Identification of bioactive phenolics from Porana sinensis Hemsl. stem by UPLC-QTOF-MS/MS and the confirmation of anti-inflammatory indicators using LPS-induced RAW264.7 cells.

To characterise bioactive phenolics and confirm anti-inflammatory indicators in Porana sinensis stem, 23 phenolics were identified by UPLC-QTOF-MS/MS from crude extract (CE) prepared optimally with 80% methanol. Further fractionalisation using D101 macroporous resin resulted in predominant enrichment of total phenols and flavonoids into Fr.II. Correspondingly, the bioactive components-enriched Fr.II exhibited the lowest IC50 for scavenging DPPH and ABTS and the highest oxygen radical absorbance capacity or ORAC followed by Fractions Fr.I + Fr.II, CE and Fr.I, implying that certain phenolics possessing lower antioxidant activity completely remained in CE. Anti-inflammatory tests with LPS-stimulated RAW264.7 cells showed that CE possessed the highest inhibition of NO-production followed by Fr.II and Fr.I, meaning that CE might contain compounds that expressed higher anti-inflammatory but lower antioxidant activities or possessed synergistic interactions but were not fractionated together. Quantitative determination of nine major phenolics revealed that caffeic acid and 3-, 4- and 5-caffeoylquinic acids were concentrated into Fr.I, whereas scopolin, scopoletin and 3,5-, 3,4- and 4,5-dicaffeoylquinic acids were enriched into Fr.II. Further experiments with three selected major phenolics reduced the proposed synergistic interactions. Anti-inflammatory tests of the nine major phenolics evidenced that caffeic acid and the six caffeoylquinic acids produced higher, and the three dicaffeoylquinic acids at 140 µΜ showed even more significant activities in suppressing NO-production and mRNA expression of iNOS, TNF-α, COX-2, and IL-6, suggesting that these three dicaffeoylquinic acids could be indicators of the anti-inflammatory potential of P. sinensis stem. These findings provided novel insights for potential use of P. sinensis or liana, as an important source of natural antioxidants, against inflammation.

Optimization of the Purification Technology of Total Flavones in Ampelopsis grossedentala with Macropo-rous Resins by Central Composite Design-Response Surface Methodology / 中国药房

OBJECTIVE:To optimize the purification technology of total flavones from Ampelopsis grossedentala with macro-porous resins. METHODS:4 kinds of macroporous resins for the purification of total flavones from A. grossedentala were screened by using drug-loading amount,desorption rate,recovery and purification rate as indicators. Single factor test and central composite design-response surface methodology were used to optimized eluant mass fraction,adsorption time,flow rate of eluant,eluant pH and other factors of purification technology,and validation test was also conducted. RESULTS:D-101 macroporous resin was the best. The optimal condition was as follows as the concentration of sample solution 2 mg(by extract weight)/ml,the volume of sam-ple solution 1.1 BV,ethanol 86.0%,adsorption time 36.7 min,flow rate of eluant 3.81 BV/h,pH 7. In validation test,mass frac-tion of total flavones increased from 66.83% to 85.00% in validation test(RSD=0.15%,n=3),and were close to predicted val-ue(85.08%). CONCLUSIONS:Central composite design-response surface methodology is feasible and stable for the optimization of purification technology of total flavones from A. grossedentala with macroporous resins.

Preparation technology of isochlorogenic acids A, B, and C / 中草药

Objective: To establish a method for separation of isochlorogenic acids A, B, and C from Lonicerae Flos. Methods: Isochlorogenic acids A, B, and C in Lonicerae Flos were isolated and purified by macroporous resin and medium-low-pressure preparative chromatography. Their structures were identified on the basis of the spectral data and physicochemical property. Results: The contents of prepared isochlorogenic acids A, B and C were 98.7%, 99.2%, and 97.6%, respectively. Conclusion: This method is economic, simple, rapid, and effective for the preparation of isochlorogenic acids A, B, and C with high purity.

Pharmacodynamic Optimization of Purification Technique for Acanthopanax Senticosus Harms / 中国药房

OBJECTIVE: To optimize the purification technique for the extract of Acanthopanax senticosus Harms. METHODS: The techniques of D- 101 macroporous resin adsorption, polyamide adsorption and n- butanol extraction were separately applied to purify the aqueous extract of Acanthopanax senticosus Harms. Syringin was detected by HPLC and total flavone was detected by UV. The anti- cerebral ischemia effects of three purified samples were compared using rat MCAO model and mouse decapitation gape model. RESULTS: The three purified samples all showed anti- cerebral ischemia effects, with the sample purified via D- 101 macroporous resin adsorption having stronger effects than the other two. The content of Syringin was correlated with anti- cerebral ischemia effects. CONCLUSIONS: D- 101 macroporous resin adsorption is a be-tter technique for purifying the aqueous extract of Acanthopanax senticosus Harms.