高效液相色谱法测定药品中的二硫苏糖醇和氧化型二硫苏糖醇
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摘要
建立了高效液相色谱法同时测定药品中二硫苏糖醇(DTT)和氧化型二硫苏糖醇(DTTox)含量。采用Waters X-Bridge C18柱(250×4.6mm,5μm)分离,流动相为体积比为30∶70的甲醇-水(含0.02%三氟乙酸),检测波长为210nm。结果表明,DTT和DTTox在选定质量浓度范围内呈良好的线性关系(r>0.999),DTT和DTTox的检出限(S/N=3)为0.02μg/mL,定量限(S/N=10)为0.07μg/mL。平均加标回收率分别为99.32%、99.49%,相对标准偏差分别为2.34%、2.23%。该方法简单、准确,灵敏度高,重复性好,适用于药品中DTT和DTTox的检测。
A method was developed for the simultaneous determination of dithiothreitol(DTT)and oxidized dithiothreitol(DTTox)in medicine by high performance liquid chromatography(HPLC).The mobile phase consisted of methanol-water(30∶70,V/V)with 0.02% trifluoroacetic acid.The chromatographic conditions were as follows:flow rate,1.0 mL/min;the detection wavelength,210 nm;the column temperature,40℃;the injection volume,100μL.Under these conditions,good linear relationships between the mass concentration and the peak areas of DTT and DTTox were observed in the selected mass range(r>0.999).The limits of detection(LODs,S/N=3)and limits of quantitation(LOQs,S/N=10)were determined as 0.02μg/mL and 0.07μg/mL,respectively.The average recoveries were 99.32%for DTT and 99.49%for DTTox,and RSDs were 2.34%(n=6)for DTT and 2.23%(n=6)for DTTox.The results showed that the developed method was sensitive,accurate and reproducible.It is suitable for the analysis of DTT and DTTox in medicine.
A method was developed for the simultaneous determination of dithiothreitol(DTT)and oxidized dithiothreitol(DTTox)in medicine by high performance liquid chromatography(HPLC).The mobile phase consisted of methanol-water(30∶70,V/V)with 0.02% trifluoroacetic acid.The chromatographic conditions were as follows:flow rate,1.0 mL/min;the detection wavelength,210 nm;the column temperature,40℃;the injection volume,100μL.Under these conditions,good linear relationships between the mass concentration and the peak areas of DTT and DTTox were observed in the selected mass range(r>0.999).The limits of detection(LODs,S/N=3)and limits of quantitation(LOQs,S/N=10)were determined as 0.02μg/mL and 0.07μg/mL,respectively.The average recoveries were 99.32%for DTT and 99.49%for DTTox,and RSDs were 2.34%(n=6)for DTT and 2.23%(n=6)for DTTox.The results showed that the developed method was sensitive,accurate and reproducible.It is suitable for the analysis of DTT and DTTox in medicine.
引文
[1]YAN Han(闫晗),DONG Jun(董君),HUANG Jing,et al(黄静等).Applied Chemical Industry(应用化工)[J],2012,03:548.
[2]QIN Li(覃莉),JIANG Li(江莉),LI Mu-jun,et al(李慕军等).Guangxi Medical Journal(广西医学)[J],2013,35(4):406.
[3]YANG Liu(杨柳),LI Hai-lin(李海林),ZHANG Zhen-qian(张振乾).Crop Research(作物研究)[J],2015,01:11.
[4]XU Ke-jing(许克静),CHEN Yun(陈云),LEI Zhen-zhen,et al(雷珍珍等).Journal of Henan Agricultural Sciences(河南农业科学)[J],2013,05:38.
[5]Esposito Edward A,Craig T J.Biol Chem[J].2004,279(43):44270.
[6]Saint-Jean Maud de,Delfosse Vanessa,Douguet Dominique.J Cell Biol[J],2011,195(6):965.
[7]MA Yong(马勇),LI Hai-long(李海龙),SHI Feng(史锋).Journal of Zhejiang University(Science Edition)(浙江大学学报(理学版))[J],2008,35(1):92.
[8]ZHUGE Wei-shan(诸葛蔚山),CHEN Ya-wen(陈桠雯),JIN Xiu-er,et al(金秀儿等).Guangdong Trace Elements Science(广东微量元素科学)[J],2011,08:33.
[9]XU Hong-wei(许红玮),LIU Ke(刘轲),ZHANG Huai-liang(张怀亮).Chinese Journal of Experimental Traditional Medical Formulae(中国实验方剂学杂志)[J],2015,07:72.
[10]TU Feng-zhang(涂逢樟),YAO Hui-mei(姚辉梅),WU Lin-hua,et al(吴粦华等).Chinese Journal of Analysis Laboratory(分析试验室)[J],2010,04:99.
[11]WU Lin(吴琳).Seek Medical and Ask the Medicine(求医问药)[J]2012,11:1048.
[12]WANG Jin-tao(王进涛),LI Xiang-peng(李祥鹏),XUAN Guangshan(玄光善).Tianjin Pharmacy(天津药学)[J],2010,01:18.
[13]Pharmacopoeia Commission of People’s Republic of China(国家药典委员会).Pharmacopoeia of People’s Republic of China.Part 1(中华人民共和国药典.一部)[M].Beijing(北京):China Medical Science and Technology Press(中国医药科技出版社),2010:140.
[2]QIN Li(覃莉),JIANG Li(江莉),LI Mu-jun,et al(李慕军等).Guangxi Medical Journal(广西医学)[J],2013,35(4):406.
[3]YANG Liu(杨柳),LI Hai-lin(李海林),ZHANG Zhen-qian(张振乾).Crop Research(作物研究)[J],2015,01:11.
[4]XU Ke-jing(许克静),CHEN Yun(陈云),LEI Zhen-zhen,et al(雷珍珍等).Journal of Henan Agricultural Sciences(河南农业科学)[J],2013,05:38.
[5]Esposito Edward A,Craig T J.Biol Chem[J].2004,279(43):44270.
[6]Saint-Jean Maud de,Delfosse Vanessa,Douguet Dominique.J Cell Biol[J],2011,195(6):965.
[7]MA Yong(马勇),LI Hai-long(李海龙),SHI Feng(史锋).Journal of Zhejiang University(Science Edition)(浙江大学学报(理学版))[J],2008,35(1):92.
[8]ZHUGE Wei-shan(诸葛蔚山),CHEN Ya-wen(陈桠雯),JIN Xiu-er,et al(金秀儿等).Guangdong Trace Elements Science(广东微量元素科学)[J],2011,08:33.
[9]XU Hong-wei(许红玮),LIU Ke(刘轲),ZHANG Huai-liang(张怀亮).Chinese Journal of Experimental Traditional Medical Formulae(中国实验方剂学杂志)[J],2015,07:72.
[10]TU Feng-zhang(涂逢樟),YAO Hui-mei(姚辉梅),WU Lin-hua,et al(吴粦华等).Chinese Journal of Analysis Laboratory(分析试验室)[J],2010,04:99.
[11]WU Lin(吴琳).Seek Medical and Ask the Medicine(求医问药)[J]2012,11:1048.
[12]WANG Jin-tao(王进涛),LI Xiang-peng(李祥鹏),XUAN Guangshan(玄光善).Tianjin Pharmacy(天津药学)[J],2010,01:18.
[13]Pharmacopoeia Commission of People’s Republic of China(国家药典委员会).Pharmacopoeia of People’s Republic of China.Part 1(中华人民共和国药典.一部)[M].Beijing(北京):China Medical Science and Technology Press(中国医药科技出版社),2010:140.