Solid-phase extraction of liquiritin and glycyrrhizin from licorice using porous alkyl-pyridinium polymer sorbent

Introduction – Liquiritin and glycyrrhizin are valuable components of licorice. An effective separation and determination procedure is needed to separate the liquiritin and glycyrrhizin from the licorice extract. Methodology – A polymer‐confined, ionic liquid sorbent was developed using a process involving polymerisation and modification. The obtained porous particles were used as a sorbent in a solid‐phase extraction process to isolate liquiritin and glycyrrhizin from licorice with different washing and elution solvents. The porous alkyl‐pyridinium polymer sorbent was compared with the C₁₈ sorbent. Results – A simple and convenient method was established to the selectively separate and determinate of liquiritin and glycyrrhizin using a porous ionic liquid‐based polymer coupled with HPLC. Additionally, this study evaluated the application of this sorbent for the detection of these two compounds in commercial medicines. Conclusion – This method was a viable tool that was compatible with the existing HPLC methods and was used to separate and analyse the content of liquiritin and glycyrrhizin in licorice. Copyright © 2010 John Wiley & Sons, Ltd.     

Sea sand disruption method (SSDM) as a valuable tool for isolating essential oil components from conifers

Essential oils are one of nature's most precious gifts with surprisingly potent and outstanding properties. Coniferous oils, for instance, are nowadays being used extensively to treat or prevent many types of infections, modify immune responses, soothe inflammations, stabilize moods, and to help ease all forms of non-acute pain. Given the broad spectrum of usage of coniferous essential oils, a fast, safe, simple, and efficient sample-preparation method is needed in the estimation procedure of essential oil components in fresh plant material. Generally, the time- and energy-consuming steam distillation (SD) is applied for this purpose. This paper will compare SD, pressurized liquid extraction (PLE), matrix solid-phase dispersion (MSPD), and the sea sand disruption method (SSDM) as isolation techniques to obtain aroma components from Scots pine (Pinus sylvestris), spruce (Picea abies), and Douglas fir (Pseudotsuga menziesii). According to the obtained data, SSDM is the most efficient sample preparation method in determining the essential oil composition of conifers. Moreover, SSDM requires small organic solvent amounts and a short extraction time, which makes it an advantageous alternative procedure for the routine analysis of coniferous oils. The superiority of SSDM over MSPD efficiency is ascertained, as there are no chemical interactions between the plant cell components and the sand. This fact confirms the reliability and efficacy of SSDM for the analysis of volatile oil components.     

Relevance of the Sea Sand Disruption Method (SSDM) for the Biometrical Differentiation of the Essential‐Oil Composition from Conifers

Sea Sand Disruption Method (SSDM) is a simple and cheap sample‐preparation procedure allowing the reduction of organic solvent consumption, exclusion of sample component degradation, improvement of extraction efficiency and selectivity, and elimination of additional sample clean‐up and pre‐concentration step before chromatographic analysis. This article deals with the possibility of SSDM application for the differentiation of essential‐oils components occurring in the Scots pine (Pinus sylvestris L.) and cypress (Cupressus sempervirens L.) needles from Madrid (Spain), Laganas (Zakhyntos, Greece), Cala Morell (Menorca, Spain), Lublin (Poland), Helsinki (Finland), and Oradea (Romania). The SSDM results are related to the analogous – obtained applying two other sample preparation methods – steam distillation and Pressurized Liquid Extraction (PLE). The results presented established that the total amount and the composition of essential‐oil components revealed by SSDM are equivalent or higher than those obtained by one of the most effective extraction technique, PLE. Moreover, SSDM seems to provide the most representative profile of all essential‐oil components as no heat is applied. Thus, this environmentally friendly method is suggested to be used as the main extraction procedure for the differentiation of essential‐oil components in conifers for scientific and industrial purposes.     

Contribution of microwave accelerated distillation in the extraction of the essential oil of Zygophyllum album L.

