There are many brands and types of liquid chromatographs, and the methods of use are different. The main reason is that you are the liquid chromatograph. When you purchase equipment, the engineers of the factory will train the method of use. The combination of high performance liquid chromatography and structural instruments is an important development direction. Liquid chromatography-mass spectrometry technology has received widespread attention, such as the analysis of carbamate pesticides and polynuclear aromatic hydrocarbons; liquid chromatography-infrared spectroscopy has also been developed rapidly as measured by environmental pollution analysis of hydrocarbons in water, non-volatile in seawater Hydrocarbons have led to new developments in environmental pollution analysis.
How to use the liquid chromatograph:
1.1 Turn on your computer.
1.2 Turn on the power to each module of the LC.
1.3 Double-click the desktop â€œInstrumentâ€”Onlineâ€ to enter the online interface.
1.4.1 Manually unscrew the flush valve at the pump (rotate about 1 turn counterclockwise).
1.4.2 Right click on the â€œPumpâ€ icon area and select the â€œMethod...â€ option to enter the pump editing screen. Set the flow rate: 5ml/min (usually 3-5ml/min) and click â€œOKâ€.
1.4.3 Right click on the â€œPumpâ€ icon, click on the â€œControl...â€ option, select â€œONâ€, click â€œOKâ€, the system will start flushing until there is no air bubble in the pipeline (from the solvent bottle to the pump inlet). For 5 minutes), switch channels continue to rinse until all channels are used without bubbles.
1.4.4 Right click on the â€œPumpâ€ icon, click on the â€œMethod...â€ option, set the flow rate: 0ml/min, and manually tighten the flush valve.
1.4.5 Right click on the â€œPumpâ€ icon, click on the â€œMethod...â€ option, and select the appropriate proportion of mobile phase according to the method requirements. Set the flow rate: 1.0ml/min.
1.4.6 Similarly, right click on the "column thermostat", "detector" icon, click the "method..." option, set the temperature, wavelength according to the method requirements, click the "control" option, "ON" to open the column oven and Detector.
2 editing method
2.1 Click on "Method" - "Edit Full Method" to start editing the full method.
2.2 Select three items except â€œData Analysisâ€ and go to the next tab.
2.3 Method Information: Add method information to the Method Comments (eg: This is for test!). Go to the next tab.
2.4 Pump parameter setting: input flow rate at â€œflow rateâ€, such as 1.0ml/min, stop time: such as 10 min (this stop time is only the time required to make a sample), select the appropriate proportion of mobile phase ratio according to requirements, For example, acetonitrile: water = 75:25, A is water, B is acetonitrile, then B: 75% can be set. Go to the next tab.
2.5 Autosampler Parameter Setting: Select â€œNeedle Injectionâ€----You can enter the injection volume and the bottle washing position to enter the next tab.
2.6 Column temperature box parameter setting: Enter the required temperature in the blank box below â€œTemperatureâ€, for example: 40 degrees. Go to the next tab.
2.7 UV Detector Parameter Setting: Enter the desired detection wavelength, such as 254 nm, in the space below â€œWavelengthâ€. Click OK.
2.8 In the â€œRuntime Options Tableâ€, select â€œData Acquisitionâ€ and click â€œOKâ€.
2.9 From the "Method" menu, select "Save Method As...", enter a method name, such as "Test", click "OK."
3 single acquisition
3.1 From the â€œRun Controlâ€ menu, select the â€œSample Informationâ€ option, select the appropriate path, select â€œPrefix/Counterâ€ in â€œData Fileâ€, enter the position of the vial, and click â€œOKâ€.
3.2 About 10 minutes after the baseline is stable, select â€œRun Methodâ€ from the â€œRun Controlâ€ menu.
4 multiple data collection
4.1 Follow step 2 to edit the full method.
4.2 Click on â€œSequenceâ€ - â€œSequence Listâ€, enter â€œsample bottleâ€, â€œsample nameâ€, â€œinjection countâ€, and select the appropriate â€œsample methodâ€
4.3 Click â€œSequenceâ€ - â€œSequence Parameterâ€, select the save path of the sequence data (the sequence will automatically generate the folder with the name â€œsequence name-timeâ€ to save the data), and the data suggestion is saved by selecting â€œprefix/counterâ€.
