Comparative studies on the camptothecin content from Nothapodytes foetida and Ophiorrhiza species

Camptothecin is an anticancer quinoline alkaloid effective against colon cancer. It acts by inhibition of the enzyme DNA topoisomerase I. A comparative study of camptothecin from the indigenous plants namely Nothapodytes foetida, Ophiorrhiza mungos and Ophiorrhiza rugosa indicated highest yields of camptothecin and 9-methoxy camptothecin in N. foetida. The other two plants O. mungos and O. rugosa contained low levels of alkaloids.     

Separation of 9-methoxycamptothecin and camptothecin from Nothapodytes foetida by semipreparative HPLC

The present work describes the isolation of camptothecin and 9-methoxycamptothecin from the aerial parts of Nothapodytes foetida by semipreparative high-performance liquid chromatography because the separation of compounds by conventional procedures is tedious and cumbersome. The purity of the isolates is determined by physicochemical data and liquid chromatography-mass spectrometry.     

Quantitative analysis of camptothecin derivatives in Nothapodytes foetida using 1H-NMR method

A quantitative analysis using (1)H-NMR has been developed for the determination of camptothecin derivatives and trigonelline in Nothapodytes foetida root, stems and leaves. In the region of delta 9.5-5.5, the signals of H-7 of camptothecin (1), H-10 of 9-methoxycamptothecin (2), H-19 of pumiloside (3) and H-2 of trigonelline (4), were well separated from each other in DMSO-d(6). The quantity of the compounds was calculated by the ratio of the intensity of each compound to the known amount of internal standard 3,4,5-trimethoxybenzaldehyde. These results were compared with the conventional HPLC method. The advantages of the method are that no reference compounds are required for calibration curves, the quantification could be directly realized on a crude extract, an overall profile of the preparation could be directly obtained, and a very significant time-gain could be achieved, in comparison to conventional HPLC methods, for instance.     

Camptothecinoids from the seeds of Taiwanese Nothapodytes foetida

Two new alkaloids, 9-methoxy-18,19-dehydrocamptothecin (1) and 5- hydroxymappicine-20-O-beta-glucopyranoside (2a/2b as a racemic mixture), together with nine known compounds: camptothecin (3), 9-methoxy-camptothecin (4), 5-hydroxycamptothecin (5a/5b racemic mixture), 5-hydroxy-9-methoxycamptothecin (6a/6b racemic mixture), diosmetin (7), apigenin (8), apigenin-7-O-glucopyranoside (9), rosin (cinnamyl-O-beta-D-glucopyranoside) (10) and amarantholidoside IV (11) were isolated from the immature seeds of Nothapodytes foetida (Wight) Sleumer. The structures were elucidated by spectroscopic analyses. In the present research, compounds 1, 3, 4, 5a/5b and 6a/6b, also showed in vitro cytotoxicity against six cancer cell lines (HepG2, Hep3B, MDA-MB- 231, MCF-7, A549, and Ca9-22). Among them, compound 1 exhibited significant cytotoxicity against these cancer cell lines, with IC(50) of 0.24-6.57 microM. Furthermore, HPLC profiles were developed for qualitative and quantitative analysis of these active constituents in different parts of this plant, including mature and immature seeds, leaves, stems and roots. The results revealed that compounds 3 and 4 have the highest concentrations, which are found in the roots part of the plant.     

Comparison of extraction techniques for extraction of bioactive molecules from Hypericum perforatum L. plant

Three methods commonly used for the extraction of bioactive molecules from natural plant material are compared. Dried Hypericum perforatum L. plant material is subjected to Soxhlet extraction, extraction by ultrasonication, and accelerated solvent extraction. The percentage of two bioactive compounds, hyperforin and hypericin, in the extracts is used as a parameter for comparison of the extraction procedure.     

HPTLC Quantitation of Camptothecin in Nothapodytes foetida (Wight) Sleumer Stem Powder

JPC – Journal of Planar Chromatography – Modern TLC - A sensitive and reliable high-performance thin layer chromato-graphic method has been developed for quantitation of camp-tothecin...     

Direct analysis of camptothecin from Nothapodytes nimmoniana by desorption electrospray ionization mass spectrometry (DESI-MS)

Desorption electrospray ionization was employed for fast and direct ambient detection of the anti-tumor drug, camptothecin, and its derivative, 9-methoxycamptothecin in Nothapodytes nimmoniana. Different parts of the plant such as leaves, stems and bark were examined. The ion intensities suggest that the concentration in bark is higher than that in the leaves and stems. The method does not require any sample preparation or preseparation. The identity of the alkaloids was further confirmed by tandem mass spectrometry.     

