Simultaneous quantification of ten key Kratom alkaloids in Mitragyna speciosa leaf extracts and commercial products by ultra‐performance liquid chromatography−tandem mass spectrometry

Kratom (Mitragyna speciosa) is a psychoactive plant popular in the United States for the self‐treatment of pain and opioid addiction. For standardization and quality control of raw and commercial kratom products, an ultra‐performance liquid chromatography?tandem mass spectrometry (UPLC?MS/MS) method was developed and validated for the quantification of ten key alkaloids, namely: corynantheidine, corynoxine, corynoxine B, 7‐hydroxymitragynine, isocorynantheidine, mitragynine, mitraphylline, paynantheine, speciociliatine, and speciogynine. Chromatographic separation of diastereomers, or alkaloids sharing same ion transitions, was achieved on an Acquity BEH C18 column with a gradient elution using a mobile phase containing acetonitrile and aqueous ammonium acetate buffer (10mM, pH 3.5). The developed method was linear over a concentration range of 1–200 ng/mL for each alkaloid. The total analysis time per sample was 22.5 minutes. The analytical method was validated for accuracy, precision, robustness, and stability. After successful validation, the method was applied for the quantification of kratom alkaloids in alkaloid‐rich fractions, ethanolic extracts, lyophilized teas, and commercial products. Mitragynine (0.7%–38.7% w/w), paynantheine (0.3%–12.8% w/w), speciociliatine (0.4%–12.3% w/w), and speciogynine (0.1%–5.3% w/w) were the major alkaloids in the analyzed kratom products/extracts. Minor kratom alkaloids (corynantheidine, corynoxine, corynoxine B, 7‐hydroxymitragynine, isocorynantheidine) were also quantified (0.01%–2.8% w/w) in the analyzed products; however mitraphylline was below the lower limit of quantification in all analyses.     

Alkaloids from Melodinus henryi with anti-inflammatory activity

One new quinolinic scandine-type monoterpeniod alkaloid, 3-oxo-scandine (1), as well as seven known ones (2–8), was isolated from the roots of Melodinus henryi. Their structures were elucidated by extensive spectroscopic analysis. All of the compounds were prepared and evaluated for their anti-inflammatory activities by measuring the inhibitory activity of nitric oxide (NO) in vitro in RAW 264.7 mouse peritoneal macrophages. Compounds 6 and 7 showed significant activities with IC₅₀ values of 8.54 and 5.19 μM, respectively.     

β-咔啉类生物碱对人胃癌SGC-7901细胞增殖、凋亡的影响及机制探讨

目的观察β-咔啉类生物碱对体外培养的人胃癌SGC-7901细胞增殖及凋亡的影响,并探讨其作用机制。方法不同浓度β-咔啉类生物碱处理人胃癌SGC-7901细胞48 h后,检测细胞增殖、凋亡情况,并以荧光定量聚合酶链反应(RTPCR)和蛋白印迹法(Western blot)分别测定细胞中抑癌基因PTEN、蛋白激酶B(AKT)基因mRNA及其蛋白表达。结果不同浓度的β-咔啉类生物碱均能抑制人胃癌SGC-7901细胞的增殖,其中40μg/m L浓度的抑制效果较其他浓度及5-Fu干预组更显著(P0.01),并可诱导细胞凋亡。不同浓度β-咔啉类生物均可引起PTEN mRNA和蛋白表达增加,AKT mRNA和蛋白表达减少,其中40μg/m L浓度的调节效果较其他浓度及5-Fu干预组更显著(P0.01)。结论β-咔啉类生物碱可抑制人胃癌SGC-7901细胞增殖,诱导凋亡,其作用机制可能是通过上调PTEN及下调AKT蛋白的表达实现。     

