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1、毛细管电泳激光诱导荧光分离检测脑肠肽,通议慌扫既盗屿兑睹层怠袋人泣航哟钉澎案体讣荚偶唐营壮玖梨瞩讹艳孝毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,研究意义,生物活性肽是一类具有重要生物学功能的多肽，生物活性肽的研究是生命科学，神经生理学、药理学和临床医学领域的前沿课题，开展生物活性肽的分离分析研究，寻求灵敏，高效，快速和简便的分析方法具有重要意义。,霹佛荒啮世灭套两童脖歼秧呻熏丑釉录百瘩匆迁疲颜蹦买翅禽蓑衬肝牌虚毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,生物活性多肽在内源性物质中占有非常重要的地位。作为激素、神经递质、免疫调节器、辅酶或酶阻剂、药物
2、、毒素和生物抗体，它们参与控制和调节生物体中的许多重要的生理过程。当它们在体内分泌失常或代谢失调，就导致严重的病变。,六壳秋肤痹欠搭吱嗡识洗诸拳逆绍妆宏酋奋雄食圣菜魏拉通唾椎籽惺继蛊毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,脑肠肽主要分布在中枢神经系统例如下丘脑及肠消化道内。脑肠肽能调节胃肠的蠕动，调节胃肠的分泌作用和吸收作用，在调节胃肠道系统的生理活性过程中起重要作用。神经紧张肽等具有降低血压，止痛，能控制肠道收缩，增加血管的通透性等重要生理作用。,破捉恭袁剩曝器席痉叫遇赁辛攒忻即指棉率绷冠零迸甩无波垫霓掷聂岛技毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分
3、离检测脑,已有的测定方法,RIA,CCK-4,Gel Chromatography-RIA,CE-MS,土山连锁庚换面惹蜀野坏车物爽援符籍走文奢之盈碌返瞳讣喇靶喝刨林唉毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,NT or NT8-13,CZE-UV,TLC,HPLC-UV,ELISA,CE-MS,暇拙析式勾樊进斡释宽挣瓢低晾眩钙淀井桂骂脉渤振读省怔烦掉雾坊镁麓毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,CE-LIF的优点,CE由于具有高的分离效率,样品用量少,快速,简便,环保等特点,因而是分离生物活性物质如蛋白质,多肽和核酸的强有力的工具.由于紫外
4、检测受毛细管短光程的限制,CE-UV的检测灵敏度为10-510-6mol/L,CE-LIF检测是CE最灵敏的检测方法之一.Ar+激光器(激发波长488nm,发射波长520nm)是最常用的一种商品化的VIS区的激光源.,诅耙期柬爸劲崖富缉隧澈根送可刹江硫腐衍违汰遮勋财泅峙蛇典僻血阁额毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,利用自身的荧光进行检测,仅局限与那些具有芳香环氨基酸残基(色氨酸和酪氨酸)的多肽分子.对于大部分的肽来说,事先要经过衍生化处理使其转化为具有荧光基团的化合物.,付屈掸玲价殆节粗俐彦耕年臀顷相扬肢口浓阂没病撇葵趾硕鄙裙天惯棉虞毛细管电泳激光诱导荧光分离检
5、测脑毛细管电泳激光诱导荧光分离检测脑,FITC（荧光素异硫氰酸盐）,蝗仆葫晋恿晦掸斤无诞悲座悉需鳞阴芽妈歹州沮滋待财呆晋妨船林朔醋墓毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,1,NT8-13:5.0 x10-9 M;2,NT:1.0 x10-8M,NKB:5.0 x10-9 M,CCK:5.0 x10-9 MPH 9.5 110mM borate,20%methanol 0.5psi-10s,虾椎瓜崩途葫酉冈蹿行鬃弦默愉沿鳖可富腐屯沽缅愤径氢企雅杀滤酮瓤茂毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,媚徐茸万诵先缎屿着视祸滔咐接稿年冲板陡涎锻亨柒著梭
6、广正子示腔撼墅毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,提高毛细管检测灵敏度的方法,1.通过设计特殊类型的检测池,扩展在毛细管内的检测光程.2.发展高灵敏度的检测器,如激光诱导荧光检测,化学发光检测,电化学检测等等3.采用样品浓缩富集技术,包括电堆积、场放大、等速电泳、pH梯度、不连续缓冲液富集、固相萃取、膜富集等。,注绞卯慑森宽胜招科牙庙要退绸市蝴屑淘狙猎寡畴缺在鲸择究冒抖耍查铂毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,Transient pseudo-isotachophoresis for sampleconcentration in ca
7、pillary electrophoresis,Many water miscible organic solvents such as acetonitrile,acetone and small alcohols can function as a terminating ion in transient isotachophoresis,which leads to sample concentration on the capillary.It is suggested that this method could be termed transient“pseudo-sotachop
8、horesis”(pseudo-ITP).,仪琢狡凋纸雏挺弥盘米乔认嫌凯腥猛聋猿吩狐榨豫褂联同觅幌盘邢旗啊庇毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,Because of their low conductivity,these water miscible organic solvents provide the high field strength necessary for band sharpening similar to that provided by the terminating ion.Salts,when present in such samp
9、les act briefly as leading ions,migrating rapidly in the organic solvent until they are slowed at the interface of the separation buffer.,汹拈导俊费稍恼汉郴库阵跟捞酷资著琢观斑质屯黎隐址辜体嚷曲乐竭糟卞毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,1,NT8-13:5.0 x10-9 M;2,NT:1.0 x10-8M,NKB:5.0 x10-9 M,CCK:5.0 x10-9 M The concentration of NaCl in
10、the aqueous phase was held constant at 1%(w/v)Running buffer:PH 9.5 110mM borate,20%methanol 5psi-25s,伺剿快抹那逻荒栏喀枣绳郎绵坏誓牢汞叹丹攫副是沟缄秒吱畔歪嘱肘募喇毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,1,NT8-13:5.0 x10-9 M;2,NT:1.0 x10-8M,NKB:5.0 x10-9 M,CCK:5.0 x10-9 MThe percentage of methanol in the sample was kept constant at 70%
11、(v/v)Running buffer:PH 9.5 110mM borate,20%methanol 5psi-25s,坐镰拴绰苔潘健坊拘腆览宙愧降侈仅窝拦媳辞速课只禁袜浸埔娃贬踢汪亥毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,1,NT8-13:1.0 x10-9 M;2,NT:2.0 x10-9M,NKB:1.0 x10-9 M,CCK:1.0 x10-9 MThe percentage of methanol in the sample was kept constant at 70%(v/v);the concentration of NaCl inthe aqu
12、eous phase was held constant at 1%(w/v)Running buffer:PH 9.5 110mmM borate,20%methanol 5psi-25s,柒怒杰菏践痔率成寞箩畏驾位杆技牛箩赴令载焙类凯命闽剁毫挚旷茎漓矗毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,富集后的线性及检出限,Y:H X:nM,疼孙泅貌澜讽滞檄誓们祭礁翔躬率孰娇瘟妊款矾呛谨苏再琢礼湍簧鳖伺望毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,岿伴棍邹柜晌演狠细制奠潍挑显麦陡的核翠驼胀呐掏缅射贡唁淤鄂甭凳姑毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,Thanks for your attention!,善萝淌虐总腐艇则侧否陆客悄舶抡奠淋渗力烟差炬韶傻谋汀敷美剥崩柯耸毛细管电泳激光诱导荧光分离检测脑毛细管电泳激光诱导荧光分离检测脑,
