The generation of specific and sensitive antibodies against small molecules is greatly dependent upon the characteristics of the hapten–protein conjugates. In the present study, we report a new fluorescence-based method for the characterization of hapten–protein conjugates. The method is based on an effect promoted by hapten–protein conjugation density upon the fluorescence intensity of the intrinsic tryptophan chromophore molecules of the protein. The proposed methodology is applied to quantify the hapten–protein conjugation density of two different class of pesticides (atrazine and 2,4-dichlorophenoxyacetic acid in this study) coupled to carrier protein. The study proved useful for monitoring the course of hapten–protein conjugation for the production of specific antibodies against small molecules. Well-characterized hapten–protein conjugates enabled obtaining highly sensitive anti-atrazine and anti-2,4-D antibodies with IC50 values equal to 12 and 70 ng mL−1 for atrazine and 2,4-D respectively. These antibodies were used for developing a fluorescence-based immunoassays format demonstrating a detection limit of atrazine and 2,4-D in standard water samples 2 and 7 ng mL−1, respectively. The developed immunoassay format could be used as convenient quantitative tools for sensitive and specific screening of pesticides in samples.
Background: Chiral purity is a critical quality attribute of pharmaceutical materials as chiral compounds may have different pharmacological and toxicological properties with their enantiomer (or diastereomers for molecules with multiple chiral centers). Getting high quality drugs to the market quickly is essential for the survival of pharmaceutical companies and in drug research and development. It is important note that about 40% of all man-made synthetic drugs are chiral, 60% of all pharmaceuticals are chiral and 45% chiral drugs are sold as racemate. So, the objective of the current review is to discuss various chromatographic techniques used for the separation of chiral compounds. Methods: Various bibliographic databases of previously published peer-reviewed research papers were explored and systematic data has been culminated in terms of various chromatographic techniques used for chiral compounds’ separation. A comparison of different techniques as well as their advantages are also discussed. Results: A comprehensive review of 130 papers including both, research and review articles, was carried out to make the article readily understandable. The analytical techniques have been discussed in detail. Apart from chromatographic techniques, other techniques such as circular dichroism, nuclear magnetic resonance, UV-visible spectroscopy using cyclodextrin derivatives have also been highlighted with proper citation of references. Conclusion: The pharmaceutical industries need analytical methods to conclude enantiomeric concentration and obtain a drug with single stereo configuration. The sensitive techniques such as HPLC, GCMS and LCMS etc. are used for identification and quantification of limited quantities of single enantiomers, specifically in drug discovery and development.
Pharmaceutical oral dosage forms are tremendously preferred by both consumers as well as pharmaceutical manufacturers owing to the plethora of benefits they offer. Lozenges (LZs) are one of the dosage forms that provide a palatable means of drug administration and have great importance with respect to their pharmaceutical applications. LZs offer additional benefits to pediatric and geriatric patients, along with people having associated problems with the gastro-intestinal tract. Dysphagia is a common problem faced by all age groups, which gives rise to the need for LZs. Moreover, the foremost merit presented by the medicated LZs includes its augmented retention time in the oral cavity that results in an enhanced bioavailability for buccal or upper gastro-intestinal disorders. Further, LZs can also be used to bypass the first-pass effect. The present review covers various aspects of LZs such as formulation, manufacturing techniques, evaluation parameters, marketed products, patents, and a compilation of research work that has been done on lozenges as a delivery system.
: Metal nanoparticular synthesis techniques essentially involve a reduction of metal ions to convert them into nanoparticles. Reducing agents originate from natural and synthetic sources. Chemical methods involve application of synthetic agents that are not healthy and eco-friendly and thus, there is a need for green methods. Green synthesis methods involve reduction of metal ions using plant-based extracts or phytoconstituents and microorganisms like bacteria, yeast, and fungi. These methods have been found to be cost-effective, more efficient and eliminate the application of hazardous chemicals. The phytoconstituents involved in the synthesis of metal nanoparticles are rich in polyols and antioxidants, which in addition to reducing properties, also offer stabilization of formed nanoparticles. Moreover, the size of nanoparticles can also be controlled based on the mechanism of reducing agent involved therein. The present review is an attempt to highlight the methods of synthesis of metallic nanoparticles with a special mention of applications of various natural antioxidants in their synthesis.
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