大孔吸附树脂法提取HK-1菌株抗菌物质

Paenibacillus brasilensis HK-1蜂房蜜蜂球菌(Melissococcus pluton)、蜂房哈夫尼菌(Hafnia alvei)、幼虫芽孢杆菌(Paenibacillus larvae)、蜜蜂球囊菌(Ascosphaera apis)等蜜蜂病原具广谱抗性。本试验通过牛津杯法测定抗菌物质的效价,研究了大孔吸附树脂对HK-1菌株产生的抗菌物质的吸附、解吸性能,筛选了解吸剂。结果表明,大孔吸附树脂X-5对抗菌物质的吸附及解吸性能最好,其饱和吸附量为5.85mg/g,最佳解吸剂为80%丙酮,以80%丙酮进行动态解吸,解吸率达95.3%。     

利用大孔吸附树脂提取吸水链霉菌BS-112菌株的抗真菌活性物质

吸水链霉菌(Streptomyces hygroscopicus)BS-112菌株产生的活性物质对白僵菌、绿僵菌、黄曲霉菌等家蚕病原真菌具有抑制作用。研究了7种大孔吸附树脂对BS-112菌株产生的抗真菌活性物质的吸附和解吸性能,筛选出对抗真菌活性物质具有良好吸附及解吸性能的X-5树脂,这种树脂对抗真菌活性物质的饱和吸附量为73.37 mg/g,最适吸附温度为25℃,饱和吸附时间为150 min,以75%乙醇进行动态解吸,解吸率可达93.68%。BS-112菌株产生的抗真菌活性物质在低浓度下即能够较好地抑制家蚕病原白僵菌的生长,96孔板法测定的最小抑菌浓度(MIC)和杀菌浓度(MFC)分别为1.56、3.13μg/mL,并且活性物质的毒性小,以2 000 mg/kg剂量的活性物质灌胃小鼠,小鼠无急性中毒表现。     

大孔树脂富集纯化藏药五脉绿绒蒿总黄酮的工艺研究

优选大孔吸附树脂富集纯化藏药五脉绿绒蒿中总黄酮的工艺条件。方法:以芦丁为对照品,采用紫外分光光度法测定总黄酮含量;以比吸附量、吸附率、比解吸量、解吸率等为指标,采用静态吸附-解吸试验,考察AB-8、NKA-9、D-101、HPD-100、DM-301等5种型号的树脂;采用单因素试验,考察静态吸附-解吸附动力学曲线,确定吸附、解吸时间,筛选解吸剂浓度、上样液浓度、流速等参数。结果:D-101型大孔树脂对五脉绿绒蒿总黄酮吸附-解吸性能较好,比吸附量492.8μg/g、吸附率68.21%,解吸率83.26%。工艺条件为:室温下,以2.0 mL/min流速循环上样动态吸附2次,后静态吸附,动态吸附和静态吸附的累计时间不小于8 h。以70%乙醇解吸,静态解吸2 h以上,后以1.0 mL/min流速进行动态解吸,静态解吸和动态解吸累计时间不小于8 h。上样液粗总黄酮质量浓度为140～150μg/mL,每100 mL上样液需装柱树脂为30～40 g。结论:经D-101大孔树脂纯化后,所得干浸膏中总黄酮纯度提高2.96倍,回收率62.23%,纯化物粘度和吸湿性降低,工艺稳定可行,五脉绿绒蒿总黄酮得到有效的富集,为后期的新药开发研究提供参考。     

紫象草原花青素的大孔吸附树脂纯化、平均聚合度及体外抗氧化活性测定

试验旨在利用大孔吸附树脂获得紫象草原花青素粗提物的纯化条件,并探究在此条件下获得纯化物的平均聚合度和体外抗氧化活性。试验采用单因素试验设计,通过比较3种大孔吸附树脂D101、HP-20、ADS-17的吸附与解吸特性,选择最优吸附树脂类型,对紫象草原花青素粗提物进行洗脱浓度、上样体积和洗脱体积的筛选,之后利用静态和动态吸附与解吸试验确定纯化条件,制备紫象草原花青素纯化物;采用盐酸-香草醛法测定纯化物原花青素的平均聚合度及其对1,1-二苯基-2三硝基肼（DPPH·）和羟自由基（·OH）的体外清除率。结果表明：采用φ2.6 cm×30.0 cm玻璃层析柱,选择D101型大孔吸附树脂、90%(V/V)浓度乙醇为洗脱剂,上样体积480 mL,洗脱体积155 mL时纯化效果较好,纯化后原花青素B2含量由纯化前10.01 mg/g提升至13.82 mg/g,提升了36.06%;紫象草原花青素粗提物经乙酸乙酯萃取后,获得水相和乙酸乙酯相的平均聚合度分别为26.87、13.63,纯化物的平均聚合度为14.67。当紫象草原花青素纯化物质量浓度为100 mg/mL时,DPPH·、·OH清除能力分别为99.5...     

