當液相色譜儀被汙染之後,就會出現數據錯誤等問題。今天,我們就和大家一起探討一下,如何判斷液相色譜儀是否被汙染以及係統 (HPLC) 中“攜帶汙染”(或俗稱“殘留”)發生的原因和減少的方法。
jixianbianhuajixianbianhuayibanbuhuihenyouguilv,ruguozaizidongliangchengxiajiankongjixianqingkuang,bukenengdedaoyuanlaiyiqilixiangzhuangtaideyouguilvdejixianzaigaopinlvshangxiafudongdebianhua,ershiyoushihouhuidedaodipinlvbianhuadefeng,kanshangquhenxiangyoudongxixituochulai,bingqiefengkuanbijiaoda,xianggongxituoxianxiangside。jixianpiaoyibuxiangyuanlainayangmanmandebianping,ershiyizhichaoyigefangxiangpiao,biruxiangshangpiao,pinghengyitianyekenengshiyizhizaipiao。zhuyabianhuazhuyabianhuayingxiangjiaoxiao,chufeishijiancechitaizang,dusaiyanzhong,huiduizhuyachanshengxiaoyu1000psi的影響(一般的用到光的檢測池也就耐這麼高的壓力),但dan這zhe種zhong情qing況kuang不bu會hui持chi續xu發fa生sheng,給gei你ni排pai查zha的de時shi間jian有you限xian,堵du的de時shi間jian長chang了le,會hui爆bao池chi,壓ya力li也ye就jiu沒mei有you影ying響xiang了le。峰feng麵mian積ji變bian化hua峰feng麵mian積ji應ying該gai是shi變bian小xiao。檢jian測ce池chi髒zang時shi會hui有you兩liang種zhong情qing形xing影ying響xiang檢jian測ce靈ling敏min度du。一yi方fang麵mian是shi基ji線xian噪zao聲sheng變bian大da,從cong整zheng體ti上shang影ying響xiang信xin噪zao比bi。另ling一yi方fang麵mian,檢jian測ce池chi髒zang了le對dui於yu光guang檢jian測ce原yuan理li的de檢jian測ce器qi來lai說shuo(UV、PDA、DAD、FLD、ELSD)shouxianhuijianruotongguojiancechideguangqiangdu,guangqiangdujiangdile,lingmindujiuhuixiajiang,xiangdangyujianceqiguangyuandenengliangjiangdi。baoliushijianyoushiyouyujianceqizangle,xitongyalizengjia,ranhoumougejiekoudedifangchuxianlouye,daozhibaoliushijianchuxianyixiebianhua,buguozhezhongkenengxingjiaoxiao。xiedaiwuranleixing、發生原因和減少方法“攜帶汙染”(或俗稱“殘留”)是用來描述一種樣品汙染的術語,它會導致分析物的色譜峰在之後的樣品分析中再次出現,而這些分析實際上並不包含樣品(例如空白樣或純溶劑)。這種汙染可持續數次連續分析,通常在每次進樣後汙染量都會減少(這是排除故障時的關鍵觀察點)。ruguotigongleshidangdeyiqipeixun,xiandaigaoxiaoyexiangsepuxitongdeshejihuishixiedaiwuranxianxiangbiandejiweihanjian,dandangxiedaixianxiangchuxianshi,wurankenengshiyouyuyixiayuanyinzaochengde:(1) 缺乏高效液相色譜維護;(2) 樣品超載,造成色譜柱殘留;(3) 洗滌瓶使用不當和/或樣品瓶選擇不當;(4) 操作員在如何設置和使用色譜係統方麵的培訓不足。注:shidangdecaozuoyuanpeixunkedadajiangdiwurandejilv,yeshizuirongyibeihushideyuanyin。yangpinchaozairuguozhurudeyangpinnongduguogao,shisepuzhuchaozai,namezaifenxijieshuhouhenchangshijiannei,sepuzhurengyoukenengjixushenchuyangpin。ruguomeiyoudingqichongxiheqingjiesepuzhu,yehuidaozhileisidewenti,yinweisuizheshijiandetuiyi,baoliudewuzhihuicongsepuzhuzhongshifangchulai。yaobimianzheyiwenti,shouxianyaojinxingzailiangyanjiu,quedingsepuzhushangnengyouxiaochengzaiduoshaowuzhi。ruguocaiyongdeshidengdufangfa,xuyaochuangjianyizhongqingxifangfa,shiyongbinindefangfagengqiangderongji(通常使用梯度),這zhe樣yang就jiu能neng在zai兩liang次ci分fen析xi之zhi間jian洗xi去qu任ren何he強qiang保bao留liu物wu質zhi,因yin為wei材cai料liao會hui保bao留liu在zai色se譜pu柱zhu上shang,必bi須xu每mei隔ge一yi段duan時shi間jian使shi用yong較jiao強qiang的de洗xi滌di液ye將jiang其qi洗xi掉diao。