질량분석기를 활용한 황칠나무 잎 추출물의 성분 프로파일 분석

Abstract

Compound profile is a kind of quantitative analysis method that uses a liquid chromatograph (LC) equipped with an ultraviolet detector (UVD) to manage the peak pattern of a chromatogram obtained as a result of analyzing a plant extract. It has been used for the purpose of quality control of herbal medicines for which index compounds have not been established. The information that can be confirmed as a result of the compound profile is limited and is not suitable for qualitative analysis. Additional information such as the molecular weight of each peak can be obtained by using LC-mass spectrometer (MS) applied to the compound profile analysis, and information that can be used to estimate the molecular structure can be obtained through LC-MS/MS analysis. About 20 kinds of active compounds present in Dendropanax morbifera leaves, which are known to have various physiological activities, have been reported through various studies. However, these compounds have not been analyzed at the same time, and information on their compounds is still insufficient compared to the number of compounds actually present in D. morbifera leaves. In this study, the MS compound profile of the compounds present in the leaf extract of D. morbifera was prepared using LC-MS to compensate for the disadvantages of the compound profile using LC-UVD. First, the online LC-DPPH radical scavenging activity was measured to prepare the antioxidant activity compound profile present in the leaf extract of D. morbifera. Based on the retention time of the active compound peak identified as the measurement result, LC-MS and MS/MS analysis were performed. Qualitative analysis of the active peak was conducted through MS data review of various literatures and standard compound analysis. Twelve radical scavenging active compound peaks were identified as 5-O-caffeoylquinic acid (neochlorogenic acid), 3-O-caffeoylquinic acid (chlorogenic acid), 4-O- caffeoylquinic acid (cryptochlorogenic acid), orientin, hesperidin, hyperoside, rutin, isoquercitrin, azelaic acid, 3,5-dicaffeoylquinic acid (isochlorogenic acid A), 3,4-dicaffeoylquinic acid (isochlorogenic acid B), caffeic acid dimer. In the MS compound profile analysis of the leaf extract of D. morbifera, which was additionally conducted, 95 compounds were reviewed. The compounds identified as having the highest number in the MS compound profile were ones containing apigenin as an aglycone, and there were a total of 19 apigenin derivative compounds whose structures could be confirmed or estimated. In addition, 6 caffeic acid derivatives, including caffeic acid and chlorogenic acid, were identified, and 5 quercetin-related compounds, including quercetin and its derivatives, were identified in the MS compound profile. Flavonoids such as hesperidin and naringin and various phenolic acid structure compounds were identified, and 8 types of hydroxy fatty acid structure compounds were identified. In order to be used in the study of the activity of D. morbifera leaves, the main physiological activity fields being studied for each compound were additionally confirmed through literature review. LC-MS/MS analysis conditions of multiple reaction monitoring (MRM) mode were set for selective analysis of radical scavenging active compounds and MS compound profile. As a result of MRM mode analysis of the compounds of the leaf extract of D. morbifera, clear and precise results were obtained compared to the general LC-UVD compound profile analysis. In a comparative analysis with 11 leaf extracts of other plants, 28 compounds specifically identified in D. morbifera leaf extract were identified. In general, it is known that gas chromatograph (GC) analysis is more advantageous than LC analysis for lipophilic compounds. In addition, using the library search tool of GC-MS has the advantage of being able to perform qualitative analysis without standard compound analysis. As a result of the scan mode analysis of GC-MS of the lipophilic extract of D. morbifera leaf, 20 peaks were detected. MS spectra of each peak were identified by library search, and 17 compounds except for 3 peaks were identified. Dendropanoxide was identified as the compound expected to have the highest content, and the contents of β- amyrin and α-amyrin were also relatively high. There were a number of triterpenoid-related compounds known to have various physiological activities, and MRM analysis conditions of GC-MS/MS were set for the compounds. Through this study, the existence of a number of active-related compounds that had not been identified in studies related to D. morbifera leaves was confirmed. In addition, the MS compound profile analysis method used in the study is useful for studying plant extracts in which numerous compounds are mixed, and the set MRM mode analysis conditions can also be used for studying the compounds of other plant extracts.