Xtimate DLC C18-5 Organic Acid Analysis Method Development Report.docx
Test Report
Chromatography conditions:
Column |
Xtimate DLC C18（4.6*250mm*5μm） |
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Mobile Phase |
Phase A: 0.05 mol/L potassium dihydrogen phosphate solution (pH=2.5) |
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Column Temperature |
30℃ |
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Detector |
UV |
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Injection Volume |
10μL |
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Flow Rate |
1.0 ml/min |
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Detect wavelength |
205 nm |
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Elution procedure |
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Note |
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Chromatogram and Data:
- 1. Blank solvent analysis spectrum
Serial number |
Peak name |
Retention time (min) |
Peak Area(mAU*min) |
Peak height mAU |
Relative peak area(%) |
Resolution |
Number of plates(EP) |
Asymmetry |
1 |
3.410 |
0.561 |
1.996 |
100.00 |
N.a. |
29111 |
1.22 |
|
Sum |
|
0.561 |
|
100.00 |
0.00 |
|
|
- 2. Overlay of Analysis Spectra for Reference Solutions of Various Substances
Serial number |
Peak name |
Retention time (min) |
Peak Area(mAU*min) |
Peak height mAU |
Relative peak area(%) |
Resolution |
Number of plates(EP) |
Asymmetry |
1 |
maleic acid |
8.025 |
971.497 |
3167.580 |
100.00 |
N.a. |
4000 |
4.86 |
Sum |
|
971.497 |
|
100.00 |
0.00 |
|
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|
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- 3. Analysis Spectrum of Sample Solution
Serial number |
Peak name |
Retention time (min) |
Peak Area(mAU*min) |
Peak height mAU |
Relative peak area(%) |
Resolution |
Number of plates(EP) |
Asymmetry |
1 |
Oxalic acid |
3.278 |
26.618 |
486.325 |
5.13 |
5.64 |
24116 |
1.15 |
2 |
Tartaric acid |
3.773 |
5.732 |
98.549 |
1.11 |
11.74 |
27279 |
1.03 |
3 |
Malic acid |
5.013 |
3.074 |
40.188 |
0.59 |
16.62 |
27664 |
1.09 |
4 |
maleic acid |
8.234 |
234.164 |
1381.263 |
45.14 |
7.72 |
15091 |
2.51 |
5 |
fumaric acid |
10.187 |
249.097 |
4634.455 |
48.03 |
N.a. |
29031 |
1.03 |
Sum |
|
518.667 |
|
100.00 |
47.71 |
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Conclusion:
Using Welch Chromatography Column Xtimate DLC C18 (4.6 × 250 mm × 5 μm) under these chromatographic conditions to analyze the sample solution, the resolution between the five organic acids is greater than 1.5, meeting the analytical requirements.