1、氣泡溢出
流(liu)(liu)動(dong)相(xiang)內有(you)(you)氣(qi)(qi)泡(pao),關(guan)閉泵(beng),打(da)(da)開(kai)(kai)泄壓(ya)閥(fa),打(da)(da)開(kai)(kai)purg鍵,清(qing)洗脫(tuo)氣(qi)(qi),氣(qi)(qi)泡(pao)不(bu)(bu)斷(duan)從(cong)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)冒出(chu),進入流(liu)(liu)動(dong)相(xiang),無論(lun)打(da)(da)開(kai)(kai)purge鍵幾(ji)(ji)次(ci),都無法清(qing)除不(bu)(bu)斷(duan)產生(sheng)的(de)氣(qi)(qi)泡(pao)。原因過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)長(chang)期沉浸(jin)于(yu)乙酸(suan)銨等緩(huan)沖(chong)(chong)液(ye)內,過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)內部由于(yu)霉菌的(de)生(sheng)長(chang)繁殖,形成菌團,阻塞了過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi),緩(huan)沖(chong)(chong)液(ye)難以(yi)流(liu)(liu)暢地通過(guo)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi),空氣(qi)(qi)在(zai)泵(beng)的(de)壓(ya)力作用下經(jing)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)進入流(liu)(liu)動(dong)相(xiang)。處(chu)理過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)浸(jin)泡(pao)于(yu)5%硝酸(suan)溶(rong)(rong)液(ye)中(zhong),超(chao)聲清(qing)洗幾(ji)(ji)分鐘即可;亦可將(jiang)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)浸(jin)泡(pao)于(yu)5%硝酸(suan)溶(rong)(rong)液(ye)中(zhong)12～36小時,輕輕震蕩(dang)幾(ji)(ji)次(ci),再將(jiang)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)用純(chun)水清(qing)洗幾(ji)(ji)次(ci),打(da)(da)開(kai)(kai)泄壓(ya)閥(fa),打(da)(da)開(kai)(kai)purge鍵清(qing)洗脫(tuo)氣(qi)(qi),如仍有(you)(you)氣(qi)(qi)泡(pao)不(bu)(bu)斷(duan)從(cong)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)冒出(chu),繼續將(jiang)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)浸(jin)泡(pao)于(yu)5%硝酸(suan)溶(rong)(rong)液(ye)中(zhong),如沒(mei)有(you)(you)氣(qi)(qi)泡(pao)不(bu)(bu)斷(duan)從(cong)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)中(zhong)冒出(chu),說明過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)內部的(de)霉菌菌團已被硝酸(suan)破(po)壞,流(liu)(liu)動(dong)相(xiang)可以(yi)流(liu)(liu)暢地通過(guo)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)。打(da)(da)開(kai)(kai)泄壓(ya)閥(fa),打(da)(da)開(kai)(kai)泵(beng),流(liu)(liu)速調至1.0～3.0ml/min,純(chun)水沖(chong)(chong)洗過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)1小時左右。即可將(jiang)過(guo)濾(lv)(lv)器(qi)(qi)(qi)(qi)清(qing)洗干凈。關(guan)閉泄壓(ya)閥(fa),純(chun)甲醇沖(chong)(chong)洗半小時即可。
