Analysis on Chemical Components for the Leaves of the Miao Medicine Toricellia Angulata Oliv. Var.Intermedia（Harms） Hu by HPLC-HESI-HRMS

OBJECTIVE To qualitatively analyze the chemical components in the leaves of the Miao Medicine Toricellia Angulata Oliv. var. intermedia(Harms) Hu by HPLC-HESI-HRMS. Methods Thermo Fisher Hypersil GOLD aQ C₁₈(100 mm×2.1 mm, 1.9 μm) column was adopted, with 0.1% formic acid water solution-methyl cyanide(0.1% formic) as the mobile phase for gradient elution; HESI ion source was used for mass spectra, the date were collected in positive and negative mode. At the same time, the difference of retention time(t_R) between 7-hydroxycoumarin, secoxyloganin, hyperin and isoquercitrin with compounds 17, 20, 24 and 25 were verified by HPLC. Compared the fluorescence characteristics of TLC with Rf values. And the characteristics of mass spectrometric fragmentation fragments of 7-hydroxycoumarin, secoxyloganin, hyperin and isoquercitrin were comprehensively verified in references. Results Thirty-eight chemical components in the leaves of the Miao Medicine Toricellia Angulata Oliv. var. intermedia(Harms) Hu were inferred by analyzing positive and negative ion spectra information of high resolution mass spectrometer with date from data base and relevant literature, including 3 amino acids, 5 alkaloids, 10 organic acids, 3 phenylpropanoids, 3 aromatic oxygen-containing derivatives, 3 terpenes, 6 flavonoids, 1 amide and 4 other compounds. The results showed that the difference of t_R of 7-hydroxycoumarin, secoxyloganin, hyperin and isoquercitrin were <0.1 min compared with compounds 17, 20,24 and 25, respectively. The same blue fluorescence spots were shown on the corresponding chromatographic positions of the medicinal sample solution, 7-hydroxycoumarin, secoxyloganin, hyperoside and isoquercitrin reference substances, and the Rf values were consistent. And the literature showed that compounds 17, 20, 24 and 25 had similar characteristics of MS fragmentation to 7-hydroxycoumarin, secoxyloganin loganin, hyperoside and isoquercitrin, respectively. Conclusion Compounds 17, 20, 24 and 25 were identified as 7-hydroxycoumarin, secoxyloganin, hyperin and isoquercitrin. It indicates that HPLC-HESI-HRMS has a scientific basis for the preliminary estimation of the chemical composition of the leaves of Toricellia Angulata Oliv. var. intermedia(Harms) Hu. It will provide basis for therapeutic material basis of the leaves of the Miao Medicine Toricellia Angulata Oliv. var. intermedia (Harms) Hu.