Hypericum perforatum
Industry request: providing a stable and standardized extract, as well as producing a new
pharmaceutical product which could be with higher bioavailability, pharmacological effect and
convenient.
Introduction
2. Fractionation
The active compounds present in hypericum perforatum
include
phloroglucinols
(hyperforin),
naphthodianthrones
(hypericin)
and
flavonoids
(quercetin), which are assumed to be responsible for the
activity of extracts in the treatment of wounds and scars.
Such compounds however are unstable when attempting
to obtain them in pure form, and they decompose over
time due to the effect of light and heat.
Pharmaceutical formulations available nowadays consist
of fresh hypericum perforatum extract as an active
ingredient and oily excipients. These oily formulations
make the skin saturated with oil and difficult to wash
when used for long term application and it is difficult to
monitor the lesion condition.
Liquid-liquid extraction was performed using solvents from low to high
polarity to obtain five fractions (hexane, dichloromethane, ethyl acetate,
n-butanol and the remaining water). The fractions yielded 21%, 3%, 11%,
14% and 51% respectively.
0.09
compound/dry fraction
[w/w %]
compound / dry fraction
[w/w %]
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0.08
0.07
0.06
0.05
0.04
0.03
0.02
0.01
0.00
0
Hexane
MeCl2
EtAc
BuOH
Hexane
MeCl2
EtAc
BuOH
Fig.2.: Concentration of hyperforin (left) and hypericin (right) in the fractions
Experiments and Results
1. Extraction
Two varieties of hypericum (hypericum perforatum and hypericum
perforatum var. Topas) were cultivated in SEKEM group farm. Hypericum
of var. Topas showed a more successful adaptation than the other one, as
it gave higher quantitative and qualitative production.
In the Joanneum Research laboratories extraction trials of hypericum
perforatum flowering tops were conducted at a laboratory scale using
Accelerated Solvent Extraction (ASE). The up-scaling of an extraction
method was performed by a pilot plant extraction unit (Dig-Maz). This
process yielded a highly concentrated dry extract. For the qualitative and
quantitative analysis of the active compounds, High Performance Liquid
Chromatography coupled with a Diode Array Detector (HPLC-DAD) was
applied and the total amount of hypericins was measured with a
spectrophotometer at λ = 590nm.
compounds / dry extract
[w/w %]
4
3.5
3
2.5
2
1.5
1
0.5
0
Fig.1.: Active compounds in dry hypericum perforatum extract
(solvent: 80% EtOH, temperature: 70°C)
MENA Supervisor:
Dr. Amira A. Motaal
Heliopolis Academy, Sekem
MENA Student:
Mohamed Adly
Student picture
3. Formulation
Hydroalcoholic gels containing 2% of the extract
were prepared and evaluated visually, by means
of HPLC-DAD and spectrophotometry, by
performing in vitro release studies (USP
dissolution test), pH measurements and stability
studies, in order obtain the best properties.
Niosomes were evaluated according to their
percentage drug entrapments, particle size
analysis, zeta potential and stability studies.
Conclusions
Fig.3.: Hyperforin (upper),
Hypericin (lower)
We were able to obtain a stable extract rich in active compounds. The
hyperforin content lies around 3%.
We recommend using hexane and ethyl acetate only as solvents for
fractionation. Our results show that a separation of hyperforin, hypericin
and polar compounds can be achieved using this method. Since the
remaining water fraction was found to be rich in flavonoids and probably
other polyphenols with antioxidant activity, this fraction could also be
incorporated in food-supplements or in cosmetics. As an alternative to
liquid extraction with hexane, one should consider performing
supercritical fluid extraction, which is a sustainable way of extracting
unpolar compounds such as hyperforin.
After comparing some products on the market we decided to choose a
formula containing 2% of hypericum perforatum extract. Further
investigations will give clarity about the stability and properties of the
hydrogels and niosomes.
European Supervisor:
Mag. Dr. Herbert Böchzelt
Joanneum Research
European Student:
Natalie Taupe
This project has been funded with support from the European
Commission.
This poster reflects the views only of the author, and the Commission
cannot be held responsible for any use which may be made of the
information contained therein.
Student picture