油菜素甾醇是一类重要的植物激素，对植物的生长发育过程起显著调节作用。已报道的油菜素甾醇分析方法存在样品前处理过程复杂和灵敏度低等问题。本研究采用C18 PEEK管填充柱为富集柱，以4-N，N-二甲氨基苯硼酸为衍生试剂，搭建了基于双泵-双六通阀的在线管内固相微萃取-超高效液相色谱-串联质谱（ UPLC-MS/MS）联用系统，对油菜素甾醇进行在线富集和原位衍生，实现植物组织中内源性油菜素甾醇的自动化分析。该系统程序化地控制了油菜素甾醇的进样、萃取、衍生、分离和检测过程，有效简化前处理过程，节省人力；在线富集步骤实现了样品溶液大体积进样，提高样品利用率；原位衍生改善了油菜素甾醇电喷雾的离子化效率，使油菜素甾醇的电喷雾质谱检出限降低至 pg/mL。该系统可在300 mg鲜重植物组织中检测到内源性油菜素甾醇，已成功用于水稻、油菜中多种油菜素甾醇的定量分析。
Brassinosteroids( BRs)are a class of naturally occurring phytohormones with poly-hydroxy steroid structure,which regulate general plant growth and many physiological proces-ses. The reported methods for BR analysis were complicated,and the detection sensitivity was relatively low. To realize the automatic analysis of trace BRs in limited plant tissues,an in-tube solid phase microextraction-ultra performance liquid chromatography-tandem mass spectrome-try( SPME-UPLC-MS/MS)system was constructed based on two valves-two pumps. Using C18 PEEK column as the trapping column and 4-(dimethylamino)phenylboronic acid(4-DMAPBA) as the derivatization reagent,an on line trapping and in situ derivatization assay method of BRs was developed. BRs could be programmed to fulfill the procedures of injection,extraction, derivatization,LC separation and MS detection in the system. The detection limits of BRs were improved more than one order of magnitude by the online trapping and in situ derivatization techniques,thus endogenous BRs could be quantified in only 300 mg plant tissues.
参考文献
| [1] | BajguzA;Tretyn A.[J].Phytochem,200362(07):1027. |
| [2] | Mitchell JW;Mandava N;Worley JF.[J].NATURE,1970225(5237):1065. |
| [3] | Grove M D;Spencer GF;Rohwedder WK.[J].NATURE,1979281(5728):216. |
| [4] | Ashraf M;Akram NA;Arteca RN.[J].CRITICAL REVIEWS IN PLANT SCIENCES,201029(03):162. |
| [5] | Clouse S D;Sasse JM.[J].Annual Review of Plant Physiology and Plant Molecular Biology,199849:427. |
| [6] | Mertens R;Deus-Neumann B;Weiler EW.[J].FEBS Letters,1983160(1/2):269. |
| [7] | Yokota T;Watanabe S;Ogino Y.[J].J Plant Growth Reg-ul,19909(1/2/3/4):151. |
| [8] | Swaczynová J;Novák O;Hauserová E.[J].Journal of Plant Growth Regulation,200726(01):1. |
| [9] | Bajguz A.[J].Journal of Plant Physiology,2009166(17):1946. |
| [10] | SchrickK;Mayer U;Martin G.[J].Plant Journal,200231(01):61. |
| [11] | Khripach VA;Zhabinskii VN;Litvinovskaya R P.Immunoassays of Brassinosteroids[A].Netherlands:Springer,2011:375. |
| [12] | Yokota T;Watanabe S;Ogino Y.[J].Journal of Plant Growth Regulation,19909(01):151. |
| [13] | Horgen PA;Nakagawa C H;Irvin R T.[J].CANADIAN JOURNAL OF BIOCHEMISTRY AND CELL BIOLOGY,198462(08):715. |
| [14] | Gupta D;Bhardwaj R;Nagar PK.[J].Plant Growth Reg-ul,200443(02):97. |
| [15] | Takatsuto S;Ying B;Morisaki M.[J].Journal of Chromatography A,1982239:233. |
| [16] | Sasse JM.[J].Physiologia Plantarum,1997100(03):696. |
| [17] | Winter J;Schneider B;Meyenburg S.[J].Phytochem,199951(02):237. |
| [18] | GamohK;Takatsuto S.[J].Analytica Chimica Acta,1989222(01):201. |
| [19] | GamohK;Sawamoto H;Kakatsuto S.[J].Journal of Chromatography A,1990515:227. |
| [20] | SvatosA;AntonchickA;Schneider B.[J].Rapid Communications in Mass Spectrometry,200418(07):816. |
| [21] | HuoF;Wang X;Han Y.[J].TALANTA,201299:420. |
| [22] | Wu Q;Wu D;Shen Z.[J].Journal of Chromatography A,20131297:56. |
| [23] | Xin P;Yan J;Fan J.[J].Plant Physiology,2013162(04):2056. |
| [24] | 张慧娟;吴剑虹;冯钰锜.[J].分析科学学报,200824(05):589. |
| [25] | Atapattu SN;Rosenfeld JM.[J].Journal of Chromatography A,20131296:204. |
| [26] | Awan MA;Fleet I;Thomas C L P.[J].Food Chemistry,2008111(02):462. |
| [27] | Ito R;Kawaguchi M;Honda H.[J].Journal of Chromatography B,2008872(1/2):63. |
| [28] | Magi E;Di Carro M;Liscio C.[J].Analytical Biochemistry397(03):1335. |
| [29] | Zhang H J;Huang JF;Lin B.[J].Journal of Chromatography A,20071160(1/2):114. |
| [30] | Park YK;ChoiK;Ahmed A Y B H.[J].Journal of Chromatography A,1917(20):3357. |
| [31] | SalvadorA;Moretton C;PiramA.[J].Journal of Chromatography A,20071145(1/2):102. |
| [32] | Risticevic S;Niri V;Vuckovic D.[J].Analytical Biochemistry,2009393(03):781. |
| [33] | Fan Y;Feng Y Q;Da S L.[J].Analytica Chimica Acta,2004523(02):251. |
| [34] | Nie J;Zhang M;Fan Y.[J].Journal of Chromatography B,2005828(1/2):62. |
| [35] | 梁炳焕;文毅;吕永宁.[J].分析测试学报,200827(01):18. |
| [36] | 黄京芳;林幸华.[J].中国药学杂志,200944(12):941. |
| [37] | Fan Y;Feng Y Q;Zhang J T.[J].Journal of Chromatography A,20051074(1/2):9. |
| [38] | WeiF;Fan Y;Zhang M.[J].ELECTROPHORESIS,200526(16):3141. |
| [39] | Lin B;Zheng M M;Ng S C.[J].ELECTROPHORESIS,200728(15):2771. |
| [40] | Ding J;Mao L J;Wang S T.[J].Phytochemical Analysis,201324(04):386. |
| [41] | Springsteen G;Wang B H.[J].Tetrahedron,200258(26):5291. |
- 下载量()
- 访问量()
- 您的评分：
-
10%
-
20%
-
30%
-
40%
-
50%