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1、HAPSITE ER/Smart Plus TrainingHAPSITE ER/Smart Plus 培 训,Advanced ER IQ/Plus IQ 高级的 ER IQ/Plus IQ,Course Outline 课程大纲,Overview 综述Instrument Setup 仪器设置Tune 调谐Troubleshooting 故障查找Advanced Data Review 高级数据处理Creating AMDIS Libraries 创建 AMDIS 库Recognizing Co-Elution 识别共淘析Chromatogram Overlay 色谱图迭加 Accessi
2、ng Quantitation Reports 查访定量报告Basic Method Writing 基本方法编写Method Editor Overview 方法编写器概述SIM Methods SIM 方法Method Sequencing 方法序列Blank Subtraction 空样扣除Communications 通讯,Instrument Setup 仪器设置,Insert Gas Canisters 插入气瓶 Turn on the Analytical Module or take the Analytical Module out of Extended Standby 将
3、分析模件电源 on 或将分析模件离开延伸的待机状态System will heat,AutoTune and run a Concentrator cleanout 系统将加热,自动调谐和运行浓缩器清理Turn on the Laptop 将计算机电源 onDouble click the ER IQ/Plus IQ icon to open the software 双击 ER IQ/Plus IQ 图标,打开软件,Tune 调谐,Tune Overview 调谐概述AutoTune 自动调谐Manual Tune 手动调谐,Tune Overview 调谐概述,Tune optimizes
4、 the performance of the MS 调谐使 MS 的性能最佳化An improperly tuned MS can result in:未正确调谐的 MS 可导致:Low sensitivity 灵敏度低Undetected compounds 未能检测到化合物Mass misalignment 质量数未对准Incorrect identifications 不正确的 ID 鉴别Incorrect Mass Ratios 不正确的质量比Low Net Fit or SI numbers 低纯拟合或 SI 数,Tune Overview 调谐概述,Ten masses are
5、used when tuning the MS 调谐 MS 使用 10 个质量数Masses are between 50-282 to encompass the full 41-300 range of mass fragments 质量数在 50-282 之间包含整个 41-300 质量分裂物范围The Internal Standard gas contains the chemicals that provide these masses used for tuning 内部标准气体包含提供这些用于调谐质量数的化合物,AutoTune 自动调谐,The HAPSITE will au
6、tomatically AutoTune:HAPSITE 将自动调谐:Upon Startup 当启动时Upon exiting Extended Standby 当退出延伸的待机状态时After 12 hours of operation运行 12 个小时后,AutoTune 自动调谐,AutoTune can be initialized by:自动调谐可由下列动作起始:Clicking Tune AutoTune on the laptop 在计算机上单击 Tune AutoTuneTouching RUN AUTOTUNE on the front panel 触摸面板上的 RUN A
7、UTOTUNE 按钮Rebooting the HAPSITE 重新启动 HAPSITE,Troubleshooting a Failed AutoTune 不成功自动调谐的故障查找,Internal Standard Canister 内部标准气瓶 Verify that the pressure is adequate 核实压强是否适当Verify the expiration date 核实是否已到使用期限Insert new IS canister if necessary 必要时插入新的 IS 气瓶Carrier Gas(Smart Plus Only)载气(仅 Smart Plus
8、)Verify pressure that the pressure is adequate 核实压强是否适当Reservoir pressure(400 kPa)贮存器压强(400 仟帕)ER does not have a reservoir ER 无贮存器Insert new Carrier Gas canister if necessary 必要时插入新的载气瓶Repeat AutoTune 重新执行自动调谐,Tune Requires Advanced Mode 调谐要求高级模式,The Access Level must be set to Advanced to manually
9、 tune the instrument 必须将 Access Level 设定于高级,手动调谐仪器Set Access Level is used to change the ER IQ/Plus IQ user mode Set Access Level 用于更改 ER IQ/Plus IQ 的用户模式 Normal Mode:Allows access to default methods and data analysis 常规模式:允许进入默认方法和数据分析Advanced Mode:Allows access to method editing,manual tuning,file
10、 transfer and file deletion 高级模式:允许进入方法编写,手动调谐,文件传送和文件删除,ER IQ/Plus IQ Set Access LevelER IQ/Plus IQ 设定查访层次,Select Set Access Level 选 Set Access LevelSelect Advanced from the drop down menu 从下拉菜单上选 AdvancedNeither access level is password protected under the default settings 默认设定查访层次无口令保护Advanced mo
