色譜柱和色譜系統(tǒng)的故障檢修ppt34-經(jīng)營管理-閱讀頁
2024-09-03 17:39本頁面
【正文】 中，高 pH溶解硅膠導(dǎo)致塌陷 2020/9/18 23 Broad Peaks Unknown “Phantom” Peaks ? Column: ExtendC18, 150 mm, 5 181。C ? Sample: 1. Phenylalanine 2. 5benzyl3,6dioxo2piperazine ? 3. Aspphe 4. Aspartame ? 10 181。l extracolumn volume 2020/9/18 26 Determining the Cause of Peak Tailing 峰拖尾原因的確定 ? Evaluate mobile phase effects alter mobile phase pH and additives to eliminate secondary interactions 評價流動相效果 改變流動相 pH和添加劑消除次級效應(yīng) /流動相中組份與柱子相互作用 ? Evaluate column choice try column with high purity silica or different bonding technology 評價柱選擇 試用高純度硅膠柱或不同鍵合技術(shù)柱 ? Reduce sample load 減小進(jìn)樣量 ? Eliminate extracolumn effects 消除柱外效應(yīng) ? Flush column and check for aging/void 沖洗柱子檢查柱子老化 /塌陷 2020/9/18 27 Retention Issues 保留時間問題 ? Retention time changes(tr) 保留時間改變 ? Capacity factor (retention) changes (k180。 ? Replacing it solved problem. 換用新的流動相就解決問題。 2020/9/18 30 Column Aging/Equilibration Causes Retention/Selectivity Changes 柱老化／平衡不夠引起保留時間／選擇性改變 ? Column 1 Initial 1 號柱 開始時 ? Column 1 Next Day 1 號柱 第二天 ? Column 1 After Cleaning with 1% H3PO4 1 號柱用 1% H3PO4 清洗 ? The primary analyte was sensitive to mobile phase aging of the column. 最初的分析對流動相老化柱子是靈敏的 ? The peak shape was a secondary issue resolved by flushing the column. 沖洗柱子第二要解決的問題是峰形 ? Retention and peak shape were as expected after cleaning. 清洗后保留時間和峰形都達(dá)到了預(yù)期效果 2020/9/18 31 Determining the Cause of Retention Changes on the same Column 同一柱上保留時間改變原因的確定 ? 1. Determine K180。, ?和 tr的確定 ? 2. Wash column 沖洗柱子 ? 3. Test new column note lot number 試驗新柱 記錄一批數(shù)據(jù) ? 4. Review column equilibration procedures 觀察柱平衡過程 ? 5. Make up fresh mobile phase4 and test 配制新鮮流動相再試驗 ? 6. Check instrument performance 檢查儀器性能 2020/9/18 32 Change in Retention/Selectivity Columnto Column 柱與柱之間保留時間 /選擇性的改變 ? Different column histories (aging) 不同的柱歷史 (老化 ) ? Insufficient/inconsistent equilibration ? 平衡不夠 /不一致 ? Poor column/mobile phase bination 柱 /流動相差 ? Change in mobile phase流動相改變 ? Change in flow rate 流速改變 ? Other instrument issues其他儀器問題 ? Slight changes in column bed volume (tr only) 柱內(nèi)體積輕微改變 2020/9/18 33 LottoLot Selectivity Change (pH) 批與批之間選擇性改變 (pH) ? pH Lot 1 pH Lot 1 ? pH Lot 2 pH Lot 2 ? pH shows selectivity change from lottolot for basic pounds pH 顯示批與批之間對堿性化合物的選擇性改變 ? pH shows no selectivity change from lottolot, indicating silanol sensitivity at pH pH ,表明在pH 2020/9/18 34 Conclusions 結(jié)論 ? HPLC column problem are evident as: 高效液相色譜柱問題明顯的為 : ? High pressure 高壓 ? Undesirable peak shape 不希望得到的峰形 ? Changes in retention/selectivity 保留時間 /選擇性的改變 ? Often these problems are not associated with the column and may be caused by instrument and experimental condition issues. ? 經(jīng)常遇到的這些問題不一定與柱有關(guān) ,而與儀器和實驗條件方面的問題有關(guān)
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