HPLC chromatographic finish

The purpose of the HPLC analysis of cleaning samples is to prove with data that the equipment and cleaning procedures work, and that the surfaces of the equipment are indeed clean. The HPLC chromatographic finish is extremely reproducible and is the easiest part of the analytical... 

HPLC is the leading Analytical procedure used for the verification of pharmaceutical cleaning validation programs. HPLC provides a linear, sensitive method for quantitating low levels of residues making the chromatographic finish the most reliable part of the cleaning verification. 

Since hot pepper is important for the food and the pharmaceutical industries, a range of different methods have been developed for the analysis of capsaicinoids in plant material and finished products. The separation of CPS (la) and nonivamide (11) is especially challenging, since these compounds have similar behavior in many chromatographic conditions. Since synthetic nonivamide is the most common adulterant of capsicum oleoresin, various strategies have been suggested to overcome this problem. Capillary GC does not require previous derivatization of capsaicinoids, but its separatory power seems lower than that of HPLC, currently the most popular technique for the quantization of capsaicinoids. GC is, however, the method of choice for the analysis of the acyl moieties of capsaicinoids as methyl esters. These can be directly produced from capsaicinoids by oxidative N-dealkylation with DDQ (2,3-dichloro-5,6-dicyanobenzoquinone), followed by alcoholysis of the resulting amides with methanol in the presence of an acidic resin (Scheme 4.4) [41]. 

A general flow chart for partly automated method development is presented in Fig. 2. From this, it is clear that the analyst repeatedly intervenes in order to evaluate intermediate results to steer the direction of development At the outset, according to the available information as well as the experience of the analyst, the chromatographic factors that are to be examined are defined and their limits are delineated. Thereafter, the HPLC system takes over the execution of the necessary experiments and/or the chromatographic runs. The analyst evaluates the runs once they have finished. The number of peaks, and their form, resolution, and sequence are important. As a result, conditions can be hypothesized that should have the highest potential for optimal separation of all of the peaks, or at least of all the relevant ones. 

---