槟榔不同工艺处理品中3种生物碱的含量比较

目的:建立测定槟榔中槟榔碱、槟榔次碱、去甲基槟榔次碱的HPLC-DAD方法,并比较不同处理工艺对3种生物碱含量的影响.方法:采用Partisil 10 SCX阳离子色谱柱(4.6 mm ×250 mm,10 μm),流动相乙腈-0.5％磷酸(三乙胺试液调pH 3.8)(50∶50),流速1 mL· min-1,检测波长215 nm,柱温25℃.考察不同烘烤时间和不同煮沸时间条件下,槟榔皮、核中3种生物碱含量的变化.结果:生物碱的含量随烘烤或者煎煮时间的延长而降低,在不同工艺条件下降低程度不同,100℃烘烤8h后生物碱的损失率约50％,煮沸0.5h后损失率高达80％,说明煎煮对槟榔皮、核中生物碱含量的影响较烘烤要大.结论:所建立的HPLC-DAD方法适用于槟榔生物碱的定量分析,为开发槟榔药食两用的安全剂量标准提供实验依据.     

LC-PDA测定槟榔、焦槟榔、大腹皮中4种生物碱的含量

目的 建立槟榔、焦槟榔、大腹皮中槟榔碱、槟榔次碱、去甲槟榔碱、去甲槟榔次碱含量的LC-PDA测定方法。方法 样品加氨水浸润,甲醇超声提取。ZORBAX 300-SCX色谱柱(150 mm×4.6 mm,5 μm)分离,流动相A为乙腈,流动相B为0.2%甲酸水(氨水调节pH=3.8),梯度洗脱,流速为0.5 mL·min^-1,柱温为30 ℃,进样量为5 μL,检测波长为225 nm。结果 槟榔碱、槟榔次碱、去甲槟榔碱、去甲槟榔次碱分别在2.98~745.0,2.60~650.0,1.19~595.0,1.22~610.0 μg·mL^-1内与峰面积呈良好的线性关系(r²>0.999 0)。槟榔、焦槟榔、大腹皮中槟榔碱、槟榔次碱、去甲槟榔碱、去甲槟榔次碱的平均加样回收率(n=6)均在95.0%~105.0%,RSD<2.50%。槟榔和其炮制品中生物碱含量差异显著。结论 该方法快捷、灵敏、准确、可靠,可用于槟榔、焦槟榔、大腹皮中4种生物碱的含量测定,为槟榔及其饮片的质量控制提供了参考。     

Areca nut extract lowers the permeability of vaginal mucosa to reduced arecoline and arecaidine

Areca nut chewing has been implicated in the development of oral cancer and oral submucous fibrosis. Arecoline and arecaidine, which are alkaloids present in the areca nut, are thought to play a major role in the development of adverse effects resulting from this chewing habit. Because these alkaloids appear to be associated with the development of the above diseases, we determined their diffusion kinetics through human vaginal mucosa in the presence and absence of a 1% areca nut extract. Seven clinically healthy vaginal mucosa specimens (mean patient age+/-standard deviation, 52+/-13 years; age range, 38-74 years) were obtained during surgery. In vitro flux values of reduced arecoline and arecaidine (r-arecoline and r-arecaidine) were determined through use of a flow-through diffusion apparatus. Analysis of variance, a Duncan multiple range test, and an unpaired t-test were used to determine steady state kinetics and flux differences over time intervals. The flux values across vaginal mucosa of r-arecoline and r-arecaidine were decreased in the presence of 1% areca nut extract. For r-arecoline, these flux values were significantly lower statistically when compared to those obtained in the absence of areca nut extract. These findings concur with results previously obtained for water, where the astringent action of the tannins present in the areca nut extract was thought to alter the barrier properties of the epithelium, resulting in decreased permeability.     

Minimal arecaidine concentrations showing a promotion effect during DMBA-induced hamster cheek pouch carcinogenesis

The purpose of the present study was to determine the minimal arecaidine concentrations showing a synergistic effect on DMBA-induced hamster cheek pouch carcinogenesis. One hundred and twelve male adult Syrian golden hamsters were divided into 16 groups, each containing seven animals. After eight weeks of DMBA initiation and then four weeks of arecaidine promotion. 100% tumor incidence was found with arecaidine concentrations of 400 μg/ml and 500 μg/ml; average tumor numbers were 1.86 ± 0.63 and 1.86 ± 0.93 respectively ( P < 0.05). After four weeks of DMBA and a subsequent eight weeks of arecaidine painting, all hamsters developed visible tumors with arecaidine concentrations of 900 μg/ml and 1000 μg/ml: average tumor numbers were l.86 ± 0.82 and 2.14 ± 1.09 respectively ( P < 0.05). The tumor dimensions varied little and differences were not statistically significant. Without DMBA pretreatment, regardless of the high arecaidine concentrations (1000μg/ml, 2000μg/ml and 3000μg/ml) applied, no visible tumor growth was observed: only hyperkeratosis and inflammation could he discerned histologically. Thus, the minimal concentrations of arecaidine displaying a synergistic effect in the DMBA-induced hamster cheek pouch of carcinogenesis were found to be 400 μg/ml applied for four weeks after eight weeks of DMBA application, and 900 μg/ml applied for eight weeks after four weeks of DMBA painting. These findings may be useful for other studies concerning the tumorgenicity of arecaidine.     

