Rapid Identification of Chemical Constituents of Pimpinella thellungiana by UHPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

Objective::Ultra high performance liquid chromatography coupled with Q-Exactive Focus hybrid quadrupole and orbitrap high resolution mass spectrometer (UHPLC-Q-Orbitrap HRMS) was applied for identification of chemical constituents in Pimpinella thellungiana. Method::Chromatographic separation was performed on a WATERS BEH C18 column (2.1 mm ×50 mm, 1.7 μm). The mobile phase consisted of 0.1% formic acid aq (A) and acetonitrile (B) with a gradient elution. Mass spectral analysis were performed on Q-Exactive Focus hybrid quadrupole and orbitrap mass spectrometer. The mass spectrometer was connected to UHPLC instrument via an ESI interface. Samples were analyzed in negative ion mode by the full-scan-dd MS 2(data-dependent MS/MS) scanning mode. Then the constituents of P. thellungiana were identified by compared HRMS data with those of the standard compounds, MS cleavage mechanism and the related literatures. Result::Based on the characteristic mass data of accurate molecular weight and fragmentation ion information, 29 chemical constituents were identified including 19 chlorogenic acids, 7 flavonoids and 3 phenolic acid. Among them, the identified components except luteolin-7-O-β-D-glucuronopyranoside and apigenin-7-O-β-D-glucuronopyranoside were reported in P. thellungiana for the first time. Conclusion::The established UHPLC-Q-Orbitrap HRMS method can be used to identify the chemical constituents of P. thellungiana quickly and accurately, providing the scientific evidence for its quality evaluation and material basis research.

Content Determination of Nine Constituents in Different Medicinal Parts of Pimpinella thellungiana / 中国实验方剂学杂志

Objective:To develop a method to quantify nine constituents in different medicinal parts of Pimpinella thellungiana, in order to compare the content difference of the nine constituents, namely protocatechuic acid,gallic acid,neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,luteolin-7-O-β-D-glucuronide,isochlorogenic acid A,apigenin-7-O-β-D-glucuronide and isochlorogenic acid C. Method:The analysis was performed on an Agilent TC-C18 column (4.6 mm×250 mm,5 μm) with acetonitrile and mixed acid solution (0.1% phosphoric acid-0.1% glacial acetic acid) as mobile phase for gradient elution. The handover detection wavelengths were at 265 and 325 nm. The column temperature was 20℃, and the flow rate was 1.0 mL·min-1. The experiment data was analyzed using the software of Markerlynx XS. Result:The nine constituents of protocatechuic acid,gallic acid,neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid, luteolin-7-O-β-D-glucuronide,isochlorogenic acid A,apigenin-7-O-β-D-glucuronide and isochlorogenic acid C had a good degree of separation and a good linearity in their respective linear ranges(r>0.999 8). The average recoveries ranged from 99.11% to 100.76%,and the RSD ranged from 0.9% to 2.0% 。The results showed that the contents of the nine constituents had significant differences in different medicinal parts of P. thellungiana. The average contents of the nine constituents were the highest in leaves,which was followed by stem,and the lowest was in root. Conclusion:The study could provide evidence for the quality control,clinical application,and scientific resources utilization of P. thellungiana.