正交試驗法優化制備用于小分子物質分離的聚(甲基丙烯酸縮水甘油酯-二乙烯基苯)型整體柱

馬 偉, 徐環昕,2, 劉坐鎮,2＊, 寧方紅,2

(1.華東理工大學生物反應器國家重點實驗室,上海200237;2.上海華震科技有限公司,上海200237)

正交試驗法優化制備用于小分子物質分離的聚(甲基丙烯酸縮水甘油酯-二乙烯基苯)型整體柱

馬 偉1, 徐環昕1,2, 劉坐鎮1,2＊, 寧方紅1,2

(1.華東理工大學生物反應器國家重點實驗室,上海200237;2.上海華震科技有限公司,上海200237)

以甲基丙烯酸縮水甘油酯(GMA)為單體,二乙烯基苯(DVB)為交聯劑,以環己醇和正十二醇為致孔劑,過氧化苯甲酰(BPO)為引發劑,直接以50mm×4.6mm色譜柱為模具,通過原位聚合制備聚(GMA-DVB)型整體柱。以GMA和DVB的體積比、環己醇和十二醇的體積比以及BPO占聚合物的質量分數為三因素,以分離苯和乙苯等小分子物質時的半峰寬分離度(R1/2)為考察指標,進行三因素三水平的正交試驗,通過測定整體柱的比表面積、孔徑和孔容分布對其進行表征。結果表明,制備整體柱的最優配方為V(GMA):V(DVB)∶V(環己醇)∶V(正十二醇)=0.825∶0.825∶1.32∶2.03,BPO的質量分數為0.7%。應用所制備的整體柱分離苯和乙苯等小分子物質,理論塔板數達到37 000塔板/m,R1/2值達到7.14,完全達到基線分離,分離時間小于10m in。該方法制備整體柱的重復性好,柱效較高,基本滿足商品化要求。

聚(甲基丙烯酸縮水甘油酯-二乙烯基苯);整體柱;制備;高效液相色譜;小分子物質

Abstract:Amonolithic column of macro porouspoly(glycidyl methacrylate-divinyl benzene)(poly(GMA-DVB))has been p repared by free radical polym erization within the confines of a Chromatographic stainless steel tube(50mm×4.6mm).For the best separation and low back pressure,orthogonal experiments were carried out with V(cyclohexanol)∶V(dodecanol),V(GMA)∶V(DVB)and BPO dosage as the three main factors.The characteristic feature of the column,including specific surface area,pore volume as well as pore diameter distribution,was studied by scanning electron microscopy(SEM),mercury intrusion porosimetry analysis and B ET analysis.The obtained optimump reparation conditions were that the volume ratio of GMA,DVB,cyclohexanol and dodecanol was0.825∶0.825∶1.32∶2.03and the B PO dosage was 0.7%(mass percentage),then it was heated at70℃for24h.U sing this m onolithic colum n,benzene and ethylbenzene and a drug of oxiracetam can be w ell separated on a high performance liquid Chromatographic(HPLC)system equipped with a ultraviolet(UV)detector at254 nm.A solution of acetonitrile-water(50∶50,v/v)for the separation of benzene and ethylbenzene,and acetonitrile-water(80∶20,v/v)for the separation of oxiracetam were used as mobile phases at a flow rate of1mL/m in.The theoretical p late number was37 000p lates/m and the resolution of peak width at half height(R1/2)was7.14.The separation time was less than10m in.The monolithic column prepared by this method is reproducible and has high column efficiency.It is an economical method to prepare monolithic column,which can be ap-plied to separate small molecules.

Key words:poly(glycidyl methacrylate-divinylbenzene);monolithic column;p reparation;high perform ance liquid Chromatography(HPLC);small molecular

整體柱(monolithic column),又稱棒柱(rod)、連續床(continuous bed)、無塞柱(fritless colum n),被人們稱為繼多聚糖、交聯與涂漬、單分散之后的第四代分離介質[1]。整體柱是利用有機或無機聚合方法在色譜柱內進行原位聚合的連續床固定相。它具有制備方法簡單、內部結構均勻、重現性好、有較高的柱效和可進行快速分離等優點,是近年來發展較快的一種色譜柱。……