Rearrangement of 4-(1-haloalkyl)- and 4-(2-haloalkyl)-2-azetidinones into methyl omega-alkylaminopentenoates via transient aziridines and azetidines

The synthesis of 4-(1-haloalkyl)-2-azetidinones and 4-(2-haloalkyl)-2-azetidinones was investigated with use of the Staudinger reaction between in situ generated ketenes and alpha-haloimines or beta-haloimines. This new class of functionalized 2-azetidinones was further evaluated for its potential use as intermediates in the synthesis of highly functionalized compounds. The reaction of 4-(1-haloalkyl)-2-azetidinones and 4-(2-haloalkyl)-2-azetidinones with sodium methoxide in methanol yielded ring-opened products, i.e., methyl 2-alkoxy-4-(alkylamino)pentenoate and methyl 5-(alkylamino)pentenoate, respectively. Further attention was paid in detail to the reaction mechanism involved in this peculiar transformation. It was proven that these reactions proceeded via intermediate aziridines or azetidines.     

Rearrangement of 2-aryl-3,3-dichloroazetidines: intermediacy of 2-azetines

An easy synthesis of 2-aryl-3,3-dichloroazetidines, a rather unexplored class of azaheterocycles, is described. The title compounds were easily obtained by reduction of the corresponding 4-aryl-3,3-dichloro-2-azetidinones with monochloroalane, which in turn were synthesized by a ketene-imine [2 + 2] cycloaddition. The reactivity of 3,3-dichloroazetidines with bases was investigated, yielding 2-[dimethoxy(aryl)methyl]aziridines by ring contraction when treated with sodium methoxide. Furthermore, reacting the 3,3-dichloroazetidines with sodium hydride in DMSO, followed by aqueous workup, afforded 1-alkyl-2-aroylaziridines, by hydrolysis of the intermediate 2-azetines and ring closure of the transient 3-amino-2-chloro-1-phenyl-1-propanone derivatives. Monitoring this reaction in an NMR tube, using sodium hydride in DMSO-d(6), allowed the characterization of the intermediate strained heterocyclic enamines, i.e., 2-azetines, by (1)H and (13)C NMR.     

Evaluation of the influence of proline, hydroxyproline or pyrrolidine in the presence of sodium nitrite on N-nitrosamine formation when heating cured meat

N-nitrosamines are meant to be probable or possible carcinogenic components, possibly formed out of a reaction between nitrite and N-containing substances such as amino acids and secondary amines. Nitrite is often used for processing meat products because of its colouring and antimicrobial properties.During this experimental setup, the influence of proline, hydroxyproline or pyrrolidine on N-nitrosamine formation in meat samples was evaluated. The N-nitrosamines concentrations were measured with gas chromatography-thermal energy analyzer. Only the concentrations of N-nitrosodimethylamine and N-nitrosopyrrolidine were found above the limit of detection in a number of tested experimental conditions. The concentration of these two N-nitrosamines was modelled as a function of temperature and nitrite concentration for different situations (presence or absence of added natural N-containing meat components). It could be concluded that proline and pyrrolidine promoted the formation of N-nitrosopyrrolidine. It could also be confirmed that the higher the temperature of the meat processing procedure and the higher the sodium nitrite amounts added, the higher were the yields of the respective N-nitrosamines.     

Recent developments in chromatographic fingerprints from herbal products: set-up and data analysis

The use of chromatographic fingerprints from herbal products where the whole chromatographic profile is applied to evaluate the quality of the investigated product. In this paper, recent developments in the set-up and data analysis of chromatographic fingerprints for herbal products are discussed. First different set-ups for fingerprint development are reviewed. Prior to fingerprint development, a suitable sample preparation, e.g. extraction, should be considered. In a second instance, this review focuses on the data analysis with regards to the different applications of fingerprints. Usually, chemometric data pretreatment is necessary. This is discussed first, followed by a short overview of the data handling techniques used in the two main application areas of herbal fingerprints, i.e. quality assurance and classification or calibration. The quality assurance, which involves the identification and quality control of the herbal products, is reviewed, followed by the use of fingerprints in classification or modelling. The different application areas are illustrated and discussed with several case studies.     