2、峰面積重復性(xing)不佳(jia)
(1)進樣閥漏液;
(2)加樣針不到位。
(3)液(ye)量不(bu)足. 處(chu)理(li)對(dui)(dui)于*種情況更(geng)換(huan)進(jin)樣(yang)(yang)閥墊(dian)圈;對(dui)(dui)于第二(er)種情況保證加樣(yang)(yang)針插到底(di),注射樣(yang)(yang)品(pin)溶液(ye)后(hou)須快(kuai)速、平穩地從LOAD狀態轉換(huan)到INJECT狀態,以保證進(jin)樣(yang)(yang)量的(de)(de)(de)準確。日(ri)常工作中(zhong),液(ye)相(xiang)色(se)譜(pu)儀(yi)的(de)(de)(de)保養(yang)非常重要(yao),如要(yao)注意不(bu)要(yao)讓空氣進(jin)入輸液(ye)系(xi)統(tong)和高(gao)壓泵中(zhong),儲(chu)液(ye)器(qi)內的(de)(de)(de)溶液(ye)如長時(shi)(shi)間未用(yong)(yong)應清洗儲(chu)液(ye)器(qi)并更(geng)換(huan)溶液(ye),每次用(yong)(yong)完(wan)色(se)譜(pu)儀(yi)后(hou)緩沖液(ye)要(yao)用(yong)(yong)純水沖洗干凈,防止無機鹽析出(chu)(chu)或沉積;樣(yang)(yang)品(pin)的(de)(de)(de)前處(chu)理(li)也很重要(yao),任何樣(yang)(yang)品(pin)都要(yao)盡可(ke)能地去除雜質,*溶解,盡量減少(shao)對(dui)(dui)色(se)譜(pu)柱(zhu)的(de)(de)(de)污(wu)染(ran),以延長色(se)譜(pu)柱(zhu)的(de)(de)(de)使用(yong)(yong)壽命(ming),同(tong)時(shi)(shi)避免注射過濃的(de)(de)(de)樣(yang)(yang)品(pin)溶液(ye),以免殘留(liu)液(ye)在(zai)進(jin)樣(yang)(yang)閥內析出(chu)(chu)固體引(yin)起堵塞;色(se)譜(pu)柱(zhu)作好標記,用(yong)(yong)于不(bu)同(tong)分(fen)析目的(de)(de)(de)的(de)(de)(de)色(se)譜(pu)柱(zhu)不(bu)要(yao)混(hun)用(yong)(yong)等。
3、柱壓高原因
(1)緩沖液鹽(yan)分如(乙酸銨等)沉積于(yu)柱內;
(2)樣品污染(ran)沉積。處理對于(yu)*種情(qing)況(kuang)先用(yong)(yong)40～50℃的(de)(de)純水(shui),低速正(zheng)向(xiang)(xiang)沖(chong)(chong)洗(xi)柱(zhu)子,待柱(zhu)壓逐(zhu)漸(jian)下(xia)降(jiang)(jiang)后,相(xiang)應提 高流速沖(chong)(chong)洗(xi),柱(zhu)壓大幅度下(xia)降(jiang)(jiang)后,用(yong)(yong)常溫純水(shui)沖(chong)(chong)洗(xi),之(zhi)后用(yong)(yong)純甲(jia)醇(chun)沖(chong)(chong)洗(xi)柱(zhu)子30分鐘;對于(yu)第二種情(qing)況(kuang),由樣品的(de)(de)沉積引起污染(ran)的(de)(de)C18柱(zhu),和純水(shui)反向(xiang)(xiang)沖(chong)(chong)洗(xi)柱(zhu)子,然后換成(cheng)甲(jia)醇(chun)沖(chong)(chong)洗(xi),接著(zhu)用(yong)(yong)甲(jia)醇(chun)+異(yi)丙醇(chun)(4+6)沖(chong)(chong)洗(xi)柱(zhu)子(沖(chong)(chong)洗(xi)時間的(de)(de)長短由樣品污染(ran)的(de)(de)情(qing)況(kuang)而定(ding)),再用(yong)(yong)換成(cheng)甲(jia)醇(chun)沖(chong)(chong)洗(xi),然后用(yong)(yong)純水(shui)沖(chong)(chong)洗(xi),zui后甲(jia)醇(chun)沖(chong)(chong)洗(xi)正(zheng)向(xiang)(xiang)沖(chong)(chong)洗(xi)柱(zhu)子30分鐘以上。
