HPLC-FLD法同时测定不同产地天麻中5个成分含量
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摘要
目的:建立高效液相色谱-荧光检测方法测定天麻中天麻素、对羟基苯甲醇、巴利森苷A、巴利森苷B、巴利森苷C的含量。方法:采用资生堂MGⅢ色谱柱(4.6 mm×250 mm,5μm),流动相为0.1%甲酸水(A)-乙腈(B),流速为1 mL·min~(-1),柱温35℃,进样量5μL,荧光检测器,激发波长为228 nm,发射波长为300 nm。结果:天麻素、对羟基苯甲醇、巴利森苷A、巴利森苷B、巴利森苷C的线性范围分别在10～5000(r=0.999 6)、10～5 000(r=0.999 9)、250～50 000(r=0.999 6)、250～50 000(r=0.999 9)、250～50 000 ng·mL~(-1)(r=0.999 7)范围内线性关系良好,平均回收率(n=6)分别为98.5%、97.2%、99.2%、98.7%、96.8%,RSD分别为5.8%、4.6%、0.86%、2.4%、4.8%;28批样品中上述5个成分的含量分别为0.5～20.0、0.1～2.5、0.5～15.4、1.6～8.2、0.7～2.1 mg·g~(-1)。结论:建立的方法杂质干扰少,分离度好,灵敏度高,线性范围宽,适于天麻成分分析及药材质量控制。
Objective:To establish a high-performance liquid chromatography with fluorescence detector method for the simultaneous determination of five components(gastrodin,4-hydroxybenzyl alcohol,parishin A,parishin B and parishin C)in Gastrodia Rhizoma.Methods:Chromatographic analysis was conducted on a Shiseido MG Ⅲcolumn(4.6 mm×250 mm,5 μm),with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as the mobile phase with gradient elution at a flow rate of 1.0 mL·min~(-1).The column temperature was 35 ℃ and the injection volume was 5 μL.The excitation wavelength and emission wavelength for fluorescence detector were 228 nm and 300 nm,respectively.Result:The linear ranges were within 10-5 000 ng·mL~(-1)(r=0.999 6)for gastrodin,10-5 000 ng·mL~(-1)(r=0.999 9)for 4-hydroxybenzyl alcohol,250-50 000 ng·mL~(-1)(r=0.999 6)for parishin A,250-50 000 ng·mL~(-1)(r=0.999 9)for parishin B and 250-50 000 ng·mL~(-1)(r=0.999 7)for parishin C.The average recoveries were 97.4%,96.9%,99.1%,98.5% and 96.3% with RSDs of 5.8%,4.6%,0.86%,2.4%and 4.8%,respectively.The contents of gastrodin,4-hydroxybenzyl alcohol,parishin A,parishin B and parishin C in 28 samples were 0.5-20.0,0.1-2.5,0.5-15.4,1.6-8.2 and 0.7-2.1 mg·g~(-1),respectively.Conclusion:The method is sensitive,accurate,and has the good repeatability in separation,which is available for the quality control of Gastrodia Rhizoma.
Objective:To establish a high-performance liquid chromatography with fluorescence detector method for the simultaneous determination of five components(gastrodin,4-hydroxybenzyl alcohol,parishin A,parishin B and parishin C)in Gastrodia Rhizoma.Methods:Chromatographic analysis was conducted on a Shiseido MG Ⅲcolumn(4.6 mm×250 mm,5 μm),with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as the mobile phase with gradient elution at a flow rate of 1.0 mL·min~(-1).The column temperature was 35 ℃ and the injection volume was 5 μL.The excitation wavelength and emission wavelength for fluorescence detector were 228 nm and 300 nm,respectively.Result:The linear ranges were within 10-5 000 ng·mL~(-1)(r=0.999 6)for gastrodin,10-5 000 ng·mL~(-1)(r=0.999 9)for 4-hydroxybenzyl alcohol,250-50 000 ng·mL~(-1)(r=0.999 6)for parishin A,250-50 000 ng·mL~(-1)(r=0.999 9)for parishin B and 250-50 000 ng·mL~(-1)(r=0.999 7)for parishin C.The average recoveries were 97.4%,96.9%,99.1%,98.5% and 96.3% with RSDs of 5.8%,4.6%,0.86%,2.4%and 4.8%,respectively.The contents of gastrodin,4-hydroxybenzyl alcohol,parishin A,parishin B and parishin C in 28 samples were 0.5-20.0,0.1-2.5,0.5-15.4,1.6-8.2 and 0.7-2.1 mg·g~(-1),respectively.Conclusion:The method is sensitive,accurate,and has the good repeatability in separation,which is available for the quality control of Gastrodia Rhizoma.