AB-8大孔树脂吸附、解吸中药复方多糖的研究

为了对中药复方免疫增强剂中的多糖进行纯化,试验采用苯酚硫酸法测定多糖含量,然后以AB-8大孔吸附树脂为分离材料,以乙醇为洗脱剂,系统研究上样液浓度、上样量、上样速率、洗脱剂乙醇浓度、洗脱速率、洗脱剂用量等参数对多糖吸附-解吸效果的影响。结果表明:上样液浓度为5.93 mg/mL,上样量为1倍量树脂体积(BV),上样液速率为每小时2倍树脂体积数;洗脱剂乙醇浓度为50%,洗脱速率为每小时3倍树脂体积数,洗脱剂用量为3倍量树脂体积;吸附率为74.3%,解吸率为93.6%。说明AB-8大孔树脂可用于纯化中药复方免疫增强剂中的多糖。     

利用大孔树脂纯化荷叶黄酮动态吸附条件的研究

本试验从荷叶中提取黄酮类化合物,通过大孔树脂对黄酮的静态吸附效果的比较,研究X-5、HP-20、AB-8及HPD-100四种大孔吸附树脂的吸附效果,然后使用通过动态吸附与解吸试验就上样流速、上样液浓度、pH、解吸曲线以及洗脱剂的浓度对大孔吸附树脂(HP-20)的吸附效果进行研究,确定荷叶黄酮的分离纯化的最佳技术参数。结果表明,HP-20分离效果最好,静态饱和吸附量可达45.84 mg,解吸量为41.20 mg,解吸率为89.9%;在提取液总黄酮含量为0.50～0.70 mg/mL,pH为提取原液4.6左右,上样流速为1.5 mL/min的条件下,HP-20对荷叶总黄酮的吸附量可以达到5.20 mg/mL,体积分数越高的乙醇水溶液洗脱效果越好。     

大孔吸附树脂纯化黄芪茎总皂苷的工艺及其抗氧化性

以总皂苷吸附率、解吸率为考察指标,探索LSA-10型大孔吸附树脂纯化黄芪茎总皂苷的工艺条件及其体外抗氧化性。采用单因素和正交试验法对黄芪茎总皂苷的纯化工艺进行优化;采用分光光度法探索了黄芪茎总皂苷样品液对DPPH·、·OH的清除活性。试验结果表明,黄芪茎总皂苷的最佳纯化工艺条件为:以生药浓度为0.015 g/ml的黄芪茎样品液进行试验,吸附时间为4 h,样品液用量/树脂质量的比值为15 ml/g,解吸液乙醇体积分数为85%,解吸液用量/树脂质量的比值为15 ml/g,解吸时间为1 h。在上述试验条件下黄芪茎总皂苷样品液的平均吸附率和解吸率分别为90.68%和93.13%,纯化后的干浸膏总皂苷含量由5.94%提高到12.66%;黄芪茎总皂苷样品液对DPPH·、·OH具有较强的清除能力。因此,LSA-10型大孔吸附树脂对黄芪茎总皂苷具有富集作用,其优化的纯化工艺条件比较稳定;黄芪茎总皂苷提取液具有一定的抗氧化活性,而且其清除活性随着质量浓度的增加而逐渐增强。     

大孔吸附树脂纯化蜂胶总黄酮的工艺研究

[目的]研究大孔树脂纯化蜂胶总黄酮的最佳工艺。[方法]以比吸附量、解析率及总黄酮回收率为指标,通过考察静态吸附及解吸附试验,筛选最佳大孔吸附树脂。并考察该最佳大孔吸附树脂的动态吸附及解吸附条件,包括原液质量浓度、吸附流量、吸附时间、洗脱剂及洗脱剂流量,从而筛选出该大孔吸附树脂分离纯化蜂胶总黄酮的最佳工艺条件。[结果]FL—1大孔树脂为四种树脂中吸附及解吸附性能最佳,因此选择FL-1为分离纯化蜂胶总黄酮的最佳树脂,其最佳工艺参数为:动态吸附质量浓度0.25～0.38mg/mL,吸附速度1mL/min;树脂柱吸附时间1h,洗脱剂为90%乙醇,洗脱流量为1mL/min。蜂胶总黄酮质量分数从13.07%提高到85.25%,总黄酮回收率达79.37%。[结论]FL-1型大孔树脂对蜂胶总黄酮具有良好的纯化效果,优选工艺合理、稳定、可行。     