Comparison of extraction techniques of robenidine from poultry feed samples

In this paper, effectiveness of six different commonly applied extraction techniques for the determination of robenidine in poultry feed has been compared. The sample preparation techniques included shaking, Soxhlet, Soxtec, ultrasonically assisted extraction, microwave – assisted extraction and accelerated solvent extraction. Comparison of these techniques was done with respect to the recovery extraction, temperature and time, reproducibility and solvent consumption. Every single extract was subjected to clean – up using aluminium oxide column (Pasteur pipette filled with 1 g of aluminium oxide), from which robenidine was eluted with 10 ml of methanol. The eluate from the clean-up column was collected in a volumetric flask, and finally it was analysed by HPLC–DAD–MS. In general, all extraction techniques were capable of isolating of robenidine from poultry feed, but the recovery obtained using modern extraction techniques was higher than that obtained using conventional techniques.

In particular, accelerated solvent extraction was more superior to other techniques, which highlights the advantages of this sample preparation technique. However, in routine analysis, shaking and ultrasonically assisted extraction is still the preferred method for the solution of robenidine and other coccidiostatics.     

Comparison of extraction techniques for the isolation of explosives and their degradation products from soil

A comparison of four extraction techniques used for the isolation of 14 explosive compounds (Method 8330-Explosives) from spiked soil samples is described. Soxhlet warm extraction (SWE), pressurized solvent extraction (PSE), microwave assisted extraction (MAE) and supercritical fluid extraction (SFE) were included. The effects of basic extraction conditions – i.e. type of extraction solvent, temperature, pressure, and extraction time – were investigated. The best extraction recovery of the monitored compounds from spiked soil was obtained using pressurized solvent extraction. Recoveries of explosives using the PSE technique were in the range from 65 to 112%. Extraction recoveries by Soxhlet warm extraction and supercritical fluid extraction reached 65–99% and 52–75%, respectively. The lowest extraction recoveries (28–65%) were obtained using microwave assisted extraction. A very low extraction recovery for tetryl was observed in all cases but the best results were achieved by pressurized solvent extraction (58%).     

Comparison of extraction techniques for quantitative analysis of pendimethalin from soil and rice grain

The paper exploits the development, optimization, and comparison of fast, efficient, quantitative analytical extraction techniques such as ultrasonic-assisted extraction (UAE) and matrix solid-phase dispersion (MSPD) for proficient extraction of pendimethalin from soil and rice samples. Residues of pendimethalin were quantified using high-performance liquid chromatography. Impact of several experimental parameters of UAE and MSPD techniques on extraction of pendimethalin from soil and rice samples was also evaluated. Under the optimized conditions, the mean percent recoveries obtained from both methods were in the range of 80.3–101.3 and 81.7–103.1, respectively, with relative standard deviation <10. Linearity was in the range of 0.003–5.0?µg?mL^?1 with limit of detection and limit of quantification as 0.001 and 0.003?µg?g^?1, respectively. MSPD method was found superior in terms of low solvent consumption, small sample size, and reduced matrix coextracts due to simultaneous extraction and cleanup steps. Both extraction methodologies were successfully applied in monitoring routine soil and rice samples, in which pendimethalin residues (0.003–0.007?µg?g^?1) were detected in few rice samples while residues in soil samples were below the quantification limit.     

Comparison of techniques for the extraction of the hypotensive drugs geniposidic acid and geniposide from Eucommia ulmoides

The extraction of geniposidic acid and geniposide from Eucommia ulmoides was carried out by Soxhlet extraction, enzyme-assisted aqueous extraction, semi-bionic extraction and supercritical fluid extraction techniques and the extracts were analyzed by high performance liquid chromatography. The results showed that the best methods for extraction of geniposidic acid and geniposide are enzyme-assisted aqueous extraction and semi-bionic extraction. Compared with various extraction methods, the semi-bionic extraction was more efficient, yielding the highest amount of geniposidic acid and geniposide. In semi-bionic extraction, no organic solvent was used. Semi-bionic extraction was performed at the same pH as that of the human body. The results indicated that semi-bionic extraction was the best method for large-scale efficient extraction of geniposidic acid and geniposide from plant materials. The supercritical fluid extraction process required more time than the other methods, and gave the lowest yields of geniposidic acid and geniposide.     

