Report on the results of method development using Ultisil PFP-GC-max.docx
Test Report
Figure 1 Customer Analysis Spectrum
Chromatography conditions:
Instrument |
Thermo Fisher Scientific Vanquish Horizon |
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Column |
Ultisil Cellu-D（4.6×250mm,5μm） |
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Mobile Phase A |
0.1% TFA water |
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Mobile Phase B |
Chromatographic grade methanol |
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Injection Volume |
5 μL |
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Flow Rate |
0.8 ml/min |
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Detect wavelength |
210 nm |
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Elution procedure |
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The analysis spectrum is shown in Figure 2
Serial number |
Retention time (min) |
Peak Area(mAU*min) |
Peak height(mAU) |
Relative Peak area(%) |
Resolution(EP) |
Number of plates(EP) |
Asymmetry(EP) |
1 |
15.658 |
112.230 |
753.192 |
32.45 |
2.30 |
73929 |
0.99 |
2 |
16.189 |
233.628 |
1554.464 |
67.55 |
N.a. |
77381 |
1.00 |
Sum |
345.858 |
|
100.00 |
2.30 |
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Figure 2 Method Development Chromatogram
Conclusion: The two peaks are separated with a resolution of 2.3.
1.2. Separation and purification process
1.2.1. Sample preparation 1
The preparation was carried out according to the following chromatographic conditions:
Instrument |
Welch Sail1000 |
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Column |
Ultisil PFP (10×250mm,5μm) |
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Mobile Phase |
0.1% TFA: methanol |
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Injection Volume |
1mg |
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Flow Rate |
5mL/min |
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Detect wavelength |
210 nm |
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Elution procedure |
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The preparation spectrum is shown in Figure 3.
Figure 3 Sample Preparation Chromatogram
This method can be presented on the preparative column, and the target fraction was collected and detected.
1.2.2. Fraction Analysis
Perform liquid chromatography analysis on the fraction collected in step 1.2.1. The specific analysis conditions are the same as those in "Step 1.1." The chromatogram is shown in the following figure:
Serial number |
Retention time (min) |
Peak Area(mAU*min) |
Peak height(mAU) |
Relative Peak area(%) |
Resolution(EP) |
Number of plates(EP) |
Asymmetry(EP) |
1 |
15.601 |
1.482 |
10.320 |
4.27 |
2.35 |
76151 |
0.98 |
2 |
16.133 |
33.240 |
222.374 |
95.73 |
N.a. |
79861 |
1.00 |
Sum |
34.722 |
|
100.00 |
2.35 |
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Figure 5 Fraction 2 Analysis Chromatogram
Conclusion: The two substances can be separated on the preparative column.
- 2. Conclusion
Using an Ultisil PFP column under the corresponding chromatographic conditions, the target peaks can be prepared and collected separately. The preparation results meet the client's requirements.