CAJ | 학술논문

采用双三元液相色谱( Dual gradient liquid chromatography,DGLC)建立了在线固相萃取技术与电喷雾串联质谱联用方法( Online SPE DGLC-ESI MS/MS),并成功应用于实际样品检测。本方法同时检测大豆不同部位中的4种酸碱性植物激素(赤霉素(GA3)、吲哚乙酸(IAA)、玉米素(ZT)和脱落酸(ABA))。通过考察固相萃取富集柱、分析色谱柱、流动相对植物激素的保留和选择性的影响,获得较高的灵敏度、回收率、稳定性及精密度。大豆样品经液氮低温研磨,以80％甲醇溶液提取,再经离心稀释过滤后,进样分析。进样后样品经在线固相萃取Hypersep Retain AX柱洗脱保留,目标分析物依次转移至分析柱Acclaim PA2色谱柱,并以0.1％甲酸和甲醇溶液作为流动相进行梯度洗脱,采用选择反应监测离子模式( SRM)同时采集正负离子通道进行定性分析,基质标准曲线外标法进行定量分析, GA3, IAA, ZT 在0.1~50μg/L 范围内线性良好,检出限为0.0002μg/g；ABA在0.5~50μg/L的范围内线性良好,其检出限为0.0010μg/g。以0.8,4.0和40μg/L分别为低、中、高浓度考察4种植物激素的回收率为76.1％~93.5％,RSD为0.8％~6.0％。结果表明,籽粒中含有的ABA浓度明显高于其它部位。本研究为快速准确地分离和测定大豆不同部位内源激素提供了有效方法。
채용쌍삼원액상색보( Dual gradient liquid chromatography,DGLC)건립료재선고상췌취기술여전분무천련질보련용방법( Online SPE DGLC-ESI MS/MS),병성공응용우실제양품검측。본방법동시검측대두불동부위중적4충산감성식물격소(적매소(GA3)、신타을산(IAA)、옥미소(ZT)화탈락산(ABA))。통과고찰고상췌취부집주、분석색보주、류동상대식물격소적보류화선택성적영향,획득교고적령민도、회수솔、은정성급정밀도。대두양품경액담저온연마,이80％갑순용액제취,재경리심희석과려후,진양분석。진양후양품경재선고상췌취Hypersep Retain AX주세탈보류,목표분석물의차전이지분석주Acclaim PA2색보주,병이0.1％갑산화갑순용액작위류동상진행제도세탈,채용선택반응감측리자모식( SRM)동시채집정부리자통도진행정성분석,기질표준곡선외표법진행정량분석, GA3, IAA, ZT 재0.1~50μg/L 범위내선성량호,검출한위0.0002μg/g；ABA재0.5~50μg/L적범위내선성량호,기검출한위0.0010μg/g。이0.8,4.0화40μg/L분별위저、중、고농도고찰4충식물격소적회수솔위76.1％~93.5％,RSD위0.8％~6.0％。결과표명,자립중함유적ABA농도명현고우기타부위。본연구위쾌속준학지분리화측정대두불동부위내원격소제공료유효방법。
There are some common analysis challenges in the hormone detection in agriculture science, including matrix interference, complicated sample preparation, poor reproducibility, trace analyte content. An automated on-line SPE and innovative fast polarity switch analysis method employing dual-gradient liquid chromatography ( DGLC ) coupled with tandem mass spectrometry ( DGLC-MS/MS ) was established and validated for the simultaneous determination of gibberellic acid ( GA3 ) , indole acetic acid ( IAA ) , zeatin ( ZT) and abscisic acid ( ABA) in the soybean plant ( leaf, grain and pod) . The method was applied in the actual sample detection successfully. In order to acquire higher sensitivity, recovery, stability and precision, some conditions including SPE column, analytical column, mobile phase, additive etc were optimized according to the selection and retain of hormone. Beans were cryogenically grinded by liquid nitrogen, extracted by 80% methanol, certrifugatel and dilluted with water, and then injected directly. Samples were transported and gradient eluted on the analytical column Acclaim PA2 by 0 . 1% formic acid in water and methanol, after retaining and separation on the SPE column Hypersep Retain AX. All analytes were detected in selection reaction monitoring ( SRM) mode in both positive and negative channels. The quantification was based on linear regression. The linear ranges of GA3, IAA and ZT were 0. 1-50 μg/L with the LOQ of 0. 0002 μg/g, and the linear of ABA was 0. 5-50 μg/L with the LOQ of 0. 0010μg/g. The recoveries of four kinds of plants hormones were 76 . 1%-93 . 5%, and RSDs were 0 . 82%-6 . 02% at low ( 0 . 8 μg/L ) , medium (4. 0μg/L) and high (40μg/L). The results noted that the content of ABA in seeds was apparently higher than others. This method could be used for the rapid and accurate detection of hormone in different parts of soya beans.