合成冰片的短期遗传毒性测试研究

目的:观察合成冰片的致突变性。方法:采用Ames试验、细胞染色体畸变试验、小鼠骨髓嗜多染红细胞微核试验。结果:合成冰片在0.4~250μg/皿范围内(+/-S9mix),Ames试验结果阴性。24h的CHL细胞IC50为85.74μg.ml-1。80、40、20、10、5(μg.ml-1)下,染色体畸变率均<5%。微核试验中各剂量组微核发生率与阴性对照组比较差异无显著性,P>0.05。结论:合成冰片未显示致突变性。     

桑叶水提液的急性毒性和遗传毒性研究

目的初步评价桑叶水提液安全性。方法通过小鼠急性毒性试验、鼠伤寒沙门氏菌试验(Ames试验)、中国仓鼠肺成纤维细胞(CHL)染色体畸变试验和小鼠骨髓嗜多染红细胞微核试验,考察桑叶水提液急性毒性和遗传毒性。结果桑叶水提液40 g/kg小鼠灌胃给药未出现死亡,体质量与正常对照组比较无明显差异(P>0.05)。经Ames试验、CHL细胞染色体畸变试验、小鼠微核试验,桑叶水提液的结果均为阴性。结论本实验条件下,桑叶水提液的小鼠灌胃给药最大给药剂量为40 g/kg,且未表现出遗传毒性。     

健脾生血颗粒遗传毒性研究

目的:考察健脾生血颗粒的遗传毒性。方法:分别采用组氨酸营养缺陷性鼠伤寒沙门氏菌TA97a、TA98、TA100、TA102、TA1535回复突变试验(Ames试验),CHO-K1细胞染色体畸变试验,以及小鼠骨髓细胞微核试验考察健脾生血颗粒的遗传毒性。结果:Ames试验结果显示,健脾生血颗粒无论在含有和不含有S9的条件下,在所有评估剂量水平的所有测试菌中均可观察到重复正常生长,且所有剂量的回复突变数与阴性对照组相比均无2倍或以上的显著上调,且无剂量反应关系;CHO-K1细胞染色体畸变试验结果显示,健脾生血颗粒650、130、26μg/m L 3个剂量组的CHO-K1细胞染色体畸变率均小于5%;小鼠骨髓细胞微核试验结果显示,健脾生血颗粒3个剂量组[2 000、1 000和500 mg/(kg·d)]与阴性对照组间微核率差异均无统计学意义(P0.05)。结论:在本试验条件下,健脾生血颗粒未见遗传毒性作用。     

风湿平胶囊的致突变性研究

目的:研究风湿平胶囊的致突变性。方法:采用鼠伤寒沙门氏菌回复突变试验(Ames试验)、小鼠骨髓细胞微核试验、CHL细胞染色体畸变试验进行观察。结果:风湿平Ames试验、小鼠骨髓细胞微核试验结果均为阴性;CHL细胞染色体畸变试验结果为阳性,经过S9代谢活化后致突变作用降低。结论:风湿平胶囊在所试条件下有潜在的致突变性,经过S9代谢活化后致突变作用降低。     

雄黄致体内外染色体畸变

目的:用仓鼠体内染色体畸变试验和中国仓鼠肺细胞CHL细胞染色体畸变试验评价雄黄的遗传毒性.方法:体内染色体畸变试验用毛足属仓鼠连续ig5 d给予33.25,66.5,133 mg·kg-1剂量雄黄悬浊液,处死前2 h ip秋水仙素.处死后取骨髓细胞制备染色体.油镜下观察每只动物骨髓细胞的有丝分裂指数和100个中期分裂相细胞的畸变类型.CHL细胞染色体畸变试验用雄黄浸出液终质量浓度为0.15,0.3,0.6g.L-1作用于CHL细胞,培养24 h或48 h,终止培养前4h加入秋水仙素.收获细胞,制备染色体,油镜下观察CHL细胞有丝分裂指数和200个中期分裂相细胞的畸变类型.结果:①雄黄265.0 mg· kg -1组和雄黄530.0 mg·kg-1组ig给药和雄黄浸出液(1.2,2.4g·L-1)作用于CHL细胞显示有明显的抑制有丝分裂作用.②与阴性对照组比较,雄黄ig给药仓鼠体内染色体畸变率和雄黄体外给药CHL细胞染色体畸变率均显著升高,差异有极显著意义,且有明显的量效关系.但体内试验的畸变率低于体外试验.③雄黄给药后CHL细胞染色体畸变试验中可见较多的染色体断片,而仓鼠体内染色体畸变试验中未发现,这可能与体内外药物作用的方式不同.结论:①雄黄能致体内外细胞染色体畸变,具有遗传毒性.②仓鼠体内染色体畸变试验可作为中药新药遗传毒性评价试验组合的方法之一.     

