高效液相色谱-串联质谱法测定蜂蜜中的5种双稠吡咯啶类生物碱

建立了强阳离子固相萃取-高效液相色谱-串联质谱法用于测定蜂蜜中野百合碱、克氏千里光宁、倒千里光碱、千里光菲啉和千里光宁等5种双稠吡咯啶类生物碱。蜂蜜样品用0.1 mol/L盐酸溶解,强阳离子交换固相萃取柱富集净化后,高效液相色谱-串联质谱进行定性和定量。以Phenomenex C₁₈柱(100 mm×4.6 mm, 2.6 μm)为分析柱,乙腈和0.1%(体积分数)甲酸-5 mmol/L乙酸铵水溶液为流动相进行梯度洗脱,在电喷雾正离子多反应监测模式下进行检测。结果表明,5种双稠吡咯啶类生物碱在1~100 μg/L范围内线性关系良好,线性相关系数均大于0.99。在1、20和50 μg/kg加标水平下,11种不同种类蜂蜜中的5种双稠吡咯啶类生物碱的平均回收率为73.1%~107.1%,相对标准偏差(RSD)为4.1%~17.0%,方法检定量限可达到1.0 μg/kg。该方法准确灵敏,可适用于不同种类进出口蜂蜜中双稠吡咯啶类生物碱的监控分析。利用该方法对来自中国8个省及自治区的洋槐蜜、葵花蜜、棉花蜜、油菜蜜、荆条蜜、枣花蜜、荞麦蜜和来自新西兰、西班牙、澳大利亚等国家的进口蜂蜜进行了筛查。结果发现,野百合碱、克氏千里光宁和倒千里光碱均未检出,而千里光菲啉和千里光宁在大多数蜂蜜中均能检出,千里光菲啉含量在11.0~31.1 μg/kg范围内,千里光宁含量在8.3~29.1 μg/kg范围内。     

Screening for pyrrolizidine alkaloids in plant materials by electron ionization RP-HPLC-MS with thermabeam interface

Plant samples from leaves of Cerinthe minor, Cynoglossum clandestinum, Echium tuberculatum (as well roots), Eritrichium rupestre, Lithospermum purpureo-coerulem, Nonnea lutea, Nonnea setosa, Onosma stellulatum and Cynoglossum amabile were screened for toxic pyrrolizidine alkaloids (PAs) with a newly elaborated procedure comprising gradient HPLC with diode array (DAD) and thermabeam electron impact mass spectrometry (EI-MS). Dried plant material was extracted with boiling 1% tartaric acid in methanol for 2 h on an electric basket and crude extracts purified with cation-exchange solid phase extraction (CE-SPE). Purified extracts containing alkaloids were separated on Zorbax SB RP18 stationary phase in gradient of 0.1% formic acid in methanol. The flow rate was 0.25 mL/min and was suitable both for DAD and EI MS detections. Applied gradient procedure permitted quite sufficient separation of PAs in various plant extracts. On the basis of EI MS spectra, toxic PAs with unsaturated 1,2-double bond in the necine moiety were found in all plant materials and in nine of them (excluding only Cynoglossum amabile) for the fi rst time. They included the following types of structures: 9- and 7-viridifloryl-retronecine monoesters, 9-angeloyl-7-viridifloryl-retronecine, 9-angeloyl-retronecine diester, 9-viridifloryl-retronecine saturated ester, 7-angeloyl-9-viridifloryl-retronecine, 7-angeloyl-9-echimidinyl-retronecine, trachelanthamine and others. Selected ion monitoring (SIM) chromatograms at m/z 119, 120 and 136 together with analysis of UV spectra from DAD detector can be applied in rapid screening for toxic PAs in new plant extracts but to obtain detailed structural information (molecular weight and stereochemistry) more expensive hyphenation is required. Consumption of all analysed plants should be avoided as carcinogenic and hepatotoxic properties of the alkaloids detected are expected.     