11、de can be password protected if desired 必要时可用口令保护高级模式Click OK 单击 OK,Set Access Level 设定查访层次,Manual Tune 手动调谐,Double click the Tune icon 双击调谐图标Select 选 default.tunClick OK 单击 OKNote:The NEG is being consumed while Manual Tune is open 注:在打开手动调谐的同时,消耗 NEG,Base Peak Gain(BPG)基峰增益(BPG),BPG is related to
12、the intensity of mass 117 BPG 与 M 117 的强度有关BPG influences sensitivity BPG 影响灵敏度ER ideal setting is 0.5 ER 理想的设定值为 0.5Range is 0.4 to 0.6 范围为 0.4 至 0.6 Smart Plus ideal setting is 1.0 Smart Plus理想的设定值为 0.1 Range is 0.8 to 2.0 范围为 0.8 至 2.0,Adjusting Base Peak Gain调整基峰增益,The BPG is adjusted using EM(E
13、lectron Multiplier)voltage 用 EM(电子倍增器)电压调整 BPGAdjust using increments of 25 V 用增量 25 伏调整 The normal operating range is 1000 to 2000 volts 常规工作范围为 1000 至 2000 伏,Adjusting Base Peak Gain(BPG)调整基峰增益(BPG),Adjusted Base Peak Gain for an ER 为 ER 调整基峰增益,Real Time Mass Calibration Table实时质量校准表,Red boxes may
14、 be present in the Real Time Mass Calibration Area 在实时质量校准区可出现红色框This indicates that resolution and/or intensity of the mass is out of range 这表明质量数的分辨率和/或强度已超出范围,Status Column 状态栏,Status Column will read OK,OK Low,OK High,Low or High 状态栏将出现 OK,OK 低,OK 高,低 或 高等读值High and Low are not acceptable 高和低是不可
15、接受的,Mass Alignment 质量对准,Perform a Mass Alignment by pressing F5 按 F5 执行质量对准Updates the Status column readings 更新状态栏读值Perform a Mass Alignment prior to and after making adjustments to the tune parameters 调整调谐参数前和后执行质量对准,Target Resolution 靶分辨率,Resolution refers to the width of the mass 分辨率与质量数的宽度有关The
16、 mass width should be 1 AMU 质量数宽度应为 1 AMUThe range for the Target Resolution is 0.85 and 1.10 靶分辨率的范围在 0.85 至 1.10 之间,Adjusting Target Resolution 调整靶分辨率,Adjust in increments of 0.05 调整的增量为 0.05The number in the Actual Percentage column is affected by the target resolution adjustments 实际百分数栏的数受靶分辨率调整
17、的影响It should fall between the Max.and Min.Targets 它应落入最大和最小靶之间If the masses cannot be brought into range by adjusting the Target Resolution,adjust the Ion Energies 如调整靶分辨率不能使质量数进入范围,调整离子能量,Ion Energies 离子能量,Adjusting the Ion Energy adjusts the intensity(height)of the mass 调整离子能量调整质量的强度(高度)Adjust the
18、 Ion Energies if the target resolution is at 0.85 and the red box continues to indicate that the mass is above the maximum target resolution 如靶分辨率在 0.85 和红框继续表示质量在最大靶分辨率以上,调整离子能量Adjust the Ion Energies if the target resolution is at 1.10 and the red box continues to indicate that the mass is below t
19、he minimum target resolution 如靶分辨率在 1.10 和红框继续表示质量在最小靶分辨率以下,调整离子能量,Ion Energies 离子能量,Adjust the Ion Energy 调整离子能量 Ion Energies should generally be in ascending order from Low to High 一般离子能量应从低至高地向上升Perform a Mass Alignment(F5)after each change until OK is reported in the Status column 每次更改后执行质量对准(F5
20、),直到状态栏内显示 OK,Save Tune 保存调谐,Verify that OK is displayed in the Status Column 核实在状态栏内显示 OKReadjustment of the BPG,Target Resolution and Ion Energies may be necessary to attain acceptable tune parameters 重新调整 BPG,靶分辨率和离子能量可能必须达到可接受的调谐参数Click Save Tune 单击保存调谐Close Manual Tune 关闭手动调谐,How to Manual Tune
21、 如何手动调谐,Verify Mass 117 is in range 核实 M 117 在范围内 Target Resolution(0.85-1.10)靶分辨率(0.85-1.10)Ion Energy(140-170)离子能量(140-170)Adjust Base Peak Gain(BPG)sensitivity,if needed 如需要,调整基峰增益(BPG)灵敏度 Adjust using EM voltage 用 EM 电压调整Range 范围0.6-0.8 for ER with 0.5 as optimal setting 对 ER 0.6-0.8,0.5为最佳设定值0.
