Discrimination and Chemical Phylogenetic Study of Four Pulsatilla Herbs Using UPLC-ESI-MS/MS Combined with Hierarchical Cluster Analysis.[Pubmed: 29236950]

A tissue-smashing based ultra-rapid extraction coupled with ultra-performance liquid chromatography tandem-mass spectrometry (UPLC-MS/MS) method was developed to determine 10 major triterpenoid saponins from Pulsatilla herbs. Compound 4 was characterized as betulinic acid glycoside 3-O-α-arabinopyranosyl-28-O-β-glucopyranosyl-23-hydroxy with HR-ESI-MS, 1H-NMR and 13C-NMR experiment. The MS spectra result showed that the ionization of compound 4 was more efficient in the positive mode. Meanwhile, the ions at m/z 789.6 and m/z 627.5 were selected as precursor and product ion for the determination, respectively. The chromatographic separation was carried out on a Phenomenex Kinetex C18 column using a gradient mobile phase system composed of 0.1% formic acid both in methanol and water at a flow rate of 0.4 mL/min. The detection was performed by multiple reaction monitoring mode, using electrospray ionization in the positive and negative mode. The total run time was 6 min. The calibration curves possessed good linearity with all coefficients higher than 0.9987. The intra- and interday precisions were no more than 4.9%, and the average recoveries were from 97.6% to 103.4% with RSD <4.7%. Moreover, hierarchical cluster analysis was performed to compare and discriminate the Pulsatilla herbs based on the quantitative data. The hierarchical cluster analysis results demonstrated that Pulsatilla chinensis, Pulsatilla cernua, Pulsatilla dahurica, Pulsatilla turczainovii samples could be easily discriminated from each other based on the contents of triterpenoid saponins and the established method is feasible for quality control of Pulsatilla herbs.

[Study on seeds after-ripening regulation and key techniques for seedling of Pulsatilla cernua].[Pubmed: 25566643]

The seeds of Pulsatilla cernua were used as tested materials for screening and establishing the main factors with different levels to control fast development of seed embryonic and seedlings of Pulsatilla cernua.

[Effects of nitrogen, phosphorus and potassium fertilizers on the yield, quality and nutrient uptake of Pulsatilla cernua].[Pubmed: 24956807]

To study the effects of nitrogen (N), phosphorus (P2O5) and potassium (K2O) fertilizers on the growth, yield total saponins content and nutrient absorption of Pulsatilla cernua and provide a theoretical basis for good agriculture practice.

Saponins with neuroprotective effects from the roots of Pulsatilla cernua.[Pubmed: 22572934]

Four new oleanene-type triterpenoid saponins together with six known saponins were isolated from the roots of Pulsatilla cernua and their structures were elucidated on the basis of spectroscopic data, including 2D NMR spectra and chemical evidence. Among these one of the aglycones (gypsogenin) is reported for the first time from this genus. Some of these compounds showed significant neuroprotective effects against the cytotoxicity induced by β-amyloid(25-35) (Aβ(25-35)) on human neuroblastoma SH-SY5Y cells.

Qualitative and quantitative determination of nine main active constituents in Pulsatilla cernua by high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry.[Pubmed: 21268254]

A novel qualitative and quantitative method using high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) was developed for simultaneous determination of the nine major active constituents in Pulsatilla cernua (Thunb.) Bercht. et Opiz., namely anemoside A3 (1), anemoside B4 (2), 23-hydroxybetulinic acid (3), cirenshenoside S (4), pulsatilloside B (5), pulsatilloside C (6), oleanolic acid (7), ajugasterone C (8) and β-ecdysterone (9), respectively. A Sapphire C18 column (250 mm × 4.6 mm, 5 μm) and gradient elution were used during the analysis. The identification and quantification of the analytes were achieved on a hybrid quadrupole linear ion trap mass spectrometer. Multiple-reaction monitoring (MRM) scanning was employed for quantification with switching electrospray ion source polarity between positive and negative modes in a single run. All calibration curves showed good linearity (r(2) > 0.9948) within the test ranges. The intra and interday variations for nine analytes were less than 3.95 and 3.78%, respectively. The developed method was successfully applied to determine the investigated compounds in 15 batches of natural and cultured samples of P. cernua. The results indicated that the method was simple, rapid, specific and reliable, which is helpful to comprehensive evaluation of quality of P. cernua.

[Optimization of extraction technics of total saponins from Pulsatilla cernua].[Pubmed: 20845794]

The extraction condition of total saponins from Pulsatilla cenua by ultrasonic wave was optimized by single factor and orthogonal experiments.

A new triterpenoid saponin from Pulsatilla cernua.[Pubmed: 20336019]

A new triterpenoid saponin was isolated from Pulsatilla cernua, along with eight known triterpenoids and triterpenoid glycosides. The new compound was identified as 3-O-beta-D-glucopyranosyl-(1-->4)-alpha-L-arabinopyranosyl-bayogenin-28-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester (1) on the basis of 1D, 2D-NMR techniques, including COSY, HMBC, and HMQC correlations, MS analysis, as well as chemical methods.

Two new triterpenoid saponins from Pulsatilla cernua (Thunb.) Bercht. et Opiz.[Pubmed: 17994387]

Two new triterpenoid saponins, named cernuaside C and D, have been isolated from Pulsatilla cernua (Thunb.) Bercht. et Opiz. The structures of the two new triterpenoid saponins were elucidated as 3-O-beta-D-xylopyranosyl(1 --> 2)-[alpha-L-rhamnopyranosyl(1 --> 3)]-alpha-L-arabinopyranosyl oleanolic acid 28-alpha-L-rhamnopyranosyl(1 --> 4)-beta-D-glucopyranosyl(1 --> 6)-beta-D-glucopyranoside. (1) and 3-O-alpha-L-arabinopyranosyl (1 --> 3)-alpha-L-rhamnopyranosyl(1 --> 2)-alpha-L-arabinopyranosyl oleanolic acid 28-O-beta-D-glucopyranoside (2) by 1D, 2D-NMR techniques, ESI-MS analysis as well as chemical methods.