摘要
建立了梯度洗脱高效液相色谱法同时测定内源激素赤霉素(GA3)、吲哚乙酸(3-IAA)、脱落酸(ABA)、细胞分裂素(6-BA)和α-萘乙酸(α-NAA)等5种植物内源激素的方法,采用Hypersil ODS C18柱和紫外检测器,以甲醇、乙腈和0.6%的乙酸为流动相梯度洗脱,分别在各组分的保留时间段设置其最佳检测波长,在12min内可将上述5种内源激素的各组分分离完全,峰形良好,该方法的回收率可达到90%~96%,线性相关系数(r)大于0.998,检出限在0.02~0.3μg/g之间。还探讨了各组分的最佳检测条件和梯度洗脱存在的问题及解决方法。
A gradient elution method was proposed for the simultaneous determination of intrinsic hormones, such as gibberellin (GA3), 3-heteroauxing (3-IAA), 6-benzylaminopurine (6-BA), abscisic acide (ABA) and a-naphthalene (a-NAA) in plant growth regulator from plants, by HPLC with a Hypersil ODS C18 column and an ultraviolet alterable wavelength detector. The mobile phase, flowing at 0.8 mL/min, was progrannned to hold, 1 - 3 min, a linear gradient elution from a mixture of 50 : 5 : 45 ( V/ V/ V, % ) methanol, acetonitrile, and 0.6% acetic acid to 48 : 3 : 49 ( V/ V/V, % ) at 3 min, then followed by isocratic elution with this mixture for 3 rain, another linear gradient elution from 6 rain to a composition of 60 : 5 : 35 ( V/ V/ V, % ) at 9 rain, and finally followed by isocratic elution with this mixture for 3 min. With above method, 5 intrinsic hormones could be simultaneously separated and detected by ultraviolet with five settings of programmable wavelength (excitation/emission). The calibration curves showed a good linearity ( r 〉 0. 998) for all components in standard samples under the concentration range of 10 - 800 μg/g. The recovery and method detection limit (MDL) were 90% - 96% and 0.02 - 0.3μg/g, respectively. This research also discussed the best detection conditions for each intrinsic hormone and the problems with gradient elution and their solutions.
出处
《分析试验室》
CAS
CSCD
北大核心
2007年第9期51-55,共5页
Chinese Journal of Analysis Laboratory
基金
江苏省高校高新技术产业发展基金(JH03-036)
南京信息工程大学基金(Y616)项目资助
关键词
梯度洗脱
高效液相色谱
内源激素
植物源调节剂
Gradient elution
HPLC
Intrinsic hormones
Plant growth regulator from plants