J. of Supercritical Fluids 61 (2012) 55 61
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The Journal of Supercritical Fluids
jou rn al h om epage: www.elsev ier .com
Anik Fe ala Semmelweis Ub Gedeon Richtc Budapest Uni ess En
a r t i c l
Article history:Received 20 JuReceived in reAccepted 5 Oc
Keywords:Alnus glutinosaPentacyclic triSupercritical GCMSLCMSRP-HPLC
on ofin orded byere idrms ole in
SFE xane ct, ret yielf trite
Importance of pharmacologically active natural compounds andplant sources has been re-evaluated in the recent years and itbecame onein low concoutstandingrials is supethe 1970s tbroadened istics [1,2].
Lupane-compoundswaxes. Amosome of thpromising toxicity andthese chemanti-HIV [6hepatoprot
Corresponnosy, 1085 Bu
treat wounds, ulcers, fever and abdominal pain . Various typesof plant secondary metabolites including anthraquinones, phenolicglycosides, avonol glycoside, terpenoids, xanthones have previ-ously been reported from the barks, buds, leaves and pollens of A.
0896-8446/$ doi:10.1016/j. of the most active research elds. They often presententration in the plants and are chemically sensitive. An
method to recover these compounds from raw mate-rcritical uid extraction (SFE). It has been applied sinceo isolate natural products and its application eld hasdue to its several well-known advantageous character-
type pentacyclic triterpenoids are naturally originated, their main sources are stem barks, leaves and fruitng them, betulin, betulinic acid and lupeol represent
e most interesting molecules due to their numerouspharmacological effects that are associated with low
high selectivity. According to the recent researches,ical agents show anti-tumoral [3,4], antiviral ,,7], antibacterial, anti-inammatory, antioxidant ,ective [9,10] and anxiolitic properties .
alder (Alnus glutinosa (L.) Gaertn., Betulaceae) is aree native to a number of countries in northern Africa,Asia and Europe. In the folk medicine it was used to
ding author at: Semmelweis University, Department of Pharmacog-dapest, lloi t. 26, Hungary. Tel.: +36 1 431 4683.ress: firstname.lastname@example.org (A. Felfldi-Gva).
glutinosa .Supercritical uid extraction has previously been used to extract
triterpenoids from various plants. Effect of different supercriticaluid extraction conditions on triterpene content and other com-ponents of chaste berry fruit (Vitex Agnus castus) and dandelionleaves (Taraxacum ofcinale Weber et Wiggers) were studied usinga 32 full factorial design. The pressure and temperature were variedover the ranges of 100450 bar and 3565 C. The extraction yield,the recovery of -sitosterol and -amyrin were compared to thoseobtained by Soxhlet extraction. Similar trends were experiencedin case of both plants. It was revealed that rather pressure thantemperature had signicant effect on recovery. By evaluation theexperiments 450 bar and 6065 C was found to be the best condi-tion within the ranges investigated where the highest yield of thesecompounds was obtained [14,15].
The inuence of modiers (methanol and dimethyl sulfoxide)on SFE of triterpenes (ginsenosides) was studied by Wood et al. onNorth American ginseng root (Panax quinquefolius). They examinedtheir effect on the total extraction yield as well as total amountand composition of extracted ginsenosides by combination ofstatic and dynamic extraction with supercritical CO2. Soxhletextraction resulted in 409 mg/g extraction yield and 75.5 mg/g ofginsenosides. Quantities obtained with pure CO2 and with dynamicextraction using modier was negligible compared to that. Several
see front matter 2011 Elsevier B.V. All rights reserved.supu.2011.10.003ritical uid extraction of Alnus glutinosa
lfldi-Gvaa,b,, Szabolcs Szarkaa, Bla Simndic, Bniversity, Department of Pharmacognosy, 1085 Budapest, lloi t 26, Hungary
er Plc, 1103 Budapest, Gymroi t 19-21, Hungaryversity of Technology and Economics, Department of Chemical and Environmental Proc
e i n f o
ne 2011vised form 4 October 2011tober 2011
(L.) Gaertn.terpenoidsuid extraction
a b s t r a c t
Supercritical carbon dioxide extractiT = 40/60 C, EtOH addition = 0/5/10%) composition of extracts were analyzcyclic triterpenes and -sitosterol wmethod over Soxhlet extraction in tecurves revealed that pressure had littco-solvent increased it. The optimumextracts was 3.81% compared to n-hewas 3.57, 2.95 and 14.33 g/100 g extraSoxhlet extraction ensures the highessubstances hence the concentration o/ locate /supf lu
zs Blazicsa, Blanka Simonc, gnes Krya
gineering, 1111 Budapest, Muegyetem rkp. 3, Hungary
Alnus glutinosa (L.) Gaertn. was performed (P = 300/450 bar,er to determine preferable process conditions. Phytochemical
means of TLC, GCMS, LCMS, RP-HPLC. Total of 11 penta-entied. The results indicated that SFE is an advantageousf yield and recovery of target compounds. Overall extractionuence on extraction yield, while temperature and amount ofcondition was 300 bar/60 C/10% EtOH, where the amount of(2.56%). Highest amount of betulin, betulinic acid and lupeolspectively, depending on the applied SFE condition. Ethanolicd (40.90%), but provides the extracts diluted with undesirablerpenes in the extract was very low.
