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Determination of nicotine, cotinine and nicotine N-Oxide in human blood, plasma, urine, semen and sperm by LC-Orbitrap MS: application to clinical study ; Bestimmung von Nikotin, Cotinin und Nikotin-N-Oxid in menschlichem Blut, Plasma, Urin, Sperma und Spermien durch LC-Orbitrap MS: Anwendung in der klinischen Studie
A high throughput, robust, simple, and economic bioanalytical method for simultaneous determination of nicotine (Nic), cotinine (Cot) and nicotine N-oxide (Nox) in human urine, plasma, semen, sperm and dried blood spot (DBS) was developed and validated by LC ESI-orbitrap-MS. The current described method was successfully developed using deuterated nicotine (nicotine-d3) as internal standard (IS) following single extraction step by trichloroacetic acid solution (TCA) for protein direct precipitation, providing top extraction recovery and short run time with accurate mass measurements. The method was then validated according to the European and American guideline for bioanalytical method validation in terms of specificity, matrix effect, linearity, sensitivity, precision, accuracy and stability. Chromatographic conditions were optimized using Kinetex-C18 column (150 × 2.1mm, 5µm) and eluted by mobile phase of methanol:water:formic acid (10:90:0.1%, v/v/v). Accurate mass measurements were obtained by ESI-orbitrap-MS at positive m/z 163.1235, 177.1028, 179.1188 and 166.1423 for Nic, Cot, Nox and IS, respectively. The established calibration range for Nic, Cot and Nox in urine analysis was linear between 50 and 3000 ng/mL, in plasma, semen and DBS was linear between 5-250 ng/mL for Nic and Nox, and between 10-500 ng/mL for Cot. The within and between days measurements precision and accuracy were all within the acceptance criteria according to the European and American guidelines for bioanalytical method validation, and the extraction recovery was higher than 95% for the studied matrixes. The ionic suppression test was achieved and discussed for the all investigated fluidic matrices. The established method was intended to investigate three objectives, including study the influence of pomegranate and licorice test drinks on Nic metabolism rate. This was done by urinary measurements for metabolic index of Nic/Cot, in addition to new introduced metabolic index Nic/Nox, where urine samples were collected from twenty four ...
Determination of nicotine, cotinine and nicotine N-Oxide in human blood, plasma, urine, semen and sperm by LC-Orbitrap MS: application to clinical study ; Bestimmung von Nikotin, Cotinin und Nikotin-N-Oxid in menschlichem Blut, Plasma, Urin, Sperma und Spermien durch LC-Orbitrap MS: Anwendung in der klinischen Studie
A high throughput, robust, simple, and economic bioanalytical method for simultaneous determination of nicotine (Nic), cotinine (Cot) and nicotine N-oxide (Nox) in human urine, plasma, semen, sperm and dried blood spot (DBS) was developed and validated by LC ESI-orbitrap-MS. The current described method was successfully developed using deuterated nicotine (nicotine-d3) as internal standard (IS) following single extraction step by trichloroacetic acid solution (TCA) for protein direct precipitation, providing top extraction recovery and short run time with accurate mass measurements. The method was then validated according to the European and American guideline for bioanalytical method validation in terms of specificity, matrix effect, linearity, sensitivity, precision, accuracy and stability. Chromatographic conditions were optimized using Kinetex-C18 column (150 × 2.1mm, 5µm) and eluted by mobile phase of methanol:water:formic acid (10:90:0.1%, v/v/v). Accurate mass measurements were obtained by ESI-orbitrap-MS at positive m/z 163.1235, 177.1028, 179.1188 and 166.1423 for Nic, Cot, Nox and IS, respectively. The established calibration range for Nic, Cot and Nox in urine analysis was linear between 50 and 3000 ng/mL, in plasma, semen and DBS was linear between 5-250 ng/mL for Nic and Nox, and between 10-500 ng/mL for Cot. The within and between days measurements precision and accuracy were all within the acceptance criteria according to the European and American guidelines for bioanalytical method validation, and the extraction recovery was higher than 95% for the studied matrixes. The ionic suppression test was achieved and discussed for the all investigated fluidic matrices. The established method was intended to investigate three objectives, including study the influence of pomegranate and licorice test drinks on Nic metabolism rate. This was done by urinary measurements for metabolic index of Nic/Cot, in addition to new introduced metabolic index Nic/Nox, where urine samples were collected from twenty four ...
Determination of nicotine, cotinine and nicotine N-Oxide in human blood, plasma, urine, semen and sperm by LC-Orbitrap MS: application to clinical study ; Bestimmung von Nikotin, Cotinin und Nikotin-N-Oxid in menschlichem Blut, Plasma, Urin, Sperma und Spermien durch LC-Orbitrap MS: Anwendung in der klinischen Studie
Awwad, Ahmad Naim Ahmad Abu (author) / Schmitz, Oliver J.
2017-02-14
Theses
Electronic Resource
English
Exposure Assessment of Nicotine and Cotinine by GC-MS
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