Introduction – The aerial parts of Zygophyllum album L. are used in folk medicine as an antidiabetic agent and as a drug active against several pathologies. In this work we present the chemical composition of Algerian essential oils obtained by microwave accelerated distillation (MAD) extraction, a solventless method assisted by microwave. Objective – Under the same analytical conditions and using GC‐FID and GC‐MS, the chemical composition of the essential oil of Zygophyllum album L. extracted by MAD was compared with that achieved using hydrodistillation (HD). Methodology – The extracted compounds were hydrosoluble, and they were removed from the aqueous solution by a liquid extraction with an organic solvent. Results – Employing MAD (100°C, 30 min), the essential oil contained mainly oxygenated monoterpenes with major constituents: carvone and α‐terpineol. However, most of the compounds present in the hydrodistilled volatile fraction were not terpene species, with β‐damascenone as a major constituent. Conclusion – The MAD method appears to be more efficient than HD: after 30 min extraction time, the obtained yields (i.e. 0.002%) were comparable to those provided by HD after 3 h extraction. MAD seems to be more convenient since the volatile fraction is richer in oxygenated monoterpenes, species that are recognised for their olfactory value and their contribution to the fragrance of the essential oil. Copyright © 2010 John Wiley & Sons, Ltd.     

Optimised separation procedures for the simultaneous assay of three plant hormones in liquid biofertilisers

Introduction – The overuse of petrochemical‐based synthetic fertilisers has caused detrimental effects to soil, water supplies, foods and animal health. This, in addition to increased awareness of organic farming, has generated considerable interest in the evaluation of renewable biofertilisers. Objective – The three objectives of the current research were: (1) to evaluate and optimise a solid phase extraction procedure for extraction of three plant hormones, IAA, GA₃ and ABA from two model biofertilisers produced from coconut shells and pineapple peels; (2) to develop an HPLC analysis procedure for the simultaneous separation and quantification of three plant hormones (IAA, GA₃ and ABA); and (3) to evaluate the changes in three plant hormones levels at four different fermentation time periods and varying number of general bacteria, lactic acid bacteria and yeast. Result – An optimised procedure for sample preparation, separation and simultaneous analysis of three plant hormones [indole‐3‐acetic acid (IAA), gibberellic acid (GA₃) and abscisic acid (ABA)] produced in liquid biofertilisers was developed. This method involves sample cleanup using a Sep‐pack Oasis^®MAX cartridge containing mixed‐mode anion‐exchange and reverse‐phase sorbents that provided optimum recovery of 85.6, 91.9 and 94.3%, respectively, for the three hormones, IAA, GA₃, and ABA. Baseline separation of three hormones was achieved using mobile phase consisting of 1% acetic acid and acetonitrile (75:25, v/v) at pH 4.0. The amounts of hormones produced in liquid biofertilisers were influenced by fruit types, fermentation time and total number of general bacteria, lactic acid bacteria and yeasts. The quantities of three plant hormones produced during fermentation correlated well with the total number of microorganisms present in the liquid biofertilisers. Conclusion – A simple and rapid sample preparation procedure followed by RP‐HPLC with UV detection was optimised and developed for simultaneous quantification and identification of three plant hormones namely, IAA, GA₃ and ABA in the liquid biofertilisers. This procedure allows quantification of the three plant hormones in their natural states without any prior derivatisation step. The results presented illustrate that the contents of the three plant hormones depended on the type of fruit wastes, fermentation time and the number of microorganisms found in liquid biofertilisers. This method can be extended to determine the quantity of three hormones in other matrices. This assay procedure will aid in the development of liquid biofertilisers, a valuable alternative fertilisers to promote plant growth. This process will help farmers to reduce production cost and pollution problems. Copyright © 2009 John Wiley & Sons, Ltd.     

An alternative method for irones quantification in iris rhizomes using headspace solid-phase microextraction

Introduction – The essential oil obtained from iris rhizomes is one of the most precious raw materials for the perfume industry. Its fragrance is due to irones that are gradually formed by oxidative degradation of iridals during rhizome ageing. Objective – The development of an alternative method allowing irone quantification in iris rhizomes using HS‐SPME‐GC. Methodology – The development of the method using HS‐SPME‐GC was achieved using the results obtained from a conventional method, i.e. a solid–liquid extraction (SLE) followed by irone quantification by CG. Results – Among several calibration methods tested, internal calibration gave the best results and was the least sensitive to the matrix effect. The proposed method using HS‐SPME‐GC is as accurate and reproducible as the conventional one using SLE. These two methods were used to monitor and compare irone concentrations in iris rhizomes that had been stored for 6 months to 9 years. Conclusion – Irone quantification in iris rhizome can be achieved using HS‐SPME‐GC. This method can thus be used for the quality control of the iris rhizomes. It offers the advantage of combining extraction and analysis with an automated device and thus allows a large number of rhizome batches to be analysed and compared in a limited amount of time. Copyright © 2010 John Wiley & Sons, Ltd.     

