二维液相色谱法在线检测哈茨木霉发酵液中2460 A含量

建立了在线检测哈茨木霉发酵液中微量2460A的二维液相色谱方法。利用Ultimate 3000双三元液相色谱仪,采用阀切换二维色谱技术,组合3根色谱柱实现2460A的在线净化、富集和含量检测。净化柱采用资生堂MF C8柱（10 mm×mm,5.0μm）,富集柱采用资生堂MGC18柱（20 mm ×mm,5.0μm）,以水-甲醇为流动相,梯度洗脱,流速2.0 mL/min；二维分析柱采用ThermoHypersil GOLD C18柱（250 mm ×mm,5.0μm）,以水-甲醇为流动相,梯度洗脱,流速1.0 mL/min；进样量1.0 mL；柱温40℃；检测波长424 nm。方法验证结果显示...

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Bibliographic Details
Published in分析化学 Vol. 42; no. 12; pp. 1828 - 1832
Main Author 山广志 周洁 左利民 姜威 刘桂霞 张洋 李元 姜蓉
Format Journal Article
LanguageChinese
Published 中国医学科学院医药生物技术研究所,北京,100050 2014
Subjects
2460A
二维液相色谱
发酵液
哈茨木霉
哈茨木霉
发酵液
Trichoderma hazianum
2460 A
2460A
Two-dimensional liquid chromatography
二维液相色谱
Fermentation liquor
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ISSN0253-3820
DOI10.11895/j.issn.0253-3820.140755

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Summary:建立了在线检测哈茨木霉发酵液中微量2460A的二维液相色谱方法。利用Ultimate 3000双三元液相色谱仪,采用阀切换二维色谱技术,组合3根色谱柱实现2460A的在线净化、富集和含量检测。净化柱采用资生堂MF C8柱（10 mm×mm,5.0μm）,富集柱采用资生堂MGC18柱（20 mm ×mm,5.0μm）,以水-甲醇为流动相,梯度洗脱,流速2.0 mL/min；二维分析柱采用ThermoHypersil GOLD C18柱（250 mm ×mm,5.0μm）,以水-甲醇为流动相,梯度洗脱,流速1.0 mL/min；进样量1.0 mL；柱温40℃；检测波长424 nm。方法验证结果显示,2460A的线性范围为0.0025～10.0 mg/L（r=0.9981, n=8）,检出限为1.2μg/L；定量限为2.5μg/L；方法回收率为88.0％～104.4％。
Bibliography:SHAN Guang-Zhi, ZHOU Jie, ZUO Li-Min, JIANG Wei, LIU Gui-Xia, ZHANG Yang, LI Yuan, JIANG Rong （ Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing 100050, China）
An on-line two dimensional liquid chromatographic （2D-LC） method was established by using an ultimate dual gradient liquid chromatography, three chromatographic columns and valve-switching technology to detect 2460A in trichoderma hazianum fermentation liquor. MF C8 column （ 10 mm×4.6 mm, 5.0μm） was used as purification column and MG C18 column （20 mm×4.6 mm, 5.0 μm） was used as enriching column. Methanol and water were used as mobile phase with a gradient elution at a flow rate of 2.0 mL/min. The sample was separated on the Thermo Hypersil GOLD C18eolumn （250 mm×4.6 mm, 5.0 μm） maintained at 40 ℃ using methanol and water. The flow rate was 1.0 mL/min and 1.0 mL sample was injected into the 2D-LC system. The detection wavelength was 424 nm. The whole analytical time was less than 60 min. The standard curve was linear ov
ISSN:0253-3820
DOI:10.11895/j.issn.0253-3820.140755