Hypericum perforatum (flowering tops)
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+ | |} | ||
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+ | =High Performance Liquid Chromatographic Identification= | ||
+ | {{Botanical | source=Indena S.p.A. | ||
+ | | companyURL=http://www.indena.com/ | ||
+ | | companyimage=Indena - Logo.png | ||
+ | |mainimage=Indena - St. John's Wort Info - hypericin hplc.jpg | ||
+ | |caption1=HYPERICUM PERFORATUM DRY EXTRACT - Total hypericins HPLC assay | ||
+ | |image2=Indena - St. John's Wort Info - hyperforin and flavonoids hplc.jpg | ||
+ | |caption2=HYPERICUM PERFORATUM DRY EXTRACT - Hyperforin and Flavonoids HPLC assay | ||
+ | | | ||
+ | | description= | ||
+ | '''Method for Hypericins''' | ||
+ | |||
+ | '''Test Sample Preparation:''' xxxxxxxxxx. | ||
+ | |||
+ | '''Column:''' 15-cm x 4.6 mm, 5-um, Waters Atlantis C18 | ||
+ | |||
+ | '''Mobile Phase:''' A mix of 39 volumes of ethyl acetate, 41 volumes of a 15.6 g/L solution of sodium dihydrogen phosphate adjusted to pH 2 with phosphoric acid and 160 volumes of methanol. | ||
+ | |||
+ | '''Column Temperature:''' 40°C | ||
+ | |||
+ | '''Elution:''' Isocratic | ||
+ | |||
+ | '''Flow rate:''' 1.0 mL/min | ||
+ | |||
+ | '''Detection:''' Vis, 590 nm | ||
+ | |||
+ | '''Injection volume:''' 20 uL | ||
+ | |||
+ | '''Method for Hyperforins and Flavonoids''' | ||
+ | |||
+ | '''Test Sample Preparation:''' xxxxxxxxxx. | ||
+ | |||
+ | '''Column:''' 15-cm x 4.6 mm, 3-um, YMC-Pack ODS-A | ||
+ | |||
+ | '''Mobile Phase:''' Water, containing 0.3 % (v/v) phosphoric acid 85% (Solution A) and acetonitrile containing 0.3 % (v/v) phosphoric acid 85% (Solution B) | ||
+ | |||
+ | '''Column Temperature:''' 25°C | ||
+ | |||
+ | '''Elution:''' Gradient, see table below | ||
+ | |||
+ | '''Detection:''' UV, 360 nm, then at 275 nm after the elution of biapigenin (about 22 min) | ||
+ | |||
+ | '''Injection volume:''' 10 uL | ||
+ | |||
+ | }} | ||
+ | ===Table: Gradient program=== | ||
+ | {| border=1 | ||
+ | | Time (min) || Solution A (%) || Solution B (%) || Flow Rate (mL/min) | ||
+ | |- | ||
+ | 0-8 || 82 || 18 || 0.8 | ||
+ | |- | ||
+ | 8-18 || 82-47 || 18-53 || 0.8 | ||
+ | |- | ||
+ | 18-18.1 || 47-3 || 53-97 || 0.8 | ||
+ | |- | ||
+ | 18.1-19 || 3 || 97 || 1.2 | ||
+ | |- | ||
+ | 19-29 || 3 || 97 || 1.2 | ||
+ | |- | ||
+ | 29-30 || 82 || 18 || 1.2 | ||
+ | |- | ||
+ | 30-31 || 82 || 18 || 0.8 | ||
+ | |- | ||
+ | 31-35 || 82 || 18 || 0.8 | ||
|} | |} | ||
Revision as of 17:14, 28 February 2014
Contents |
Nomenclature
Hypericum perforatum L. Clusiaceae
Standardized common name (English): St John's wort
Botanical Voucher Specimen
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Organoleptic Characteristics
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Macroscopic Characteristics
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Microscopic Characteristics
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High Performance Liquid Chromatographic Identification
'Method for Hypericins
Test Sample Preparation: xxxxxxxxxx.
