Centrifugal partition chromatography (CPC) coupled online with high-performance liquid chromatography (HPLC) with diode-array detection (DAD) and mass spectrometry (MS) is presented in this work. This strategy offers the possibility to obtain simultaneously CPC fractionation of natural extracts, the HPLC fingerprint of separated fractions and structural information on molecules contained in each fraction. This new approach was applied to the fractionation and purification of xanthones from Garcinia mangostana (Clusiaceae) pericarp. A biphasic solvent system of heptane/ethyl acetate/methanol/water (2:1:2:1, v/v) was used for the CPC separation of 175?mg crude ethanolic extract. The HPLC analysis was conducted with a reversed-phase monolithic column allowing fast and repeatable separation. This combined CPC-HPLC-DAD-MS method led to isolation of 33?mg α-mangostin and 6?mg γ-mangostin at 98?% and 98.5?% purity, respectively, in 140?min. Furthermore, in the same time a total of 16 other xanthones were detected in the extract, and ten of them were identified on the basis of their UV and MS spectra. 相似文献
The robustness study of the reversed-phase liquid chromatographic method developed for the quantitative analysis of carboxylic acids is a real asset to prepare method transfer because it provides an indication of its reliability during routine use. Indeed, it was possible to predict the consequences of small variations in operating conditions on the responses. The design of experiments approach was applied to model the effects and interactions of a high number of factors varying simultaneously with a limited number of runs. First we identified the factors which potentially affect the chromatographic responses used for carboxylic acids quantitation: detection wavelength (λ), column temperature (T), acetonitrile ratio in mobile phase (Me), duration of the plateau before the gradient (L) and gradient slope (S). Then we estimated the order of magnitude of realistic variations to assign factor levels. Finally a central composite design was carried out around the nominal conditions defined during method optimization. The statistical treatment of responses (retention factors, and concentrations) showed that the column temperature, the acetonitrile ratio in the mobile phase, the duration of the plateau before the gradient and the gradient slope were the most influent factors. The building of the robust domain from response-surfaces allowed us to give tolerance limits for the factors (216 nm < λ < 222 nm, 49.3 °C < T < 51.4 °C, 4.90% < Me < 5.18%, v/v, 4.5 min < L < 5.4 min, 9% < S < 11%) for which the performances of the method were maintained. 相似文献
A simple and reliable method based on capillary electrophoresis with electrochemical detection (CE–ED) was applied to study the effect of aerobic exercises on creatinine and uric acid concertration in saliva and urine. The pH value, the running buffer concentration, the SDS concentration, separation voltage, injection time and the potential applied to the working electrode were investigated to find the optimum conditions. The detection limits (S/N = 3) for creatinine and uric acid were 3.6 μmol L−1 and 0.86 μmol L−1, respectively. This method was successfully used in the rapid analysis of creatinine and uric acid in saliva samples. After aerobic exercises, creatinine concentration decreased, and uric acid concentration increased in saliva. In urine, the concentrations of creatinine and uric acid both increased after exercise.