22、8-2.0 for Smart Plus with 1.0 as optimal setting 对 Smart Plus 0.8-2.0,1.0为最佳设定值Adjust peak width resolutions,if needed如需要,调整峰宽分辨率Adjust using Target Resolution 用靶分辨率调整Range 0.85-1.10 范围 Adjust mass ratio intensity,if needed如需要,调整质量比强度Adjust using Ion Energy 用离子能量调整Refer to Ion Energy range chart 参阅离
23、子能量范围图表Save tune as default.tun 保存调谐为 default.tun,Tune Worksheet 调谐工作表,Advanced Manual Tune Worksheet 高级的手动调谐工作表,Troubleshooting 故障查找,Common Problems 常见问题High Pressure Troubleshooting 高压强故障查找,Common Problems:High Internal Standard TIC常见问题:内部标准 TIC 高,Possible causes 可能原因Blocked air flow in probe 探头中空
24、气流堵塞Sample bag closed 样品袋关闭Plugged exhaust 排气阻塞 Note:The air peak is not present in the chromatogram 注:色谱图中未出现空气峰,Common Problems:Shifted Retention Time常见问题:移位的滞留时间,A blank may show that the Retention time has shifted 空样可表示滞留时间已移位Smart Plus BPFB range is 8:00(+/-10 seconds)Smart Plus BPFB 的范围为 8:00(
25、+/-10 秒)ER BPFB range is 3:45(+/-5 seconds)ER BPFB 的范围为 3:45(+/-5 秒)Adjusting the GC column pressure will adjust the Retention time 调整 GC 柱压强将调整滞留时间GC Column Pressure settings are unique to each HAPSITE 每台 HAPSITE 的 GC 柱压强是独特的,Common Problems:Shifted Retention Time 常见问题:移位的滞留时间,Double click on the S
26、tatus icon 双击状态图标Click 单击 Pressure Flow and Offsets Increase kPa to decrease RT 增大仟帕来减小RT1 kPa change 3 second RT change 更改1 仟帕 RT更改3 秒Re-run the blank to verify that RT for BPFB is corrected 重新运行空样,证实BPFB的RT已改正,Common Problems:Multiple Internal Standard Peaks 常见问题:多个内部标准峰,Occurs when the previous r
27、un is aborted 当先前的运行被中止时发生 The Internal Standards were unable to elute内部标准不能淘析Continue running blanks until the extra peaks are no longer present 继续运行空样直到额外的峰不再出现,Common Problems:GC Column Hardware Error常见问题:GC 柱硬件误差,Possible causes include:可能的原因包括:Empty or low nitrogen canister 氮瓶空或压力低Insert a new,
28、full can of nitrogen 插入新的,满的氮瓶A cracked or chipped Concentrator 浓缩器破裂或碎裂Uninstall the Concentrator to verify its condition 卸下浓缩器,核实它的情况Incorrect ferrule orientation 套圈方向不正确Orient ferrules as shown in picture 套圈方向如图所示Ferrules are old or warped 套圈老化或弯曲Verify the condition of the ferrules 核实套圈的情况The Co
29、ncentrator is improperly seated 浓缩器就位不正确Reinstall the Concentrator 重新安装浓缩器The Concentrator nuts are not fully tightened 浓缩器螺帽未完全拧紧,Pump Failure:High Pressure Warning泵损坏:高压强警告,A high pressure warning appears when the total MS pressure is higher than acceptable 当 MS 的总压强高于可接受的压强时出现高压强警告Acceptable pres
30、sure is less than 6 E-3 Pa可接受的压强低于6 E-3 帕The HAPSITE will no longer run methods HAPSITE不再运行方法The source of the high pressure can be isolated using the Service Module 可用服务模件隔离高压强源Attach and pump down for a couple of hours连接和抽空数小时If the HAPSITE runs a method,the cause is the NEG Pump 如 HAPSITE 运行方法,原因
31、在于 NEG 泵If the HAPSITE does not run a method,the cause is the Ion Pump 如 HAPSITE 不运行方法,原因在于离子泵,Pump Overview 泵概述,Ion Pump离子泵Works in conjunction with the NEG to maintain vacuum in the Mass Spectrometer 与 NEG 一起工作维持质谱仪 的真空Removes the noble gases 清除惰性气体Average lifetime of an Ion Pump is 2000 to 4000 h
