UPLC同時測定墨旱蓮藥材中8種成分的含量
發(fā)布時間:2019-08-29 來源: 感恩親情 點擊:
[摘要]建立一種超高效液相色譜法同時測定墨旱蓮中的異槲皮苷、木犀草苷、異去甲蟛蜞菊內(nèi)酯、異綠原酸A、異綠原酸C、木犀草素、蟛蜞菊內(nèi)酯、芹菜素8種成分含量的方法。采用Waters Acquity UPLC BEH C18色譜柱(2.1 mm ×100 mm,1.7 μm),流動相A為乙腈,B為0.1%的甲酸水,梯度洗脫條件:0~4 min,10%~13% A;4~10 min,13%~16% A;10~13 min,16%~25% A;13~17 min,25%~28% A;17~20 min,28%~40% A;20~25 min,40%~95% A。流速:0.3 mL·min-1;柱溫35 ℃;檢測波長350 nm。研究結(jié)果表明各指標成分之間分離度良好,在選定的濃度范圍內(nèi)線性關(guān)系良好(r≥0.999 0),平均加樣回收率(n=6)在96.60%~103.4%,RSD為0.86%~2.4%。所建立的UPLC同時測定墨旱蓮藥材中8種指標成分的含量的方法回收率較好,重復(fù)性、穩(wěn)定性良好,為墨旱蓮的鑒別及質(zhì)量評價提供了科學(xué)依據(jù)。
[關(guān)鍵詞]墨旱蓮; 超高液相色譜法; 多指標成分; 含量測定
[Abstract]To establish an UPLC method for the simultaneous determination of 8 compounds in Eclipta Herba, such as isoquercitrin, luteoloside, demethylwedelolactone, isochlorogenic acid A, isochlorogenic acid C, luteolin, wedelolactone and apigenin. The experiment was performed with a Waters Acquity UPLC BEH C18 (2.1 mm×100 mm, 1.7 μm) column by gradient elution of 0.1% formic acid in water and acetonitrile: 0-4 min,10%-13% A; 4-10 min, 13%-16% A; 10-13 min, 16%-25% A; 13-17 min, 25%-28% A; 17-20 min,28%-40% A;20-25 min,40%-95% A. The flow rate was 0.3 mL·min-1 .The condition of was the colum temperature was maintained at 35 ℃ and the detected wavelength was set at 350 mm. 8 components were separated clearly by this method. Also a good linearity was obtained between the chosen concentration(r≥0.999 0). The measured data showed that the recovery rate range from 96.60%-103.4% (n=6) and their RSD values were 0.86%-2.4%. The method has high recovery rate, good reproducibility and stability. It provides a scientific basis for the identification and quality evaluation of Eclipta Herba.
[Key words]Eclipta Herba; UPLC; multi-component quantification; content determination
doi:10.4268/cjcmm20162116
墨旱蓮為菊科植物鱧腸Eclipta prostrate L.干燥地上部分[1],廣泛分布于河南、安徽、江蘇、江西、浙江、福建、廣東、廣西、湖南、湖北等地[2]。其始載于《新修本草》,有滋陰益腎涼血止血之功效,F(xiàn)代藥理研究證明,墨旱蓮具有保肝益腎[3-4]、調(diào)節(jié)免疫[5-6]、抗氧化[7-8]、抗菌抗炎[9]、抗瘧疾[10]及抗腫瘤[11-12]等作用,其主要化學(xué)成分主要有香豆草醚類、黃酮類、酚酸類等[13]。目前《中國藥典》2015年版對墨旱蓮藥材的質(zhì)量評價依賴于蟛蜞菊內(nèi)酯單一化學(xué)成分的含量測定[1]。然而傳統(tǒng)中藥的療效是多成分共同作用的結(jié)果,單一活性成分并不能代表藥材整體的質(zhì)量。近年來多指標評價墨旱蓮的活性成分的報道較少,且多為2種成分同時定量[14-15],運用方法多為HPLC,大多集中在某一類別化學(xué)成分的評價[16-18]。然而HPLC耗時長,其靈敏度較低,面對傳統(tǒng)中藥中低含量成分的檢測問題時常常束手無策。
UPLC具有耐高壓、快速分離、高靈敏度、高分離度等特點,能克服HPLC靈敏度低,分析時間長等缺點,在中藥等復(fù)雜體系的分離分析上具有明顯優(yōu)勢[18],被廣泛地應(yīng)用于中藥復(fù)雜體系的研究[19-20]。本實驗首次建立了UPLC同時測定墨旱蓮藥材中8種活性成分含量的測定方法,該方法經(jīng)試驗認證安全可靠簡便易行,重現(xiàn)性好,溶劑消耗少,為建立墨旱蓮藥材的質(zhì)量控制與評價標準提供借鑒。
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