Plant extracts sample preparation

In the TLC analysis of dry extracts prepared from medicinal plants, the sample preparation is performed in a different way from that prescribed in the monographs for the drugs in the pharmacopoeias. Also, there is no binder in the recommended solvent system, and in these cases a validation of the new in-house method is certainly necessary. 

Analysis of methyl parathion in sediments, soils, foods, and plant and animal tissues poses problems with extraction from the sample matrix, cleanup of samples, and selective detection. Sediments and soils have been analyzed primarily by GC/ECD or GC/FPD. Food, plant, and animal tissues have been analyzed primarily by GC/thermionic detector or GC/FPD, the recommended methods of the Association of Official Analytical Chemists (AOAC). Various extraction and cleanup methods (AOAC 1984 Belisle and Swineford 1988 Capriel et al. 1986 Kadoum 1968) and separation and detection techniques (Alak and Vo-Dinh 1987 Betowski and Jones 1988 Clark et al. 1985 Gillespie and Walters 1986 Koen and Huber 1970 Stan 1989 Stan and Mrowetz 1983 Udaya and Nanda 1981) have been used in an attempt to simplify sample preparation and improve sensitivity, reliability, and selectivity. A detection limit in the low-ppb range and recoveries of 100% were achieved in soil and plant and animal tissue by Kadoum (1968). GC/ECD analysis following extraction, cleanup, and partitioning with a hexane-acetonitrile system was used. 

Preparative planar chromatography is a very important step in the complicated procedures of isolation of group of compounds or pure substances from complex matrices. The method gives additional possibilities of using various adsorbents and eluent systems to achieve complete separation of stracmral analogs. The method also enables combining the various methods of sample application, plate development, and derivatization to achieve satisfactory separation of isolated plant extracts components. 

Sample preparation techniques vary depending on the analyte and the matrix. An advantage of immunoassays is that less sample preparation is often needed prior to analysis. Because the ELISA is conducted in an aqueous system, aqueous samples such as groundwater may be analyzed directly in the immunoassay or following dilution in a buffer solution. For soil, plant material or complex water samples (e.g., sewage effluent), the analyte must be extracted from the matrix. The extraction method must meet performance criteria such as recovery, reproducibility and ruggedness, and ultimately the analyte must be in a solution that is aqueous or in a water-miscible solvent. For chemical analytes such as pesticides, a simple extraction with methanol may be suitable. At the other extreme, multiple extractions, column cleanup and finally solvent exchange may be necessary to extract the analyte into a solution that is free of matrix interference. 

Because of their polymeric forms, alkylenebis(dithiocarbamates) are insoluble in water and most organic solvents. Additionally, they form strong complexes with different metal ions No extraction and chromatographic procedure has been reported for the parent compound of this chemical class. These compounds decompose readily under acidic conditions, for example by contact with the fruit or plant juice generated during sample preparation. 

An internal standard method gives more reliable results when elaborate sample preparation is required, as in extraction of a drug substance from biological fluids, or extraction of pesticides and herbicides from soil and plant matter. The addition of internal standard (IS) to the sample and standard acts as a marker to give accurate values of the recovery of the desired compound(s). Since the determination of wt% involves the ratio of the detector responses in the two chromatograms, the injection volume is not critical as in an external standard method. 

These samples are prepared by either wet or dry ashing. Many of the metals can be determined in aqueous solution, but for the more trace ones, solvent extraction procedures similar to those described above are resorted to. Similar sample preparation procedures apply to plants. The elements... 

Plant extraction has been carried out under the auspices of BGVS, the "in-country" member of the group. To date, a total of 3352 extracts have been prepared from 2053 plant samples (two extracts per sample). The extraction procedure used has been to first make an... 

The ultrasound-assisted extraction of freeze-dried plant-based materials normally takes 2 hr. If the extract is evaporated to dryness, a total of 3 hr is necessary. As an additional sample preparation step, 2 to 4 days should be allotted for freeze-drying fresh plant materials, depending on the quantity of the material. Homogenizer-assisted extraction of fresh fruit takes <4 hr. 

This unit describes procedures for extraction, purification, and identification by MALDI-MS of fiavonol glycosides from a plant source. The extraction and purification protocols are not meant to be comprehensive, but rather to offer guidelines for sample preparation prior to a MALDI-MS analysis. The MALDI-MS technique is suggested as a complement to other analytical methods such as HPLC or NMR. Its strength lies in the ability to rapidly screen a number of samples for the presence of fiavonol glycosides, which can be identified on the basis of their molecular weights. 

The robustness of a sample preparation technique is characterized by the reliability of the instrumentation used and the variability (precision) of the information obtained in the subsequent sample analysis. Thus, variations in controlled parameters and sequences are to be avoided. In sample preparation methods employing supercritical fluids as the extracting solvents, it has been our experience that minimal variations in efficient analyte recoveries are possible using a fully automated extraction system. The extraction solvent operating parameters under automated control are temperature, pressure (thus density), composition and flow rate through the sample. The precision of the technique will be discussed by presenting replicability, repeatability, and reproducibility data for the extraction of various analytes from such matrices as sands and soils, river sediment, and plant and animal tissue. Censored data will be presented as an indicator of instrumental reliability. 

Direct analysis of the enantiomers in biological fluids is very important because it reduces both analysis time and sample preparation time. Indeed, when there is risk of the quick racemization of the enantiomer, direct analysis is essential. It has been observed that CD-based CSPs employ mobile phases that are generally compatible with biological samples, hence can be used for the direct analysis of the enantiomers in biological fluids [67,80]. Stalcup et al. [58] employed coupled column chromatography to isolate scopolamine from a plant extract and found that the extent of racemization depends on the isolation... 

Usually there are two main components deemed responsible for the decrease in extraction efficiency, that is the lipid content and the cell wall in the case of samples of plant or fungal origin that may remain partly intact during sample pretreatment. This fact prompted sample preparation including also cell wall degrading and/or lipolytic enzymes, as reported soon after the initiation of the second phase of enzymatic sample preparation techniques in the publication by Gilon et al. [79]. 

In what many consider to be a landmark publication on metabolomics, Fiehn et al. (2000) state it is crucial to perform unbiased (metabolite) analyses in order to define precisely the biochemical function of plant metabolism. The authors argue that for metabolomics/metabolite profiling to become a robust and sensitive method suited to automation, a mature technology such as gas chromatography-mass spectrometry (GC-MS) is required as an analytical technique. The authors go on to describe a simple sample preparation and analysis regime that allowed for the detection and quantification of more than 300 compounds from a single-leaf sample extract. 

Fahmy, T.M., Pulaitis, M.E., Johnson, D.M., McNally, M.E.P., Modifier effects in the supercritical fluid extraction of solutes from clay, soil, and plant materials. Anal. Chem., 65 (10), 1462-1469,1993. Langenfeld, J.J., Hawthorne, S.B., Miller, D.J., Pawliszyn, J., Role of modifiers for analytical scale supercritical fluid extraction of environmental samples. Anal. Chem., 66(6), 909-916,1994. Hawthorne, S.B., Methodology for off-line supercritical fluid extraction. In Supercritical Fluid Extraction and Its Use in Chromatographic Sample Preparation, Westwood S.A. (Ed.), Blackie Academic and Professional, 39-64, 1993. 

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