Prospecting for Camptothecines from Nothapodytes nimmoniana in the Western Ghats, South India: identification of high-yielding sources of camptothecin and new families of camptothecines

Camptothecin (CPT), a monoterpene alkaloid, is an important anti-cancer compound obtained from several plant sources including Camptotheca acuminta (from China) and Nothapodytes nimmoniana (from India). Currently, by far the highest levels of CPT (approximately 0.3% w/w) are reported from Nothapodytes nimmoniana, a small tree distributed in the Western Ghats, India. In recent years because of the heavy demand, there has been a serious threat of extinction of the populations of the tree in the Western Ghats forest of south India. Several studies have chemically profiled populations of the species in the Western Ghats to identify sources of high yield and therefore to enable the sustainable production and harvesting of CPT. In this study, using both high-performance liquid chromatography and liquid chromatography-mass spectrometry, we report for the first time the identification of trees that produce at least 5- to 8-fold more CPT than hitherto reported. Furthermore, we show for the first time the production of a few minor camptothecines, including 10-hydroxy camptothecin, in the stem and root bark extracts of the tree. These results have important implications for not only harnessing the high-yielding individuals for clonal multiplication but also for exploiting some of the minor camptothecines, which also have been shown to have important anti-cancer and anti-viral activity.     

Comparison of extraction techniques for isolation of steroid oestrogens in environmentally relevant concentrations from sediment

The comparison of four extraction techniques for isolation of five native and one labelled steroid oestrogens from sediment was described. The three conventional extraction techniques Soxhlet warm extraction (SWE), accelerated solvent extraction (ASE), microwave-assisted extraction (MAE) and a promising technique QuEChERS were tested for isolation of low environmentally relevant oestrogen concentrations using different extraction conditions. The least expensive and time-consuming method QuEChERS provided the best extraction recoveries (53–84%) from all techniques. MAE achieved the highest recovery from conventional techniques for less polar oestrogens using dichloromethane: acetone 3:1 mixture as an extraction solvent (50–71%), but for extraction of the whole group of oestrogens including more polar estriol acetone or methanol must be used. ASE provided higher extraction recoveries using dichloromethane at 60°C (53–74%) for less polar oestrogens. However, the repeatability of results was unsatisfactory and recoveries using other extraction conditions were lower than for MAE. The most time-consuming SWE achieved the worst extraction recoveries and for isolation of low oestrogen concentrations from sediments, it is completely unsuitable.     

Comparison of extraction techniques and modeling of accelerated solvent extraction for the authentication of natural vanilla flavors

Accelerated solvent extraction (ASE) of vanilla beans has been optimized using ethanol as a solvent. A theoretical model is proposed to account for this multistep extraction. This allows the determination, for the first time, of the total amount of analytes initially present in the beans and thus the calculation of recoveries using ASE or any other extraction technique. As a result, ASE and Soxhlet extractions have been determined to be efficient methods, whereas recoveries are modest for maceration techniques and depend on the solvent used. Because industrial extracts are obtained by many different procedures, including maceration in various solvents, authenticating vanilla extracts using quantitative ratios between the amounts of vanilla flavor constituents appears to be unreliable. When authentication techniques based on isotopic ratios are used, ASE is a valid sample preparation technique because it does not induce isotopic fractionation.     

Comparison of various techniques for the extraction and determination of antioxidants in plants

The following extraction techniques have been used for extracting antioxidants (apigenin, coumarin, esculetin, umbelliferone, bergapten, quercetin, rutin, scopoletin and xanthotoxin) from plant material: supercritical fluid extraction, pressurized liquid extraction, extraction by means of Soxhlet apparatus, ultrasonic extraction in ultrasonic bath, and by means of ultrasonic probe. The analytical method based on HPLC?UV detection for the determination of selected antioxidants was developed. For all extracts the antioxidant capacity based on the reduction of free 2,2‐diphenyl‐1 ‐picrylhydrazyl radical was also determined. Comparing all results the ultrasonic probe method using 0.75 g of sample extracted by 50 mL of acetonitrile in water (30%, v/v) for 25 min at room temperature and with amplitude at 60% (equal to 90 W) without pulsation was evaluated as the best tool. The most significant indicator demonstrating this statement is the antioxidant capacity expressed as gallic acid equivalent where the ultrasonic probe method showed the best results in 10 of 16 samples. Also the operability of ultrasonic probe extraction method compared to other tested methods is more favorable.     