4.4 From the "Sequence" menu, select "Save Sequence As...", enter a sequence name, such as "Test", and click "OK."
4.5 Select Run Sequence from the Run Control menu.
5 data analysis (offline use)
5.1 Double-click the offline screen accessed by the "Instrument - Offline" icon.
5.2 From the â€œViewâ€ menu, click â€œData Analysisâ€ to enter the data analysis screen.
5.3 Select â€œCall Signalâ€ from the â€œFileâ€ menu and select your data file name. Click "OK" and the data will be recalled. (If you pre-establish a standard curve, you should first open the lower concentration standard map.)
5.4 Doing Spectral Optimization: Select Signal Options from the Graphics menu. Select "Full Scale" or "Auto Range" from the "Range" and the appropriate time range or select the "Custom Range" adjustment. Repeat until the scale of the graph is appropriate. Click "OK."
6.1 Select the â€œIntegration Eventâ€ option from the â€œIntegrationâ€ menu and select the appropriate â€œSlope Sensitivityâ€, â€œPeak Widthâ€, â€œMinimum Peak Areaâ€, â€œMinimum Peak Heightâ€. Click to automatically load the integration parameters.
6.2 Click the â€œâˆšâ€ icon on the left to save the integration parameters into the method and exit the â€œintegration eventâ€.
6.3 If the result of the integration is not satisfactory, modify the corresponding integral parameters until satisfactory.
7 standard curve
7.1 Click â€œCalibrationâ€ - â€œCalibration Settingsâ€ and enter â€œContent Unitâ€.
7.2 Click â€œCalibrationâ€ - â€œNew Calibration Tableâ€ and click OK. Enter "Compound Name" and "Content", click "OK" and follow the prompts to delete other components.
7.3 At this point, complete the single-level calibration. To increase the calibration level, select â€œCall Signalâ€ from the â€œFileâ€ menu, select your data file name (second standard), click â€œCorrectionâ€ - â€œAdd Levelâ€, and click OK. , enter "content" and increase the correction level in turn.
8 Print report
8.1 Select the â€œSet Reportâ€ option from the â€œReportâ€ menu, click the black triangle to the right of â€œQuantityâ€ in the â€œQuantitative Resultsâ€ box, select â€œExternal Standard Methodâ€, the other options remain unchanged, and click â€œOKâ€.
8.2 Select â€œPrint Reportâ€ from the â€œReportâ€ menu, the report results will be printed on the screen. If you want to output to the printer, click the â€œPrintâ€ button at the bottom of â€œReportâ€.
8.3 Click â€œFileâ€ - â€œSave Asâ€ - â€œMethodâ€ to save the data analysis method. The next analysis can be called up directly under â€œFileâ€ - â€œCallâ€ - â€œMethodâ€. (The method called includes integral method, standard curve method and print report method)
9.1 Before shutting down, first turn off the UV lamp and rinse the system thoroughly with the appropriate solvent (methanol or acetonitrile) for about 30 minutes. (The column should eventually be stored in methanol or acetonitrile)
9.2 Exit the ChemStation, turn off the computer by following the prompts to turn off the pump, and other windows.
9.3 Turn off the power switches of each module of the Agilent 1260.
10 Other considerations
10.1 Do not open the front of the autosampler while the sample is running, otherwise the injection process will stop.
10.2 When the system leaks, the machine will detect and stop the injection, and the status indicator will be red. Check the dry and place the leak, dry the leak sensor, click the ON button, and the system re-initializes.
10.3 Pay attention to the life of the UV lamp. Do not switch the UV lamp back and forth.
High performance liquid chromatography requires only a sample to be made into a solution, independent of the volatility of the sample, a wide range of mobile phases, and a wide variety of stationary phases, thus separating thermally unstable and non-volatile, dissociated sums. Non-dissociated and various molecular weight ranges of substances. In combination with the sample pretreatment technology, the high resolution and high sensitivity achieved by HPLC make it possible to separate and simultaneously measure substances of similar nature, which can separate trace components in complex phases. With the development of the stationary phase, it is possible to complete its separation HPLC under the condition of fully maintaining the activity of biochemical substances as the most promising method for solving the problem of biochemical analysis. Because HPLC has high resolution, high sensitivity, fast speed, reusable column, easy collection of effluent components, it is widely used in various fields such as biochemistry, food analysis, medical research, environmental analysis, inorganic analysis, etc. .
Thank you for your interest in Shanghai Jiapeng's products. In addition to this product introduction, the company also has three kinds of UV analysis, nucleic acid protein detector, gel imaging analysis system, photochemical reactor, constant current pump, automatic partial collector, etc. Introduction, if you are interested in Shanghai Jiapeng products, please contact us. Thank you!
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