DM-2型大孔树脂分离纯化沙枣多糖的工艺研究

为了确定DM-2型大孔树脂分离、纯化沙枣多糖的最佳工艺条件,试验以吸附率和解吸率为评价指标,通过动态试验研究各因素对分离、纯化效果的影响。结果表明:在沙枣多糖样品溶液浓度为0.7 mg/mL、pH值为10、上样速率为2.5 BV/h、上样量为3 BV、洗脱剂浓度为55%、洗脱速率为1 BV/h、洗脱剂用量为3 BV时,吸附率和解吸率分别达到84.05%和97.3%。说明在同时满足上述条件的情况下,DM-2型大孔树脂分离、纯化沙枣多糖的效率较高。     

盐穗木活性成分的提取及抑菌效果研究

为了解盐穗木的药用性能,试验将盐穗木全草经乙醇提取得到总浸膏,结合药敏试验对其进行效价测定。结果表明:盐穗木总浸膏对大肠埃希杆菌、金黄色葡萄球菌、枯草芽孢杆菌有一定的抑菌活性,对酵母菌、链球菌、普通变形杆菌无抑菌作用;不同季节盐穗木总浸膏的抑菌活性无差异。进一步提取得到有机酸、生物碱、酚类3种物质进行试验,结果有机酸与酚类抑菌作用不明显,生物碱有一定的抑菌作用。利用不同有机溶剂对生物碱进行分离,结果乙酸乙酯相抑菌活性明显,其他分离相抑菌效果均不明显。乙酸乙酯对金黄色葡萄球菌和枯草芽孢杆菌的最小抑菌浓度为7.8 mg/mL,对大肠埃希杆菌的最小抑菌浓度为15.6 mg/mL。总浸膏对大肠埃希杆菌、金黄色葡萄球菌和枯草芽孢杆菌的最小抑菌浓度为31.2 mg/mL。     

大孔吸附树脂分离纯化贯叶连翘中的金丝桃素

以贯叶连翘粗提浸膏为材料,考察六种大孔吸附树脂分离纯化金丝桃素的性能。采用静态、动态吸附方法筛选树脂,以中压分离方法确定分离纯化的条件。结果表明:HZ-801树脂以其高吸附率、高洗脱率成为优选的分离填料,其最佳分离纯化条件为:进样液质量浓度为0.1125 g/m L,进样速度为5.0 m L/min、洗脱速度为20 m L/min,0~95%乙醇溶液梯度洗脱,金丝桃素的纯度为79.11%。HZ-801可以较好分离纯化金丝桃素,纯化工艺具有较大实践性及参考价值。     

Florfenicol pharmacokinetics in lactating cows after intravenous, intramuscular and intramammary administration

Pharmacokinetics of florfenicol 30% injectable solution was determined in lactating cows after intravenous, intramammary and intramuscular administration. Serum concentration-time data generated in the present study were analysed by non-compartmental methods based on statistical moment theory. Florfenicol half-life was 176 min, mean residence time 129 min, volume of distribution at steady-state 0.35 L/kg, and total body clearance 2.7 mL/min·kg after intravenous administration at 20 mg/kg. The absorption after intramuscular administration appeared slow and the kinetic parameters and the serum concentration vs. time curve were characteristic of absorption rate-dependent elimination. The absorption after intramammary administration of florfenicol at 20 mg/kg was good (53.9%) and resulted in serum concentrations with apparent clinical significance. The intramammary administration resulted in serum florfenicol concentrations that were significantly higher than the respective serum concentrations following Intravenous administration 4 h after administration and thereafter. Florfenicol absorption was faster from the mammary gland than from the muscle. The maximum serum concentrations ( C _max) were 6.9 μg/mL at 360 min after intramammary administration and 2.3 μg/mL at 180 min after intramuscular administration. The bioavailability of florfenicol was 54% and 38% after intramammary and intramuscular administration, respectively. The C _max in milk was 5.4 μg/mL at 180 min after intravenous and 1.6 μg/mL at 600 min after intramuscular administration.     

高效液相色谱法测定卡巴匹林钙产品与反应液中尿素的含量

建立了一种用于测定卡巴匹林钙产品及反应液中尿素含量的高效液相色谱法。确立了最佳色谱条件,色谱柱为Waters Xselect HSS T3 C18(4.6 mm×250 mm,5μm),流动相为甲醇-水(5∶95,V/V),检测波长为195 nm,流速为0.5 mL/min,柱温30℃,进样量10μL。结果显示,尿素浓度在0.12~120μg/mL范围内具有良好的线性关系(r>0.999);尿素的检出限为0.05μg/mL,定量限为0.1μg/mL;分离度R>1.5,重复性的相对标准偏差<1.0%(n=6);卡巴匹林钙产品中尿素的回收率范围为98.10%~100.46%,反应液中尿素的回收率范围为95.32%~101.20%。本方法操作简便、准确度高、重复性好、检测速度快,在合成工艺中对卡巴匹林钙产品及反应液中尿素的质量控制具有重要意义。     

