Ultisil LP-C18-Human Insulin Analysis Method Development Report
Test Report
Chromatography conditions:
Column |
Welch Utisil LP-C18（4.6×250mm,5μm） |
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Mobile Phase |
0.2mol/L sulfate buffer (pH=2.3)-acetonitrile=74:26 |
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Column Temperature |
40℃ |
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Detector |
UV |
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Injection Volume |
20μL |
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Flow Rate |
1.0 ml/min |
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Detect wavelength |
214 nm |
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Elution procedure |
|
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Note |
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Chromatogram and Data:
- 1) Blank solvent analysis spectrum
- 2) Analysis spectrum of injection sample solution:
Serial number |
Peak name |
Retention time (min) |
Peak Area(mAU*min) |
S/N |
Relative peak area(%) |
Resolution |
Number of plates(EP) |
Asymmetry |
1 |
API |
9.817 |
545.456 |
1829.6 |
100.00 |
N.a |
12878 |
1.07 |
Sum |
545.456 |
|
100.00 |
0.00 |
|
|
- 3) Analysis spectrum of frozen liquid sample solution:
Serial number |
Peak name |
Retention time (min) |
Peak Area(mAU*min) |
S/N |
Relative peak area(%) |
Resolution |
Number of plates(EP) |
Asymmetry |
1 |
API |
9.607 |
0.359 |
0.0 |
100.00 |
N.a |
21081 |
1.14 |
Sum |
0.359 |
|
100.00 |
0.00 |
|
|
- 4) Multiple analysis superposition of injection sample solution:
Serial number |
Peak name |
Retention time (min) |
Peak Area(mAU*min) |
S/N |
Relative peak area(%) |
Resolution |
Number of plates(EP) |
Asymmetry |
1 |
API |
9.817 |
546.653 |
9737.7 |
100.00 |
N.a |
12881 |
1.07 |
Sum |
546.653 |
|
100.00 |
0.00 |
|
|
Conclusion:
Welch column Ultisil LP-C18 (4.6×250mm, 5μm) was used to analyze the relevant solutions under these chromatographic conditions. The retention time of the target human insulin was 9.82 min and the retention time was stable after multiple injections, which met the analysis requirements.