花叶型与艾叶型乌头叶片基因组DNA甲基化修饰的MSAP分析

建立乌头叶片DNA甲基化MSAP分析方法,并对不同形状的乌头叶片DNA甲基化修饰位点进行MSAP分析。以花叶型、艾叶型乌头不同部位叶片为实验材料,考察乌头叶片MSAP双酶切反应时间,筛选适宜的选择性扩增引物,并通过计算6%丙烯酰胺凝胶电泳图泳道和条带统计分析叶片甲基化修饰差异。结果 150 ng乌头叶片DNA MSAP双酶切反应体系在37℃条件下,酶切16 h较充分;使用25对筛选出的适宜性引物进行乌头基因组的MSAP选择性扩增,总共获得273个电泳条带,其中无甲基化或单链内甲基化228条,双链内甲基化条带27条,单链外甲基化条带18条,总甲基化率达16.48%;花叶型与艾叶型乌头总甲基化率稍有差异,分别为15.36%,14.34%,并各自获得8条、6条基因组甲基化修饰差异的核苷酸片段,占其总条带数的3.00%,2.26%。通过研究可为乌头植株的分子鉴定、良种选育与栽培,以及乌头的遗传演化等提供新思路。     

HPLC-FLD免疫亲和柱-光衍生化法测定胖大海中黄曲霉毒素的残留量

目的：建立长柱十大功劳生物碱一测多评方法,对生物碱含量进行动态分析,确定长柱十大功劳的最佳采收期。方法：采用HPLC,以盐酸小檗碱为参照物,建立该组分与盐酸药根碱、盐酸巴马汀的相对校正因子,测定盐酸小檗碱含量,用相对校正因子计算盐酸药根碱、盐酸巴马汀含量;同时用外标法测定3种生物碱的绝对含量,比较计算值与测量值,评价一测多评的准确性;采用一测多评模式测定长柱十大功劳不同采收期的生物碱含量。结果：建立了长柱十大功劳生物碱一测多评方法,计算值与测量值无显著性差异;总碱含量4~8月逐渐升高,8月达峰值,9~12月降低。结论：该法可简便、准确测量长柱十大功劳不同生物碱含量;最佳采收期为8月。     

关白附的HPLC指纹图谱

目的：对东北产10个批号的关白附药材指纹图谱进行了比较.方法：应用RP-HPLC法,Diamonsil C18色谱柱,流动相为2 mg/mL浓度的庚烷磺酸钠水溶液（含0.2%三乙胺并以H3PO4调pH至3.0）-乙腈二元梯度洗脱,流速为1.0 mL/min,检测波长205 nm.结果：不同批号的关白附药材所含生物碱成分均得到很好的分离,确定了指纹图谱中10个共有峰,共有峰峰面积之和均大于各总峰面积的90%.结论：该方法可用于关白附的真伪鉴别和质量评价.     

生物碱类化合物抗病毒活性及其机制研究进展

生物碱是普遍存在于植物体内的含氮次生代谢产物,具有广泛的生物活性。研究表明,生物碱类化合物对多种病毒表现出良好的抑制作用和广谱的抗病毒活性,成为近年来抗病毒小分子化合物研究的新焦点。研究证实,生物碱对甲型流感病毒、乙型肝炎病毒、丙型肝炎病毒、单纯疱疹病毒、艾滋病毒、寨卡病毒、柯萨奇病毒和烟草花叶病毒等表现出显著的抑制作用,其抗病毒机制主要包括抑制病毒吸附和进入、减少病毒RNA和蛋白质合成、促进细胞凋亡和自噬、调节各种病毒相关蛋白以及通路等。根据生物碱的抗病毒种类进行分类,以氧化苦参碱、麻黄碱、石斛碱等为例,综述了常见生物碱抗病毒作用特点和机制,为生物碱类抗病毒小分子化合物的研究提供参考。     

Advances in cocaine signature methodology: Alkaloid and isotope profiles of coca grown in Puno,Peru

Drug Enforcement Administration's (DEA) Cocaine Signature Program previously identified 19 coca-growing regions within South America and developed methodology to geo-source cocaine using a combination of trace cocaine alkaloids, stable isotopes, and multivariate statistics. Twenty-nine coca leaf samples collected in 2016 and 2019 from a previously unanalyzed coca-growing region located in Puno, Peru, were analyzed with this methodology. Trace cocaine alkaloids and stable isotopes were compared with other Peruvian regions. Minor differences were observed in the extracted cocaine alkaloid profiles when compared with samples collected from the Ucayali-Huallaga Valley and Cusco-Apurimac regions while the stable isotopes of δ²H (−177.1‰) and δ¹⁸O (23.8‰) were enriched. Puno's alkaloid and stable isotope results are presented in this publication to assist forensic laboratories and enhance their cocaine geo-sourcing capabilities.     

New dianthramide and cinnamic ester glucosides from the roots of Aconitum carmichaelii

Four new compounds N-salicyl-3-hydroxyanthranilic acid methyl ester (1), N-(2′-dehydroxysalicyl)-3-hydroxyanthranilic acid methyl ester (2), methyl-4-β-D-allopyranosyl-ferulate (3), and methyl-4-β-D-gulopyranosyl-cinnamate (4), along with six known compounds (5–10), were isolated from the roots of Aconitum carmichelii Debx. Their structures were elucidated on the basis of spectral data analysis, including 1D, 2D-NMR, and HR-ESI-MS. Compounds 1 and 2 showed the inhibition of nitric oxide (NO) production with IC₅₀ values of 9.13 and 19.94 μM, respectively.     

Four new C20-diterpenoid alkaloids from Aconitum rotundifolium

Four new C₂₀-diterpenoid alkaloids, rotundifosines D-G (1–4), along with eight known ones (5–12) were isolated from the whole plant of Aconitum rotundifolium Kar. & Kir. The structures of the compounds were elucidated on the basis of spectroscopic analyses, including HR-ESI-MS and 1D, 2D NMR. Rotundifosine F (3) is a rare C₂₀-diterpenoid alkaloid with quaternary ammonium salt. Alkaloids 1–4, 5, 6, 9, and 12 were evaluated for cytotoxicity against MCF-7, HCT-116 and HepG2 human cancer cell lines.     

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.     

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.     

无花果内生真菌FL10中吲哚二酮哌嗪类生物碱的研究

目的:研究无花果内生真菌FL10的次级代谢产物成分.方法:采用反复硅胶柱色谱法、Sephadex LH-20凝胶色谱法等进行分离纯化,从无花果内生真菌FL10的发酵液中分离得到6个吲哚二酮哌嗪类生物碱.结果:通过理化常数测定和光谱分析,6个吲哚二酮哌嗪类生物碱鉴定分别为verruculogen(1),cyclotryprostatins B(2),fumitremorgin C(3),cyclotryprostatin A(4),tryprostatin A(5),tryprostatin B(6).结论:这6个吲哚二酮哌嗪类生物碱是首次从无花果内生真菌中分离得到,目前研究表明,无花果内生真菌Aspergillus tamarii可以作为一种可产生吲哚二酮哌嗪类生物碱的新来源.