Column: 15-cm x 4.6 mm, 5-um, Waters Atlantis C18
Mobile Phase: A mix of 39 volumes of ethyl acetate, 41 volumes of a 15.6 g/L solution of sodium dihydrogen phosphate adjusted to pH 2 with phosphoric acid and 160 volumes of methanol.
Column Temperature: 40°C
Elution: Isocratic
Flow rate: 1.0 mL/min
Detection: Vis, 590 nm
Injection volume: 20 uL
Method for Hyperforins and Flavonoids
Test Sample Preparation: xxxxxxxxxx.
Column: 15-cm x 4.6 mm, 3-um, YMC-Pack ODS-A
Mobile Phase: Water, containing 0.3 % (v/v) phosphoric acid 85% (Solution A) and acetonitrile containing 0.3 % (v/v) phosphoric acid 85% (Solution B)
Column Temperature: 25°C
Elution: Gradient, see table below
Detection: UV, 360 nm, then at 275 nm after the elution of biapigenin (about 22 min)
Injection volume: 10 uL
Source: Indena S.p.A. [16]
Table: Gradient program
0-8 || 82 || 18 || 0.8 8-18 || 82-47 || 18-53 || 0.8 18-18.1 || 47-3 || 53-97 || 0.8 18.1-19 || 3 || 97 || 1.2 19-29 || 3 || 97 || 1.2 29-30 || 82 || 18 || 1.2 30-31 || 82 || 18 || 0.8 31-35 || 82 || 18 || 0.8
Time (min) | Solution A (%) | Solution B (%) | Flow Rate (mL/min) |
High Performance Thin Layer Chromatographic Identification
St. John's Wort (herb) (Hypericum perforatum) Lane Assignments Lanes, from left to right (Track, Volume, Sample):
Reference materials used here have been authenticated by macroscopic, microscopic &/or TLC studies according to the reference source cited below held at Alkemists Laboratories, Costa Mesa, CA. Stationary Phase Silica gel 60, F254, 10 x 10 cm HPTLC plates Mobile Phase ethyl acetate: Acetic acid: HCOOH: H2O [10/1.1/1.1/2.4] Sample Preparation Method 0.3 g + 3 ml 70% grain EtOH sonicated + heated @ 50° C ~ 1 hr Detection Method Natural Product Reagent + PEG -> UV 365 nm Reference see British Pharmacopoeia, 2003
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Supplementary Information
Sources
- ↑ Tropicos.org. Missouri Botanical Garden. 28 Feb 2014 <http://www.tropicos.org/Image/61762>
- ↑ Botanical Voucher Specimen Library, Alkemists Laboratories http://www.alkemist.com
- ↑ WHO monographs on selected medicinal plants, Vol.2, World Health Organization, Geneva, 1999
- ↑ WHO monographs on selected medicinal plants, Vol.2, World Health Organization, Geneva, 1999
- ↑ Roy Upton, American Herbal Pharmacopoeia® http://www.herbal-ahp.org/
- ↑ American Herbal Pharmacopoeia® http://www.herbal-ahp.org/
- ↑ American Herbal Pharmacopoeia® http://www.herbal-ahp.org/
- ↑ American Herbal Pharmacopoeia® http://www.herbal-ahp.org/
- ↑ WHO monographs on selected medicinal plants, Vol.2, World Health Organization, Geneva, 1999
- ↑ Elan M. Sudberg, Alkemist Laboratories http://www.alkemist.com
- ↑ Elan M. Sudberg, Alkemist Laboratories http://www.alkemist.com
- ↑ Claudia Borst, PhytoLab http://www.phytolab.com/en.html
- ↑ Claudia Borst, PhytoLab http://www.phytolab.com/en.html
- ↑ Claudia Borst, PhytoLab http://www.phytolab.com/en.html
- ↑ Claudia Borst, PhytoLab http://www.phytolab.com/en.html
- ↑ Indena S.p.A. http://www.indena.com/
- ↑ Elan M. Sudberg, Alkemist Laboratories http://www.alkemist.com