清qing洗xi瓶ping的de使shi用yong和he/或huo樣yang品pin瓶ping的de選xuan擇ze許xu多duo老lao式shi自zi動dong進jin樣yang器qi設she計ji使shi用yong低di壓ya進jin樣yang器qi,這zhe些xie進jin樣yang器qi需xu要yao單dan獨du的de清qing洗xi瓶ping,因yin為wei它ta們men不bu需xu要yao持chi續xu清qing洗xi。清qing洗xi瓶ping中zhong應ying注zhu入ru流liu動dong相xiang或huo溶rong液ye,以yi溶rong解jie係xi統tong中zhong可ke能neng殘can留liu的de任ren何he物wu質zhi。如ru果guo使shi用yong的de是shi現xian代dai高gao壓ya“直通式 ”自(zi)動(dong)進(jin)樣(yang)器(qi),則(ze)很(hen)少(shao)會(hui)出(chu)現(xian)攜(xie)帶(dai)汙(wu)染(ran)問(wen)題(ti),因(yin)為(wei)這(zhe)些(xie)現(xian)代(dai)進(jin)樣(yang)器(qi)使(shi)用(yong)高(gao)壓(ya)泵(beng)將(jiang)樣(yang)品(pin)吸(xi)入(ru)並(bing)直(zhi)接(jie)注(zhu)入(ru)流(liu)動(dong)流(liu)路(lu),從(cong)而(er)減(jian)少(shao)了(le)對(dui)任(ren)何(he)清(qing)洗(xi)階(jie)段(duan)的(de)需(xu)求(qiu)。注(zhu):對dui於yu多duo次ci穿chuan刺ci的de小xiao瓶ping,必bi須xu更geng換huan隔ge膜mo或huo使shi用yong密mi封feng時shi間jian足zu夠gou長chang的de隔ge膜mo材cai料liao,以yi減jian少shao這zhe種zhong影ying響xiang。擦ca拭shi隔ge膜mo針zhen可ke消xiao除chu部bu分fen汙wu染ran。這zhe個ge問wen題ti會hui造zao成cheng兩liang種zhong汙wu染ran。(a)當針頭浸入同樣有一個大隔膜開口的樣品瓶(相同或不同的樣品瓶)時,針頭會帶著一些樣品沉積到新的樣品瓶(或樣品瓶的隔膜上)。(b)wuranwuhaikenengshunzhezhentoubenshenliuxia,zaijinyangshididaozhenzuoshang,zaochengzhenzuohuoyangbenwuran。jianshaozhentouwaibuwurandezuijiandandejiejuefanganzhiyi,jiushizaixipingzhongjiaruyizhongrongye,zhezhongrongyejingguoyouhua,keyikuaisujiangyangpinrongjiedaorongyezhong。zhetingqilaihenjiandan,danxuduosepufenxirenyuanxuanzedeqingxiyegenbenwufatigaoxizhendeqingjiedu。lirutai、蛋白質、脂肪、油類和/或脂類等樣品可能很麻煩,因為它們的溶解性可能與所選的流動相不符;流動相通常是理想的,但在某些情況下卻不起作用。這些清洗瓶必須經常更換(隻需在係統中設置多個清洗瓶位置即可)。此外,每次使用清洗瓶時都應將瓶蓋關閉,以減少吸取汙染(此步驟至關重要)。缺乏 HPLC 維護大多數自動進樣器閥門,依靠旋轉密封將樣品從針環路轉移到係統的流路中。這些閥門內的部件會磨損,應至少每6個月檢查一次,必要時更換。此外,還應經常檢查針座和針是否有磨損或泄漏的跡象。注:查(zha)看(kan)進(jin)樣(yang)器(qi)是(shi)否(fou)有(you)泄(xie)漏(lou)跡(ji)象(xiang)。泄(xie)漏(lou)總(zong)是(shi)表(biao)明(ming)存(cun)在(zai)問(wen)題(ti),應(ying)立(li)即(ji)解(jie)決(jue)。有(you)泄(xie)漏(lou)時(shi)不(bu)要(yao)進(jin)行(xing)進(jin)樣(yang)。您(nin)的(de)方(fang)法(fa)和(he)獲(huo)得(de)的(de)數(shu)據(ju)將(jiang)無(wu)效(xiao)。應(ying)更(geng)換(huan)任(ren)何(he)磨(mo)損(sun)的(de)部(bu)件(jian)並(bing)測(ce)試(shi)係(xi)統(tong)性(xing)能(neng)。造(zao)成(cheng)樣(yang)品(pin)攜(xie)帶(dai)汙(wu)染(ran)的(de)最(zui)常(chang)見(jian)原(yuan)因(yin)之(zhi)一(yi)是(shi):yangpinzhurufaxuanzhuanmifengmosun。mosundemifengjianhuishiyangpinzailiangcijinyangzhijianzhiliuzaiqikongjianheaocaozhong,daozhizaisuihoudefenxizhongchuxianyangpinsepufeng。ciwai,huanchongyanyehuizhiliuzaimifengjianzhijian,zaochengxielouhuoxiedaiwuran。lixingde HPLC 維護以及每天衝洗所有緩衝液/鹽(如適用)可消除這些問題。操作員培訓不足良好的色譜分析需要全麵了解所使用的硬件和HPLC的基本原理。您必須能夠對係統的完整流路和方法開發中使用的色譜概念進行故障排除,通過指導,采用合理的步驟來學習。
文章來源:網絡
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