CN101034080B - 适合便携式微芯片毛细管电泳设备的简易进样方法 - Google Patents
适合便携式微芯片毛细管电泳设备的简易进样方法 Download PDFInfo
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Abstract
一种适合便携式微芯片毛细管电泳设备的简易进样方法,它是将微芯片平放,将样品溶液注入样品池中,将磷酸盐(PBS)缓冲溶液注入样品废液池和缓冲液池中,缓冲液废液池中也注入缓冲溶液,并且与缓冲液池的液面高度基本一致,然后将微芯片倾斜进样,倾斜的角度为10°-25°,倾斜的方向是使样品池高,样品废液池低,进样40秒钟,将进样后的微芯片放平,同时施加分离电压进行电泳分离。本发明的进样方法简单易行,进样效果好,基本上不需要进样设备。
Description
技术领域
本发明涉及适合便携式芯片毛细管电泳设备的简易进样方法。具体地说,通过倾斜微芯片,样品池中的液面高度高于样品废液池的液面高度,于是产生液压差,实现了液压驱动进样。
背景技术
自从Manz等设计使用了平面玻璃芯片[参见:(a)D.J.Harrison,A.Manz,Z.Fan,H.Luedi and H.M.Widmer,Anal.Chem.,1992,64,1926-1932.(b)A.Manz,D.J.Harrison,E.M.J.Verpoorte,J.C.Fettinger,A.Paulus,H.Ludi and H.M.Widmer,J.Chromatogr.A,1992,593,253-258.],微芯片毛细管电泳就被广泛地研究应用[参见:(a)D.Mijatovic,J.C.T.Eijkel and A.van den Berg,Lab Chip,2005,5,492-500.(b)T.Vilkner,D.Janasek andA.Manz,Anal.Chem.,2004,76,3373-3386.(c)P.S.Dittrich,K.Tachikawa,and A.Manz,Anal.Chem.,2006,78,3887-3907.]。成功地进行分析的先决条件是精确地实现微量样品进样。科研工作投入了大量的精力和时间研究和改进进样方法,在所有已报道的进样方式中,基于电渗流驱动的电动进样最为主要法,但这种进样方式有两个缺点,进样状况受管道表面性质的影响非常大和对于不同淌度的组分进样存在偏差(bias)[参见:(a)J.P.Alarie,S.C.Jacobson and J.M.Ramsey,Electrophoresis,2001,22,312-317.(b)B.E.Slentz,N.A.Penner and F.Regnier,Anal.Chem.,2002,74,4835-4840.]。为了避免这种偏差进样,压力驱动进样得以应用,这种进样方式不受管壁表面和样品的性质的影响。Bai等提出了使用多通进样阀和三个注射泵组成的压力夹流进样方式,进样量可以达到纳升级[参见:X.Bai,H.J.Lee,J.S.Rossier,F.Reymond,H.Schafer,M.Wossner and H.H.Girault,Lab Chip,2002,2,45-49.]。Solignac和Gijs建立了一种利用机械驱动器,在储液池的隔膜上施加一个压力脉冲,实现了样品流体动力进样[参见:D.Solignac and M.A.M.Gijs,Anal.Chem.,2003,75,1652-1657.]。Zhang等建立了一种方法用于微芯片毛细管电泳的定量准确、没有偏差的皮升级的样品进样,样品的驱动是利用注射泵产生的负压、电渗流以及芯片储液池中不同液面高度差产生的液压共同作用[参见:L.Zhang,X.Yinand Z.Fang,Lab Chip,2006,6,258-264.]。Lin课题组报道了在微流控芯片上,通过液压与电动力相结合的进样方式[参见:H.Gai,L.Yu,Z.Dai,Y.Ma and B.C.Lin,Electrophoresis,2004,25,1888-1894.],这种进样方式主要的缺点在于存在峰形拖尾,拖尾的影响有时甚至对分离是致命的,使得样品不能被分离或定量。