高速逆流色谱法分离制备吴茱萸中的柠檬苦素、吴茱萸碱和吴茱萸次碱

目的:制备柠檬苦素、吴茱萸碱和吴茱萸次碱化学对照品.方法:采用高速逆流色谱法分离纯化吴茱萸二氯甲烷萃取物,溶剂系统为正己烷-乙酸乙酯-甲醇-水(5:5:5:5),上相做固定相,下相做流动相,流速2.0mL·min<'-1>,仪器转速855rpm,进样量400mg,所得样品由高效液相色谱法检测纯度.结果:一次分离可制备6.2mg柠檬苦素、10.4mg吴茱萸碱、10.5mg吴茱萸次碱,其纯度分别为96.2%、98.1%和95.4%.结论:该方法操作简单,可作为柠檬苦素、吴茱萸碱和吴茱萸次碱化学对照品的制备分离方法.     

高速逆流色谱法分离制备吴茱萸中的柠檬苦素、吴茱萸碱和吴茱萸次碱

目的:制备柠檬苦素、吴茱萸碱和吴茱萸次碱化学对照品。方法:采用高速逆流色谱法分离纯化吴茱萸二氯甲烷萃取物,溶剂系统为正己烷-乙酸乙酯-甲醇-水(5∶5∶5∶5),上相做固定相,下相做流动相,流速2.0mL.min-1,仪器转速855rpm,进样量400mg,所得样品由高效液相色谱法检测纯度。结果:一次分离可制备6.2mg柠檬苦素、10.4mg吴茱萸碱、10.5mg吴茱萸次碱,其纯度分别为96.2%、98.1%和95.4%。结论:该方法操作简单,可作为柠檬苦素、吴茱萸碱和吴茱萸次碱化学对照品的制备分离方法。     

妇科千金胶囊对CHL细胞株染色体畸变影响的研究

目的：探讨妇科千金胶囊对CHL细胞株染色体畸变的影响,对其遗传安全性进行研究。方法：采用CHL细胞培养技术,设妇科千金胶囊不同剂量对CHL细胞进行体外染毒,观察其致CHL细胞株染色体数目及结构变化。结果：在5000、2500、1 250μg/mL不同测试剂量浓度下,无论24h及48h收集细胞,并在＋S9与-S9两种测试条件下,进行染色体畸变分析,各剂量组、溶剂对照染色体畸变率均在正常范围。结论：妇科千金胶囊在本试验所确定的5000μg/mL测试剂量浓度内,初步认为无诱发CHL染色体畸变作用。     

蜂胶遗传毒性研究

目的探讨蜂胶的遗传毒性,为其应用提供安全性毒理学评价依据。方法用鼠伤寒沙门细菌营养缺陷型突变株TA97(a)、TA98、TA100和TA102,采用平板掺入法进行Ames试验,将实验分为加和不加代谢激活系统S9 2组平行试验。受试物蜂胶设5个剂量组(0.005、0.025、0.250、1.000、5.000 mg/皿)。应用小鼠骨髓嗜多染红细胞微核试验,检测小鼠骨髓嗜多染红细胞微核率;利用小鼠精子畸形试验,观察不同浓度的蜂胶致小鼠精子畸形的数目。结果在Ames试验中,蜂胶各剂量组引起的回变菌落数未超过对照组自发回变菌落数的1倍以上;小鼠骨髓嗜多染红细胞微核试验显示,蜂胶3个剂量组的微核发生率均在正常范围内,与阴性对照组比较无显著性差异(P>0.05),与阳性对照组比较差异显著(P<0.05);小鼠精子畸形试验可见,蜂胶3个剂量组的精子畸形率均在正常范围内,与阴性对照组比较无显著性差异(P>0.05),与阳性对照组比较差异显著(P<0.05)。结论蜂胶对所试菌株、小鼠体细胞及生殖细胞无诱变性。     

神昌滴丸的遗传毒性研究

目的:研究神昌滴丸的遗传毒性,为其应用提供安全性毒理学评价依据.方法:采用Ames试验、小鼠骨髓细胞染色体畸变试验、小鼠微核试验,考察神昌滴丸对鼠伤寒沙门茵和细胞的遗传毒性.结果:经Ames试验、小鼠骨髓细胞染色体畸变试验和小鼠微核试验,神昌滴丸的结果均呈阴性.结论:神昌滴丸无遗传毒性.     