苦豆子总生物碱对小鼠乳腺癌4T1细胞体外增殖抑制及诱导凋亡作用的研究

目的研究苦豆子总生物碱对小鼠乳腺癌4T1细胞体外增殖抑制及诱导凋亡作用。方法苦豆子总生物碱提取并纯化,处理成澄清的药物溶液后用于细胞给药。CCK-8法测定不同浓度的苦豆子总生物碱作用24h后4T1细胞存活率。流式细胞术检测给药24h后4T1细胞凋亡率。结果与对照组比较,实验组药物浓度为5.00mg/mL时,4T1细胞存活率为63.05%;药物浓度为10.00mg/mL时,4T1细胞存活率为20.30%,随着给药浓度的增加,细胞存活率呈明显下降趋势。实验组药物浓度为6.25mg/mL时,细胞凋亡率为11.4%;药物浓度为10.00mg/mL时,细胞凋亡率为41.7%,随着给药浓度的增加,细胞凋亡率明显增加。结论苦豆子总生物碱能诱导细胞凋亡,对小鼠乳腺癌4T1细胞的增殖有明显抑制作用。     

生物碱对动物吗啡依赖性的戒断作用

目的：探讨一种可食用植物提取物——生物碱对成瘾动物依赖性的戒断作用。方法：50只雄性小鼠，分为5组各10只，第1—7天，A组小鼠尾静脉注射生理盐水200μl／只，B、C、D组注射吗啡150pg／只，E组注射生物碱1．7μg／只，每日2次；第7天时各组冉皮下注射纳络酮0．1mg／只催瘾。第8—14天，A、B组尾静脉注射生理盐水200μl／a，C与D组分别注射生物碱1．7与0．85μg／只，E组注射吗啡150μg／只。第15天再用纳络酮催瘾，观察并比较各时间段各组小鼠症状和体重的变化。结果：第7天时，B、C、D组小鼠有强烈成瘾表现；A、E组无异常。第15天时，B、E组出现明显成瘾症状；D组成瘾症状较轻；A、C组无异常。结论：可食用植物生物碱无成瘾性，对动物及人体依赖性的戒断作用有重要参考价值。     

Treatment of opioid use disorder with ibogaine: detoxification and drug use outcomes

Background: Ibogaine is a monoterpene indole alkaloid used in medical and nonmedical settings for the treatment of opioid use disorder. Its mechanism of action is apparently novel. There are no published prospective studies of drug use outcomes with ibogaine. Objectives: To study outcomes following opioid detoxification with ibogaine. Methods: In this observational study, 30 subjects with DSM-IV Opioid Dependence (25 males, 5 females) received a mean total dose of 1,540 ± 920 mg ibogaine HCl. Subjects used oxycodone (n = 21; 70%) and/or heroin (n = 18; 60%) in respective amounts of 250 ± 180 mg/day and 1.3 ± 0.94 g/day, and averaged 3.1 ± 2.6 previous episodes of treatment for opioid dependence. Detoxification and follow-up outcomes at 1, 3, 6, 9, and 12 months were evaluated utilizing the Subjective Opioid Withdrawal Scale (SOWS) and Addiction Severity Index Composite (ASIC) scores, respectively. Results: SOWS scores decreased from 31.0 ± 11.6 pretreatment to 14.0 ± 9.8 at 76.5 ± 30 hours posttreatment (t = 7.07, df = 26, p < 0.001). At 1-month posttreatment follow-up, 15 subjects (50%) reported no opioid use during the previous 30 days. ASIC Drug Use and Legal and Family/Social Status scores were improved relative to pretreatment baseline at all posttreatment time points (p < .001). Improvement in Drug Use scores was maximal at 1 month, and subsequently sustained from 3 to 12 months at levels that did not reach equivalence to the effect at 1 month. Conclusion: Ibogaine was associated with substantive effects on opioid withdrawal symptoms and drug use in subjects for whom other treatments had been unsuccessful, and may provide a useful prototype for discovery and development of innovative pharmacotherapy of addiction.     