32、ours离子泵的平均寿命为 2000 至 4000 小时To view the Ion Pump hours:查看离子泵的小时数Double click the Status icon双击状态图标Select Maintenance 选维护,Pump Overview 泵概述,NEG Pump NEG 泵Maintains vacuum 维护真空NEG can last over 200 hours when stored in Extended Standby 当贮存于延伸的待机模式时,NEG 可在 200 小时以上 Storing in Extended Standby improves
33、NEG life 贮存于延伸的待机模式可延长 NEG寿命To view the NEG hours 查看 NEG 的小时数Double click Status icon 双击状态图标Click on the NEG Status Tab,High Pressure Troubleshooting 高压强故障查找,NEG bakeout NEG 烘烤NEG bakeout can be used if a NEG has less than 100 hours of“On Time”如 NEG 的“On Time”时间少于 100 小时,可使用 NEG 烘烤The NEG is heated
34、to 700C for 2 hours 将 NEG 加热至 700C,保持 2 个小时A bakeout may prolong NEG life,but is not a guarantee 烘烤可 延长 NEG 的寿命,但不保证,NEG Bakeout NEG 烘烤,Double click the Status Icon 双击状态图标Click on Service 单击服务Click on Bakeout NEG 单击烘烤 NEG,High Pressure in the HAPSITE,Attach to Service Module and pump down for 2 hour
35、s,Run a method.Does the method run?,High pressure was caused by ion pump failure,High pressure is caused by NEG pump failure,Are there a lot of hours on the NEG pump?,Replace the NEG pump,Bakeout the NEG pump,YES,NO,NO,YES,Flowchart for High Pressure Troubleshooting,高压强在 HAPSITE 中,连接至 服务模件 抽空 2 个小时,
36、运行方法.方法是否 运行?,高压强的原因 是由于 离子泵故障,高压强的原因是由于 NEG 泵故障,NEG 泵是否还有许多 小时数?,更换 NEG 泵,烘烤 NEG 泵,是,否,否,是,高压强故障查找流程图,Advanced Data Review 高级数据处理,Creating AMDIS Libraries 创建 AMDIS 库Using New AMDIS Library 使用新的 AMDIS 库Recognizing Co-Elution 识别共淘析Chromatogram Overlay 色谱图迭加Chromatogram Subtract 色谱图扣除,Creating AMDIS L
37、ibraries 创建 AMDIS 库,To create an AMDIS library 为创建 AMDIS 库 Select a spectrum from the NIST library 从 NIST 库选谱图Export the spectrum to a new folder 输出谱图至新文件夹Create library file 创建库文件Add library to AMDIS 添加库至 AMDIS,Creating AMDIS Libraries 创建 AMDIS 库,In this example,we will create a library that will i
38、nclude only CWAs 本例中,我们将创建仅包含 CWAs 的库We will add the following compounds:将添加下列化合物:Ethylmethylphosphonofluoridate(VX)乙基甲基膦酰基氟化物(VX)Isobutylmethylphosphonofluoridate(R-33)异丁基甲基膦酰基氟化物(R-33)Sarin 沙林Soman 束曼Tabun 塔崩Mustard Gas(Sulfur Mustard)芥子气(硫芥子气)Cyclohexylmethylphosphonofluoridate(Cyclosarin)环己基甲基膦酰
39、基氟化物(环沙林),Creating AMDIS Libraries 创建 AMDIS 库,Open NIST 打开 NISTEnter the desired compound 键入所需的化合物EnterEthylmethylphosphonofluoridate(as much as will fit)键入乙基甲基膦酰基氟化物(尽管多将拟合)Right-click on the highlighted name 右击点亮的名称Click Export Selected 单击输出已选,Creating AMDIS Libraries 创建 AMDIS 库,Enter VX as the fi
40、le name 在文件名中键入 VXThis is the file that will be entered into the new library 这是将输入新库的文件Click Save 单击 保存,Creating AMDIS Libraries 创建 AMDIS 库,Type Isobutylmethylphosphonofluoridate 键入异丁基甲基膦酰基氟化物 Right-click on the name,and select Export Selected右击名称,并选输出已选,Creating AMDIS Libraries 创建 AMDIS 库,Click on
