Encyclopedia of Life http://eol.org/data_objects/2447928
  7. ↑ Encyclopedia of Life http://eol.org/data_objects/24932881
  8. ↑ Natural Remedies Pvt Ltd http://www.naturalremedy.com/
  9. ↑ Encyclopedia of Life http://eol.org/data_objects/19163752
  10. ↑ Natural Remedies Pvt Ltd http://www.naturalremedy.com/
  11. ↑ Natural Remedies Pvt Ltd http://www.naturalremedy.com/
  12. ↑ Natural Remedies Pvt Ltd http://www.naturalremedy.com/
  13. ↑ Natural Remedies Pvt Ltd http://www.naturalremedy.com/
  14. ↑ Natural Remedies Pvt Ltd http://www.naturalremedy.com/
  15. ↑ AHPA Practical, CAMAG HPTLC http://www.camag.com/index.php
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