超高效液相色谱法测定不同产地甘草中甘草苷和甘草酸的含量
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摘要
目的建立超高效液相色谱法(ultraperformanceliquidchromatography,UPLC)同时测定甘草中甘草苷和甘草酸的含量,并对比不同产地甘草中甘草苷和甘草酸的含量。方法采用C_(18)柱(3.0mm×100mm,1.9μm)为分析柱,乙腈-0.05%磷酸水溶液为流动相梯度洗脱,流速0.4 mL/min,进样量2μL;柱温30℃,检测波长237 nm。结果甘草苷在8.7～217.4μg/mL浓度范围内与峰面积值呈良好的线性关系(r~2=1.0000),平均加样回收率为98.2%,相对标准偏差(relative standard deviation, RSD)为1.21%;甘草酸在8.0～200.7μg/mL浓度范围内与峰面积值呈良好的线性关系(r~2=0.9999),平均加样回收率为97.8%,RSD为0.94%。不同产地甘草中甘草苷和甘草酸含量有较大差异。结论该方法快速、准确、可靠,能有效分离甘草中杂质,可用于甘草中甘草苷和甘草酸的含量测定。
Objective To establish a method for simultaneous determination of liquiritin and glycyrrhizic acid in Gancao by ultra performance liquid chromatography(UPLC), and compare the content of liquiritin and glycyrrhizic acid in Gancao from different place. Methods The chromatographic column was C_(18)(3.0 mm×100 mm, 1.9 μm), the mobile phase was ACN-0.05% H3PO4 at the flow of 0.4 mL/min, the injection volume was 2 μL, the column temperature was 30 ℃ and the detection wavelength was 237 nm. Results Liquiritin had good linear relationships with the peak areas in the range of 8.7-217.4 μg/mL(r~2=1.0000), and the average recovery was 98.2%, with the relative standard deviation(RSD) of 1.21%. Glycyrrhizic acid had good linear relationships with the peak areas in the range of 8.0～200.7 μg/mL(r~2=0.9999), and the average recovery was 97.8%, with the RSD of 0.94%. The content of liquiritin and glycyrrhizic acid in Gancao from different place was quite different. Conclusion This method is rapid, accurate and reliable, and can effectively separate impurities from Gancao, which can be used for the determination of liquiritin and glycyrrhizic acid in Gancao.
Objective To establish a method for simultaneous determination of liquiritin and glycyrrhizic acid in Gancao by ultra performance liquid chromatography(UPLC), and compare the content of liquiritin and glycyrrhizic acid in Gancao from different place. Methods The chromatographic column was C_(18)(3.0 mm×100 mm, 1.9 μm), the mobile phase was ACN-0.05% H3PO4 at the flow of 0.4 mL/min, the injection volume was 2 μL, the column temperature was 30 ℃ and the detection wavelength was 237 nm. Results Liquiritin had good linear relationships with the peak areas in the range of 8.7-217.4 μg/mL(r~2=1.0000), and the average recovery was 98.2%, with the relative standard deviation(RSD) of 1.21%. Glycyrrhizic acid had good linear relationships with the peak areas in the range of 8.0～200.7 μg/mL(r~2=0.9999), and the average recovery was 97.8%, with the RSD of 0.94%. The content of liquiritin and glycyrrhizic acid in Gancao from different place was quite different. Conclusion This method is rapid, accurate and reliable, and can effectively separate impurities from Gancao, which can be used for the determination of liquiritin and glycyrrhizic acid in Gancao.
引文
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[2]姜雪,孙森凤,王悦,等.甘草药理作用研究进展[J].化工时刊,2017,31(7):25-28.Jiang X,Sun SF,Wang Y,et al.Research progress on pharmacological effects of licorice[J].Chem Ind Time,2017,31(7):25-28.
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[4]Eidi A,Nateghi Z,Mortazavi P,et al.Effects of ethanol extracts in licorice root(Glycyrrhiza glabra L.)on activity of liver enzymes in normal and alloxan-induced diabetic rats[J].?ibb-i Junūb,2016,19(4):526-535.
