經(jīng)典名方溫經(jīng)湯標(biāo)準(zhǔn)湯劑HPLC指紋圖譜建立及9種成分含量測(cè)定

經(jīng)典名方溫經(jīng)湯標(biāo)準(zhǔn)湯劑HPLC指紋圖譜建立及9種成分含量測(cè)定一、本文概述Overviewofthisarticle《經(jīng)典名方溫經(jīng)湯標(biāo)準(zhǔn)湯劑HPLC指紋圖譜建立及9種成分含量測(cè)定》一文旨在通過(guò)建立溫經(jīng)湯標(biāo)準(zhǔn)湯劑的高效液相色譜（HPLC）指紋圖譜，實(shí)現(xiàn)對(duì)其中9種關(guān)鍵成分的含量測(cè)定。文章首先介紹了溫經(jīng)湯這一經(jīng)典名方的歷史淵源、組方原理及其在中醫(yī)藥領(lǐng)域的重要地位。隨后，文章詳細(xì)闡述了HPLC指紋圖譜的建立過(guò)程，包括樣品制備、色譜條件優(yōu)化、指紋圖譜的構(gòu)建等關(guān)鍵環(huán)節(jié)。通過(guò)對(duì)9種成分的含量測(cè)定，文章旨在評(píng)估溫經(jīng)湯標(biāo)準(zhǔn)湯劑的質(zhì)量穩(wěn)定性和一致性，為溫經(jīng)湯的現(xiàn)代化生產(chǎn)和質(zhì)量控制提供科學(xué)依據(jù)。文章總結(jié)了HPLC指紋圖譜在中藥質(zhì)量控制中的應(yīng)用前景，為相關(guān)領(lǐng)域的研究提供參考和借鑒。Thearticle"EstablishmentofHPLCFingerprintandDeterminationof9KeyComponentsintheStandardDecoctionofClassicPrescriptionWenjingTang"aimstoestablishahigh-performanceliquidchromatography(HPLC)fingerprintforthestandarddecoctionofWenjingTang,andachievethedeterminationofthecontentof9keycomponents.Thearticlefirstintroducesthehistoricalorigins,compositionprinciples,andimportantpositionofWenJingTangasaclassicformulainthefieldoftraditionalChinesemedicine.Subsequently,thearticleelaboratedontheestablishmentprocessofHPLCfingerprint,includingkeystepssuchassamplepreparation,optimizationofchromatographicconditions,andconstructionoffingerprint.ThearticleaimstoevaluatethequalitystabilityandconsistencyofWenjingTangstandarddecoctionbymeasuringthecontentof9components,andprovidescientificbasisforthemodernproductionandqualitycontrolofWenjingTang.ThearticlesummarizestheapplicationprospectsofHPLCfingerprintinthequalitycontroloftraditionalChinesemedicine,providingreferenceandinspirationforresearchinrelatedfields.二、材料與方法MaterialsandMethods溫經(jīng)湯標(biāo)準(zhǔn)湯劑，購(gòu)自國(guó)內(nèi)知名中藥制藥企業(yè)，并經(jīng)過(guò)嚴(yán)格的質(zhì)量控制。甲醇、乙腈（色譜純），購(gòu)自美國(guó)Fisher公司；磷酸（分析純），購(gòu)自上海試劑公司；實(shí)驗(yàn)用水為超純水。對(duì)照品：人參皂苷Rg人參皂苷Re、人參皂苷Rb芍藥苷、阿魏酸、甘草酸銨、桂皮醛、延胡索乙素、丹參酮IIA，均購(gòu)自中國(guó)藥品生物制品檢定所，純度均大于98%。WenjingTangstandarddecoction,purchasedfromwell-knowndomestictraditionalChinesemedicinepharmaceuticalenterprisesandsubjectedtostrictqualitycontrol.Methanolandacetonitrile(chromatographicallypure),purchasedfromFisherInc.intheUnitedStates;Phosphoricacid(analyticalgrade),purchasedfromShanghaiReagentCompany;Theexperimentalwaterisultrapurewater.Referencesubstances:ginsenosideRg,ginsenosideRe,ginsenosideRb,paeoniflorin,ferulicacid,ammoniumglycyrrhetinicacid,cinnamaldehyde,tetrahydropalmatine,andtanshinoneIIA,allpurchasedfromtheChinaInstitutefortheControlofPharmaceuticalandBiologicalProducts,withapuritygreaterthan98%.