QuEChERS method combined to liquid chromatography high-resolution mass spectrometry for the accurate and sensitive simultaneous determination of pyrrolizidine and tropane alkaloids in cereals and spices

Within the last decades, in the EU, there has been an increasing interest in toxic plant alkaloids as food contaminants, especially after the continuous and growing consumption of plant-based foods compared with food of animal origin. In this regard, the once neglected presence of these tropane alkaloids (TAs) and pyrrolizidine alkaloids (PAs) has recently been reconsidered by the European Food Safety Authority, highlighting the lack of data and the need to develop risk assessment strategies. For this reason, the emphasis has been placed on detecting their occurrence in food through the development of accurate and sensitive analytical methods to achieve the determination of these compounds. The present study aims to elaborate and validate an analytical method based on QuEChERS sample preparation approach, exploiting the UHPLC coupled to the HRMS to simultaneously identify and quantify 21 PAs and two TAs in cereals and spices. For TAs, the obtained limit of detection (LOD) is 0.1 μg·kg⁻¹ and the limit of quantification (LOQ) is 0.4 μg·kg⁻¹, while for PAs, the LODs values ranging between 0.2 to 0.3 μg·kg⁻¹ and the LOQ, between 0.4 and 0.8 μg·kg⁻¹, ensuring compliance with the recently established European Regulations. Several commercial samples were analysed to further verify the applicability of this comprehensive analytical approach.     

Screening and identification of metabolites of two kinds of main active ingredients and hepatotoxic pyrrolizidine alkaloids in rat after lavage Farfarae Flos extract by UHPLC‐Q‐TOF‐MS mass spectrometry

Farfarae Flos, the dried flower buds of Tussilago farfara L., is usually used to treat coughs, bronchitic and asthmatic conditions as an important traditional Chinese medicine. Tussilagone and methl butyric acid tussilagin ester are seen as representatives of two kinds of active substances. In addition, the pyrrolizidine alkaloids, mainly senkirkine and senecionine, present in the herb can be hepatoxic. In this study, a rapid and sensitive ultra‐high‐performance liquid chromatography coupled with hybrid triple quadrupole time‐of‐flight mass spectrometry method was successfully applied to identify the metabolites of tussilagone, methl butyric acid tussilagin ester, senkirkine and senecionine. A total of 35, 37, 18 and nine metabolites of tussilagone, methl butyric acid tussilagin ester, senkirkine and senecionine in rats were tentatively identified. Hydrolysis, oxidation, reduction and demethylation were the major metabolic reactions for tussilagone and methl butyric acid tussilagin ester. The main biotransformation routes of senkirkine and senecionine were identified as demethylation, N‐methylation, oxidation and reduction. This study is the first reported analysis and characterization of the metabolites and the proposed metabolic pathways might provide further understanding of the metabolic fate of the chemical constituents after oral administration of Farfarae Flos extract in vivo.     

Supercritical fluid extraction of oxindole alkaloids from Uncaria tormentosa

Supercritical fluid extraction of oxindole alkaloids from Uncaria Tormentosa is described. The extraction was performed with supercritical carbon dioxide alone and with supercritical carbon dioxide modified with 10% metanol, and the extracts were analyzed by GC/MS and HPLC/MS.     

Determination of total retronecine esters‐type hepatotoxic pyrrolizidine alkaloids in plant materials by pre‐column derivatization high‐performance liquid chromatography

A pre‐column derivatization high‐performance liquid chromatography method with diode array detection was developed and validated to determine the total retronecine esters‐type hepatotoxic pyrrolizidine alkaloids (RET‐HPAs) in herbs. The RET‐HPAs reacted with o‐chloranil in methanolic solution heated for 3 h, and an oxidative derivative was produced that could be detected at a maximal absorption of 223 nm. The analysis was performed using a C₁₈ column with an isocratic elution of methanol and aqueous 0.01% triethylamine (adjusted to pH 4 with formic acid), and the detection was carried out with DAD at 223 nm. The validation of the method included linearity, sensitivity, recovery and stability. It showed a good linear regression (r² > 0.9900) in the range of 2.5–250 µm with a limit of detection (S/N = 3) of 0.5 µm . The method provided desirable repeatability with overall intra‐ and inter‐day variations of less than 4.6%. The obtained recoveries for both of the extraction and derivatization process were between 94.6 and 100.7% (n = 3). Copyright © 2009 John Wiley & Sons, Ltd.     

Structural elucidation and NMR assignments of a new pyrrolizidine alkaloid from Crotalaria vitellina Ker Gawl

A new pyrrolizidine alkaloid, named crotavitelin, was isolated from fruits of Crotalaria vitellina, Fabaceae (Papilionoideae). The structure was established by spectroscopic techniques such as one‐dimensional and two‐dimensional NMR, IR, and MS. Copyright © 2013 John Wiley & Sons, Ltd.     