甾体生物碱定量分析方法的研究进展

甾体生物碱是天然甾体的含氮衍生物,在化学特性和生物活性方面具有甾体类和生物碱类化合物的双重属性。甾体生物碱是贝母、黄杨、藜芦、龙葵等传统中药中的主要药效成分,并且也存在于马铃薯、蕃茄、茄等常用食物中。现代药理学研究表明,甾体生物碱类成分在抗肿瘤、心血管系统、抗菌、杀虫等方面具有很强的活性,同时也具有较强的毒性。甾体生物碱因其结构的特殊性,对其定量分析造成一定难度。对1998—2017年相关文献中甾体生物碱类成分的定量分析方法进行综述,其中应用于甾体生物碱总含量的测定方法主要有分光光度法-酸性染料比色法、酶联免疫吸附法、滴定法等;特定甾体生物碱的含量普遍采用高效液相色谱法,主要包括HPLC-UV,HPLC-ELSD,HPLC-MS和HPLC-CAD等。本文对近20年甾体生物碱定量分析的相关研究进行了归纳,并对各种含量测定方法的优势和局限性加以总结,以促进对相关中药质量以及相关食物安全性控制方法的改进和提升,同时也为甾体生物碱类成分以及相关中药的进一步开发利用提供分析方法方面的参考。     

荜茇在中医、蒙医中的应用概况及研究进展

荜茇为胡椒科胡椒属植物荜茇Piper longum的未成熟果穗,虽然中医、蒙医均用荜茇治疗胃肠道疾病,但在配伍及治疗疾病种类方面具有较大的差异。对荜茇在中医、蒙医中的用法、性味、功能、临床配伍等研究进行了对比分析,整理出荜茇在中医、蒙医本草中的记载情况,再结合计算机软件分析,对荜茇在蒙医临床中的使用频率及用药规律进行分析,并简要介绍荜茇在中医临床中的配伍及其化学成分和药理活性研究进展,为荜茇在中医药、蒙医药领域的应用提供依据。     

夏天无与延胡索的比较分析及其质量标志物预测

夏天无与延胡索药材的源植物同为紫堇属植物,均收载于《中国药典》2015年版一部,性味功效相近,但实际临床应用侧重点有所不同。通过查阅文献,从传统功效、化学成分、药理作用等方面比较分析夏天无与延胡索的异同;并基于质量标志物(Q-marker)的核心概念,从生源途径、药效、药动学和体内过程以及传统药性药效等方面对夏天无质量标志物进行预测分析,为建立和完善其药材质量标准提供理论依据。     

多种统计方法比较黄连姜制工序中影响生物碱含量的因素

目的：测定黄连不同姜制品中6种生物碱的含量,采用多种统计方法对数据进行分析,找出姜制工序中影响姜黄连质量的因素。方法：采用高效液相色谱法测定黄连不同姜制品中药根碱、非洲防己碱、表小檗碱、黄连碱、巴马汀及小檗碱的含量。运用主成分分析、聚类分析、LSD（Least-Significant Difference）检验、独立t检验对结果进行分析。结果：黄连经姜炙后,表小檗碱、黄连碱含量略有上升,药根碱、非洲防己碱、巴马汀、小檗碱含量略有下降,传统姜炙品中各成分含量均高于烘制品。聚类分析发现榨汁组和煮汁组可以聚为一类;烘法和炙法具有明显不同的聚类,对黄连姜制品中的6种生物碱之和进行LSD检验,亦表明烘法与炙法具有显著性差异。对榨汁组进行独立t 检验,结果表明姜汁与黄连饮片拌匀或闷润至透心在6种生物碱上无显著性差异,LSD与独立t 检验的结果与聚类分析的结果一致。结论：采用同一批次黄连炮制所得的姜制品中,姜的来源、不同方法制备的姜汁及闷润过程对6种生物碱影响不大,加热方式可能是影响姜黄连质量的因素。考虑到润至透心需加入大量水稀释姜汁,炒干所需工时较长,故辅料姜汁与黄连饮片拌匀吸尽即可。