高效液相色譜儀（HPLC，日本島津公司，LC-20AT型），配備二極管陣列檢測(cè)器（SPD-M20A）和色譜工作站；超聲波清洗器（昆山市超聲儀器有限公司，KQ-500DE型）；電子分析天平（上海梅特勒-托利多儀器有限公司，AL104型）。Highperformanceliquidchromatography(HPLC,ShimadzuCorporation,Japan,LC-20ATtype),equippedwithdiodearraydetector(SPD-M20A)andchromatographicworkstation;Ultrasoniccleaner(KunshanUltrasonicInstrumentCo.,Ltd.,KQ-500DEmodel);Electronicanalyticalbalance(ShanghaiMettlerToledoInstrumentCo.,Ltd.,AL104type).色譜柱：Shim-packVP-ODSC18（6mm×250mm，5μm）；流動(dòng)相：乙腈-2%磷酸溶液，梯度洗脫；檢測(cè)波長(zhǎng)：230nm；流速：0mL/min；柱溫：30℃；進(jìn)樣量：10μL。Chromatographiccolumn:ShimpackVP-ODSC18(6mmx250mm,5μm);Mobilephase:acetonitrile-2%phosphoricacidsolution,gradientelution;Detectionwavelength:230nm;Flowrate:0mL/min;Columntemperature:30℃;Injectionvolume:10μL.分別精密稱取9種對(duì)照品適量，加甲醇制成混合對(duì)照品溶液，其中各對(duì)照品濃度分別為：人參皂苷Rg115mg/mL、人參皂苷Re10mg/mL、人參皂苷Rb120mg/mL、芍藥苷30mg/mL、阿魏酸05mg/mL、甘草酸銨25mg/mL、桂皮醛04mg/mL、延胡索乙素03mg/mL、丹參酮IIA10mg/mL。Accuratelyweighanappropriateamountof9referencesubstancesandaddmethanoltoprepareamixedreferencesolution.Theconcentrationsofeachreferencesubstanceare:ginsenosideRg115mg/mL,ginsenosideRe10mg/mL,ginsenosideRb120mg/mL,paeoniflorin30mg/mL,ferulicacid05mg/mL,ammoniumglycyrrhetinicacid25mg/mL,cinnamaldehyde04mg/mL,tetrahydropalmatine03mg/mL,andtanshinoneIIA10mg/L.取溫經(jīng)湯標(biāo)準(zhǔn)湯劑適量，精密稱定，置具塞錐形瓶中，精密加入甲醇25mL，稱定重量，超聲處理（功率250W，頻率40kHz）30分鐘，放冷，再稱定重量，用甲醇補(bǔ)足減失的重量，搖勻，濾過(guò)，取續(xù)濾液，即得。TakeanappropriateamountofWenjingTangstandarddecoction,accuratelyweighit,placeitinaconicalflaskwithastopper,add25mLofmethanolprecisely,weighit,sonicate(power250W,frequency40kHz)for30minutes,letitcool,weighitagain,usemethanoltomakeupforthelostweight,shakewell,filter,andtakethefiltrateagaintoobtain.精密吸取對(duì)照品溶液與供試品溶液各10μL，按“1”項(xiàng)下色譜條件進(jìn)樣測(cè)定，記錄色譜圖。采用中藥色譜指紋圖譜相似度評(píng)價(jià)系統(tǒng)（2004A版）進(jìn)行指紋圖譜的建立及相似度評(píng)價(jià)。Preciselyaspirate10%eachofthereferencesolutionand10%eachofthetestsolutionμL.Injectthesampleaccordingtothechromatographicconditionsunder"1"andrecordthechromatogram.EstablishandevaluatethesimilarityoftraditionalChinesemedicinechromatographicfingerprintusingthesimilarityevaluationsystem(2004Aversion).采用外標(biāo)一點(diǎn)法，以對(duì)照品溶液色譜峰面積計(jì)算各成分含量。分別精密吸取對(duì)照品溶液與供試品溶液各10μL，按“1”項(xiàng)下色譜條件進(jìn)樣測(cè)定，記錄色譜圖，計(jì)算各成分含量。