Table 3 Selected operation and validation parameters of the developed UHPLC-MS method for analysis of amino acids
From: Determination of immunogenic proteins in biopharmaceuticals by UHPLC–MS amino acid analysis
Mass (m/z) | tR (min) | Range (µM) | Calibration line | Linearity (r2) | LOD (µM) | LOQ (µM) | |
|---|---|---|---|---|---|---|---|
Glycine | 246.1 | 2.06 | 1–500 | y = 0.0190x + 1.0335 | 0.9995 | 0.009 | 1 |
Alanine | 260.1 | 2.56 | 1–500 | y = 0.0196x + 0.0057 | 0.9998 | 0.185 | 1 |
Serine | 276.1 | 1.99 | 1–500 | y = 0.0205x − 0.0088 | 0.9993 | 0.263 | 1 |
Proline | 286.1 | 2.91 | 1–500 | y = 0.0193x − 0.0140 | 0.9998 | 0.551 | 1 |
Valine | 288.1 | 4.60 | 1–500 | y = 0.0197x + 0.0151 | 0.9998 | 0.168 | 1 |
Threonine | 290.1 | 2.33 | 1–500 | y = 0.0202x + 0.0237 | 0.9991 | 0.287 | 1 |
Cysteine | 581.0 | 4.13 | 2.5–250 | y = 0.0226x − 0.1034 | 0.9979 | 0.822 | 2.5 |
Isoleucine | 302.1 | 5.49 | 1–500 | y = 0.0214x + 0.0333 | 0.9998 | 0.218 | 1 |
Leucine | 302.1 | 5.56 | 1–500 | y = 0.0222x − 0.0107 | 0.9997 | 0.205 | 1 |
Asparagine | 303.1 | 1.90 | 1–500 | y = 0.0951x + 1.3291 | 0.9918 | 0.656 | 1 |
Aspartic acid | 304.1 | 2.13 | 1–500 | y = 0.0197x + 0.1095 | 0.9990 | 0.532 | 1 |
Glutamine | 317.1 | 2.01 | 1–500 | y = 0.0152x + 0.0115 | 0.9992 | 0.735 | 1 |
Lysine | 487.1 | 4.25 | 1–500 | y = 0.0147x − 0.0021 | 0.9994 | 0.368 | 1 |
Glutamic acid | 318.1 | 2.24 | 1–500 | y = 0.0210x − 0.0184 | 0.9996 | 0.204 | 1 |
Methionine | 320.1 | 4.47 | 1–500 | y = 0.0229x + 0.0056 | 0.9998 | 0.331 | 1 |
Histidine | 326.1 | 1.79 | 1–500 | y = 0.0141x − 0.0121 | 0.9995 | 0.127 | 1 |
Phenylalanine | 336.1 | 5.72 | 1–500 | y = 0.0166x − 0.0287 | 0.9995 | 0.153 | 1 |
Arginine | 345.1 | 1.96 | 1–500 | y = 0.0208x − 0.0114 | 0.9990 | 0.291 | 1 |
Tyrosine | 352.1 | 4.36 | 1–500 | y = 0.0161x − 0.0157 | 0.9998 | 0.128 | 1 |
Tryptophan | 375.1 | 5.86 | 1–500 | y = 0.0223x + 0.0663 | 0.9997 | 0.108 | 1 |