Methods of alkaloid analysis

These analytical dilemmas interfere with the methods of alkaloid analysis. Each group of alkaloids has its own methods of extraction, isolation and crystallization, as well as detection in structure, molecule and dynamicity. Not all these stages are still possible in the majority of alkaloids. In recent years, many techniques have been used in alkaloid detection. There are atomic and molecular electronic spectroscopy, vibration spectroscopy and electron and nuclear spin orientation in magnetic fields, mass spectroscopy, chromatography, radioisotope and electrochemical techniques. Although important developments in methodology and methods of alkaloid analysis have occurred over the last 200 years, the most efficient methods are still awaited. The oldest parts of these methods seen to be the extraction and isolation stages.

2.9.1. Methods in history

The first method of alkaloid analysis was developed in 1805, in the case of morphine. This method of isolation, with minor and major variations, is still used today. By this method, the first quinolizidine alkaloids were also extracted: sparteine in 1851, lupinine in 1865 andlupanine 2 years later. At the beginning of the 20th century, the extraction and determination of total quinolizidine alkaloids in the same analysis (common) was carried out by Jurkowski270, Nowotnowna271, Trier272, Ivanov273 280, Sengbusch274,275,276,277, Lukaszewicz278, Wuttke279. Reifer and Niziolek281 and Wiewiorowski and Skolik282 initiated research in which the sum of the contents of the different and separate alkaloids is the total alkaloid content. The method of isolation of quinolizidine alkaloids was developed next by Wysocka et al.249 250 and Wysocka and Przybyl244 245.

2.9.2. Basic methods and instruments

The first step in the development of methods was the evidence that molecules synthesized and degradated, and that intermediate compounds existed. Initial methods have provided molecule isolation, and subsequently the place in the metabolic chains. The basic methods of alkaloid determination developed historically as follows: iodine, taster, seed colour, Dragendorf reagent, fluorescence, calorimetry, photometry, electrophotometry, spectrometry, paper chromatography, thin layer chromatography, high-performance liquid chro-matography, gas chromatography, gas liquid chromatography-mass spectrome-try, nuclear magnetic resonance, X-ray, enzyme-like immunosorbent assay, radio immuno assay and scintillation proximity assay methods4. The most effective method for establishing a metabolic pathway is the use of isotopes in radiotracing and mass spectrometry methods. The basic instruments that have been developed are photometers, calorimeters, analyzers, spectrometers, chromatographs and different mass spectrometers. These instruments have subsequently been improved to be more exact, and have been through many generations in their development by many different producers.

2.9.3. From iodine to enzyme

Alkaloid analytical methods were developed by applications based on different hypotheses, from the simple to the very complicated. Subsequently, corresponding instruments were developed. This development can be seen by considering the example of quinolizidine alkaloids.

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