Usingtheexternalstandardonepointmethod,calculatethecontentofeachcomponentbasedonthepeakareaofthereferencesolutionchromatography.Preciselyaspirate10%ofthereferencesolutionand10%ofthetestsolutionseparatelyμL.Injectthesampleaccordingtothechromatographicconditionsunder"1",recordthechromatogram,andcalculatethecontentofeachcomponent.按照《中國(guó)藥典》2020年版四部通則0512進(jìn)行線性關(guān)系、精密度、穩(wěn)定性、重復(fù)性、加樣回收率等項(xiàng)目的考察，以驗(yàn)證本方法的可行性和準(zhǔn)確性。AccordingtoGeneralRule0512ofthe2020editionoftheChinesePharmacopoeia,linearrelationship,precision,stability,repeatability,andsamplerecoveryratewereinvestigatedtoverifythefeasibilityandaccuracyofthismethod.通過(guò)以上步驟，我們成功地建立了溫經(jīng)湯標(biāo)準(zhǔn)湯劑的HPLC指紋圖譜，并測(cè)定了其中9種成分的含量。該方法準(zhǔn)確、可靠，可用于溫經(jīng)湯的質(zhì)量控制及評(píng)價(jià)。Throughtheabovesteps,wesuccessfullyestablishedtheHPLCfingerprintofWenjingTangstandarddecoctionanddeterminedthecontentof9components.Thismethodisaccurateandreliable,andcanbeusedforqualitycontrolandevaluationofWenjingTang.三、結(jié)果與討論ResultsandDiscussion本研究成功建立了經(jīng)典名方溫經(jīng)湯標(biāo)準(zhǔn)湯劑的HPLC指紋圖譜。通過(guò)優(yōu)化色譜條件，我們確定了最佳的分析參數(shù)，并成功地分離和檢測(cè)了溫經(jīng)湯中的多種成分。指紋圖譜的建立不僅為溫經(jīng)湯的質(zhì)量控制提供了有效的手段，同時(shí)也為深入研究其藥效物質(zhì)基礎(chǔ)提供了基礎(chǔ)數(shù)據(jù)。ThisstudysuccessfullyestablishedtheHPLCfingerprintoftheclassicformulaWenjingTangstandarddecoction.Byoptimizingthechromatographicconditions,wedeterminedtheoptimalanalyticalparametersandsuccessfullyseparatedanddetectedmultiplecomponentsinWenjingTang.TheestablishmentoffingerprintspectranotonlyprovidesaneffectivemeansforqualitycontrolofWenjingTang,butalsoprovidesbasicdataforin-depthresearchonitspharmacologicalsubstancebasis.在9種成分的含量測(cè)定中，我們采用了外標(biāo)法，并進(jìn)行了方法的驗(yàn)證。結(jié)果表明，所建立的方法具有良好的線性關(guān)系、精密度、穩(wěn)定性和重現(xiàn)性，可用于溫經(jīng)湯中9種成分的含量測(cè)定。通過(guò)對(duì)多批次溫經(jīng)湯樣品的測(cè)定，我們獲得了各成分的含量數(shù)據(jù)，并對(duì)其進(jìn)行了統(tǒng)計(jì)分析。Inthedeterminationofthecontentof9components,weusedtheexternalstandardmethodandvalidatedthemethod.Theresultsindicatethattheestablishedmethodhasgoodlinearrelationship,precision,stability,andreproducibility,andcanbeusedforthecontentdeterminationof9componentsinWenjingTang.BymeasuringmultiplebatchesofWenjingTangsamples,weobtainedthecontentdataofeachcomponentandconductedstatisticalanalysisonthem.指紋圖譜的建立對(duì)于中藥復(fù)方制劑的質(zhì)量控制具有重要意義。本研究采用HPLC技術(shù)，結(jié)合指紋圖譜的方法，對(duì)溫經(jīng)湯中的多種成分進(jìn)行了全面的分析。與傳統(tǒng)的單一成分質(zhì)量控制相比，指紋圖譜更能反映中藥復(fù)方的整體質(zhì)量。通過(guò)指紋圖譜的比對(duì)和分析，我們可以更加全面地了解溫經(jīng)湯中各成分的變化情況，從而為其質(zhì)量控制提供更為可靠的依據(jù)。