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  • Title: Simultaneous determination of 11 alkaloids in rat plasma by LC-ESI-MS/MS and a pharmacokinetic study after oral administration of total alkaloids extracted from Naucleaofficinalis.
    Author: Wang G, Wang H, Lin Z, Hou L, Wang JY, Sun L.
    Journal: J Ethnopharmacol; 2022 Jan 10; 282():114560. PubMed ID: 34454053.
    Abstract:
    ETHNOPHARMACOLOGICAL RELEVANCE: Nauclea officinalis, a widely used Li medicine, has been used for the treatment of cold, fever, bronchitis, pneumonia, acute tonsillitis, and other ailments. Modern pharmacological studies have demonstrated that the most abundant and active components in N. officinalis are alkaloids, which possess various biological properties such as antibacterial and antitumor activities. AIM OF THE STUDY: To investigate the phytochemical profile of a selected group of alkaloids from the N. officinalis total alkaloids, and to determine the chemical profile of the alkaloids extracted from rat plasma. Further investigation was conducted to determine the pharmacokinetic behaviors of 11 selected major alkaloids, including pumiloside, naucleoxoside A, naucleoxoside B, nauclefine, angustidine, angustoline, (3S,19S)-3,14-dihydroangustoline,[α]D20: (-)191°, (3S,19R)-3,14-dihydroangustoline, [α]D20: (-) 294.7°, strictosamide, angustine, and 3,14-dihydroangustine. MATERIALS AND METHODS: N. officinalis total alkaloids were extracted with 79% ethanol and enriched with AB-8 macroporous resin. The phytochemical profile of alkaloids from the N. officinalis total alkaloids and the chemical profile of the alkaloids extracted from rat plasma were first analyzed by UPLC-Q-TOF-MS/MS. A simple, convenient, and sensitive LC-ESI-MS/MS method was subsequently developed and validated for the simultaneous determination of major active alkaloids in rat plasma after oral administration of N. officinalis total alkaloids. After addition of an internal standard (verapamil), plasma samples were pretreated first by protein precipitation with methanol and then underwent liquid-liquid extraction with ethyl acetate. Chromatographic separation was achieved using a Waters BEH C18 column (2.1 mm × 100 mm, 1.7 μm) at 30 °C, with gradient elution using a mobile phase consisting of 0.1% formic acid aqueous solution (A) and acetonitrile (B), a flow rate of 0.2 mL/min, and a total run time of 30 min. The detection was performed using an electrospray ionization triple quadrupole tandem mass spectrometer with multiple reaction monitoring and positive ionization mode. RESULTS: Based on the fragmentation patterns of 11 authentic alkaloids and previous reports, 55 alkaloids were identified or tentatively identified in the N. officinalis total alkaloids. Among them, 25 alkaloids were absorbed through the gastrointestinal tract in rats after administration of the N. officinalis total alkaloids. The 11 alkaloids were selected for quantitative analysis. The established quantitative method was fully validated and proved to be sensitive and specific. Satisfactory linearity of the 11 alkaloids obtained in the respective concentration ranges (r > 0.9931). The lower limits of quantification for strictosamide was 20.86 ng/ml, and the other ten alkaloids were all less than 4.47 ng/ml in rat plasma. The intra-and inter-day precision was less than 15% for all 11 alkaloids in terms of relative standard deviation, and the accuracies ranged from -11.4% to 11.1% in terms of relative error. Extraction recovery, matrix effect, and stability were within the required limits in rat plasma. CONCLUSION: The validated method was successfully applied to investigate the pharmacokinetics of the 11 alkaloids in rat plasma after oral administration of N. officinalis total alkaloids. Eleven alkaloids were rapidly absorbed to achieve a maximum plasma concentration with Tmax from 0.25 h to 1.5 h after oral administration. The pharmacokinetic parameters and plasma concentration-time profiles will prove valuable in pre-clinical and clinical investigations on the disposition of N. officinalis total alkaloids.
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