中药中理化常数检测方案

智能文字提取功能测试中

2 Results and DiscussionVanquish UHPLC-UV MethodA Thermo Fisher Scientific BrandAB71212-EN 0714S To develop an improved method to resolve many different analyteswithin a mangosteen pericarp sample in a finite time period withimproved throughput， better resolution and enhanced peak capacityusing the Thermo ScientificVanquishUHPLC platform Introduction There is considerable interest in botanical supplementsdue to their purported health benefits. Mangosteen(Garcinia mangostana L) is a tropical fruit that isindigenous to Southeast Asia， where it has been histori-cally used to treat abdominal pain， diarrhea， dysentery，inflammation， wound infection， suppuration， and chroniculcer. Recently mangosteen has been proposed as ahomeopathic therapy in the treatment of Parkinson’sdisease. Such therapeutic benefits have been mostlyattributed to a unique family of compounds referred toas xanthones that are most abundant in the pericarp ofthe fruit. The structures of the five major xanthones，including a-mangostin， 3-isomangostin， gartanin，9-hydroxycalabaxanthone， and 8-desoxygartanin， arepresented in Figure 1. H.CCH. Figure 1. Structures of selected xanthones found in mangosteen. The chromatographic analysis of the xanthones and otherpotentially important analytes contained in this supple-ment presents a huge challenge. Although reversed phaseHPLC with UV detection is widely used for the analysis ofxanthones， such methods lack analyte resolution and/orrequire exceedingly long analysis time.4.5 These issues canbe improved， somewhat， by using UHPLC but even thensuch methods still require >25 mins to complete since theflow rates required to achieve optimal column efficiencygenerate exceedingly high back pressures， well beyondthose that can be used with the typical UHPLC system. Inorder to address this situation the new Vanquish UHPLCsystem， was developed. The Vanquish platform consists ofa binary parallel pump capable of operating at pressuresup to 1500 bar， autosampler and diode-array detection.Consequently， UHPLC columns containing smallerparticle size can be operated at high flow rates to improveanalyte resolution and sample throughput. The analyticalpower of the Vanquish system is typified by the improve-ments in analyte resolution and shorted analysis timeshown for the analysis of mangosteen pericarp. Methods Sample Preparation Equipment and Materials： · Thermo ScientificDionexASE 350 AcceleratedSolvent Extractor system · 10 mL stainless extraction cells · Cellulose filters Clear collection vials， 60 mL ·Thermo ScientificDionexASE PrepDE Diatomaceous Earth Accelerated Solvent Extraction Conditions Solvent： 95% ethanol lemperature： 80°C Static Time： 5 min Static Cycle： 4 Flush： 60% Purge： 90 s Extraction： Weigh 0.5 g of each mangosteen pericarp powder sampleand mix with diatomaceous earth. Transfer the mixtureinto a separate 10 mL stainless steel cell (equipped withtwo cellulose filters on the bottom) to nearly fill the cell.Extract the loaded cells with the above conditions andtransfer the extracts into separate 25 mL volumetric flasksto volume with 95% ethanol. Filter the extract with a0.2 um filter and dilute 50-fold with 50% acetonitrilebefore analysis. Liquid Chromatography Vanquish UHPLC Method Vanquish UHPLC system including： Binary Pump H (P/N VH-P10-A) ·Split Sampler HT (P/N VH-A10-A) ·Column Compartment H (P/N VH-C10-A) ·Diode Array Detector HL， 320 nm (P/N VH-D10-A) Column： Thermo ScientificAcclaim" 120 C18， 2.2 um， 2.1×250 mm Still Air Temperature： 45℃ Mobile Phase A： Water Mobile Phase B： Acetonitrile Injection Volume： 3.0pL Flow Cell： LightPipe， 10 mm Vanquish Gradient Method： Time Flow[mL/min1 %B Curve [min] -2.5 1.0 50 5 0.1 1.0 50 5 9.0 1.4 90 5 11.0 1.4 90 5 12.0 1.0 50 5 Data Analysis Thermo ScientificDionexChromeleonChromatography Data System software， 6.8 Minutes Figure 2. UV chromatogram showing separation of the extract of mangosteen pericarp powder sample and detection of xanthones using the Vanquishsystem with an Acclaim RSLC C18 column. The extracted sample was separated using an Acclaim120 C18， 2.2 um， 2.1×250 mm column on the Vanquishplatform. The analysis was now completed in 11 minutesusing gradient conditions with a flow ramp from 1.0 mL/min to 1.4 mL/min over the course of the chromatogramas illustrated in Figure 2. As the viscosity of the solventsdecreased the flow rate could be increased constantly togain additional speed for the analysis without a significantimpact on resolution. The pressure trace observed usingthese conditions exceeded 1100 bar during the run. Morethan 60 peaks were determined with an average peakwidth at half height of 2.21 seconds. This indicates thatan approximate peak capacity of 300 was obtained usingthe Vanquish system with the Acclaim 2.2 micron fullyporous UHPLC column. Conclusions This application describes an improved method achievedon the Vanquish platform that provides enhancedthroughput with better resolution and peak capacity.The analysis time was completed in 11 minutes on theVanquish platform using gradient conditions with a flowramp from 1.0 mL/min to 1.4 mL/min over the courseof the analysis. · When using the Acclaim 120 C18 column withthe Vanquish platform more than 60 peaks weredetermined with an average peak width at half heightof 2.21 seconds. This indicates that an approximatepeak capacity of 300 is achieved in 11 minutes. With the increased focus on the quality of analyticaldata and the need for valid authentication of rawmaterials and ingredients， methods that offerhigh resolution and fast throughput are extremelyimportant. Important actionable decisions concer-ning product quality can be made sooner so thatunacceptable products never leave the factory.Newer chromatographic tools such as the onesreported in this application note provide a quickway to verify product quality of complex samples. Acknowledgements The authors are grateful to Prof. Douglas Kinghorn(Department of Medicinal Chemistry and Pharmacognosy，College of Pharmacy， University of Illinois at Chicago) forhis generous donation of o-mangosteen. ( References ) ( 1.Pedraza-Chaverri，J； Cardenas-Rodriguez，N.； Orozco- Ibarra，M.； Perez-Rojas， J.M. M edicinal properties ofmangosteen (Garcinia mangostana). Food and Chemi-cal Toxicology. 2008， 46， 3227-3239. ) ( 2. Jung， H.A.； Su， B.N.；Keller， W . J.； Mehta， R.G.； andKinghorn， A.D. Antioxidant x a nthones from the pericarp of Garcinia mangostana (Mangosteen).J. Agric Food Chem 2006： 54，2077-2082. ) 3. Kosem， N.； Youn-Hee， H.； and Moongkarndi， P.Antioxidant and cytoprotective activities of methanolicextract from Garcinia mangostana hulls. Sci Asia. 2007，33， 83-292 4. Walker， E.B. HPLC analysis of selected xanthones inmangosteen fruit. J. Sep Sci. 2007， 30， 1229-34. ( 5. Pothitirat， W. and Gritsanapan， W.HPLC quantitative analysis for the determination of o-mangostin in mangosteen fruit rind extract. Thai J. 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