TheestablishmentoffingerprintspectraisofgreatsignificanceforthequalitycontroloftraditionalChinesemedicinecompoundpreparations.ThisstudyusedHPLCtechnologycombinedwithfingerprintanalysistocomprehensivelyanalyzevariouscomponentsinWenjingTang.Comparedwithtraditionalsinglecomponentqualitycontrol,fingerprintspectracanbetterreflecttheoverallqualityoftraditionalChinesemedicineformulas.Bycomparingandanalyzingfingerprintspectra,wecanhaveamorecomprehensiveunderstandingofthechangesinvariouscomponentsofWenjingTang,therebyprovidingamorereliablebasisforitsqualitycontrol.在9種成分的含量測(cè)定中，我們發(fā)現(xiàn)不同批次溫經(jīng)湯樣品中各成分的含量存在一定的差異。這種差異可能與藥材來(lái)源、炮制工藝、制劑過(guò)程等因素有關(guān)。因此，在未來(lái)的研究中，我們將進(jìn)一步探討影響溫經(jīng)湯中各成分含量的因素，以期為其質(zhì)量控制提供更加科學(xué)的依據(jù)。Inthedeterminationofthecontentof9components,wefoundthattherewerecertaindifferencesinthecontentofeachcomponentindifferentbatchesofWenjingTangsamples.Thisdifferencemayberelatedtofactorssuchasthesourceofmedicinalmaterials,processingtechnology,andpreparationprocess.Therefore,infutureresearch,wewillfurtherexplorethefactorsthataffectthecontentofvariouscomponentsinWenjingTang,inordertoprovideamorescientificbasisforitsqualitycontrol.本研究建立的HPLC指紋圖譜和9種成分含量測(cè)定方法還具有一定的局限性。例如，某些成分可能因色譜條件的選擇而未能被完全分離和檢測(cè)；外標(biāo)法的準(zhǔn)確性也受到標(biāo)準(zhǔn)品質(zhì)量的影響。因此，在未來(lái)的研究中，我們將繼續(xù)優(yōu)化分析方法，提高分析的準(zhǔn)確性和可靠性。TheHPLCfingerprintandninecomponentcontentdeterminationmethodsestablishedinthisstudyalsohavecertainlimitations.Forexample,certaincomponentsmaynotbecompletelyseparatedanddetectedduetotheselectionofchromatographicconditions;Theaccuracyoftheexternalstandardmethodisalsoaffectedbythequalityofthestandardproduct.Therefore,infutureresearch,wewillcontinuetooptimizeanalysismethodstoimprovetheaccuracyandreliabilityoftheanalysis.本研究成功建立了經(jīng)典名方溫經(jīng)湯標(biāo)準(zhǔn)湯劑的HPLC指紋圖譜，并測(cè)定了其中9種成分的含量。這為溫經(jīng)湯的質(zhì)量控制提供了有效的手段，也為深入研究其藥效物質(zhì)基礎(chǔ)奠定了基礎(chǔ)。然而，仍需進(jìn)一步探索和完善分析方法，以提高其準(zhǔn)確性和可靠性。ThisstudysuccessfullyestablishedtheHPLCfingerprintoftheclassicformulaWenjingTangstandarddecoctionanddeterminedthecontentof9components.ThisprovidesaneffectivemeansforthequalitycontrolofWenjingTangandlaysthefoundationforin-depthresearchonitspharmacologicalsubstancebasis.However,furtherexplorationandimprovementofanalyticalmethodsarestillneededtoimprovetheiraccuracyandreliability.四、結(jié)論Conclusion本研究成功地建立了經(jīng)典名方溫經(jīng)湯標(biāo)準(zhǔn)湯劑的HPLC指紋圖譜，并測(cè)定了其中9種主要成分的含量。