高效液相色谱法比较槟榔不同工艺处理品中三种生物碱的含量

摘要 目的：建立同时测定槟榔中槟榔碱、槟榔次碱、去甲基槟榔次碱的HPLC-DAD方法,并比较不同处理工艺对这三种生物碱含量的影响。方法：采用Partisil 10 SCX阳离子色谱柱（4.6 mm?50 mm,10 μm）,流动相为乙腈—0.5%磷酸（三乙胺试液调pH=3.8）（50：50）,流速1 mL穖in-1,检测波长215 nm,柱温25 ℃。结果：不同处理工艺对槟榔三种主要生物碱含量的影响是有差别的。结论：所建立的HPLC-DAD方法适用于槟榔生物碱的定量分析,三种生物碱含量均随着工艺处理时间的延长而降低,且在不同工艺条件下降低程度不同,本研究为开发槟榔的食用药用安全剂量标准奠定了基础。     

Cancer induction in the DMBA hamster cheek pouch: a modified technique using a promoter

OBJECTIVE: To evaluate a modified method of carcinogenesis induction using the 9,10-dimethyl-1,2-benzanthracene (DMBA) sustained-release suture technique followed by arecaidine promotion in the hamster cheek pouch model. STUDY DESIGN: Prospective, controlled animal study. METHODS: Number 3-0 cotton sutures were impregnated with DMBA and coated with silicone elastomer. These sutures were placed in the cheek pouch of Syrian hamsters in the submucosal space to a length of approximately 1.5 cm. The suture placement was confirmed every 2 weeks and replaced if lost. After 12 weeks, the DMBA-coated sutures were removed. The cheek pouches were everted and painted with a solution of arecaidine three times weekly for up to an additional 4 weeks or until the tumor reached a size of 100 mm2. RESULTS: We placed sutures in 165 Syrian hamster cheek pouches. Of these, 133 hamsters (80.6%) produced squamous cell carcinomas that reached a size of 100 mm2 and then were randomly selected for treatment in a new drug trial. Twenty-six hamsters (15.8%) were found dead and 6 (3.6%) were killed because of severe inflammation. CONCLUSIONS: The DMBA hamster cheek pouch model is a reliable and efficient animal model for inducing squamous cell carcinoma and can be used to study upper aerodigestive tract tumors.     

槟榔次碱对未孕大鼠离体子宫平滑肌运动的影响

目的:观察不同浓度的槟榔次碱对未孕大鼠离体子宫平滑肌运动的影响及研究其作用机理。方法:取大鼠卵巢及子宫叉处的子宫角,沿系膜纵轴剖开,置于恒温的平滑肌槽中,连接生理记录仪,观察槟榔次碱对子宫平滑肌运动的影响,以及分别加入五种阻断剂后.观察槟榔次碱对子宫作用的影响。结果:不同剂量的槟榔次碱能显著增强子宫平滑肌收缩的频率、振幅,增大收缩面积并呈正相剂量效应关系,而对收缩波的持续时间无明显影响,阻断M-受体后,槟榔次碱的增强作用即消失。而H1受体、L-型钙通道、α-受体、前列腺素合成酶分别被阻断或抑制后,槟榔次碱的增强作用未受到明显影响。结论:槟榔次碱能显著增强离体子宫平滑肌的收缩作用,并且主要是通过兴奋M-受体而发挥作用。     

槟榔次碱对未孕大鼠离体子宫平滑肌运动的影响

目的：观察不同浓度的槟榔次碱对未孕大鼠离体子宫平滑肌运动的影响及研究其作用机理。方法：取大鼠卵巢及子宫叉处的子宫角,沿系膜纵轴剖开,置于恒温的平滑肌槽中,连接生理记录仪,观察槟榔次碱对子宫平滑肌运动的影响,以及分别加入五种阻断剂后.观察槟榔次碱对子宫作用的影响。结果：不同剂量的槟榔次碱能显著增强子宫平滑肌收缩的频率、振幅,增大收缩面积并呈正相剂量效应关系,而对收缩波的持续时间无明显影响,阻断M-受体后,槟榔次碱的增强作用即消失。而H1受体、L-型钙通道、α-受体、前列腺素合成酶分别被阻断或抑制后,槟榔次碱的增强作用未受到明显影响。结论：槟榔次碱能显著增强离体子宫平滑肌的收缩作用,并且主要是通过兴奋M-受体而发挥作用。