HPLC 및 LC-MS/MS를 이용한 산마늘과 박새의 판별법 개발

Abstract

In the recent past, there is an increasing trend in society to use natural and pollution free food in view to maintain good health. Every year in the spring season, a variety of plants are collected from mountains and agriculture fields to consume as food for good taste and refresh lives. However, food poisoning is continuously reported in relation to intake of spring herbs and the majority of these cases are due to mistakes in selection of wild edible plants due to their similar appearance with toxic plants. The wild edible plant Allium victorialis is often confused with plant Veratrum patulum, due to similarity in physical appearance. V. patulum causes poisoning symptoms such as hypotension, vomiting, dizziness, and abdominal pain. Thus to avoid any confusion of V. patulum with A. victorialis; an accurate method is very much required at the time of harvesting plants. This study is therefore designed to develop an analytical method for identification of A. victorialis and to prevent its possible adultration due to V. patulum. First the elemental contents were analyzed and applied to identify V. patulum contained in A. victorialis. Then a discriminant method for analyzing selected indicator was carried out, after method validation and blind tests confirmation. Finally, the developed method was applied to the processed foods in the markets to confirm the indicator compound for any adultration. The HPLC-DAD and LC-MS/MS were used as analytical techniques in this study. The HPLC-DAD screening test was performed with the samples solutions in which the polarity index was sequentially extracted using a soxhelt apparatus; to check the indicator component. Peak was confirmed as luteolin-7-O-glucuronide and analyzed using LC-MS/MS for the confirmed peak, and was established as the final indicator component of V. patulum after comparative analysis with the standard. For efficient analysis, this study developed an efficient extraction method too for analytical analysis of luteolin-7-O-glucuronide contained in V. patulum. A methanol reflux extract under the condition of 2 hours was found as optimum extraction method. Proper method validation was carried out to obtain the reliability of the developed method and effectiveness of the obtained results. In order to confirm V. patulum contained in A. victorialis, the luteolin-7-O-glucuronide analysis method was applied to 19 processed foods commonly consumed in South Korea, and were found that there is no contamination of V. patulum and safe from causing aforementioned symptoms of poisoning. From this research, the indicator compound of luteolin-7-O-glucuronide was developed and the analysis method so designed was applied successfully to determine whether there is any mixing of V. patulum with A. victorialis in processed foods from open markets of South Korea. As a result of this research, it will be possible to minimize damage by prompt response to the cause of food poisoning caused by V. patulum in A. victorialis. This will improve the reliability foods especially made of spring herb A. victorialis.