Chemical Composition,Enantiomeric Analysis,AEDA Sensorial Evaluation and Antifungal Activity of the Essential Oil from the Ecuadorian Plant Lepechinia mutica Benth (Lamiaceae)

This study describes the GC‐FID, GC/MS, GC‐O, and enantioselective GC analysis of the essential oil hydrodistilled from leaves of Lepechinica mutica (Lamiaceae), collected in Ecuador. GC‐FID and GC/MS analyses allowed the characterization and quantification of 79 components, representing 97.3% of the total sample. Sesquiterpene hydrocarbons (38.50%) and monoterpene hydrocarbons (30.59%) were found to be the most abundant volatiles, while oxygenated sesquiterpenes (16.20%) and oxygenated monoterpenes (2.10%) were the minor components. In order to better characterize the oil aroma, the most important odorants, from the sensorial point of view, were identified by Aroma Extract Dilution Analysis (AEDA) GC‐O. They were α‐Pinene, β‐Phellandrene, and Dauca‐5,8‐diene, exhibiting the characteristic woody, herbaceus, and earthy odors, respectively. Enantioselective GC analysis of L. mutica essential oil revealed the presence of twelve couples and two enantiomerically pure chiral monoterpenoids. Their enantiomeric excesses were from a few percent units to 100%. Moreover, the essential oil exhibited moderate in vitro activity against five fungal strains, being especially effective against M. canis, which is a severe zoophilic dermatophyte causal agent of pet and human infections.     

Determination of Ro 23-7637 in dog plasma by multidimensional ion-exchange—reversed-phase high-performance liquid chromatography with ultraviolet detection

Ro 23-7637 (I) is a new drug under development for the treatment of metabolic diseases. A high-performance liquid chromatographic—ultraviolet detection (HPLC—UV) analytical procedure for its analysis in dog plasma was developed and is reported here. Following C₁₈ solid-phase extraction, the sample is applied to a strong cation-exchange column in the first dimension. The analyte and internal standard, Ro 24-4558 (II), are transferred to a base-deactivated C₁₈ reversed-phase column in the second dimension (orthogonal separation mechanism), with UV detection at 254 nm. The reversed-phase solid-phase extraction provides a gross isolation of the drug, based on hydrophobicity. The first-dimension ion-exchange separation allows neutral species and anions to elute with the column void volume, while separating basic species according to pK_a. The second dimension provides a high-resolution separation dependent upon the hydrophobicity of the sample species. The rationale for using orthogonal multidimensional chromatography was that an exhaustive examination of reversed-phase and normal-phase columns invariably resulted in co-elution of I with endogenous plasma components, which limited the sensitivity of the method. We have utilized C₁₈ solid-phase extraction, followed by multidimensional HPLC—UV, to develop an accurate and precise analytical method whose limit of quantitation, 5 ng/ml using 0.5 ml of plasma, is determined by inherent detector sensitivity. Increased sensitivity can be readily achieved by using more sample or by using microbore HPLC on the second dimension.     

Comprehensive two‐dimensional gas chromatography–mass spectrometry analysis of volatile constituents in Thai vetiver root oils obtained by using different extraction methods

Introduction – Vetiver root oil is known as one of the finest fixatives used in perfumery. This highly complex oil contains more than 200 components, which are mainly sesquiterpene hydrocarbons and their oxygenated derivatives. Since conventional GC‐MS has limitation in terms of separation efficiency, the comprehensive two‐dimensional GC‐MS (GC × GC‐MS) was proposed in this study as an alternative technique for the analysis of vetiver oil constituents. Objective – To evaluate efficiency of the hyphenated GC × GC‐MS technique in terms of separation power and sensitivity prior to identification and quantitation of the volatile constituents in a variety of vetiver root oil samples. Methodology – Dried roots of Vetiveria zizanioides were subjected to extraction using various conditions of four different methods; simultaneous steam distillation, supercritical fluid, microwave‐assisted, and Soxhlet extraction. Volatile components in all vetiver root oil samples were separated and identified by GC‐MS and GC × GC‐MS. The relative contents of volatile constituents in each vetiver oil sample were calculated using the peak volume normalization method. Results – Different techniques of extraction had diverse effects on yield, physical and chemical properties of the vetiver root oils obtained. Overall, 64 volatile constituents were identified by GC‐MS. Among the 245 well‐resolved individual components obtained by GC × GC‐MS, the additional identification of 43 more volatiles was achieved. Conclusion – In comparison with GC‐MS, GC × GC‐MS showed greater ability to differentiate the quality of essential oils obtained from diverse extraction conditions in terms of their volatile compositions and contents. Copyright © 2009 John Wiley & Sons, Ltd.     

