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A Review of Modern Sample-preparation Techniques for the Extraction and Analysis of Medicinal Plants

A Review of Modern Sample-preparation Techniques for the Extraction and Analysis of Medicinal Plants

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Sample preparation is the crucial first step in the analysis of herbs. In recent years there has been increasing interest worldwide in the use of alternative/herbal medicine for the prevention and treatment of various illnesses. Currently, however, quality-related problems (lack of consistency, safety, and efficacy) seem to be overshadowing the potential genuine health benefits of various herbal products. Thus, the development of “modern” sample-preparation techniques with significant advantages over conventional methods for the extraction and analysis of medicinal plants is likely to play an important role in the overall effort of ensuring and providing high-quality herbal products to consumers worldwide. In this article, recent developments and applications of modern sample-preparation techniques for the extraction, clean-up, and concentration of analytes from medicinal plants or herbal materials are reviewed. These modern techniques include solidphase microextraction, supercritical-fluid extraction, pressurized-liquid extraction, microwave-assisted extraction, solid-phase extraction, and surfactant-mediated extraction.
Sample preparation is the crucial first step in the analysis of herbs. In recent years there has been increasing interest worldwide in the use of alternative/herbal medicine for the prevention and treatment of various illnesses. Currently, however, quality-related problems (lack of consistency, safety, and efficacy) seem to be overshadowing the potential genuine health benefits of various herbal products. Thus, the development of “modern” sample-preparation techniques with significant advantages over conventional methods for the extraction and analysis of medicinal plants is likely to play an important role in the overall effort of ensuring and providing high-quality herbal products to consumers worldwide. In this article, recent developments and applications of modern sample-preparation techniques for the extraction, clean-up, and concentration of analytes from medicinal plants or herbal materials are reviewed. These modern techniques include solidphase microextraction, supercritical-fluid extraction, pressurized-liquid extraction, microwave-assisted extraction, solid-phase extraction, and surfactant-mediated extraction.

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Published by: Ahmad Abdullah Najjar on Aug 05, 2008
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05/09/2014

 
Abstract
Sample preparation is the crucial first step inthe analysis of herbs. In recent years there has been in-creasing interest worldwide in the use of alternative/herbalmedicine for the prevention and treatment of various ill-nesses. Currently, however, quality-related problems (lack of consistency, safety, and efficacy) seem to be overshad-
owing the potential genuine health benefits of various herbal
products. Thus, the development of “modern” sample-prep-
aration techniques with significant advantages over con-ventional methods for the extraction and analysis of me-dicinal plants is likely to play an important role in theoverall effort of ensuring and providing high-quality herbalproducts to consumers worldwide. In this article, recentdevelopments and applications of modern sample-prepa-ration techniques for the extraction, clean-up, and concen-tration of analytes from medicinal plants or herbal materi-als are reviewed. These modern techniques include solid-phase microextraction, supercritical-fluid extraction, pres-surized-liquid extraction, microwave-assisted extraction,solid-phase extraction, and surfactant-mediated extrac-tion.
