TABLE 1

Identification of NBP metabolites in human plasma and urine after a single oral administration of 200-mg NBP soft capsules using UPLC-UV/Q-TOF mass spectrometry

NameDescriptionm/zm/z [M-H]−FormulaRetention Time (minutes)MatrixFragment Ions
[M+H]+[M+Na]+ESI(+)ESI(−)
M0Parent191.106213.093−C12H14O218.0plasma173.098, 145.103, 135.047, 131.052, 117.072, 105.074, 103.057, 91.057−
M1Phthalide-3-acetic acid193.050215.033191.035C10H8O44.2urine133.029147.045, 103.056
M210-Keto-NBP205.087227.071−C12H12O311.5plasma, urine187.074, 145.063, 117.071−
M3-13-Hydroxylation207.104229.080205.086C12H14O311.8plasma, urine189.094, 171.085, 161.099, 147.049, 145.104, 143.089, 133.032, 117.073161.094
M3-210-Hydroxylation207.103229.083205.088C12H14O312.1plasma, urine189.091, 171.079, 153.071, 145.102, 143.086, 133.031, 128.064, 117.072161.092
M3-38- or 9-Hydroxylation207.106229.086−C12H14O313.1plasma189.092, 171.081, 153.070, 149.061, 131.050, 103.055−
M3-48- or 9-Hydroxylation207.098229.083−C12H14O313.8plasma189.092, 171.080, 153.070, 149.059, 131.049, 103.055−
M4Hydrolysis−−207.101C12H16O311.1plasma, urine−163.110
M5-1Hydroxylation of M2−243.060219.067C12H12O45.8plasma, urine203.072, 185.063201.052, 175.078, 157.068, 147.046, 131.055, 119.051, 103.058
M5-2NBP-11-oic acid221.093243.074219.066C12H12O47.8plasma, urine203.081, 185.070, 175.080, 159.052, 157.075, 143.059, 131.058, 103.060175.078, 157.068, 131.089
M6-1Hydroxylation of M5-2237.080259.057235.063C12H12O55.9plasma, urine219.065, 173.067, 143.116, 133.029217.046, 191.075, 175.040, 173.065, 147.084, 131.053, 103.056, 91.059
M6-2Hydroxylation of M5-2237.080259.059235.063C12H12O56.3plasma, urine219.065, 173.059, 145.063, 133.029, 117.069217.053, 191.075, 173.063, 145.068, 129.073, 91.057
M7-1Sulfation of M3-1−−285.048C12H14O6S9.5urine−205.091, 161.064
M7-2Sulfation of M3-2−−285.044C12H14O6S12.7plasma, urine−205.087
M8N -acetylcysteine conjugation352.123374.099350.099C17H21NO5S14.5urine189.095, 171.086, 147.047, 133.032, 117.074, 91.061221.065, 177.076, 148.037
M9-1Glucuronidation of M3-2383.139405.118381.119C18H22O98.5plasma, urine207.105, 189.093, 171.082, 153.073, 145.104, 143.087, 133.031, 128.066363.103, 319.118, 205.087, 187.076, 175.025, 157.013, 113.024, 85.029
M9-2Glucuronidation of M3-2383.136405.121381.119C18H22O98.7plasma, urine207.105, 189.093, 171.083, 153.072, 145.105, 143.089, 133.032363.103, 319.118, 205.087, 187.076, 175.025, 157.013, 113.024, 85.029
M9-3Glucuronidation of M3-3383.130405.122381.118C18H22O99.3plasma, urine207.105, 189.092, 171.079, 153.067, 149.060, 131.052, 103.056363.123, 319.127, 205.088, 193.038, 175.029, 157.014, 113.026, 85.031
M9-4Glucuronidation of M3-3383.144405.119381.114C18H22O99.8plasma, urine207.105, 189.091, 171.076, 153.072, 149.061, 131.050, 103.057363.108, 205.089, 193.035, 175.025, 131.035, 113.025, 85.029
M9-5Glucuronidation of M3-4383.139405.123381.119C18H22O910.3plasma, urine207.105, 189.094, 171.081, 149.061, 131.055363.119, 205.088, 193.036, 175.030, 157.016, 131.036, 113.026, 85.030
M9-6Glucuronidation of M3-1383.144405.116381.122C18H22O911.3urine207.102, 189.090, 171.082, 161.097, 147.043, 143.086, 133.030, 117.070205.088, 161.099, 113.025, 85.033
M10Glucuronidation of M5-2397.121419.106395.093C18H20O108.0urine221.082, 203.074, 185.064, 175.080, 159.044, 157.068, 143.052, 131.052219.064, 193.034, 175.074, 145.061, 113.023
103.058, 91.057
M11-1Dihydroxylation + glucuronidation399.121421.106397.114C18H22O106.7urine223.092, 205.086, 187.076, 159.085, 149.066, 131.053, 103.056, 91.054221.078
M11-2Dihydroxylation + glucuronidation399.129421.115397.111C18H22O107.1urine223.103, 205.092, 187.083, 159.088, 149.066, 131.055, 103.058, 91.058221.084

  • NBP, 3-n-butylphthalide; Q-TOF, quadruple time-of-flight; UPLC-UV, ultraperformance liquid chromatography-UV.