Variation in Essential Oil and Bioactive Compounds of Curcuma kwangsiensis Collected from Natural Habitats

The chemical compositions of essential oils (EOs) extracted from Curcuma kwangsiensis rhizomes collected from six natural habitats in P. R. China were evaluated using gas chromatography/mass spectrometry (GC/MS). Fifty‐seven components were identified from the six EOs, and their main constituents were 8,9‐dehydro‐9‐formyl‐cycloisolongifolene (2.37 – 42.59%), germacrone (6.53 – 22.20%), and l ‐camphor (0.19 – 6.12%). The six EOs exhibited different DPPH radical‐scavenging activities (IC₅₀, 2.24 – 31.03 μg/ml), with the activity of most of EOs being much higher than that of Trolox C (IC₅₀, 10.49 μg/ml) and BHT (IC₅₀, 54.13 μg/ml). Most EOs had potent antimicrobial effects against the tested bacteria and fungus. They also exhibited cytotoxicity against B16 (IC₅₀, 4.44 – 147.4 μg/ml) and LNCaP cells (IC₅₀, 73.94 – 429.25 μg/ml). The EOs showed excellent anti‐inflammatory action by significantly downregulating expression of pro‐inflammatory cytokines, cyclooxygenase‐2, and tumor necrosis factor‐α. This study provides insight into the interrelation among growth location, phytoconstituents, and bioactivities, and the results indicate the potential of C. kwangsiensis as natural nutrients, medicines, and others additives.     

Matrix solid-phase dispersion as a valuable tool for extracting contaminants from foodstuffs

This review updates our knowledge on matrix solid-phase dispersion (MSPD), a sample treatment procedure that is increasingly used for extracting/purifying contaminants from a variety of solid, semi-solid, viscous, and liquid foodstuffs. MSPD is primarily used because of its flexibility, selectivity, and the possibility of performing extraction and cleanup in one step, this resulting in drastically shortening of the analysis time and low consumption of toxic and expensive solvents. Technical developments and parameters influencing the extraction yield and selectivity are examined and discussed. Experimental results for the analysis of pesticides, veterinary drugs, persistent environmental chemicals, naturally occurring toxicants, and surfactants in food are reviewed.     

Phytochemical profile of aerial parts and roots of Wachendorfia thyrsiflora L. studied by LC-DAD-SPE-NMR

Hyphenated liquid chromatography – diode array detection – solid phase extraction – nuclear magnetic resonance spectroscopy (LC-DAD-SPE-NMR) was used to investigate the phytochemical composition of aerial parts and roots of Wachendorfia thyrsiflora (Haemodoraceae). Eleven phenylphenalenones and related compounds were identified in the aerial parts of the plant, ten compounds were found in the roots, and four additional compounds occurred in both plant parts. Twelve compounds are previously unreported natural products including five alkaloids (phenylbenzoisoquinolinones) are described here for the first time. In the work presented here, phenylphenalenones with an intact C₁₉ core structure were found only in the roots. Oxa analogs with a C₁₈O scaffold occurred both in the roots and in the aerial plant parts, while most of the aza analogs with a C₁₈N scaffold were detected in the aerial plant parts. This distribution pattern suggests that phenylphenalenones form in the roots, then the intact C₁₉ skeleton is converted into oxa analogs in the roots, translocated into the leaves and further reacted with amines or amino acids to form aza analogs (phenylbenzoisoquinolin-1,6-dione alkaloids).     

Determination of fumonisins in corn: Evaluation of two purification procedures

The performance of two solid phase extraction (SPE) purification procedures, used in the determination of fumonlsin B₁ (FB₁), B₂ (FB₂) and B₃ (FB₂) In corn, was evaluated using both thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC). Fewer interferences were observed In extracts prepared using the strong anion exchange (SAX) media, in contrast to those purified on C₁₈ media, where on occasions, visual discernment of the TLC bands was hampered by the presence of interfering compounds. Precipitate formation, resulting In the blocking of SPE cartridges was also encountered when using the C₁₈ procedure. HPLC analyses of extracts prepared by both media indicated that they gave comparable fumonlsin recoveries from naturally contaminated corn samples. The results suggest that the C₁₈ procedure, originally developed for the TLC analyses of FB₁ in mixed feeds, may also be applied to the determination of FB₂ and FB₂. However, where TLC is used quantitatively for fumonlsin levels <1 μg/g, purification of sample extracts on SAX media is recommended.     