[5]Zhao H,Zhao M,Wang Y,et al.Glycyrrhizic acid prevents sepsis-induced acute lung injury and mortality in rats[J].J Histochem Cytochem,2016,64(2):125.
[6]Fang F,Xing W,Wang S,et al.Recent progress of medicinal value and method of extraction,separation of licorice[J].Mechan Electr Inf,2017,(5):39-47.
[7]Lim C,Lim S,Lee YJ,et al.Effects of glycyrrhizin PRE-treatment on transient ischemic brain injury in mice[J].African J Tradit Com Alt Med,2017,14(5):113-120.
[8]Ekanayaka SA,Mcclellan SA,Barrett RP,et al.Glycyrrhizin reduces HMGB1 and bacterial load in pseudomonas aeruginosa keratitis[J].Investigat Opthalmol Visual Sci,2016,57(13):5799-5809.
[9]Huang X,Wang Y,Ren K.Protective effects of liquiritin on the brain of rats with alzheimer’s disease[J].West Indian Med J,2016,64(5):468-472.
[10]Hosseinzadeh H,Nassiri-Asl M.Pharmacological effects of Glycyrrhiza spp.and its bioactive constituents:update and review[J].Phy Res,2015,29(12):1868-1886.
[11]赵月然,卢丹.HPLC法同时测定复方甘草片中甘草苷、异甘草苷、甘草素、甘草酸[J].中成药,2014,36(4):867-869.Zhao Y,Lu D.Determination of liquiritin,isoliquiritin,liquritigenin and glycyrrhizic acid in compound liquorice tablets by HPLC[J].Chin Tradit Patent Med,2014,36(4):867-869.
[12]魏宁,郎伟君.甘草中甘草酸和甘草苷的提取工艺研究[J].哈尔滨商业大学学报(自然科学版),2015,31(2):143-145.Wei N,Lang WJ.Study on extraction technology for glycyrrhizic acid and liquorice glycosides from Glycyrrhiza uralensis[J].J Harbin Univ Commerce(Natl Sci Ed),2015,31(2):143-145.
[13]朱金霞,郑国保,孔德杰,等.甘草中甘草苷、甘草酸及总黄酮的测定及其含量相关性研究[J].北方园艺,2014,(3):149-151.Zhu JX,Zheng GB,Kong DJ,et al.Study on the determination of liquiritin,glycyrrhizic acid and total flavonoids in licoric and their relationships[J].Northern Hortic,2014,(3):149-151.
[14]Chang SS,Jin-Ah L,Dayoung J,et al.Simultaneous determination of liquiritin,hesperidin,and glycyrrhizin by HPLC-photodiode array detection and the anti-inflammatory effect of Pyungwi-san[J].Arch Pharm Res,2011,34(2):203-210.
[15]Zhu T,Tian ML,Kyung HR.Comparison of adsorption equilibrium of glycyrrhizic acid and liquiritin on C18 column[J].J Ind Eng Chem,2010,16(6):929-934.
[16]胡汉芳,阎正,鲁杰,等.用比色法测定不同产地甘草中甘草酸的含量[J].河北大学学报(自然科学版),1998,(4):369-371.Hu HF,Yan Z,Lu J,et al.Determination of glycyrrhizic acid in glycyrrhiza uralensis from different origins by colorimetry[J].J Hebei Univ(Nat Sci Ed),1998,(4):369-371.
[17]谷会岩,宫立冬.不同产地栽培甘草中甘草酸含量的测定与比较[J].林业研究(英文版),2002,(2):141-143.Gu HY,Gong LD.Determination and comparison of glycyrrhizic acid content in cultivated licorice from different habitats[J].J Forest Res,2002,(2):141-143.
[2]姜雪,孙森凤,王悦,等.甘草药理作用研究进展[J].化工时刊,2017,31(7):25-28.Jiang X,Sun SF,Wang Y,et al.Research progress on pharmacological effects of licorice[J].Chem Ind Time,2017,31(7):25-28.
[3]王兵,王亚新,赵红燕,等.甘草的主要成分及其药理作用的研究进展[J].吉林医药学院学报,2013,34(3):215-218.Wang B,Wang YX,Zhao HY,et al.Research progress of the major components and the pharmacological effect of Glycyrrhiza uralensis fisch[J].J Jilin Med Coll,2013,34(3):215-218.
