Comparison of different clean-up methods for simultaneous HPLC determination of carotenoids and polyacetylenes in carrot roots

Carrot roots and leaves contain various poly acetylenes such as falcarinol, falcarindiol and falcarindiol acetate with putative beneficial effects on human health, however, these poly acetylenes are also made responsible for the occasional unpleasant bitter taste of carrot juice. Therefore, the aim of this study was to develop an accelerated sample preparation method for simultaneous extraction of carotenoids and poly acetylenes followed by fast and reliable routine HPLC-DAD and HPLC-MS quantification. The individual recoveries of the analytes obtained by ASE-HPLC were compared to those obtained by Soxhlet extraction and solid phase extraction methods, respectively. It has been found that ASE show significantly better extraction efficiency in comparison to the other clean-up procedures. Furthermore, identical ASE parameters can be used for the clean-up of carotenoids and polyacetylenes. In order to further speed up sample preparation a subsequent concentration step using a rotary vacuum concentrator (Christ, sterode, Germany) was performed. The novel extraction procedure was found to be a very useful tool especially applicable for routine analysis, e.g. in plant breeding and evaluation of genetic resources or quality control in food industry. The combination of ASE and rotary vacuum concentration allows high-throughput analysis also ensuring high sensitivity and reproducibility by using HPLC-DAD and HPLCMS methods for subsequent identification and quantification of the individual analytes