手性化合物中纯化检测方案(制备液相色谱)

Released December 9， 2020Teledyne IscoP.O. Box 82531，Lincoln，Nebraska， 68501 USAToll-free： (800) 228-4373·Phone： (402) 464-0231·Fax： (402) 465-3091E-mail： Iscolnfo@teledyne.com Chiral Purifications with theChromatography Application NoteACCQPrep HP150 AN107 Abstract When there is a need to purify chiral compounds， theACCQPrep HP150 can be equipped with a chiral columnand can easily perform this type of purification. In thisapplication trans-stilbene oxide is used as a model forother chiral compounds since it is a standard for chiralpurification. Preparing the ACCQPrep HP150 To avoid possible damage to the chiral columns frominappropriate solvents， follow the steps below beforeattaching your column. Read the instructions for yourcolumn to determine which solvents are compatiblewith your column： When configuring the chiral column， set themaximum pressure limit in the ACCQPrepconfiguration as per the instructions that camewith the column. Setting this pressure limitprevents possible damage to the column. Since the ACCQPrep is generally used for reversephase separations， it is a good idea to prime thesystem with an intermediate solvent such as2-propanol first. This solvent is typically misciblewith both reverse phase solvents and the solventsmost often used for chiral. Remove the reverse phase column and replace thecolumn with a simple union so the columnposition can be bypassed and the entire fluid pathcan be flushed with 2-propanol. Use manual control to flush the injection loop andcolumn tubing. Remove and dry the solvent lines and then placethe lines into the solvents to be used for the chiralpurification. Prime the ACCQPrep with the solvents used for thepurification. Use manual control to flush the injection loop andtubing leading to the column with the solventsused for the chiral purification. If the system is equipped with an AutoSampler， fillthe wash station with methanol. Verify the Bracketed Injection Tube Station， ifused， contains a solvent compatible with thechiral column used (system equipped with theAutoSampler only). Experimental and Results All solvents were HPLC grade. The column was aChiralPak AS column， 21x250 mm， 20 p particle size. Thecolumn was run at 18 mL/min with a mobile phase of10% 2-propanol in hexanes.The trans-stilbene oxide wasdissolved in hexanes containing 10% 2-propanol. For thefirst run， 3.9 mg was injected while the second run used52.0 mg sample. Detection was 230 and 210 nm. All runsused an ACCQPrep HP150 system and AutoSampler (PN68-5230-097). Run 1： 3.9 mg injection Run 2： 52.0 mg injection Chromatography Application Note AN107 Both isomers were easily resolved with theACCQPrep HP150 system. Resolution betweencompounds determines the load capacity for a column.Columns， solvent systems and compounds willdetermine the actual load for any experiment. Conclusion The ACCQPrep HP150 can accurately deliver solvent，even at the low back-pressures generated by manychiral columns， to generate accurate isocraticpurifications. The ability to set a pressure limit fordifferent columns allows a mix of columns to run on thesystem， but prevents damage to the column in the caseof a clog or other over-pressure event. The AutoSampleroption allows unattended automated injection ofsamples， allowing many runs to be made to compensatefor the relatively low loading allowed by chiral columns.The active solvent and waste management also allowsfor unattended operations since the system willautomatically pause the run if a solvent is low or thewaste bottle is full. Active solvent management preventsloss of compound or damage to the column， while activewaste management prevents spills from over-filledwaste containers. ( Teledyne Isco is continually improving its products and reserves the right to change product specifications， replacement parts，schematics， and instructions without notice. ) 摘要当需要纯化手性化合物时，ACCQPrep HP150可以配备手性柱，可以很容易地进行这种类型的纯化。 由于反式二苯乙烯氧化物是手性纯化的标准物，因此它被用作其他手性化合物的模型。 ACCQPrep HP150的准备为了避免不合适的溶剂对手性色谱柱可能造成的损害，在安装色谱柱之前，请遵循以下步骤。 阅读柱的说明，以确定哪些溶剂与柱兼容: 配置手性色谱柱时，根据色谱柱附带的说明，在ACCQPrep配置中设置最大压力限制。 设置这个压力限制可以防止对柱造成可能的损害。  由于ACCQPrep通常用于反相分离，使用中间溶剂来启动系统是一个好主意，例如丙胺。 这种溶剂通常与反相溶剂和最常用的手性溶剂混相。  移除反相柱，用一个简单的接头替换柱，这样就可以绕过柱的位置，并可以用2-丙醇冲洗整个流体通道。  使用手动控制冲洗注入回路和柱状油管。  去除并干燥溶剂管路，然后将管路放入用于手性纯化的溶剂中。  在ACCQPrep上涂上用于净化的溶剂。  使用手动控制，用用于手性纯化的溶剂冲洗通向色谱柱的注射回路和管道。  如果系统配备了自动采样器，将清洗工位装满甲醇。  验证支架注射管站，如果使用，包含与所使用的手性柱兼容的溶剂(系统只配备自动进样器)。实验与结果所有溶剂均为HPLC级。 色谱柱为ChiralPak AS柱，尺寸为21x250mm，粒度为20 μ g。 色谱柱流速为18 mL/min，流动相为10%的2-丙醇正己烷。 反式二苯乙烯氧化物溶于含10% 2-丙醇的己烷中。 第一组注射3.9 mg，第二组注射52.0 mg。 检测波长为230、210 nm。 所有运行均使用ACCQPrep HP150系统和自动采样器(PN 68-5230-097)。 Run 1: 3.9 mg injectionRun Time(minutes)Run 2: 52.0 mg injectionRun Time(minutes)两种异构体均可通过ACCQPrep HP150系统轻松分离。 化合物之间的分辨率决定了柱的承载能力。 色谱柱，溶剂系统和化合物将决定任何实验的实际负载。结论ACCQPrep HP150可以准确地交付溶剂，即使在许多手性色谱柱产生的低背压，以产生准确的等压纯化。 为不同的色谱柱设置压力限制的能力允许混合色谱柱在系统上运行，但防止在堵塞或其他超压事件的情况下损坏色谱柱。 自动进样器允许无人值守的自动进样，允许多次下入，以补偿手性色谱柱允许的相对较低的负载。 活性溶剂和废物管理还允许无人值守操作，因为如果溶剂低或废物瓶满，系统将自动暂停运行。 积极的溶剂管理防止化合物的损失或损坏柱，而积极的废物管理防止溢出充满废物容器。 END