Assessment of mutagenicity in Parthenium hysterophorus L

The mutagenic potential of a crude extract of Parthenium hysterophorus L. was assessed in the Salmonella/microsome (Ames) assay and the mouse bone marrow micronucleus test. Results in the bacterial mutagenicity assay were negative for the five strains employed, e.g. TA 1535, TA1537, TA 98, TA 100 and TA 102, while cytotoxicity was evident in all cases at 5000 microg per plate, the highest concentration assayed. A decrease in toxicity was observed with exogenous mammalian metabolic activation (S9) or glutathione (5 micromol per plate). When mutagenicity was monitored after column chromatography fractionation of the crude, fraction 1 was mutagenic in strain TA 98 (+S9). Besides, cytotoxicity was found in fraction 5, where parthenin was eluted. The micronucleus test was negative in mice upon oral administration, at doses up to 96 mg of crude per kg. Bone marrow toxicity was not observed. The crude extract exhibited some in vitro pro-oxidant activity. It also inhibited lipid peroxidation (IC(50)=4.1 microg/ml) but failed to act as .OH scavenger.     

液相色谱-质谱法同时测定吴茱萸水提物中吴茱萸碱和吴茱萸次碱的含量

目的:建立同时测定吴茱萸水提物中吴茱萸碱(Evodiamine,EVO)和吴茱萸次碱(Rutaecarpine,RUT)的LC/MS方法。方法:液相色谱采用Welch Materials Xtimate-C18色谱柱(2.1 mm×150 mm,3μm),以甲醇-10 mmol/L乙酸铵水溶液(85∶15)为流动相,流速0.2 mL/min,柱温30℃。质谱采用正离子全扫描模式,m/z:0~1000,电喷雾离子化源(ESI)。雾化气为氮气,雾化压力为40 psi;喷雾电压4000 V,源温度为100℃;去溶剂气为氮气,温度350℃,流速为10 L/min。结果:吴茱萸碱在2.02~504 ng/mL(r=0.9992),吴茱萸次碱在1.97~493.33 ng/mL(r=0.9999)线性关系良好,回收率分别为87.8%~97.04%和86.35%~98.22%,日内、日间精密度均10%。结论:该方法简便、可靠、灵敏度高,可用于同时测定吴茱萸水提物中吴茱萸碱和吴茱萸次碱的含量及药代动力学研究。     

Mutagenicity and antioxidant assessment of Stachitarpheta jamaicensis (L.) Vahl.

Stachitarpheta jamaicensis (L.) Vahl. is a member of the Verbenaceae commonly used in Cuba, mainly as vermifugue and against diarrhoea. The mutagenic potential of a hydroalcohol extract of its aerial parts was assessed in vitro using the Salmonella/microsome assay and in vivo in the mouse bone marrow micronucleus test. No positive response was observed in a battery of four Salmonella typhimurium strains employed: TA 1535, TA 1537, TA 98 and TA 100, when exposed to concentrations up to 5 mg/plate, with and without mammalian metabolic activation. In the same way, no increase in the micronucleus frequency in mitotic erythropoietic tissue was observed when animals were administered the extract orally in doses of 500, 1000 and 2000 mg/kg. The extract inhibited lipid peroxidation in the rat liver microsomal fraction (IC(50) = 3.6 microg/mL) but it does not seem to be an effective.OH radical scavenger (IC(50) = 76.7 microg/mL). Noteworthy, it increased in a dose dependent way the level of revertant colonies in E. coli IC 203, a strain sensitive to oxidative mutagenesis, when assayed together with hydrogen peroxide and ferrous sulphate, which suggests a pro-oxidant action.     

正交试验法优选姜制吴茱萸的炮制工艺

目的:对姜制吴茱萸的炮制工艺进行优选研究,为规范吴茱萸的炮制工艺提供技术参数。方法:采用L9(34)正交试验设计,以吴茱萸碱、吴茱萸次碱及柠檬苦素的含量为指标,考察姜用量、闷润时间、炒制温度、炒制时间4因素对姜制吴茱萸炮制工艺的影响。结果:姜用量、炒制温度、炒制时间对柠檬苦素含量有显著影响,药材与干姜比例为100∶7.5,闷润4 h,160℃炒制8 min为最佳炮制条件。结论:为规范姜制吴茱萸炮制工艺提供了部分科学依据。     