41、VX 单击VXClick on Save 单击保存,Creating AMDIS Libraries 创建 AMDIS 库,Click Append 单击 添加This will save the selected compound to the same library that VX is saved to 这将保存所选的化合物至与保存 VX 相同的库Repeat the previous steps until all desired compounds have been added 重复先前的步骤直到全部所需的化合物已经被添加Make sure to click on VX each
42、 time 务必每次单击 VX,Creating AMDIS Libraries 创建 AMDIS 库,Open the data file to be analyzed 打开要分析的数据文件In the Basic Training Chromatograms-ER folder,open the data file labeled Agents 在基本培训色谱图-ER 文件夹中,打开标签为 Agents 的数据文件,Creating AMDIS Libraries 创建 AMDIS 库,Open AMDIS 打开 AMDISRight-click in the chromatogram 在
43、色谱图上右击Highlight Peaks 点亮峰Highlight Switch to AMDIS 点亮 切换至 AMDIS,Creating AMDIS Libraries 创建 AMDIS 库,Click Confirm 单击 确认This will open up the Confirmation page(see next slide)这将开启确认页面(见下一个幻灯片),Creating AMDIS Libraries 创建 AMDIS 库,Creating AMDIS Libraries 创建 AMDIS 库,In the Menu bar select Library,then
44、select Library Transfer 在菜单栏中选 库,然后选 库传送,Creating AMDIS Libraries 创建 AMDIS 库,Click on Files 单击 文件Click on Load Library 单击 加载库,Creating AMDIS Libraries 创建 AMDIS 库,Change the File Type to.msp 将文件类型更改为.msp,Creating AMDIS Libraries 创建 AMDIS 库,Find the file saved earlier from NIST 从 NIST 寻找早期保存的文件Click t
45、he Up One Level button to get to the main NIST folder 单击 返回上一级文件夹 按钮,进入主 NIST 文件夹,Creating AMDIS Libraries 创建 AMDIS 库,Double-click the MSSEARCH folder 双击 MSSEARCH 文件夹Highlight the compound to add to the library 点亮化合物,将它添加至库中This example is VX 本例为 VXClick Open 单击 打开,Creating AMDIS Libraries 创建 AMDIS
46、库,Click Create New Library 单击 创建新库,Creating AMDIS Libraries 创建 AMDIS 库,In File Name,type CWA Library.msl 在文件名中,键入 CWA Library.mslThis is the name of the new library that is being created 这是创建的新库名Click OK 单击 OKClick Close in the Files window 单击 在文件窗口中关闭,Creating AMDIS Libraries 创建 AMDIS 库,Hold Ctrl b
47、utton on the keyboard and click each compound to highlight 按住键盘上的 Ctrl 按钮,单击每个化合物,点亮它们Click Transfer to add to the new CWA Library 单击 传送,添加至新的 CWA 库中,Creating AMDIS Libraries 创建 AMDIS 库,Click Exit 单击 退出This will save the spectra to the new library 这将保存谱线至新库中,Change to New AMDIS Library 变换至新 AMDIS 库,
48、To change to the new AMDIS Library 为变换至新 AMDIS 库In the Menu bar click File,then Go to Results 在菜单栏中单击文件,接着单击 前往结果This will bring up the Results page 这将调出结果页面,Change to New AMDIS Library 变换至新 AMDIS 库,Click on Analyze 单击 分析,Change to New AMDIS Library 变换至新 AMDIS 库,Click on Target Library 单击 靶库,Change
49、to New AMDIS Library 变换至新 AMDIS 库,Click on Select New 单击 选新的,Change to New AMDIS Library 变换至新 AMDIS 库,Click on the Up One Level button until the C:directory appears 单击返回上一级文件夹按钮直到出现 C:目录,Change to New AMDIS Library 变换至新 AMDIS 库,Double-click on the NIST08 folder 双击 NIST08 文件夹Double-click on the MSSEA
50、RCH folder 双击 MSSEARCH 文件夹,Change to New AMDIS Library 变换至新 AMDIS 库,Click on Files of type,and select MS Library(*.MSL)单击文件类型,并选 MS 库(*.MSL),Change to New AMDIS Library 变换至新 AMDIS 库,Click on CWA Library.msl 单击 CWA Library.msl Click Open 单击 打开,Change to New AMDIS Library 变换至新 AMDIS 库,Click Save 单击 保存