4、既(ji)無壓力(li)指示,又無液體(ti)流(liu)過(guo)
(1)泵密封墊(dian)圈(quan)磨(mo)損;
(2)大量氣泡進入泵體。處理對于*種情況,更換(huan)密封墊圈;對于第(di)二種情況,在泵作用(yong)的同時,用(yong)一個50ml的玻璃針筒在泵的出口(kou)處幫助(zhu)抽出空氣。
5、壓力波動(dong)大,流量不(bu)穩定
原因系統(tong)中有(you)(you)空(kong)氣(qi)或(huo)者單向閥(fa)(fa)(fa)的寶石球和(he)閥(fa)(fa)(fa)座(zuo)之間(jian)夾有(you)(you)異物,使得兩者不能密封。處(chu)理工作中注意觀察流(liu)動相的量,保證(zheng)不銹(xiu)鋼濾器沉入(ru)儲液器瓶底,避免吸入(ru)空(kong)氣(qi),流(liu)動相要充分脫氣(qi)。如為單向閥(fa)(fa)(fa)和(he)閥(fa)(fa)(fa)座(zuo)之間(jian)夾有(you)(you)異物,拆下(xia)單向閥(fa)(fa)(fa),放(fang)入(ru)盛(sheng)有(you)(you)丙(bing)酮(tong)的燒杯用超聲波清洗。
6、出峰不佳(jia),峰分叉(cha)
(1)色譜柱(zhu)被污染(ran);
(2)柱(zhu)(zhu)(zhu)(zhu)頭填(tian)(tian)(tian)料(liao)(liao)塌陷。處理對(dui)(dui)于*種(zhong)情況(kuang)(kuang),先用(yong)純(chun)水(shui)(shui)反(fan)向沖洗(xi)柱(zhu)(zhu)(zhu)(zhu)子(zi),然后換(huan)成(cheng)甲(jia)(jia)醇(chun)沖洗(xi),接著(zhu)用(yong)甲(jia)(jia)醇(chun)+異丙醇(chun)(4+6)沖洗(xi)柱(zhu)(zhu)(zhu)(zhu)子(zi)(沖洗(xi)時間的長短(duan)由樣品(pin)污染的情況(kuang)(kuang)而定),再(zai)換(huan)成(cheng)甲(jia)(jia)醇(chun)沖洗(xi),然后用(yong)純(chun)水(shui)(shui)沖洗(xi),zui后甲(jia)(jia)醇(chun)沖洗(xi)正向沖洗(xi)柱(zhu)(zhu)(zhu)(zhu)子(zi)30分(fen)鐘(zhong)(zhong)以(yi)上。如沖洗(xi)后依然出峰不佳,則考慮第二種(zhong)情況(kuang)(kuang)。對(dui)(dui)于第二種(zhong)情況(kuang)(kuang),擰(ning)開柱(zhu)(zhu)(zhu)(zhu)頭,檢查柱(zhu)(zhu)(zhu)(zhu)填(tian)(tian)(tian)料(liao)(liao)是否硬(ying)結或塌陷。去除硬(ying)結部(bu)分(fen)(污染的填(tian)(tian)(tian)料(liao)(liao)),裝入(ru)新填(tian)(tian)(tian)料(liao)(liao),滴一滴甲(jia)(jia)醇(chun),填(tian)(tian)(tian)料(liao)(liao)下(xia)陷,再(zai)填(tian)(tian)(tian),用(yong)與柱(zhu)(zhu)(zhu)(zhu)內(nei)徑相同的頂端平(ping)(ping)滑的不銹鋼桿(gan)壓緊,再(zai)填(tian)(tian)(tian)平(ping)(ping),滴甲(jia)(jia)醇(chun),再(zai)壓緊反(fan)復(fu)幾次,直至裝滿填(tian)(tian)(tian)平(ping)(ping)。柱(zhu)(zhu)(zhu)(zhu)頭用(yong)甲(jia)(jia)醇(chun)沖洗(xi)干凈,擦凈柱(zhu)(zhu)(zhu)(zhu)外(wai)壁的填(tian)(tian)(tian)料(liao)(liao),擰(ning)緊柱(zhu)(zhu)(zhu)(zhu)頭,用(yong)純(chun)甲(jia)(jia)醇(chun)沖洗(xi)30分(fen)鐘(zhong)(zhong)以(yi)上。