GC‐MS of amaryllidaceous galanthamine‐type alkaloids

Galanthamine‐type alkaloids produced by plants of the Amaryllidaceae family are potent acetylcholinesterase inhibitors. One of them, galanthamine, has been marketed as a hydrobromide salt for the treatment of Alzheimer's disease. In the present work, gas chromatography with electron impact mass spectrometry (GC‐EIMS) fragmentation of 12 reference compounds isolated from various amaryllidaceous plants and identified by spectroscopic methods (1D and 2D nuclear magnetic resonance, circular dichroism, high‐resolution MS (HRMS) and EIMS) was studied by tandem mass spectrometry (GC‐MS/MS) and accurate mass measurements (GC‐HRMS). The studied compounds showed good peak shape and efficient GC separation with a GC‐MS fragmentation pattern similar to that obtained by direct insertion probe. With the exception of galanthamine‐N‐oxide and N‐formylnorgalanthamine, the galanthamine‐type compounds showed abundant [M]^+. and [M‐H]⁺ ions. A typical fragmentation pattern was also observed, depending on the substituents of the skeleton. Based on the fragmentation pathways of reference compounds, three other galanthamine‐type alkaloids, including 3‐O‐(2′‐butenoyl)sanguinine, which possesses a previously unelucidated structure, were identified in Leucojum aestivum ssp. pulchelum, a species endemic to the Balearic islands. GC‐MS can be successfully applied to Amaryllidaceae plant samples in the routine screening for potentially new or known bioactive molecules, chemotaxonomy, biodiversity and identification of impurities in pharmaceutical substances. Copyright © 2012 John Wiley & Sons, Ltd.     

Mass spectrometry-guided isolation of two new benzoquinoline alkaloids from Macleaya cordata

Two new alkaloids, named 2,3-methylenedioxy-7,10-dimethyl-7,8,9,10-tetrahydro-benzoquinoline (1) and 2,3-methylenedioxy-7,10-dimethyl-8-carboxyl-benzoquinoline (2), were detected primarily from the fruits of Macleaya cordata by their different fragmentation pathways. And then isolation of the two compounds was performed by column chromatography and preparative HPLC under the guiding of mass spectrometry. Finally, their structures were determined by spectroscopic analysis.     

Molecularly imprinted polymer solid‐phase microextraction coupled with ultra high performance liquid chromatography and tandem mass spectrometry for rapid analysis of pyrrolizidine alkaloids in herbal medicine

Pyrrolizidine alkaloids are the most widely distributed natural toxins, and pyrrolizidine alkaloid‐containing herbal medicines are probably the most common poisonous plants affecting humans. We reported pyrrolizidine alkaloid‐molecularly imprinted polymer solid‐phase microextraction for the selective adsorption of toxic pyrrolizidine alkaloids from herbal medicine. A sulfonic compound, sodium allylsulfonate, was chosen as the functional monomer to interact with pyrrolizidine alkaloids through strong ionic interaction. To avoid template leakage and for the aim of cost saving, a relatively cheap dummy template was used for the fabrication of molecularly imprinted polymer‐solid‐phase microextraction fibers. The obtained fibers showed selective adsorption ability for four pyrrolizidine alkaloids, including europine, echimidine, lasiocarpine, and heliotrine. The extraction parameters, such as extraction time, extraction temperature, shaking speed, elution solvent and elution time, were optimized. Then ultra high performance liquid chromatography with mass spectrometry coupled with molecularly imprinted polymer‐solid‐phase microextraction method was developed for the fast and efficient analysis of four pyrrolizidine alkaloids from the model herbal plant Farfarae Flos. The established method was validated and exhibited satisfactory accuracy and precision. The present method provides an innovative and fast analytical strategy for the determination of trace toxic pyrrolizidine alkaloids in complicated samples.     