Rapid determination of the anti-cancer drug chlorambucil (Leukeran™) and its phenyl acetic acid mustard metabolite in human serum and plasma by automated solid-phase extraction and liquid chromatography–tandem mass spectrometry

A bioanalytical method for the determination of the anticancer drug chlorambucil (Leukeran™) and its phenyl acetic acid mustard metabolite in human serum and plasma is described. Automated solid-phase extraction of the analytes is carried out with C₁₈ sorbent packed in a 96 well format microtitre plate using a robotic sample processor. The extracts are analysed by isocratic reversed-phase liquid chromatography using pneumatically and thermally assisted electrospray ionisation (TurboIonspray) with selected reaction monitoring. The method is specific and sensitive, with a range of 4–800 ng/ml in human serum and plasma for both parent drug and metabolite (sample volume 200 μl). The method is accurate and precise with intra-assay and inter-assay precision (C.V.) of <15% and bias <15% for both analytes. The automated extraction procedure is significantly faster than manual sample pre-treatment methods, a batch of 96 samples is extracted in 50 min allowing for faster sample turnaround. The method has been used to provide pharmacokinetic support to biocomparability studies of Leukeran™ following single doses of oral tablet formulations.     

Matrix solid phase dispersion (MSPD)

A review of the many uses of matrix solid phase dispersion (MSPD) in the extraction and analysis of a variety of compounds from a range of samples is provided. Matrix solid phase dispersion (MSPD) has found particular application as a somewhat generic analytical process for the preparation, extraction and fractionation of solid, semi-solid and/or highly viscous biological samples. Its simplicity and flexibility contribute to it being chosen over more classical methods for these purposes. MSPD is based on several simple principles of chemistry and physics, involving forces applied to the sample by mechanical blending to produce complete sample disruption and the interactions of the sample matrix with a solid support bonded-phase (SPE) or the surface chemistry of other solid support materials. These principles are discussed as are the factors to be considered in conducting a MSPD extraction.     

Volatile compounds with characteristic odour in moso‐bamboo stems (Phyllostachys pubescens Mazel ex Houz. De ehaie)

Introduction – Moso‐bamboo (Phyllostachys pubescens) is well known as an edible shoot in Asia, and the stems of moso‐bamboo are used as tableware due to its characteristic odour. Despite the pleasant odour of bamboo stems, no detailed analysis of the volatile compounds has been reported. Objective – To clarify the potent odourants contributing to the characteristic aroma of the bamboo, the aroma extract dilution analysis (AEDA) method was performed through gas chromatography olfactometry (GC‐O) analysis. In addition, relative flavour activity (RFA) was calculated, in which both the flavor dilution (FD) factor and weight percentage of each compound are involved. Results – Eighty‐nine compound in bamboo stems oil were identified by GC and GC‐MS. The main components of the oil were palmitic acid (16.5%), (E)‐nerolidol (10.2%) and indole (8.1%). In sensory analysis, 18 aroma‐active compounds were detected by aroma extract dilution analysis (AEDA). The most intense aroma‐active compounds were eugenol (sweet, clove‐like, green) and (E)‐2‐nonenal (green). Conclusion – The results of the sniffing test, RFA and FD factor indicated that (E)‐2‐nonenal and eugenol were estimated to have a bamboo‐like aroma, and aldehyde compounds, such as a phenylacetaldehyde (floral) and C9–C10 unsaturated aldehydes, make the aroma of bamboo. Copyright © 2010 John Wiley & Sons, Ltd.     

Determination and pharmacokinetic study of chiisanogenin in rat plasma by ultra performance liquid chromatography-tandem mass spectrometry

