Analysis of chlorogenic acids in Helianthus tuberosus Linn leaves using high performance liquid chromatography-mass spectrometry
To identify chlorogenic acids in Helianthus tuberosus Linn leaves, a method of high performance liquid chromatography-ultraviolet-mass spectrometry (HPLC-UV-MS) was developed. HPLC analysis was performed on an Inertsil ODS-3 column (250 mm x 4.6 mm, 5 microm). The mobile phase consisted of 1% acetic acid (A) and methanol (B). A gradient program was adopted as follows: 0 - 10 min, 20% B to 35% B; 10 - 25 min, 35% B to 50% B; 25 - 35 min, 50% B to 80% B. The flow rate was set at 1.0 mL/min and the column temperature was 35 degrees C. HPLC chromatogram was extracted at 327 nm. The mass spectrometer used was a TSQ triple quadrupole MS equipped with an electrospray ionization (ESI) interface. Initially, the mass spectrometer was programmed to perform full scan ranging from m/z 250 - 1 200. For MS/MS, negative ion monitoring mode was used and the collision energy was set at 10 - 45 eV. By analyzing UV characteristics and MS fragmentation patterns, 7 chlorogenic acids were assigned to be three categories, i. e., three caffeoylquinic acids, one feruloylquinic acid and three dicaffeoylquinic acids. As the results of the method development efforts, an effective and fast method for the qualitative identification of the chlorogenic acids in Helianthus tuberosus Linn leaves was established.
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- "The phenolic compounds in H. tuberosus leaves extracts were identified by comparing their UV-Vis absorption spectra, matching their molecular ions (m/z) obtained by ESI-MS and ESI-MS/MS chromatographic characteristics with the literature data reported [5, 6] or with available reference standards. The external standard method was used for the quantification of main phenolic acids. "
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ABSTRACT: Plant phenolics can have applications in pharmaceutical and other industries. To identify and quantify the phenolic compounds in Helianthus tuberosus leaves, qualitative analysis was performed by a reversed phase high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) and quantitative analysis by HPLC. Ten chlorogenic acids (CGAs) were identified (3-o-caffeoylquinic acid, two isomers of caffeoylquinic acid, caffeic acid, p-coumaroyl-quinic acid, feruloylquinic acid, 3,4-dicaffeoyquinic acid, 3,5-dicaffeoylquinic acid, 1,5-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid) by comparing their retention times, UV-Vis absorption spectra, and MS/MS spectra with standards. In addition, four other phenolic compounds, including caffeoyl glucopyranose, isorhamnetin glucoside, kaempferol glucuronide, and kaempferol-3-o-glucoside, were tentatively identified in Helianthus tuberosus leaves for the first time. The 3-o-caffeoylquinic acid (7.752 mg/g DW), 4,5-dicaffeoylquinic acid (5.633 mg/g DW), and 3,5-dicaffeoylquinic acid (4.900 mg/g DW) were the major phenolic compounds in leaves of Helianthus tuberosus cultivar NanYu in maturity. The variations in phenolic concentrations and proportions in Helianthus tuberosus leaves were influenced by genotype and plant growth stage. Cultivar NanYu had the highest concentration of phenolic compounds, in particular 3-o-caffeoylquinic acid and 4,5-dicaffeoylquinic acid compared with the other genotypes (wild accession and QingYu). Considering various growth stages, the concentration of total phenolics in cultivar NanYu was higher at flowering stage (5.270 mg/g DW) than at budding and tuber swelling stages. Cultivar NanYu of Helianthus tuberosus is a potential source of natural phenolics that may play an important role in the development of pharmaceuticals.
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ABSTRACT: An HPLC-ESI-MS/MS method was established to identify the absorbed components and metabolites in rat plasma after oral administration of Rhizoma Chuanxiong decoction (RCD), a well-known traditional Chinese medicine. By comparing the extracted ion chromatograms (EICs) obtained from dosed rat plasma, blank rat plasma and RCD, a total of 25 compounds were detected in dosed rat plasma. Among them, 13 compounds were absorbed into rat plasma in prototype and identified as ferulic acid, senkyunolide J, senkyunolide I, senkyunolide D or 4,7-dihydroxy-3-butylphthalide, senkyunolide F, senkyunolide M, senkyunolide Q, senkyunolide A, E-butylidenephthalide, E-ligustilide, neocnidilide, Z-ligustilide, levistolide A, according to the retention times, UV, MS, MS/MS spectra. In addition, 12 conjugated metabolites including 6 senkyunolide I-related metabolites, 4 senkyunolide J-related metabolites and 2 butylidenephthalide-related metabolites were also detected and identified by comparing their MS, MS/MS spectra with that of corresponding original components. Conjugated with glutathione, cysteine, glucuronic acid and sulphuric acid were the main metabolic reactions of phthalides. Finally the in vivo metabolic pathways of chemical constituents of Chuanxiong in rat plasma were proposed in this study.
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ABSTRACT: The total phenolic content and radical scavenging activities of Jerusalem artichoke (Helianthus tuberosus L.) leaves were investigated. Results indicated that the ethyl acetate fraction contained the highest total phenolic content (266.69±2.51mgGAE/g dry extract) accompanied with strongest free radical scavenging abilities. Following an in vitro radical scavenging activity-guide fractionation procedure, six phenolic compounds which strongly quenched free radicals were separated from ethyl acetate fraction. Among them, 3-O-caffeoylquinic acid and 1,5-dicaffeoylquinic acid played a dominant role due to their strong free radical scavenging abilities and their high contents. The content of 3-O-caffeoylquinic acid in n-butanol fraction was 74.58±1.05mg/g, while 1,5-dicaffeoylquinic acid in ethyl acetate fraction was 104.51±2.86mg/g. The results imply that the leaves of Jerusalem artichoke might be a potential source of natural antioxidants.
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