Keywords
Sample preparation · Extraction · Qualitycontrol · Medicinal plants · Herbs
Introduction
Plants are naturally gifted at the synthesis of medicinalcompounds. The extraction and characterization of activecompounds from medicinal plants have resulted in thediscovery of new drugs with high therapeutic value [1, 2].A classic example is aspirin, which was initially discov-ered as salicylic acid in willow bark and leaves [1]; an-other noted example is taxol, recently proven to be effec-tive against breast and ovarian cancers, which was ini-tially discovered in bark of yew trees [2].The use of medicinal plants (herbs) has a long historythroughout the world and herbal preparations, includingherbal extracts, can be found in the pharmacopoeias of numerous countries [3]. In recent years there have been arenaissance of interest in natural or herbal remediesworldwide, partly because of the realization that modernmedicine is not capable of providing a “cure-all” solutionagainst human diseases and that the presence of unwantedside-effects is almost unavoidable. Unlike modern drugsthat invariably comprise a single active species, herb ex-tracts and/or prescriptions contain multiple active con-stituents. Interestingly, natural compounds contained inthese “herbal cocktails” can act in a synergistic mannerwithin the human body, and can provide unique therapeu-tic properties with minimal or no undesirable side-effects[4].A key factor in the widespread acceptance of natural oralternative therapies by the international community in-volves the “modernization” of herbal medicine. In otherwords, the standardization and quality control of herbalmaterials by use of modern science and technology is crit-ical. At present, however, quality-related problems (lack of consistency, safety, and efficacy) seem to be overshad-owing the potential genuine health benefits of variousherbal products, and a major cause of these problemsseems to be related to the lack of simple and reliable ana-lytical techniques and methodologies for the chemicalanalysis of herbal materials [5, 6].Sample preparation is the crucial first step in the analy-sis of herbs, because it is necessary to extract the desiredchemical components from the herbal materials for fur-ther separation and characterization. Thus, the develop-ment of “modern” sample-preparation techniques withsignificant advantages over conventional methods (e.g.reduction in organic solvent consumption and in sampledegradation, elimination of additional sample clean-upand concentration steps before chromatographic analysis,improvement in extraction efficiency, selectivity, and/orkinetics, ease of automation, etc.) for the extraction and
Carmen W. Huie
 A review of modern sample-preparation techniques for the extraction and analysis of medicinal plants
Anal Bioanal Chem (2002) 373:23–30DOI 10.1007/s00216-002-1265-3Received: 12 November 2001 / Revised: 12 February 2002 / Accepted: 14 February 2002 / Published online: 3 April 2002
SPECIAL ISSUE PAPER
C.W. Huie (
)Department of Chemistry, Hong Kong Baptist University,Kowloon Tong, Hong Konge-mail: cwhuie@net1.hkbu.edu.hk ©Springer-Verlag 2002
 
analysis of medicinal plants is likely to play an importantrole in the overall effort of ensuring and providing high-quality herbal products to consumers worldwide.In this article recent developments and applications of modern sample-preparation techniques for the extraction,clean-up, and concentration of analytes from medicinalplants or herbal materials are reviewed. These moderntechniques include solid-phase microextraction, supercrit-ical-fluid extraction, pressurized-liquid extraction, mi-crowave- assisted extraction, solid-phase extraction, andsurfactant-mediated extraction. Emphasis is placed onbrief description of the unique capabilities and advantagesand disadvantages of each modern sample-preparationtechniques and of how these techniques were exploited toimprove the extraction and analysis of a variety of medic-inal plants. More detailed description of the basic princi-ples of these modern sample-preparation techniques forthe extraction of solid materials in general is available innumber of excellent review articles recently appeared inthe literature [7, 8, 9, 10].
Modern techniques for sample preparationin the analysis of medicinal plants