2011 Elsevier B.V. All rights reserved.
56 A. Felfldi-Gva et al. / J. of Supercritical Fluids 61 (2012) 55 61
experimental conditions were studied using CO2 + MeOH(20.748.3 MPa, 50110 C, 530% MeOH mole percentage) incombined extraction. Modier usage was found to have the mostsignicant effect on the quantity of ginsenosides extracted. Onlya small amamount (1at 30% Memately 90%ginsenoside
Some au(Calendula extraction The extracdiol esters (T = 29931oresin by Can increasinincrease in complex dusure that rephysical meevaluated tmodels. Thelogistic mod
SFE alsoous oilseedroselle seedide extractiof 200400ical uid in roselle scholesterolvariable withe extractephytosterol40 C, a highrate of 20 m
Betulin by Zhang etlike modiand extractmum betul1.5 mL g1 bthe extracti
In our pan alternatiin addition plant in varus a good poof differenttion of targSolvent extable SFE conthe plant, fo
The aldenia. Plant sdeposited inversity, Buddry and dar(Hungary).
from Carlo Erba (Italy) and acetic acid of LC grade was fromFluka (Switzerland). Methanol of LC super gradient grade wasfrom SigmaAldrich (Germany). All other chemicals of analytical-reagent grade were obtained from Reanal (Hungary). Water used
partto thre dacopainede divalu00 e0 = 1
of 0tulinrt RC
Soxh extrient illede cotracless. tion
Supe rawre pred b00 g (essund 6
10%n wraturout and
on used t
mentountount of ginsenosides was extracted at low modier2 mg/g at 1 g modier/g ginseng). On the contrary,
OH mole percentage (4.1 g mod/g ginseng) approxi-, while with DMSO (3.6 g mod/g ginseng) 64% of thes obtained in Soxhlet extraction were extracted .thors investigated SFE on triterpenoids of marigold
ofcinalis). Hamburger et al. applied supercritical uidfor purication of faradiol esters under 500 bar/50 C.tion yield was 5% and approximately 85% of fara-were extracted. Also, SFE under different conditions3 K, P = 1220 MPa) was investigated on marigold ole-ampos et al. According to their results, pressure hadg effect on the yield at constant temperature due to
solvent density. However, the effect of temperature wase to the combined effect of density and vapour pres-sulted crossover of the yield isotherms at 15 MPa. Thechanisms involved in the mass transfer process werehrough the application of ve different mathematical
best t to the experimental data was obtained for theel [17,18].
has been used to extract -sitosterol from vari-s and other products. Recovery of phytosterol froms (Hibiscus sabdariffa L.) via supercritical carbon diox-on modied with ethanol was investigated at pressures
bar, temperatures from 40 to 80 C and at supercrit-ow rates from 10 to 20 mL min1. The major sterolseed oil were -sitosterol, campesterol, stigmasterol,
and 5-avenasterol. Pressure was determined to be theth the highest inuence on phytosterol composition ind seed oil. The highest extraction yield and the highest
composition were obtained at the low temperature of pressure of 400 bar and a high supercritical uid owL min1 .was extracted by SFE from bark of Betula platyphylla
al. The authors investigated and analyzed parameterser dosage (12 mL), extraction pressure (1535 MPa)ion temperature (3575 C). It was found that the opti-in recovery is achieved when the modier dosage wasark powder, the extractive pressure was at 20 MPa, andve temperature was at 55 C .revious work, we described A. glutinosa (L.) Gaertn. asve source of lupane-type triterpenes and phytosterolsto Betula species . As these compounds occur in theious chemical forms like alcohols, carbonic acids, it gavessibility to use as a model plant investigating the effects
SFE conditions on the extraction yield and concentra-et compounds compared to those obtained by classicalraction. Objective of this work was to determine prefer-dition and to identify valuable bioactive compounds incusing on pentacyclic triterpenoids.
ls and methods
r bark was collected