Characteristic ion clusters as determinants for the identification of pyrrolizidine alkaloid N‐oxides in pyrrolizidine alkaloid–containing natural products using HPLC–MS analysis

Pyrrolizidine alkaloid (PA)–containing plants are widely distributed in the world. PAs are hepatotoxic, affecting livestock and humans. PA N‐oxides are often present together with PAs in plants and also exhibit hepatotoxicity but with less potency. HPLC–MS is generally used to analyze PA‐containing herbs, although PA references are unavailable in most cases. However, to date, without reference standards, HPLC–MS methodology cannot distinguish PA N‐oxides from PAs because they both produce the same characteristic ions in mass spectra. In the present study, the mass spectra of 10 PA N‐oxides and the corresponding PAs were systemically investigated using HPLC–MS to define the characteristic mass fragment ions specific to PAs and PA N‐oxides. Mass spectra of toxic retronecine‐type PA N‐oxides exhibited two characteristic ion clusters at m/z 118–120 and 136–138. These ion clusters were produced by three unique fragmentation pathways of PA N‐oxides and were not found in their corresponding PAs. Similarly, the nontoxic platynecine‐type PA N‐oxides also fragmented via three similar pathways to form two characteristic ion clusters at m/z 120–122 and 138–140. Further application of using these characteristic ion clusters allowed successful and rapid identification of PAs and PA N‐oxides in two PA‐containing herbal plants. Our results demonstrated, for the first time, that these characteristic ion clusters are unique determinants to discriminate PA N‐oxides from PAs even without the availability of reference samples. Our findings provide a novel and specific method to differentiate PA N‐oxides from PAs in PA‐containing natural products, which is crucial for the assessment of their intoxication. Copyright © 2012 John Wiley & Sons, Ltd.     

Qualitative and quantitative analysis of pyrrolizidine alkaloids for the entire process quality control from Senecio scandens to Senecio scandens‐containing preparations by high performance liquid chromatography‐tandem mass spectrometry

Senecio scandens as a commonly used traditional Chinese medicine that is used alone or in combination with other herbs in preparations such as QianBai BiYan tablets has attracted much attention because of its hepatotoxic pyrrolizidine alkaloids. Nowadays, most studies for pyrrolizidine alkaloids are only performed on herbs or a preparation, however, production of preparations is a dynamic process, control of toxic impurities for raw materials, or finished products cannot monitor the production process dynamically. Thus, in this study, qualitative and quantitative analysis of pyrrolizidine alkaloids for the entire process quality control from S. scandens to its preparations was carried out with HPLC‐MS/MS for the first time, which was more comprehensive and dynamic than the previous single‐layer analysis. First, the species of pyrrolizidine alkaloids in S. scandens were analyzed, and the characteristic fragmentation rules of pyrrolizidine alkaloids containing common parent nucleus were found, which can be used to identify these components rapidly in the future. Then, a quantitative method for S. scandens to QianBai BiYan tablets and other nine S. scandens‐containing preparations was established, and after the medication safety speculation, all of them met the relevant safety requirements. After that, in order to ensure the stability and controllable of drug quality, the limit of pyrrolizidine alkaloids in preparations was determined according to the safe dosage that is stipulated to be the same as raw materials. Finally, the factors causing the content change of pyrrolizidine alkaloids in S. scandens from different source were studies, which can provide theoretical basis for selecting suitable raw materials for production.     

Chemistry of pyrrolizidine alkaloids revisited—semi-synthetic microwave and continuous-flow approaches toward Crotalaria-alkaloids

Efficient and rapid syntheses of various pyrrolizidine alkaloids via microwave synthetic protocols using monocrotaline from a natural source (Crotalaria spectabilis) are reported. Retronecine, hydroxydanaidal, and acetylated hydroxydanaidal (utilized in the chemical communication system of various insect species) were obtained. In addition, a continuous flow hydrogenolysis/hydrogenation approach to directly access platynecine, retronecanol, and desoxyretronecine was realized.     

A rapid cleanup method for the isolation and concentration of pyrrolizidine alkaloids in comfrey root

Preparations from comfrey (Symphytum officinale and S. x uplandicum) root and leaf contain varying levels of the hepatotoxic pyrrolizidine alkaloids (PAs). Reference compounds for comfrey are not commercially available, and there is currently no rapid extraction or analytical method capable of determining low levels in raw materials or as adulterants in commercially available extracts. A solid-phase extraction (SPE) method was developed using an Ergosil cleanup column that specifically binds the PAs. With this method, powdered comfrey root was extracted by sonication and shaking with basic chloroform. The extract was applied to the cleanup column under vacuum, washed with 2 mL acetone-chloroform (8 + 2, v/v) followed by 2 mL petroleum ether to remove excess chloroform. The column was dried under vacuum, and the PAs were eluted with 2 successive 1 mL aliquots methanol. Percent recoveries of the PAs following Ergosil SPE had an overall average of 96.8%, with RSD of 3.8% over a range of 1.0 to 25.0 g extracted in 100 mL. Average precision of the method (n = 3 over 4 extraction concentrations) gave an overall RSD of 6.0% for the 5 alkaloids, with a range of 0.8% (5 g in 100 mL) to 11.2% (25 g in 100 mL). Recovery optimization testing showed that 1.0 g comfrey root extracted in 100 mL yielded the greatest recovery (% dry weight) of the PAs, with an extraction efficiency and accuracy of 94.2%, and RSD of 1.7% (n = 9). The unique properties of the Ergosil cleanup column provide rapid sample cleanup, volume reduction, and concentration of PAs from comfrey extracts, and allow the eluant to be analyzed directly by traditional chromatographic methods.     