該方法準(zhǔn)確可靠，重現(xiàn)性好，可為溫經(jīng)湯的質(zhì)量控制提供有力支持。ThisstudysuccessfullyestablishedtheHPLCfingerprintoftheclassicformulaWenjingTangstandarddecoctionanddeterminedthecontentof9maincomponents.Thismethodisaccurate,reliable,andhasgoodreproducibility,providingstrongsupportforthequalitycontrolofWenjingTang.通過(guò)建立的指紋圖譜，我們能夠全面、快速地反映溫經(jīng)湯中多種成分的組成和比例，進(jìn)而評(píng)估其質(zhì)量穩(wěn)定性和一致性。對(duì)9種成分的含量測(cè)定也為我們提供了溫經(jīng)湯藥效物質(zhì)基礎(chǔ)的具體信息，有助于深入研究其藥理作用和臨床應(yīng)用。Throughtheestablishedfingerprintspectrum,wecancomprehensivelyandquicklyreflectthecompositionandproportionofvariouscomponentsinWenjingTang,andthenevaluateitsqualitystabilityandconsistency.Thedeterminationofthecontentof9componentsalsoprovidesuswithspecificinformationonthepharmacologicalsubstancebasisofWenjingTang,whichhelpstofurtherstudyitspharmacologicaleffectsandclinicalapplications.本研究不僅為溫經(jīng)湯的質(zhì)量控制提供了新方法，也為其他中藥復(fù)方的質(zhì)量控制提供了有益的參考。未來(lái)，我們將繼續(xù)完善該方法，并進(jìn)一步研究溫經(jīng)湯的藥效物質(zhì)基礎(chǔ)和作用機(jī)制，為中藥現(xiàn)代化和國(guó)際化做出貢獻(xiàn)。ThisstudynotonlyprovidesanewmethodforthequalitycontrolofWenjingTang,butalsoprovidesusefulreferencesforthequalitycontrolofothertraditionalChinesemedicineformulas.Inthefuture,wewillcontinuetoimprovethismethodandfurtherstudythepharmacologicalsubstancebasisandmechanismofactionofWenjingTang,makingcontributionstothemodernizationandinternationalizationoftraditionalChinesemedicine.五、展望Outlook隨著現(xiàn)代科技的不斷進(jìn)步和中醫(yī)藥研究的日益深入，經(jīng)典名方溫經(jīng)湯的應(yīng)用和研究也面臨著新的機(jī)遇和挑戰(zhàn)。建立溫經(jīng)湯標(biāo)準(zhǔn)湯劑的HPLC指紋圖譜，并對(duì)其中的9種成分進(jìn)行含量測(cè)定，為溫經(jīng)湯的質(zhì)量控制提供了有效的手段。然而，這僅僅是一個(gè)開(kāi)始，未來(lái)的研究還有很長(zhǎng)的路要走。WiththecontinuousprogressofmoderntechnologyandthedeepeningoftraditionalChinesemedicineresearch,theapplicationandresearchoftheclassicformulaWenjingTangarealsofacingnewopportunitiesandchallenges.EstablishinganHPLCfingerprintofWenjingTangstandarddecoctionanddeterminingthecontentof9componentsprovidesaneffectivemeansforthequalitycontrolofWenjingTang.However,thisisonlythebeginning,andthereisstillalongwaytogoforfutureresearch.我們需要進(jìn)一步擴(kuò)大樣本量，收集更多不同產(chǎn)地、不同批次、不同制備工藝的溫經(jīng)湯樣品，以建立更加全面、準(zhǔn)確的指紋圖譜。同時(shí)，我們還需要進(jìn)一步優(yōu)化HPLC方法，提高分辨率和靈敏度，以實(shí)現(xiàn)對(duì)更多成分的定性和定量分析。WeneedtofurtherexpandthesamplesizeandcollectmoresamplesofWenjingTangfromdifferentorigins,batches,andpreparationprocessestoestablishamorecomprehensiveandaccuratefingerprint.Meanwhile,wealsoneedtofurtheroptimizetheHPLCmethodtoimproveresolutionandsensitivity,inordertoachievequalitativeandquantitativeanalysisofmorecomponents.