Headspace analysisThe medicinal properties of plants can be related in part tothe presence of volatile constituents (e.g. essential oils) inthe plant matrix, and gas chromatography (GC) is fre-quently used for determination of the volatile compositionof plant materials. Because the sample to be injectedshould be free from non-volatile components, a fractiona-tion step is necessary before GC analysis. The disadvan-tages of commonly used sample-preparation techniquessuch as distillation and liquid solvent extraction are thatthey usually require large amounts of organic solvents andmanpower; these methods also tend to be destructive innature (i.e. significant artifact formation can occur owingto sample decomposition at high temperatures) [11].Headspace (HS) sampling is well suited for the frac-tionation of volatile compounds from complex solid ma-trices such as plant materials. Sanz and co-workers [12]have shown that reproducible and rapid identification of volatile compounds in aromatic plants can be achievedwhen static HS sampling is coupled with GC–mass spec-trometry (MS), with the advantages of eliminating the ex-traction or fractionation step, reducing artifact formation,and providing on-line capacity. More recently the sameresearch group has extended the capabilities of automatedHS sampling in the GC–MS analysis of volatile com-pounds from
Origanum vulgare
[13].Another example of herb analysis by automatic HS–GCis recent work by Stuppner and Ganzera [14] on determi-nation of the safrole content from different
 Asarum
spe-cies from China and Europe. These
 Asarum
species con-tain up to 5.5% essential oils with methyleugenol as themajor constituent and several minor components, e.g. saf-role, which is known to have mutagenic and carcinogeniceffects. The HS–GC results for safrole were found to be ingood agreement with those obtained by “classical” GCanalysis, i.e., using an organic solvent (dichloromethane)for extraction.A novel and effective approach, known as solid-phasemicroextraction (SPME), was recently developed by Paw-liszyn and co-workers [15] as a solvent-free samplingtechnique. In this approach, the analytes from the sampleare adsorbed directly onto an adsorbent coated fused-sil-ica fiber (which fits inside a syringe needle) and then ei-ther thermally desorbed directly into a GC injection portor high-performance liquid chromatography (HPLC) in- jection valve. By placing the fiber in head space at equi-librium with a sample (i.e. the technique of HS-SPME), arapid and inexpensive method for isolation of volatile or-ganic analytes for subsequent GC–MS analysis has beensuccessfully demonstrated for the quality control of herbalmedicine and other formulations containing herb extracts,such as terpenoids, peppermint, rosemary, sage, and thyme[16]. In addition to endogenous active components in me-dicinal plants, the usefulness of SPME as a sampling toolhas also been recently demonstrated by Hwang and Lee[17] for the GC–MS analysis of toxic contaminants, i.e.,
organochlorine pesticide residues, present in Chinese herbal
materials.In a recent paper Pawliszyn and co-workers [18] re-ported successful demonstration of the feasibility andunique advantages of SPME for the characterization andquantification of the biogenic volatile organic compounds(e.g. isoprene and terpenoid compounds) emitted by liv-ing plants (leaves of 
 Eucalyptus citriodora
). By use of coated SPME fibers they were able to identify 33 com-pounds emitted by the living plant and, using diffusion-based SPME quantitation, it was possible to quantify sub-parts-per-billion amounts of isoprene after a very shortextraction time (15 s).The effect of the fiber coating in HS-SPME–GC analy-sis of aromatic and medicinal plants has been investigatedin depth by Bicchi and co-workers [19]. Interestingly, itwas found that fibers consisting of two components, a liq-uid polymeric coating for the less polar analytes and asolid polymeric coating for the more polar analytes, weremore effective for HS-SPME analysis of the volatile frac-tion of the four aromatic and medicinal plants investigated(rosemary, sage, thyme, and valerian). More recently, re-placement of the fiber needle with a stir-bar, a new sam-pling technique known as stir-bar sorptive extraction(SBSE), has been described by Bicchi and co-workers[20] for the headspace sampling of the same four aro-matic/medicinal plants. Because of the larger amounts of trapping material (e.g. polydimethylsiloxane) coated onthe stir-bar, the concentration capability is significantlygreater than that of SPME.Supercritical- and subcritical-fluid extractionSupercritical fluid extraction (SFE) has been used formany years for the extraction of volatile components, e.g.
