Establishment of UPLC Fingerprint of Pyrrosia petiolosa from Southwest China and Content Determination of Phenolic Acids Component / 中国药房

OBJECTIVE：To esta blish a UPLC fingerprint of Pyrrosia petiolosa from southwest China ，and to determine the contents of 4 kinds of phenolic acids （neochlorogenic acid ，caffeic acid ，chlorogenic acid and cryptochlorogenic acid ）. METHODS：The determination was performed on Waters Cortecs T 3 C18 column（100 mm×2.1 mm，1.6 μm）with mobile phase consisted of methanol- 0.1％ phosphoric acid （gradient elution ）at the flow rate of 0.35 mL/min. The detection wavelength was set at 326 nm. The column temperature was 30 ℃，and injection volume was 1 μL. UPLC method was used to establish the UPLC fingerprint of P. petiolosa in combination with the Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition）. Cluster analysis and principle component analysis （PCA）were performed by using SPSS 20.0 software. The contents of 4 kinds of phenolic acids in 20 batches of P. petiolosa were determined by external standard method. RESULTS ：There were 9 common peaks for the UPLC fingerprint of P. petiolosa . Peaks 1，3，4，5 and 9 were identified as neochlorogenic acid ，caffeic acid，chlorogenic acid ，cryptochlorogenic acid and isochlorogenic acid C ，respectively. RSDs of the relative retention time of each peak in different batches of P. petiolosa were 0-0.68％，and the RSDs of the relative peak area were 0-62.35％. The similarities between the fingerprint of 20 batches of medicinal materials and the control chromatogram were not less than 0.990. The result of cluster analysis showed that P. petiolosa from different regions could be sorted into three species. Results of PCA showed the differences among P. petiolosa from different regions. The linear range of neochlorogenic acid ，caffeic acid ，chlorogenic acid and cryptochlorogenic acid were 0.61-61.41，0.18-17.60，2.00-200.11，0.62-61.51 μ g/mL （R2＞0.999 9）. RSDs of precision ， reproducibility and stability tests were all lower than 2.00％. The recoveries were 96.23％-98.17％（RSD＝0.96％-2.28％， n＝6）. Among 20 batches of samples ，the contents of above 4 kinds of phenolic acids were 0.385 3-1.891 9，0.018 0-0.129 5，2.569 5-10.676 0，0.563.5-1.860 5 mg/g. CONCLUSIONS ： The established UPL C fingerprint could reflect the main chemical constituents of P. pedunculata . Phenolic acids could be used as the main evaluation indexes for the quality of P. petiolosa . The quality order of P. petiolosa from southwest China was Chongqing product＞Sichuan product ＞Guizhou product.

Molecular structure and phylogenetic analyses of the complete chloroplast genomes of three original species of Pyrrosiae Folium / 中国天然药物

Pyrrosia petiolosa, Pyrrosia lingua and Pyrrosia sheareri are recorded as original plants of Pyrrosiae Folium (PF) and commonly used as Chinese herbal medicines. Due to the similar morphological features of PF and its adulterants, common DNA barcodes cannot accurately distinguish PF species. Knowledge of the chloroplast (cp) genome is widely used in species identification, molecular marker and phylogenetic analyses. Herein, we determined the complete cp genomes of three original species of PF via high-throughput sequencing technologies. The three cp genomes exhibited a typical quadripartite structure with sizes ranging from 158 165 to 163 026 bp. The cp genomes of P. petiolosa and P. lingua encoded 130 genes, whilst that of P. sheareri encoded 131 genes. The complete cp genomes were compared, and five highly divergent regions of petA-psbJ, matK-rps16, ndhC-trnM, psbM-petN and psaC-ndhE were screened as potential DNA barcodes for identification of Pyrrosia genus species. The phylogenetic tree we obtained indicated that P. petiolosa and P. lingua are clustered in a single clade and, thus, share a close relationship. This study provides invaluable information for further studies on the species identification, taxonomy and phylogeny of Pyrrosia genus species.

Comparative pharmacognosy of Pyrrosia petiolosa and Pyrrosia davidii

Pyrrosia petiolosa (Christ) Ching, Polypodiaceae, is an important medicinal pteridophyte used for the treatment of nephritis and bronchitis, while P. davidii (Giesenhagen. ex Diels) Ching, Polypodiaceae, often substitutes medicinal Pyrrosia in clinic. The present study was aimed to compare the pharmacognosy of P. petiolosa and P. davidii, including plant morphology, microscopic characteristics, physico-chemical parameters, UV and IR spectrum, and HPLC fingerprint. It was revealed that the two herbs had basically similar pharmacognostical characteristics but with certain differences. The present study contributes to the standardization and verification of these medicinal materials.

Analysis on the Essential Oil of the Leaves of Pyrrosia petiolosa vs.Pyrrosia subfurfuracea / 中国药房

OBJECTIVE: To analyze the essential oils from the leaves of Pyrrosia petiolosa or Pyrrosia subfurfuracea.METHODS: The essential oils from the leaves of P.petiolosa or P.subfurfuracea were analyzed by head-space solid microextraction coupled with GC-MS(HS-SPME-GC-MS) on capillary column HP-5 MS (30.0 m?250 ?m?0.25 ?m).The carrier gas was helium(with a high purity of 99.999%) at a flow rate of 1.0 mL?min-1.The ionization mode: EI ionization source with electron energy of 70 eV.RESULTS: For the first time,39 constituents were identified from the leaves of P.petiolosa and 29 constituents were identified from the leaves of P.subfurfuracea.Of which,12 constituents were mutual in both samples.CONCLUSION: The components of the essential oil in leaves of the two Pyrrosia species bear some similarities,but the essential components of the two Pyrrosia species are different from each other fundamentally.Butylated hydroxytoluene in the leaves of P.petiolosa and caryophyllene in the leaves of P.subfurfuracea have medicinal value.