Xanthones are well known for their interesting phytochemical properties, which make them attractive to the pharmaceutical and medicinal industry. We have therefore developed a method to analyse the major xanthones in Garcina mangostana. The xanthones were extracted by pressurized liquid extraction with ethanol and separated at the semi-preparative scale by centrifugal partition chromatography (CPC) with a biphasic solvent system consisting of heptane/ethyl acetate/methanol/water (2:1:2:1, v/v/v/v). A CPC-electrospray ionisation MS coupling was performed and used to simultaneously separate and identify the compounds. Thanks to a variable flow splitter and an additional stream of ethanol/1 mol L−1 ammonium acetate (95:5, v/v), all the compounds were ionised, detected and monitored whatever the solvents used in mobile phase for the CPC separation. The dual mode or elution–extrusion which are less solvent-consuming and faster than the elution mode were used without loss of ionisation and detection. 相似文献
Complexation of aluminium(III) with the fluorogenic ligand chromotropic acid (4,5-dihydroxynaphthalene-2,7-disulfonic acid) has been revisited with the aim of using enhancement of the fluorescence intensity as an analytical tool. Complexation at the optimum pH4 was shown to lead to a 1:1 complex with a stability constant log110=18.4±0.7. The fluorogenic effect was thoroughly investigated. Nearly selective excitation of the chelate rather than the ligand could be achieved at wavelengths longer than 360 nm. For analytical purposes the main interfering ion was Ga3+. The strongest competing ligand was shown to be citric acid. Competitive complexation by acetate or formate ions can also make their use in a buffer at the usual concentration, 0.2 mol L–1, questionable, whereas a 10–2 mol L–1 formic acid buffer was shown to be a good alternative. The calibration plot showed that the dependence of response on Al(III) concentration was linear up to 500 g L–1; the detection limit was 0.65 g L–1 (3SDblank, n=10, SD=±1.4% at 10 g L–1 and ±0.8% at 100 g L–1). The analytical procedure was successfully applied to several samples of tap water and the results were in good agreement with those from AAS determination. 相似文献
A microfabricated device has been developed for fluorimetric detection of potassium ions without previous separation. It is
based on use of a fluorescent molecular sensor, calix–bodipy, specially designed to be sensitive to and selective for the
target ion. The device is essentially made of a Y-shape microchannel moulded in PDMS fixed on a glass substrate. A passive
mixer is used for mixing the reactant and the analyte. The optical detection arrangement uses two optical fibres, one for
excitation by a light-emitting diode, the other for collection of the fluorescence. This system enabled the flow-injection
analysis of the concentration of potassium ions in aqueous solutions with a detection limit of 0.5 mmol L−1 and without interference with sodium ions. A calibration plot was constructed using potassium standard solutions in the range
0–16 mmol L−1, and was used for the determination of the potassium content of a pharmaceutical pill.
Figure Photography of the microfluidic channel showing the ridges in the PDMS substrate at the top of the channel 相似文献
Centrifugal Partition Chromatography (CPC), a liquid–liquid preparative chromatography using two immiscible solvent systems, benefits from numerous advantages for the separation or purification of synthetic or natural products. This study presents the on-line hyphenation of CPC-Evaporative Light Scattering Detector (CPC-ELSD) with High Performance Liquid Chromatography-UV (HPLC-UV) for the fractionation of flavonols from a solvent-free microwave extract of sea buckthorn (Hippophaë rhamnoides L., Elaeagnaceae) berries. An Arizona G system was used for the fractionation of flavonoids by CPC and a fused core Halo C18 column allowed the on-line analyses of collected fractions by HPLC. The on-line CPC/HPLC procedure allowed the simultaneous fractionation step at preparative scale combined with the HPLC analyses which provide direct fingerprint of collected fractions. Thus the crude extract was simplified and immediate information on the composition of fractions could be obtained. Furthermore, this methodology reduced the time of post-fractionation steps and facilitated identification of main molecules by Mass Spectrometry (MS). Rutin, isorhamnetin-3-O-rutinoside, isorhamnetin-3-O-glucoside, quercetin-3-O-glucoside, isorhamnetin-rhamnoside, quercetin and isorhamnetin were identified. CPC-ELSD/HPLC-UV could be considered as a high-throughput technique for the guided fractionation of bioactive natural products from complex crude extracts. 相似文献
Countercurrent chromatography (CCC) is an attractive separation method because the analytes are partitioned between two immiscible liquid phases avoiding problems related to solid stationary phase. In recent years, this technique has made great progress in separation power and detection potential. This review describes coupling strategies involving high speed CCC (HSCCC) or centrifugal partition chromatography (CPC). It includes on-line extraction–isolation, hyphenation with mass spectrometry (MS) and nuclear magnetic resonance (NMR) detectors, multidimensional CCC (MDCCC), two-dimensional CCC (2D-CCC), on-line coupling with liquid chromatography (LC), and biological tests, and innovative off-line developments. The basic principles of each method are presented and applications are summarized. 相似文献