Dynamic pH junction-sweeping capillary electrophoresis for online preconcentration of toxic pyrrolizidine alkaloids in Chinese herbal medicine

There is a need to develop simple yet effective preconcentration methods to enhance concentration sensitivity for CE analysis of trace level analytes in real samples, particularly when commonly available but less sensitive detection methods, e.g., UV detection, are used. In this report, a hyphenated online preconcentration strategy combining dynamic pH junction with sweeping (i.e., dynamic pH junction-sweeping) was employed for the analysis of four toxic pyrrolizidine alkaloids (PAs) of senkirkine, senecionine, retrorsine, and seneciphylline in Chinese herbal medicine (Kuan donghua). Direct electrokinetically focusing of a large sample volume injection (up to 20% of capillary length) on the capillary was performed using the dynamic pH junction-sweeping method. A sample matrix consisting of 10 mM phosphate with 20% methanol at pH 4.0 and a BGE containing 20 mM borate, 30 mM SDS, and 20% methanol at pH 9.1 were utilized to realize dynamic pH junction-sweeping for PAs. This online preconcentration strategy resulted in sensitivity enhancement factors ranging from 23.8- to 90.0-fold for the four toxic PAs, giving an LOD as low as 30 ppb for the PAs. Critical factors such as sample matrix type, pH, and salt concentration were also examined to achieve higher sensitivity enhancement, shorter analysis time, and better resolution. The results indicate that the proposed dynamic pH junction-sweeping technique is a powerful alternative approach for identification and determination of trace levels of these toxic PAs and other hydrophobic, protonatable compounds in real samples.     

Characterization of secondary metabolites of leaf and stem essential oils of Achillea fragrantissima from central region of Saudi Arabia

In this paper, a detailed study on chemical characterization of essential oils (EOs) constituents of leaves and stems of Achillea fragrantissima were carried out using GC-FID and GC–MS analysis employing two different stationary phase columns. In the studied plant which is collected from the central region of Saudi Arabia, trans-sabinyl acetate and trans-sabinol have been identified as the major components. To the best of our knowledge, these components are being reported for the first time as the major constituents in the EOs of A. fragrantissima. The results showed that chief chemical components of both (leaves and stems) oils were found to be almost same, however, their contents varied noticeably from each other. Among 108 identified components in the leaves oil, the major components were trans-sabinyl acetate (20.7 ± 0.00), trans-sabinol (14.9 ± 0.13), artemisia ketone (12.7 ± 0.46), santolina alcohol (10.1 ± 1.30), β-sesquiphellandrene (5.5 ± 0.01), β-thujone (5.1 ± 0.11). Whereas, in the stem oil 85 components were identified and trans-sabinyl acetate (24.0 ± 0.19), trans-sabinol (19.2 ± 0.01), artemisia ketone (16.3 ± 0.74), santolina alcohol (10.4 ± 1.50), and β-sesquiphellandrene (4.8 ± 0.01) were found to be the major components. Among the identified components form both oils, 23 components were specific to only leaves oil, whereas 85 components were found to be common in both oils.     

Optimization of solid-phase microextraction conditions for the determination of triclosan and possible related compounds in water samples

A solid-phase microextraction (SPME) method for the determination of triclosan, methyl triclosan, 2,4-dichlorophenol and 2,3,4-trichlorophenol (considered as possible triclosan metabolites) in water samples was optimised. Analytes were first concentrated on a SPME fibre, directly exposed to the sample, and then triclosan and the two chlorinated phenols on-fibre silylated using N-methyl-N-(tert.-butyldimethylsilyl)-trifluoroacetamide (MTBSTFA). Methyl triclosan remained unaffected during the derivatization step. Compounds were determined using gas chromatography in combination with mass spectrometry (GC-MS). Influence of different factors on the efficiency of extraction and derivatization steps was systematically investigated. Using a polyacrylate (PA) fibre quantification limits below 10 ng/l, and acceptable relative standard deviations, were obtained for all compounds after an extraction time of 30 min. On-fibre silylation was carried out in only 10 min. Moreover, the efficiency of the procedure was scarcely affected by the type of water sample. The method was applied to several samples of treated and raw wastewater, triclosan was found in all samples, at concentrations from 120 to 14,000 ng/l, and 2,4-dichlorophenol in most of them, at levels up to 2222 ng/l.     

