Pure Chromatography
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Amino Acids (9-Fluorenylmethyl-Chloroformate Derivatives) on Raptor ARC-18
| Peaks | tR (min) | Conc. (µmole/mL) | |
|---|---|---|---|
| 1. | Arginine (Arg) | 5.145 | 0.4 |
| 2. | Asparagine (Asn) | 5.518 | 0.4 |
| 3. | Glutamine (Gln) | 5.630 | 0.4 |
| 4. | trans-4-Hydroxyproline (Hyp) | 5.878 | 0.4 |
| 5. | Serine (Ser) | 6.139 | 0.4 |
| 6. | Aspartic acid (Asp) | 6.256 | 0.4 |
| 7. | Glutamic acid (Glu) | 6.696 | 0.4 |
| 8. | Threonine (Thr) | 7.027 | 0.4 |
| 9. | Glycine (Gly) | 7.466 | 0.4 |
| 10. | Alanine (Ala) | 8.227 | 0.4 |
| 11. | Proline (Pro) | 8.803 | 0.4 |
| 12. | Methionine (Met) | 9.324 | 0.4 |
| 13. | Valine (Val) | 9.537 | 0.4 |
| 14. | Norvaline | 9.656 | 0.4 |
| 15. | Tryptophan (Trp) | 9.910 | 0.4 |
| 16. | Phenylalanine (Phe) | 10.120 | 0.4 |
| 17. | Isoleucine (Ile) | 10.214 | 0.4 |
| 18. | Leucine (Leu) | 10.285 | 0.4 |
| 19. | Cystine | 10.749 | 0.2 |
| 20. | Histidine (His) | 12.045 | 0.4 |
| 21. | Lysine (Lys) | 12.174 | 0.4 |
| 22. | Cysteine (Cys) | 12.463 | 0.4 |
| 23. | Tyrosine (Tyr) | 12.801 | 0.4 |
Amino acids are analyzed as 9-fluorenylmethyl-chloroformate derivatives.
LC_FF0539
| Column | Raptor ARC-18 (cat.# 9314A1E) | ||||||||||||||||||||||||||||||||
| Dimensions: | 100 mm x 3 mm ID | ||||||||||||||||||||||||||||||||
| Particle Size: | 2.7 µm | ||||||||||||||||||||||||||||||||
| Temp.: | 30 °C | ||||||||||||||||||||||||||||||||
| Sample | |||||||||||||||||||||||||||||||||
| Diluent: | 0.1 N HCl | ||||||||||||||||||||||||||||||||
| Conc.: | 0.4 μmole/mL for each amino acid (0.2 μmole/mL for cystine) | ||||||||||||||||||||||||||||||||
| Inj. Vol.: | 1 µL | ||||||||||||||||||||||||||||||||
| Mobile Phase | |||||||||||||||||||||||||||||||||
| A: | 0.1% Formic acid + 20 mM ammonium formate in water | ||||||||||||||||||||||||||||||||
| B: | 0.1% Formic acid + 10 mM ammonium formate in 90:10 acetonitrile:water | ||||||||||||||||||||||||||||||||
| |||||||||||||||||||||||||||||||||
| Detector | UV/Vis @ 265, 4.8 nm | ||||||||||||||||||||||||||||||||
| Instrument | Waters ACQUITY UPLC H-Class | ||||||||||||||||||||||||||||||||
| Notes | Derivatization reaction: 50 μL amino acid mix + 100 μL 0.2 N borate buffer (pH 10.0) + 50 μL 15 mM 9-fluorenylmethyl-chloroformate solution + 50 μL acetonitrile The injection can be performed after 5 minutes of reaction time. |
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