Hypericum perforatum (flowering tops)
Line 118: | Line 118: | ||
'''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. | '''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. | ||
− | |||
− | |||
'''Elution:''' Isocratic | '''Elution:''' Isocratic | ||
+ | |||
+ | '''Column Temperature:''' 40°C | ||
'''Flow rate:''' 1.0 mL/min | '''Flow rate:''' 1.0 mL/min | ||
Line 136: | Line 136: | ||
'''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) | '''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) | ||
− | |||
− | |||
'''Elution:''' Gradient, see table below | '''Elution:''' Gradient, see table below | ||
+ | |||
+ | '''Column Temperature:''' 25°C | ||
'''Detection:''' UV, 360 nm, then at 275 nm after the elution of biapigenin (about 22 min) | '''Detection:''' UV, 360 nm, then at 275 nm after the elution of biapigenin (about 22 min) |
Revision as of 18:35, 28 February 2014
Contents |
Nomenclature
Hypericum perforatum L. Clusiaceae
Standardized common name (English): St John's wort
Botanical Voucher Specimen
|
|
Organoleptic Characteristics
|
Macroscopic Characteristics
|
Microscopic Characteristics
|
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.
Elution: Isocratic
Column Temperature: 40°C
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)
Elution: Gradient, see table below
Column Temperature: 25°C
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
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 |
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
|
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