HPLC法同时测定金叶清瘟散中绿原酸和咖啡酸的含量

为了建立一种同时测定金叶清瘟散中绿原酸和咖啡酸的高效液相色谱法,选用Agilent C18色谱柱（250 mm×4.6 mm,5μm）,乙腈∶0.4%磷酸溶液（12∶88）等度洗脱,进样量10μL,流速1 mL/min,波长为328 nm进行检测。结果显示,被测物质能与样品中杂质有效分离,绿原酸和咖啡酸分别在5.0~500μg/mL和2.5~250μg/mL的范围内呈良好线性关系,两种药物的添加回收率均高于95%,RSD小于2%。该方法简便、快速、准确,适用于金叶清瘟散中绿原酸和咖啡酸的同时检测。     

大孔树脂纯化苜蓿总黄酮及纯化前后抗氧化能力比较

为了探讨苜蓿(Medicago sativa L.)总黄酮的大孔树脂纯化工艺及其抗氧化特性,本研究以苜蓿干粉为试验材料,采用静态和动态试验,考察树脂种类、上样液pH值和体积、上样液总黄酮质量浓度对苜蓿总黄酮吸附和解吸附性能的影响,确定最佳纯化工艺条件,比较纯化后苜蓿总黄酮的抗氧化能力。结果表明,D101大孔树脂纯化苜蓿总黄酮效果较好,上样液pH值为3、体积为2 BV、质量浓度为0.8mg·mL^-1纯化参数条件下,吸附率和解析率分别为92.72%和75.99%,此条件下苜蓿总黄酮纯度为31.46%。纯化后的苜蓿总黄酮质量浓度为0.8mg·mL^-1时,对二苯基-2-三硝基苯肼自由基、2,2’-联氮-双-3-乙基苯并噻唑啉-6-磺酸、羟自由基和超氧自由基的清除率分别为92.46%,92.23%,87.79%和86.73%,羟自由基的清除能力高于Vc,其它3种自由基清除能力较Vc有所降低,纯化后苜蓿总黄酮抗氧化能力明显增强。     

Pharmacokinetics and the effect of application site on a novel, long-acting transdermal fentanyl solution in healthy laboratory Beagles

Application of transdermal drugs to different anatomical sites can result in different absorption characteristics. The pharmacokinetics (PKs) and bioequivalence of a single 2.6 mg/kg (50 μL/kg) dose of a novel, long-acting transdermal fentanyl solution were determined when applied topically to the ventral abdominal or dorsal interscapular skin of 40 healthy laboratory Beagles. The PKs were differentiated by a more rapid initial absorption of fentanyl from the dorsal application site. Mean plasma fentanyl concentrations remained above 0.6 ng/mL from 4 to 96 h in the dorsal application group and from 8 to 144 h in the ventral application group. Bioequivalence analysis demonstrated that the sites were not equivalent; the 90% confidence intervals of the ratio of the geometric means for both the maximum concentration (C(max)) and the area under the curve (AUC) were not contained within the 80-125% interval. The C(max) was 2.34 ± 1.29 (mean ± standard deviation) and 2.02 ± 0.84 ng/mL for the ventral and dorsal application groups, respectively. The terminal elimination half-lives (t(1/2)) for both groups were similar with values of 137 ± 58.9 and 117 ± 59.6 h for the ventral and dorsal application site groups, respectively. A mean absorption rate of ≥ 2 μg · kg/h was maintained from 2 to 144 h following dorsal application and from 2 to 264 h following ventral application. These results suggest that transdermal fentanyl solution could be applied as a single dose to the dorsal scapular area 2-4 h prior to surgery with analgesia lasting a minimum of 4 days.     

Removal of Endotoxin from Swine Visfatin Recombinant Lipoprotein Solution

The purpose of this experiment was to explore the influence of the sample loading volume,flow rate and protein concentration of protein on the removal efficiency of endotoxin and protein loss in porcine visfatin recombinant lipoprotein solution at 4 ℃.In this study,commercial and modified polymyxin B was used as the medium to specifically remove endotoxin from the protein solution.At 4 ℃,different loading volume (6,8,10,12 mL),flow rate (0.125-0.75 mL/min) and protein concentration (0.9,1.2,1.3,1.7 mg/mL) were used for endotoxin removal,the content of endotoxin was determined by a quantitative method using endotoxin reagents,and the BCA protein concentration assay kit was used to determine the protein concentration.The results showed that at 4 ℃ endotoxin removal was the best when protein loading volume was 12 mL and flow rate was 0.125-0.25 mL/min,the removal rate was 99.5%,the residual amount of endotoxin (1 EU/mL) was within the biosafety range,and the protein recovery rate was as high as 90% to 95%.The experimental results showed that the endotoxin was successfully removed from the solution of porcine lipin recombinant protein,which provided references for the related functional research and clinical application of the protein.