引文
[1]中华人民共和国药典2015年版.一部[S].2015:58ChP 2015.VolⅠ[S].2015:58
[2]王莉,肖红斌,梁鑫淼.天麻化学成分研究(Ⅰ)[J].中草药,2003,34(7):584WANG L,XIAO HB,LIANG XM.Studies on chemical constituents of Gastrodia elata[J].Chin Tradit Herb Drugs,2003,34(7):584
[3]李云,王志伟,刘大会,等.天麻化学成分研究进展[J].山东科学,2016,29(4):24LI Y,WANG ZW,LIU DH,et al.Research advances in chemical constituents of Gastrodia elata Blume[J].Shandong Sci,2016,29(4):24
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[5]杜伟锋,陈琳,丛晓东,等.天麻化学成分及质量控制研究进展[J].中成药,2011,33(10):1785DU WF,CHEN L,CONG XD,et al.Research progress on chemical composition and quality control of Gastrodia elata[J].Chin Tradit Patent Med,2011,33(10):1785
[6]唐大轩,杨文宝,梁娅君,等.天麻提取物对神经系统作用的药理研究[J].四川中医,2010,28(05):64TANG DX,YANG WB,LIANG YJ,et al.Pharmacological study of the action of Gastrodia elata extract on nervous system[J].Sichuan Tradit Chin Med,2010,28(5):64
[7]HAYASHI J,SEKINE T,DEGUCHI S,et al.Phenolic compounds from Gastrodia rhizome and relaxant effects of related compounds on isolated smooth muscle preparation[J].Phytochemistry,2002,59(5):513
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[9]周楠楠,朱燃,赵雪梅,等.天麻素抑制小鼠大脑内Aβ斑块形成及其作用机制[J].药学学报,2016,51(4):588ZHOU NN,ZHU R,ZHAO XM,et al.Effect and mechanism of gastrodin inhibitingβ-amyloid plaques in brain of mice[J].Acta Pharm Sin,2016,51(4):588
[10]王艳.天麻素对脂多糖诱导PC12细胞氧化应激保护作用的研究[D].昆明:昆明医科大学,2012WANG Y.The Rresearch on the Protective Function of Gastrodin to The Oxidative Stress Injury Induced by LPS in PC12 cells[D].Kunming:Kunming Medical University,2012
[11]LIN JH,LIU YC,HAU JP,et al.Parishins B and C from rhizomes of Gastrodia elata[J].Phytochemistry,1996,42(2):549
[12]谢淼,邵明莎,翟庆超,等.天麻中巴利森苷类成分研究进展[J].广东化工,2016,43(22):93XIE M,SHAO MS,ZHAI QC,et al.Research progress on rarishins from Gastrodia Elata[J].Guangdong Chem Ind,2016,43(22):93
[13]ZHAO Y,GONG XJ,ZHOU X,et al.Relative bioavailability of gastrodin and parishin from extract and powder of Gastrodiae rhizoma in rat[J].J Pharm Biomed Anal,2014,100(21):309
[14]TANG C,WANG L,LIU X,et al.Pharmacokinetic study of Gastrodia elata in rats[J].Anal Bioanal Chem,2015,407(29):8903
[15]陈琛,李新生,周建军,等.天麻素提取纯化及检测技术研究进展[J].陕西理工学院学报(自然科学版),2013,29(3):69CHEN C,LI XS,ZHOU JJ,et al.Research progress on extraction,purification and determination technology for gastrodin[J].JShaanxi Univ Technol(Nat Sci),2013,29(3):69
[16]刘玉红,易进海,陈燕,等.RP-HPLC法测定天麻药材中游离天麻素和总天麻素的含量[J].药物分析杂志,2010,30(1):30LIU YH,YI JH,CHEN Y,et al.RP-HPLC determination of free gastrodin and total gastrodin in Gastrodia elata[J].Chin J Pharm Anal,2010,30(1):30
[2]王莉,肖红斌,梁鑫淼.天麻化学成分研究(Ⅰ)[J].中草药,2003,34(7):584WANG L,XIAO HB,LIANG XM.Studies on chemical constituents of Gastrodia elata[J].Chin Tradit Herb Drugs,2003,34(7):584
