Experimental report on testing vitamin B6 in milk powder by Ultisil AQ-C18
Test Report
Chromatography conditions:
Column |
Welch Ultisil®AQ-C18（4.6×150 mm, 5 μm） |
Mobile Phase |
2 g/L sodium octane sulfonate solution (containing 8 mL triethylamine, glacial acetic acid to adjust pH to 3.0): methanol (95:5) |
Column Temperature |
30℃ |
Detector |
Fluorescence detector |
Injection Volume |
10μL |
Flow Rate |
1.0 ml/min |
Detect wavelength |
Excitation 293 nm Emission 395 nm |
Elution procedure |
Isocratic elution |
Note |
/ |
Chromatogram and Data:
1) Mixed standard sample detection chart
Peak name |
Retention time (min) |
Peak Area(mAU*min) |
Relative peak area(%) |
Peak height |
Resolution(EP) |
Tailing factor |
Number of plates(EP) |
Pyridoxal |
5.147 |
39.87 |
32.42 |
4.53 |
|
1.01306 |
8149.96003 |
Pyridoxine |
6.725 |
39.72 |
32.29 |
3.76 |
6.27385 |
0.98832 |
9583.46935 |
Pyridoxamine |
12.085 |
43.39 |
35.28 |
2.49 |
14.75834 |
0.96770 |
11427.94435 |
2) Sample solution detection chart:
Serial number |
Peak name |
Retention time (min) |
Peak Area(mAU*min) |
Relative peak area(%) |
Peak height |
Resolution (EP) |
Tailing factor |
Number of plates(EP) |
1 |
2.462 |
32.73 |
40.01 |
3.67 |
|
1.17602 |
2803.16143 |
|
2 |
Pyridoxal |
5.139 |
11.62 |
14.21 |
1.32 |
13.00446 |
0.91175 |
8284.20582 |
|
Pyridoxine |
6.703 |
37.45 |
45.78 |
3.46 |
6.26047 |
1.05075 |
9629.68748 |
Sum |
81.81 |
|
|
|
|
|
Conclusion:
Using the Welch Ultisil® AQ-C18 (4.6×150 mm, 5 μm) chromatographic column under this chromatographic condition, pyridine, pyridine alcohol, and pyridine amine can be separated. Pyridine amine can be separated from impurity peaks in milk powder, and no pyridine amine is detected in milk powder samples, meeting the customer's separation requirements.