[4]Eidi A,Nateghi Z,Mortazavi P,et al.Effects of ethanol extracts in licorice root(Glycyrrhiza glabra L.)on activity of liver enzymes in normal and alloxan-induced diabetic rats[J].?ibb-i Junūb,2016,19(4):526-535.
[5]Zhao H,Zhao M,Wang Y,et al.Glycyrrhizic acid prevents sepsis-induced acute lung injury and mortality in rats[J].J Histochem Cytochem,2016,64(2):125.
[6]Fang F,Xing W,Wang S,et al.Recent progress of medicinal value and method of extraction,separation of licorice[J].Mechan Electr Inf,2017,(5):39-47.
[7]Lim C,Lim S,Lee YJ,et al.Effects of glycyrrhizin PRE-treatment on transient ischemic brain injury in mice[J].African J Tradit Com Alt Med,2017,14(5):113-120.
[8]Ekanayaka SA,Mcclellan SA,Barrett RP,et al.Glycyrrhizin reduces HMGB1 and bacterial load in pseudomonas aeruginosa keratitis[J].Investigat Opthalmol Visual Sci,2016,57(13):5799-5809.
[9]Huang X,Wang Y,Ren K.Protective effects of liquiritin on the brain of rats with alzheimer’s disease[J].West Indian Med J,2016,64(5):468-472.
[10]Hosseinzadeh H,Nassiri-Asl M.Pharmacological effects of Glycyrrhiza spp.and its bioactive constituents:update and review[J].Phy Res,2015,29(12):1868-1886.
[11]赵月然,卢丹.HPLC法同时测定复方甘草片中甘草苷、异甘草苷、甘草素、甘草酸[J].中成药,2014,36(4):867-869.Zhao Y,Lu D.Determination of liquiritin,isoliquiritin,liquritigenin and glycyrrhizic acid in compound liquorice tablets by HPLC[J].Chin Tradit Patent Med,2014,36(4):867-869.
[12]魏宁,郎伟君.甘草中甘草酸和甘草苷的提取工艺研究[J].哈尔滨商业大学学报(自然科学版),2015,31(2):143-145.Wei N,Lang WJ.Study on extraction technology for glycyrrhizic acid and liquorice glycosides from Glycyrrhiza uralensis[J].J Harbin Univ Commerce(Natl Sci Ed),2015,31(2):143-145.
[13]朱金霞,郑国保,孔德杰,等.甘草中甘草苷、甘草酸及总黄酮的测定及其含量相关性研究[J].北方园艺,2014,(3):149-151.Zhu JX,Zheng GB,Kong DJ,et al.Study on the determination of liquiritin,glycyrrhizic acid and total flavonoids in licoric and their relationships[J].Northern Hortic,2014,(3):149-151.
[14]Chang SS,Jin-Ah L,Dayoung J,et al.Simultaneous determination of liquiritin,hesperidin,and glycyrrhizin by HPLC-photodiode array detection and the anti-inflammatory effect of Pyungwi-san[J].Arch Pharm Res,2011,34(2):203-210.
[15]Zhu T,Tian ML,Kyung HR.Comparison of adsorption equilibrium of glycyrrhizic acid and liquiritin on C18 column[J].J Ind Eng Chem,2010,16(6):929-934.
[16]胡汉芳,阎正,鲁杰,等.用比色法测定不同产地甘草中甘草酸的含量[J].河北大学学报(自然科学版),1998,(4):369-371.Hu HF,Yan Z,Lu J,et al.Determination of glycyrrhizic acid in glycyrrhiza uralensis from different origins by colorimetry[J].J Hebei Univ(Nat Sci Ed),1998,(4):369-371.
[17]谷会岩,宫立冬.不同产地栽培甘草中甘草酸含量的测定与比较[J].林业研究(英文版),2002,(2):141-143.Gu HY,Gong LD.Determination and comparison of glycyrrhizic acid content in cultivated licorice from different habitats[J].J Forest Res,2002,(2):141-143.