Comparison of different extraction techniques for the determination of polychlorinated organic compounds in sediment

The performance of various enhanced extraction techniques, such as accelerated solvent extraction, microwave-assisted extraction, Soxhlet extraction, ultrasonic extraction and fluidized-bed extraction for the determination of polychlorinated organic compounds in dry sediment was investigated in two laboratories. The results of the two laboratories were in good agreement. The extraction yields from the batch extraction methods were lower than those from the dynamic techniques. Accelerated solvent extraction, especially, exhibited higher extraction efficiency than the standard procedure, Soxhlet extraction, whereas the results of fluidized-bed extraction were comparable.     

Comparison of various extraction techniques for the determination of polycyclic aromatic hydrocarbons in worms

Two less laborious extraction methods, viz. (i) a simplified liquid extraction using light petroleum or (ii) microwave-assisted solvent extraction (MASE), for the analysis of polycyclic aromatic hydrocarbons (PAHs) in samples of the compost worm Eisenia andrei, were compared with a reference method. After extraction and concentration, analytical methodology consisted of a cleanup of (part) of the extract with high-performance gel permeation chromatography (HPGPC) and instrumental analysis of 15 PAHs with reversed-phase liquid chromatography with fluorescence detection (RPLC-FLD). Comparison of the methods was done by analysing samples with incurred residues (n=15, each method) originating from an experiment in which worms were exposed to a soil contaminated with PAHs. Simultaneously, the performance of the total lipid determination of each method was established. Evaluation of the data by means of principal component analysis (PCA) and analysis of variance (ANOVA) revealed that the performance of the light petroleum method for both the extraction of PAHs (concentration range 1-30 ng/g) and lipid content corresponds very well with the reference method. Compared to the reference method, the MASE method yielded somewhat lower concentrations for the less volatile PAHs, e.g., dibenzo[ah]anthracene and benzo[ghi]perylene and provided a significant higher amount of co-extracted material.     

Comparison of alternative and conventional extraction techniques for the determination of zearalenone in corn

Naturally contaminated corn samples of different origin were extracted using two conventional techniques (blending and shaking) and three alternative approaches (ultrasonic extraction, accelerated solvent extraction, and microwave-assisted extraction). Use of the same extraction mixture for all trials enabled the efficiency of the various extraction techniques to be compared. Extracts were filtered and directly analyzed by LC–ESI–MS, without further clean-up. The yield from the alternative extraction techniques showed efficiency to be higher than for conventional techniques. In particular, microwave-assisted extraction was slightly superior to other techniques.     

Comparison of different extraction techniques for the determination of chlorinated pesticides in animal feed

The performances of Soxhlet extraction, dive-in Soxhlet extraction, microwave-assisted extraction (MAE), accelerated solvent extraction (ASE), fluidized-bed extraction (FBE), and ultrasonic extraction (UE) for the analysis of organochlorine pesticides in animal feed have been investigated. ASE and MAE provided significantly better extraction efficiency than Soxhlet extraction. The concentrations were 126.7 and 114.8%, respectively, of the values obtained by classical Soxhlet extraction, whereas the results from FBE and dive-in Soxhlet were comparable with those from the standard Soxhlet procedure. The reproducibility of FBE was the best, with RSDs ranging from 0.3 to 3.9%. Under the investigated operation conditions UE was not efficient, with the recoveries of target compounds being about 50% less than Soxhlet. Additionally, the performances of Soxhlet, dive-in Soxhlet, MAE, ASE and FBE were validated by determination of the certified reference material BCR-115. The results from the extraction techniques were in good agreement with the certified values.     

A process for preparation of anticancer drug 9-nitro camptothecin

Camptothecin and some of its semisythetic derivatives such as topotecan, irinotecan and 9-nitrocamptothecin, have exhibited strong antitumor activity against various experimental tumor model. The water insoluble 9-nitrocamptothecin demonstrated very potent antitumor activity against many different types of human cancers and HIV. Literature procedures for preparing 9-nitrocamptothecin are direct nitration of camptothecin using a concentrated nitric/concentrated sulfuric acid system or a combin…