[3]李云,王志伟,刘大会,等.天麻化学成分研究进展[J].山东科学,2016,29(4):24LI Y,WANG ZW,LIU DH,et al.Research advances in chemical constituents of Gastrodia elata Blume[J].Shandong Sci,2016,29(4):24
[4]李艳,蒋石,郭营营,等.天麻成分对羟基苯甲醇抗实验性脑血栓形成及抗炎作用研究[J].昆明医科大学学报,2015,36(1):28LI Y,JIANG S,GUO YY,et al.Anti-thrombotic and antiinflammatory effects of 4-hydroxybenzyl alcohol from Gastrodia elata[J].J Kunming Med Univ,2015,36(1):28
[5]杜伟锋,陈琳,丛晓东,等.天麻化学成分及质量控制研究进展[J].中成药,2011,33(10):1785DU WF,CHEN L,CONG XD,et al.Research progress on chemical composition and quality control of Gastrodia elata[J].Chin Tradit Patent Med,2011,33(10):1785
[6]唐大轩,杨文宝,梁娅君,等.天麻提取物对神经系统作用的药理研究[J].四川中医,2010,28(05):64TANG DX,YANG WB,LIANG YJ,et al.Pharmacological study of the action of Gastrodia elata extract on nervous system[J].Sichuan Tradit Chin Med,2010,28(5):64
[7]HAYASHI J,SEKINE T,DEGUCHI S,et al.Phenolic compounds from Gastrodia rhizome and relaxant effects of related compounds on isolated smooth muscle preparation[J].Phytochemistry,2002,59(5):513
[8]黄俊华,王桂莲.天麻注射液及天麻苷药理作用的初步研究[J].中国医学科学院学报,1985,7(5):399HUANG JH,WANG GL.Studies on some phamacologieal effects of Gastrodia injection and gastrodin[J].Acta Acad Med Sin,1985,7(5):399
[9]周楠楠,朱燃,赵雪梅,等.天麻素抑制小鼠大脑内Aβ斑块形成及其作用机制[J].药学学报,2016,51(4):588ZHOU NN,ZHU R,ZHAO XM,et al.Effect and mechanism of gastrodin inhibitingβ-amyloid plaques in brain of mice[J].Acta Pharm Sin,2016,51(4):588
[10]王艳.天麻素对脂多糖诱导PC12细胞氧化应激保护作用的研究[D].昆明:昆明医科大学,2012WANG Y.The Rresearch on the Protective Function of Gastrodin to The Oxidative Stress Injury Induced by LPS in PC12 cells[D].Kunming:Kunming Medical University,2012
[11]LIN JH,LIU YC,HAU JP,et al.Parishins B and C from rhizomes of Gastrodia elata[J].Phytochemistry,1996,42(2):549
[12]谢淼,邵明莎,翟庆超,等.天麻中巴利森苷类成分研究进展[J].广东化工,2016,43(22):93XIE M,SHAO MS,ZHAI QC,et al.Research progress on rarishins from Gastrodia Elata[J].Guangdong Chem Ind,2016,43(22):93
[13]ZHAO Y,GONG XJ,ZHOU X,et al.Relative bioavailability of gastrodin and parishin from extract and powder of Gastrodiae rhizoma in rat[J].J Pharm Biomed Anal,2014,100(21):309
[14]TANG C,WANG L,LIU X,et al.Pharmacokinetic study of Gastrodia elata in rats[J].Anal Bioanal Chem,2015,407(29):8903
[15]陈琛,李新生,周建军,等.天麻素提取纯化及检测技术研究进展[J].陕西理工学院学报(自然科学版),2013,29(3):69CHEN C,LI XS,ZHOU JJ,et al.Research progress on extraction,purification and determination technology for gastrodin[J].JShaanxi Univ Technol(Nat Sci),2013,29(3):69
[16]刘玉红,易进海,陈燕,等.RP-HPLC法测定天麻药材中游离天麻素和总天麻素的含量[J].药物分析杂志,2010,30(1):30LIU YH,YI JH,CHEN Y,et al.RP-HPLC determination of free gastrodin and total gastrodin in Gastrodia elata[J].Chin J Pharm Anal,2010,30(1):30