还没有文献报道使用一种不需要外加设备且进样效果理想的简单进样方法。
发明内容
本发明的目的是提供一种适合便携式微芯片毛细管电泳设备的简易进样方法。
本发明的技术方案如下:
一种适合便携式微芯片毛细管电泳设备的简易进样方法,它由下列步骤组成:
步骤1.将微芯片平放,将样品溶液注入样品池中,将磷酸盐(PBS)缓冲溶液注入样品废液池和缓冲液池中,缓冲液废液池中也注入缓冲溶液,并且与缓冲液池的液面高度基本一致,
步骤2.然后将步骤1的微芯片倾斜进样,倾斜的角度为10°-25°,倾斜的方向是使样品池高,样品废液池低,进样40秒钟,
步骤3.将步骤2进样后的微芯片放平,同时施加分离电压进行电泳分离。
上述的进样方法,所述的步骤2,可以用一个固定角度的楔将微芯片倾斜。
本发明的进样方法简单易行,进样效果好,基本上不需要额外的进样设备。
附图说明
图1为本发明中测量电渗流时所应用的微芯片示意图,其中:A为缓冲液池,B为分离管道出口,C为样品池,D为样品废液池,E为进样管道和分离管道交叉口,F为缓冲液废液池,WE为电化学工作电极。
图2为本发明的实施例中500μM多巴胺和500μM儿茶酚的混合溶液实施液压进样后芯片电泳的分离电泳图,图中5纳安标尺说明峰高的强度,A为多巴胺,B为儿茶酚。
具体实施方式
实施例1.利用本发明的进样方式,以多巴胺和儿茶酚混合样品实施微芯片毛细管电泳分离测定。
利用500μM多巴胺和500μM儿茶酚的标准溶液来检验本发明的进样方法的效果。
将微芯片平放,将13.5μL的500μM多巴胺和500μM儿茶酚混合样品溶液放入样品池(C)中(液面高度大约为0.27cm),15μL20mM的磷酸盐(PBS)缓冲溶液放入样品废液池(D)和缓冲液池(A)中(液面高度大约为0.30cm),缓冲液废液池(F)中也放入20mM的PBS缓冲溶液,并且与缓冲液池(A)的液面高度基本一致。然后将微芯片倾斜进样,用一个固定角度17°的楔使微芯片倾斜17°。进样40秒后,将微芯片放平,同时施加分离电压800V进行电泳分离,在检测电位+1.4V(vs.Ag/AgCl电极)进行检测。得到电泳图,见图2。根据峰形和分离效果来看,进样效果完全满足芯片毛细管电泳的要求。
实施例2.
除了将楔的角度改成10°,使微芯片倾斜10°以外,其他都同实施例1,其分离效果与实施例1的分离效果相同。
实施例3.
除了将楔的角度改成25°,使微芯片倾斜25°以外,其他都同实施例1,其分离效果与实施例1的分离效果相同。
Claims (1)
1.一种适合便携式微芯片毛细管电泳设备的简易进样方法,其特征是它由下列步骤组成:
将微芯片平放,将13.5μL的500μM多巴胺和500μM儿茶酚混合样品溶液放入样品池(C)中,液面高度为0.27cm,15μL 20mM的磷酸盐缓冲溶液放入样品废液池(D)和缓冲液池(A)中,液面高度为0.30cm,缓冲液废液池(F)中也放入20mM的磷酸盐缓冲溶液,并且与缓冲液池(A)的液面高度基本一致,然后将微芯片倾斜进样,用一个固定角度17°的楔使微芯片倾斜17°,进样40秒后,将微芯片放平,同时施加分离电压800V进行电泳分离,以Ag/AgCl电极为参比电极,在检测电位+1.4V进行检测。
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凌云扬等.在线衍生微流控芯片电泳和激光诱导荧光法测定单细胞中谷胱甘肽.高等学校化学学报25.2004,25167-168. * |
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