Volatiles of Helichrysum italicum (Roth) G. Don from Croatia

Helichrysum italicum (Roth) G. Don is a flowering plant of the family Asteraceae. It is rich in oil that is used for different medicinal purposes and in fragrance industry. Volatile profile of four populations of H. italicum, collected from natural habitat in Dalmatia (Croatia), was analysed by capillary GC–MS. Sample from Bra? Island had α-trans-bergamotene (10.2%) and β-acoradiene (10.1%) as the majors, whereas sample collected on Biokovo Mt. was rich in neryl acetate (8.1%). β-Acoradiene was also the main constituent of sample collected near Tijarica, whereas rosifoliol (8.5%) was the most abundant constituent in sample collected near Makarska. Presented results show the influence of environmental conditions on chemical differentiation of the volatiles of H. italicum from Croatia.     

Detection and validated quantification of toxic alkaloids in human blood plasma--comparison of LC-APCI-MS with LC-ESI-MS/MS

Poisonings with toxic plants may occur after abuse, intentional or accidental ingestion of plants. For diagnosis of such poisonings, multianalyte procedures were developed for detection and validated quantification of the toxic alkaloids aconitine, atropine, colchicine, coniine, cytisine, nicotine and its metabolite cotinine, physostigmine, and scopolamine in plasma using LC-APCI-MS and LC-ESI-MS/MS. After mixed-mode solid-phase extraction of 1 ml of plasma, the analytes were separated using a C8 base select separation column and gradient elution (acetonitrile/ammonium formate, pH 3.5). Calibration curves were used for quantification with cotinine-d(3), benzoylecgonine-d(3), and trimipramine-d(3) as internal standards. The method was validated according to international guidelines. Both assays were selective for the tested compounds. No instability was observed after repeated freezing and thawing or in processed samples. The assays were linear for coniine, cytisine, nicotine and its metabolite cotinine, from 50 to 1000 ng/ml using LC-APCI-MS and 1 to 1000 ng/ml using LC-ESI-MS/MS, respectively, and for aconitine, atropine, colchicine, physostigmine, and scopolamine from 5 to 100 ng/ml for LC-APCI-MS and 0.1 to 100 ng/ml for LC-ESI-MS/MS, respectively. Accuracy ranged from -38.6 to 14.0%, repeatability from 2.5 to 13.5%, and intermediate precision from 4.8 to 13.5% using LC-APCI-MS and from -38.3 to 8.3% for accuracy, from 3.5 to 13.8%, for repeatability, and from 4.3 to 14.7% for intermediate precision using LC-ESI-MS/MS. The lower limit of quantification was fixed at the lowest calibrator in the linearity experiments. With the exception of the greater sensitivity and higher identification power, LC-ESI-MS/MS had no major advantages over LC-APCI-MS. Both presented assays were applicable for sensitive detection of all studied analytes and for accurate and precise quantification, with the exception of the rather volatile nicotine. The applicability of the assays was demonstrated by analysis of plasma samples from suspected poisoning cases.     

Binding behaviour and solubilisation of p-sulfonatocalixarenes to cinchona alkaloids

In this study, we investigated the binding behaviours of three water-soluble p-sulfonatocalixarenes with four cinchona alkaloids in aqueous and phosphate buffer solutions (pH 7.2 and 2.0). The complexation stability constants obtained by fluorescence titrations were comparatively discussed from several aspects: host cavity, pH effect and ionic strength. Among three hosts, p-sulfonatocalix[4]arene (SC4A) forms the most stable complexes with cinchona alkaloids, especially in acidic aqueous conditions. Furthermore, SC4A was elected as model drug carrier for cinchona alkaloids, where solubilisation by the complexation of SC4A and mimic release from the calixarene cavity in the presence of negatively charged micelles were initially studied.