24
 
essential oils and aroma compounds from plant materials,on an industrial scale [21]. Recently, the application of this technique on an analytical scale has started to attractwide interest for sample preparation before chromato-graphic analysis [22]. The potential advantages includethe ability to perform rapid (often less than 30 min) ex-tractions, to reduce the use of hazardous solvents (i.e. car-bon dioxide is commonly used as the extraction solvent),and to couple the extraction step with gas, liquid, or su-percritical-fluid chromatography.An important advantage of applying SFE to the extrac-tion of active compounds from medicinal plants is thatdegradation as a result of lengthy exposure to elevatedtemperatures and atmospheric oxygen are avoided. Forexample:1.Smith and Burford [23] showed that the active com-pound of feverfew, i.e. the sesquiterpene lactoneparthenolide (well known to be unstable and to de-grade during storage), can be efficiently and selec-tively extracted from dried feverfew by SFE withoutthermal degradation;2.Bartley and Foley [24] demonstrated that althoughvolatile compounds (essential oils) in ginger were ex-pected to be strongly influenced by heat treatment,possibly as a result of hydrolysis, oxidation, and re-arrangement, a very low concentration of gingeroldegradation products were, however, found after SFEof Australian-grown ginger, which attested to the mildnature of the extraction procedure; and3.in the determination of the enantiomeric purity of at-ropine (a tropane alkaloid with significant medical in-terest found in plants of the Solanaceae family), Ma-teus et al. [25] found that SFE induced less racemiza-tion than classical liquid–solid extraction procedures.In SFE several experimental conditions can be adjusted tooptimize the recovery, kinetics, and selectivity of the ex-traction. For example:1.during optimization of SFE conditions for extractionof active ingredients from
Curcuma zedoaria
, Ma andco-workers [26] found that the density of the CO
2
andthe fluid volume passing through the plant matrix arethe most important factors affecting extraction effi-ciency, whereas increasing the temperature has littleeffect;2.when using SFE to extract indirubin (the active ingre-dient in the herbs
Strobilanthes cusia
,
Isatis tinctoria
,and
Polygonum tinctorium
) Li et al. [27] found thatfaster extraction kinetics were achieved by employingfactorial design to optimize experimental variablessuch as temperature, pressure, modifier concentration,static extracting time, and CO
2
dynamic extracting vol-ume; and3.it is interesting to note from another study byHawthorne and co-workers [28] that the rate of extrac-tion of volatile active components (essential oils) fromplant matrices by SFE seemed to be governed by ana-lyte–matrix interaction rather than the bulk solubilityof the analyte in pure CO
2
, because the extraction rateswere found to increase greatly due to an addition of or-ganic modifiers.In another fundamental study Fahmy and co-workers [29]clearly demonstrated that swelling of the plant matrix, asa result of interaction between modifiers and matrix, wasan important factor in enhancing extraction recovery andkinetics in SFE. Also, by use of different conditions of pressure and temperature, Reglero and co-workers [30]found that SFE conditions can be fined tuned for selectiveextraction of an antioxidant fraction with almost no resid-ual aroma from rosemary plants. It has also been shown,by operating under sub-critical temperature and pressureconditions, that CO
2
fluid can be used as solvent for theselective extraction of essential oils [31] and diterpeneglycosides [32] from plants of medicinal interest. Morerecent examples of the application of SFE for the opti-mum extraction of analytes from medicinal plants underdifferent extraction conditions are listed in Table 1.The on-line coupling of SFE with supercritical-fluidchromatography (SFC) has recently been shown to affordenhanced speed and sensitivity, as a result of the ability touse this technique to perform consecutive extraction, con-centration, and separation of the constituents of herbalmaterials. For example, using an amino column for trap-ping and separation, Suto and co-workers [44] demon-strated that the on-line SFE–SFC analysis of the activecompounds (magnolol and honokiol) in Magnoloae cortexcan be completed within 5 min. Also, by using silica gelas the trapping and separation column and sodium sulfo-succinate as the counter-ion for ion-pair formation, thesame research group recently showed that the on-line cou-pling of ion-pair SFE–SFC enabled the rapid analysis(within 10 min) of berberine and palmatine (positivelycharged active species) in Phellodendri cortex [45].Another interesting recent development is the on-linecoupling of SFE to a uterotonic bioassay by Sewram andco-workers [46]. In South Africa, the use of traditionalmedicine is popular and the ingestion of plant extractsduring pregnancy to provide health supplements or to in-duce labor is common. In the on-line SFE uterotonicbioassay system [46] SFE extracts from four local medic-inal plants were transferred directly to a uterus musclechamber to identify the active fractions (i.e. the fractionscapable of inducing muscle contraction can be determinedrapidly, safely, and sensitively). This novel on-line SFEbioassay method could also be adapted for screeningplants with other therapeutic properties, e.g. those usedfor treatment of diabetes mellitus and hypertension. An-other novel approach involving the off-line coupling of SFE to GC in such a way that the glass liner of a pro-grammed temperature vaporizer is placed after the separa-tion vessels of the SFE extraction module has been re-cently demonstrated by Blanch et al. [47] to be effectivefor the sensitive and selective analysis of complex plantmatrices.
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