Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, 1200 Cailun Road, Shanghai, 201203, China.
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, 1200 Cailun Road, Shanghai, 201203, China.
J Pharm Biomed Anal. 2020 Jul 15;186:113232. doi: 10.1016/j.jpba.2020.113232. Epub 2020 Mar 20.
Zanthoxylum nitidum (Roxb.) DC. (ZN) belongs to the genus Zanthoxylum of Rutaceae and has various chemical ingredients and pharmacologic effects. Alkaloids are its main active constituents responsible for diverse pharmacologic effects, such as anti-tumor, anti-bacterial, anti-inflammatory, and analgesic activities. The chemical and pharmacological effects of ZN are well reported, but the in vivo pharmacokinetic profiles of its main active alkaloids are poorly investigated. This study aims to elucidate the absorbed constituents and pharmacokinetic behavior of main active ingredients in rat plasma after the oral administration of ZN extract. The absorbed constituents in rat plasma were qualitatively analyzed using ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Ultra-high-performance liquid chromatography with triple quadrupole mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous determination and pharmacokinetic studies of dihydrochelerythrine (DHCHE), nitidine chloride (NIT), chelerythrine (CHE), sanguinarine (SAN), liriodenine (LIR), skimmianine (SKI), γ-fagarine (FAG), and dictamnine (DIC) in rat plasma. Eighteen prototypes and metabolites were identified according to exact mass, characteristic diagnostic fragment ions, and reference standards. The established UPLC-MS/MS quantitative method met the requirements of FDA for biological analysis methods. Method validation showed that this method has good linearity (r ≥ 0.9910), precision (RSD ≤ 18.63 %), accuracy (88.11 %-117.50 %), and stability. The limit of detection (LOD) could reach 1 ng/mL, and the limit of quantitation could reach 2 ng/mL. The plasma drug concentration of benzophenanthridine alkaloids, such as NIT, CHE, and DHCHE, were still low even after dose differences were deducted. For the furan quinoline alkaloids (such as SKI, FAG, and DIC), only SKI showed high plasma drug concentration, although SKI content comprised only approximately 1/6 of benzophenanthridine alkaloids. This study is the first to simultaneously determine the above-mentioned active alkaloids in rat plasma and would contribute to the comprehensive understanding of in vivo pharmacokinetic behavior on active alkaloids in ZN extract.
荜澄茄（ZN）属于芸香科花椒属，含有多种化学成分和药理作用。生物碱是其主要活性成分，具有多种药理作用，如抗肿瘤、抗菌、抗炎和镇痛活性。ZN 的化学和药理学作用已有相关报道，但体内主要活性生物碱的药代动力学特征研究甚少。本研究旨在阐明 ZN 提取物灌胃给药后大鼠血浆中吸收成分和主要活性成分的药代动力学行为。采用超高效液相色谱-四极杆飞行时间质谱（UPLC-Q-TOF-MS）对大鼠血浆中的吸收成分进行定性分析。建立了同时测定和研究大鼠血浆中二氢白屈菜红碱（DHCHE）、盐酸荜茇宁（NIT）、盐酸花椒碱（CHE）、血根碱（SAN）、白屈菜红碱（LIR）、斯可吉宁（SKI）、γ-崖椒碱（FAG）和蝙蝠葛碱（DIC）的超高效液相色谱-三重四极杆质谱（UPLC-MS/MS）方法。根据精确质量、特征诊断碎片离子和对照品，鉴定了 18 个原型和代谢产物。所建立的 UPLC-MS/MS 定量方法符合 FDA 对生物分析方法的要求。方法学验证表明，该方法具有良好的线性（r≥0.9910）、精密度（RSD≤18.63%）、准确度（88.11%-117.50%）和稳定性。检测限（LOD）可达 1ng/mL，定量限可达 2ng/mL。即使扣除剂量差异，苯并菲啶生物碱（如 NIT、CHE 和 DHCHE）的血药浓度仍较低。对于呋喃喹啉生物碱（如 SKI、FAG 和 DIC），仅 SKI 显示出较高的血药浓度，尽管 SKI 含量约为苯并菲啶生物碱的 1/6。本研究首次同时测定了大鼠血浆中的上述活性生物碱，有助于全面了解 ZN 提取物中活性生物碱的体内药代动力学行为。