Identification of poliovirions and subviral particles by capillary electrophoresis

Poliovirions, purified from infected cell extracts with anion-exchange chromatography, can be analyzed and identified by CE in untreated fused silica capillaries using UV detection. Other subviral particles can be eluted as well from the same infected cell extract using a higher salt concentration buffer on the ion-exchange chromatography. Virions can be identified because of their conversion into empty capsids upon heating at 56°C. As a result of heating, the viral genome is released from the capsid. Here, we show that during this incubation some intermediate particles were found. The latter were identified by enzymatic peak shift analysis. The high salt concentration eluate subviral particles were analyzed with preincubation affinity CE together with their sensitivity for RNase and proteinase K treatment. Electropherograms of the higher salt concentration eluate display a mixture of at least four different subviral particles. One particle proved to have an [N1, H] antigenicity and was resistant to RNase and proteinase K digestion. The remaining particles were all sensitive to proteinase K treatment. This CE method proved to be valuable in the detection, identification and analysis of poliovirions and poliovirus particles offering an alternative powerful, cheap, fast and easy analysis method.     

Quality control of Citri reticulatae pericarpium: exploratory analysis and discrimination

Extracts of Citri reticulatae pericarpium (PCR) are commonly used in the Traditional Chinese Medicine. The quality control of PCR is currently performed by single marker analysis, which can hardly describe the complexity of such natural samples. In this study, a fingerprint methodology for PCR based on high-performance liquid chromatography (HPLC) was developed and validated. A total of 69 fingerprints of authenticated PCR samples, commercial PCR samples, mixed peel samples, and other Citrus peels were recorded. Exploratory data analysis allowed optimizing the extraction procedure and detecting mixed peel samples. Once the optimizations were performed and the method validated, discrimination between the authentic PCR samples and all other samples was performed by p-Discriminant Partial Least Squares. The established model was able to differentiate between classes with a high reliability for each sample. Furthermore, evaluation of the score and loading plots of the model indicated nobiletin, tangeretin, naringin and hesperidin as important markers for the quality control of PCR.     

Chromatographic separation techniques and data handling methods for herbal fingerprints: a review

As herbal medicines have an important position in health care systems worldwide, their current assessment and quality control are a major bottleneck. Over the past decade, major steps were taken not only to improve the quality of the herbal products but also to develop analytical methods ensuring their quality. Nowadays, chromatographic fingerprinting is the generally accepted technique for the assessment and quality control of herbal products. This paper briefly considers the evolution of the regulations and guidelines on the quality control of herbal medicines, and reviews the established analytical techniques for herbal fingerprinting with an emphasis on the most recent developments, such as miniaturized techniques, new stationary phases, analysis at high temperatures and multi-dimensional chromatography. Accessory to the new analytical techniques, the chemometric data handling techniques applied are discussed. Chemometrics provide scientists with useful tools in understanding the huge amounts of data generated by the analytical advances and prove to be valuable for quality control, classification and modelling of, and discrimination between herbal fingerprints.     

Supersaturated designs: set-ups,data interpretation,and analytical applications

For screening purposes, two-level screening designs, such as fractional factorial (FF) and Plackett–Burman (PB) designs, are usually applied. These designs enable examination of, at most, N−1 factors in N experiments. However, when many factors need to be examined, the number of experiments still becomes unfeasibly large. Occasionally, in order to reduce time and costs, a given number of factors can be examined in fewer experiments than with the above screening designs, by using supersaturated designs. These designs examine more than N _SS−1 factors in N _SS experiments. In this review, different set-ups to construct supersaturated designs are explained and discussed, followed by several possible data interpretations of supersaturated design results. Finally, some analytical applications of supersaturated designs are given.     

Validation of a fluorimetric assay for 4-aminophenol in paracetamol formulations

4-aminophenol (4-AP) is the primary degradation product of paracetamol (PARA). According to the European Pharmacopoeia, 50 ppm 4-AP/PARA is the specification limit of 4-aminophenol in paracetamol drug substance. For drug products, often higher specification limits, such as 1000 ppm 4-AP/PARA are applied. This paper describes a fluorimetric method to quantify the low amount of this degradant (50 ppm) in a pharmaceutical preparation, i.e. in paracetamol tablets. The fluorimetric method was validated and the linearity, precision, trueness, range, limit of detection and limit of quantification were determined. They were found acceptable to assay the low amounts of 4-aminophenol in paracetamol tablets.     

Ruggedness and robustness testing

Due to the strict regulatory requirements, especially in pharmaceutical analysis, analysis results with an acceptable quality should be reported. Thus, a proper validation of the measurement method is required. In this context, ruggedness and robustness testing becomes increasingly more important. In this review, the definitions of ruggedness and robustness are given, followed by a short explanation of the different approaches applied to examine the ruggedness or the robustness of an analytical method. Then, case studies, describing ruggedness or robustness tests of high-performance liquid chromatographic (HPLC), capillary electrophoretic (CE), gas chromatographic (GC), supercritical fluid chromatographic (SFC), and ultra-performance liquid chromatographic (UPLC) assay methods, are critically reviewed and discussed. Mainly publications of the last 10 years are considered.