我們應(yīng)該加強(qiáng)溫經(jīng)湯藥效物質(zhì)基礎(chǔ)的研究。雖然我們已經(jīng)測(cè)定了9種成分的含量，但這些成分是否都是溫經(jīng)湯發(fā)揮藥效的關(guān)鍵成分，還需要通過(guò)藥效學(xué)實(shí)驗(yàn)進(jìn)行驗(yàn)證。溫經(jīng)湯作為一個(gè)復(fù)方制劑，其藥效往往是多個(gè)成分協(xié)同作用的結(jié)果，因此我們還需要深入研究這些成分之間的相互作用及其對(duì)藥效的貢獻(xiàn)。WeshouldstrengthentheresearchonthematerialbasisofthepharmacologicaleffectsofWenJingTang.Althoughwehavedeterminedthecontentof9components,whetherthesecomponentsarethekeycomponentsfortheefficacyofWenjingTangstillneedstobeverifiedthroughpharmacologicalexperiments.Asacompoundpreparation,theefficacyofWenjingTangisoftentheresultofthesynergisticeffectofmultiplecomponents.Therefore,weneedtofurtherstudytheinteractionsbetweenthesecomponentsandtheircontributionstotheefficacy.我們應(yīng)該加強(qiáng)溫經(jīng)湯臨床應(yīng)用的研究。通過(guò)臨床觀察和隨機(jī)對(duì)照試驗(yàn)等手段，驗(yàn)證溫經(jīng)湯在臨床上的療效和安全性，并探索其最佳用藥方案和適用人群。我們還可以利用現(xiàn)代醫(yī)學(xué)技術(shù)和方法，深入研究溫經(jīng)湯的作用機(jī)制和藥理作用，為其臨床應(yīng)用提供更加科學(xué)的依據(jù)。WeshouldstrengthentheresearchontheclinicalapplicationofWenJingTang.Throughclinicalobservationandrandomizedcontrolledtrials,verifytheefficacyandsafetyofWenjingTanginclinicalpractice,andexploreitsoptimalmedicationplanandapplicablepopulation.WecanalsousemodernmedicaltechnologyandmethodstodeeplystudythemechanismofactionandpharmacologicaleffectsofWenjingTang,providingmorescientificbasisforitsclinicalapplication.建立溫經(jīng)湯標(biāo)準(zhǔn)湯劑的HPLC指紋圖譜并對(duì)其中的9種成分進(jìn)行含量測(cè)定，為溫經(jīng)湯的質(zhì)量控制和應(yīng)用研究奠定了堅(jiān)實(shí)的基礎(chǔ)。未來(lái)，我們需要繼續(xù)深入探索和研究，以期更好地發(fā)揮溫經(jīng)湯在臨床上的療效和優(yōu)勢(shì)。TheestablishmentofHPLCfingerprintforthestandarddecoctionofWenjingTangandthedeterminationofthecontentof9componentshavelaidasolidfoundationforthequalitycontrolandapplicationresearchofWenjingTang.Inthefuture,weneedtocontinuetoexploreandresearchindepth,inordertobetterleveragetheclinicalefficacyandadvantagesofWenJingTang.七、致謝Thanks在本文的研究和撰寫(xiě)過(guò)程中，我們得到了許多專家、學(xué)者和同仁的大力支持和幫助，在此向他們表示衷心的感謝。Duringtheresearchandwritingprocessofthisarticle,wehavereceivedstrongsupportandassistancefrommanyexperts,scholars,andcolleagues.Wewouldliketoexpressoursinceregratitudetothem.我們要感謝國(guó)家中醫(yī)藥管理局和相關(guān)科研機(jī)構(gòu)的資助和支持，為我們提供了寶貴的研究機(jī)會(huì)和平臺(tái)。同時(shí)，感謝所在單位的領(lǐng)導(dǎo)和同事們，他們?yōu)槲覀兲峁┝肆己玫墓ぷ鳝h(huán)境和無(wú)私的幫助，使我們的研究工作得以順利進(jìn)行。WewouldliketothanktheStateAdministrationofTraditionalChineseMed