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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
  1. LOD, limit of detection; LOQ, limit of quantification; tR, retention time