Introduction – Chiisanogenin existing in many Acanthopanax species has been reported to possess anti‐inflammatory, antibacterial and antiplatelet aggregatory activities. Objective – To develop and validate a rapid and sensitive ultra performance liquid chromatography‐tandem mass spectrometry method for the determination of chiisanogenin in rat plasma and to investigate its pharmacokinetics after oral administration of chiisanogenin or the extract of Acanthopanax sessiliflorus fruits. Methodology – The sample pretreatment involved a one‐step extraction of 0.2 mL plasma with diethyl ether. Acetaminophen was used as the internal standard. The separation was carried out on an ACQUITY UPLC? BEH C₁₈ column with a mobile phase of acetonitrile‐5 mM ammonium acetate (90:10, v/v) at a flow rate of 0.2 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode via electrospray ionization (ESI) source. Results – A high sample throughput was achieved with an analysis time of 1.1 min per sample. The calibration curve was linear (r² ≥ 0.99) over the concentration range of 5–500 ng/mL with a lower limit of quantification (LLOQ) of 5 ng/mL. The intra‐day and inter‐day precision (relative standard deviation, R.S.D.) values were below 11% and the accuracy (relative error, R.E.) was within 8% at all three quality control (QC) levels. Conclusion – The method was successfully applied to the pharmacokinetic study of chiisanogenin in rat after oral administration of chiisanogenin and the extract of Acanthopanax sessiliflorus fruits. Other constituents in the extract affected the pharmacokinetic behavior of chiisanogenin. Copyright © 2010 John Wiley & Sons, Ltd.     

Impact of soil types on chemical composition of essential oil of purple basil

Purple basil is among the most important basil varieties and its essential oil is used for several purposes including medicinal and aromatic uses. Soil types may impact the plant growth, development, and essential oil composition. Hence, it is important to find the most suitable soil type which may produce basil plants having essential oil with the best composition and concentration. For this reason, plant samples of purple basil that were grown in areas with clay, loamy sand, and sandy-clay loam soil types were collected and evaluated to determine the changes in the yield and essential oil components. Essential oil contents were determined with the Clevenger Device, and essential oil compositions were determined by using GC and GC/MS analysis. The highest essential oil yield according to soil types was obtained from the plant samples that were grown in the loamy sand soil. It was also found that the main compounds present in Arapgir town purple basil were methylcinnamate and linalool that was also present in all Turkish purple basil under all types of soil. According to the soil types, the highest concentration (46.03%) of methylcinnamate was observed in loamy sand soils, and the lowest (42.33%) was obtained from sandy-clay loam soils and found to be significantly different. Data regarding correlations between soil types and essential oil ratios showed that organic matter and P₂O₅ had a significant negative correlation with methylcinnamate. The present study will help researchers and farmers to choose the most suitable soil type to achieve maximum essential oil production from purple basil.     

Simple high-performance liquid chromatography determination of ampicillin in human serum using solid-phase extraction disk cartridges

A simple and reproducible method for the analysis of ampicillin in human serum was developed. Serum samples were extracted using solid-phase extraction disk cartridges containing a sorbent of styrene divinyl/benzene. Extracts were separated by reversed-phase C₁₈ high-performance liquid chromatography with UV detection at 220 nm. The mobile phase consisted of acetonitrile–10 mM NaH₂PO₄ (6.5:93.5, v/v). Using this extraction procedure, recovery from serum was 98.4±5.6%. The quantitation limit was 0.19 μg/ml using 0.5 ml of serum. The calibration curves from 0.19 to 9.41 μg/ml were linear with correlation coefficients of 0.999. This method is suitable for therapeutic drug monitoring of ampicillin (ABPC) after oral administration of lenampicillin hydrochloride.     

Study on the extraction, purification and quantification of jasmonic acid, abscisic acid and indole-3-acetic acid in plants

Introduction  – Jasmonic acid (JA), abscisic acid (ABA) and indole‐3‐acetic acid (IAA) are important plant hormones. Plant hormones are difficult to analyse because they occur in small concentrations and other substances in the plant interfere with their detection. Objective  – To develop a new, inexpensive procedure for the rapid extraction and purification of IAA, ABA and JA from various plant species. Methodology  – Samples were prepared by extraction of plant tissues with methanol and ethyl acetate. Then the extracts were further purified and enriched with C₁₈ cartridges. The final extracts were derivatised with diazomethane and then measured by GC‐MS. The results of the new methodology were compared with those of the Creelman and Mullet procedure. Results  – Sequential elution of the assimilates from the C₁₈ cartridges revealed that IAA and ABA eluted in 40% methanol, while JA subsequently eluted in 60% methanol. The new plant hormone extraction and purification procedure produced results that were comparable to those obtained with the Creelman and Mullet's procedure. This new procedure requires only 0.5 g leaf samples to quantify these compounds with high reliability and can simultaneously determine the concentrations of the three plant hormones. Conclusion  – A simple, inexpensive method was developed for determining endogenous IAA, ABA and JA concentrations in plant tissue. Copyright © 2008 John Wiley & Sons, Ltd.