联苯双酯的致突变性研究

 联苯双酯为一新的治疗乙型病毒肝炎药物，本文对其致突变性进行了评价。当联苯双酯的浓度在１０，５０，１００和５００μｇ／皿，无论加或不加Ｓ＿９混合物对ＴＡ＿（９８）、ＴＡ＿（１００）、ＴＡ＿（１５３５）和ＴＡ＿（１５３７）菌株的回复突变作用均为阴性；当给昆明种小鼠剂量为３０００，１５００和７５０ｍｇ／ｋｇ的联苯双酯时，骨髓嗜多染细胞的微核率分别为３．３‰，３．３‰和１．７‰；当浓度为５００，２５０和１２５μｇ／ｍｌ对中国仓鼠肺细胞的染色体畸变率为３‰，２‰和２‰，上述结果表明，联苯双酯没有致突作用。     

关于当归、吴茱萸药对提取及含量测定的研究

目的:当归和吴茱萸药对是古方温经汤治疗月经不调、痛经的主要成分,确定当归和吴茱萸药对的提取方法。方法:采用紫外可见分光光度法对药对中总生物碱进行含量测定研究;采用HPLC法对药对中的阿魏酸、吴茱萸碱和吴茱萸次碱进行含量测定研究。结果:文章所提供的方法准确、重复性线性关系等符合要求,测定结果稳定、可靠,为当归、吴茱萸药对配伍的提取方法研究提供了依据。结论:单煎合并比合煎所得到的阿魏酸及吴茱萸碱含量高,本法可以为确定药对提取的方法提供科学可靠的依据。     

Simultaneous Analysis of 13 Major Bioactive Components in Evodiae rutaecarpa by HPLC-DAD-MS

Objective To control the quality of Evodia rutaecarpa better. Methods An HPLC-DAD-MS/MS method was established for the rapid and efficient identification of bioactive constituents and for simultaneous quantitative analysis of four bioactive ingredients including evodiamine, rutaecarpine, dehydroevodiamine, and evodin in E. rutaecarpa, which was applied to evaluating eight samples of E. rutaecarpa and its varieties from different areas. Results Thirteen potentially bioactive constituents including one flavonoid glycoside, one limonin, four indoloquinazoline alkaloids, and seven quinolone alkaloids were identified in all samples and the contents of dehydroevodiamine, evodine, evodiamine, and rutaecarpine varied widely from 0.10% to 0.51%, 0.49% to 3.12%, 0.07% to 1.56%, and 0.10% to 0.69%, respectively. Conclusion This method is found to be convenient, fast, accurate, and it is facilitated to improve the quality control standard of E. rutaecarpa and related products.     

UFLC-PDA同时快速测定藏药翼首草中5种化学成分的含量

目的：建立同时快速测定藏药翼首草药材中绿原酸、马钱苷、獐牙菜苷、吴茱萸苷、大花双参苷A含量的方法。方法：采用超快速液相色谱仪、Agilent Poroshell 120 SB-C18色谱柱（100 mm＆#215;4.6 mm,2.7μm）,柱温30益,流速1.0 mL·min-1,进样量4μL,流动相为：乙腈-0.2%磷酸水溶液,梯度洗脱,检测波长237 nm、325 nm。结果：绿原酸、马钱苷、獐牙菜苷、吴茱萸苷、大花双参苷 A分别在8.72-218.0、1.52-38.0、2.44-61.0、29.36-734.0、3.00-75.0μg·mL-1范围内（r〉.9996,n =6）呈良好线性关系,平均加样回收率分别为99.46%、99.41%、100.14%、98.89%、99.42%,RSD 分别为0.69%、0.66%、0.60%、1.21%、0.64%（n=9）。结论：该方法简便、准确、重复性好,可同时快速测定藏药翼首草中5种化学成分的含量。     

Antimutagenic effect of broccoli flower head by the ames salmonella reverse mutation assay

A study was performed to investigate the antimutagenic effect of broccoli flower head by the Ames Salmonella reverse mutation assay. Broccoli flower head being the most highly edible part in the plant was analysed for its antimutagenic effect. Without isolating the phytomolecules, the crude ethanol extract of broccoli flower head was tested for suppressing the mutagenic effect induced by certain chemical mutagens. Three strains - TA 98, TA102 and TA 1535 were used in the study. The tester strains were challenged with their respective mutagens. These were challenged with the ethanol extract of broccoli flower head at concentrations of 23 and 46 mg/plate. The plates were incubated for 72 h and the revertant colonies were counted. The crude extract did not prove to be promutagenic. The ethanol extract of the broccoli flower head at 46 mg/plate suppressed the mutagenic effect induced by the corresponding positive mutagens on all the three tester strains used in this study. The crude extract of broccoli flower head alone was not cytotoxic even at the maximum concentration tested (46 mg/plate). In conclusion, the ethanol extract of broccoli at 46 mg/plate suggests their diverse antimutagenic potential against the mutagenic chemicals employed in this study.