This work was supported by the Korea Ministry of Environment (MOE) as part of the "The advancement of scientific research and technological development in environmental science program (E315-00015-0414-2)."

1.在管理集管的制备注意:正执行切割步骤，以消除对药物吸附在管的长度差异的影响。 <ol><li>使用标记标注与管型的管子的端部（ 例如，聚氯乙烯，聚氨酯（PU），和聚烯烃（PO））。 </li><li>删除所有可拆卸的配件，如连接器和针头盖。 </li><li>用锋利的剃刀，以确保清洁的边缘，从滴注腔的连接切断的管，以1米的长度。 </li></ol> 2.药物注射稀释注:使用玻璃瓶（1L）作为容器用于注射药液。执行精确的稀释步骤。验证市售药物产物的组成，并使用相同的批号为一个整体的实验组。 <ol><li>标签与瓶药品名称（ 例如，地西泮或他克莫司）。 </li><li>稀释药物注射s的5％葡萄糖溶液，从5毫克/毫升到100微克/毫升为地西泮注射（2毫升在100 5毫升％葡萄糖溶液西泮注射的），并从5毫克/毫升至10微克/毫升为他克莫司注射（ 200μL的在100毫升的5％葡萄糖溶液）他克莫司注射。 注:设置基于临床使用的药物和溶剂的测试浓度。 </li><li>轻轻旋动，以获得均匀的药液混合在瓶稀释液。 </li><li>在使用玻璃琥珀色小瓶收集稀释液（1-10）溶液量筒。 </li><li>验证的浓度，如在步骤4中描述。 注:使用药物这些浓度作为出发点的浓度。 </li></ol> 3.吸附动力学研究使用输液泵注:确认使用前的吸附试验的泵管依赖性流量由于管的硬度。在精确的时间点，利用GLA采集样品SS瓶和小瓶，以防止在储存期间另外的药物吸附。 如图2进行测试。保护药物溶液对轻若该药具有光敏性。一式三份进行实验。 <ol><li>而不产生气泡，预加载药物的稀释液到使用注射器的管。 <ol><li>该管的一端连接到注射器。 </li><li>把管子的另一端插入瓶装药液。 </li><li>拉回注射器柱塞直到管被完全填充有药液。 </li></ol></li><li>安装管插入的输液泵。 <ol><li>打开输液泵的门，推释放杆。 </li><li>将预装管进入输液泵并保持平直。 </li><li>在安装完毕后的管端取下注射器。 </li><li>放管的一端插入一个耐化学性的硼硅玻璃量筒收集药物SOLUT离子穿过管后。 </li></ol></li><li>置（PVC，PU，或PO）的基础上管中的给药装置的类型的流量和药物（ 例如，用于安定1毫升/分钟和用于他克莫司为10mL /小时）。 </li><li>在不同时间点收集样品到琥珀色小瓶，在室温下。 <ol><li>收集1毫升西泮样品在0.05，0.30，0.55，和1.05小时。 </li><li>收集10毫升的他克莫司样品在1.05，2.05，3.05，4.05和小时。 </li></ol></li></ol>使用药物4.分析高效液相色谱法（HPLC） 注:建议在药物分析的HPLC方法在参考文献1，8，9描述。使用串联质谱法（MS / MS）和免疫测定法的样品制备作为可供选择的方法10，11之后。一式三份进行实验。<ol><li>权衡药物和在作为储备溶液1mg / mL的浓度溶解它们的有机溶剂。 <ol><li>使用甲醇作为安定原液的溶剂，由于西泮在5％葡萄糖中的低溶解度。 </li><li>使用乙腈作为用于他克莫司原液的溶剂，由于他克莫司在5％葡萄糖中的低溶解度。 </li></ol></li><li>通过稀释储备溶液制备标准溶液。 <ol><li>用甲醇稀释安定储备溶液至0.3125，0.625，1.25，2.5，5.0，10.0和20.0微克/毫升。 </li><li>用5％的右旋糖稀释他克莫储备溶液至2.5，5.0，10.0，15.0，和20.0微克/毫升。 </li></ol></li><li>分析使用带有UV检测1，8，9 HPLC法的标准。 注意:使用适当的检测方法（紫外，荧光等 ）进行药物分析。 <ol><li>斯坦注射10μLdards成用装有C18柱（1.5毫米×250毫米，5微米）进行UV检测的HPLC系统。 表1为分析条件。 </li><li>确认的特异性和线性1，8，9。 <ol><li>对于特异性，监测药物峰（ 即，是否它是从其他峰在色谱分离），以确定该药物。 </li><li>确认在校准范围的线性度（即峰面积结果是否成正比的浓度）。 </li></ol></li><li>得到校正曲线。 <ol><li>创建基于浓度对从标准1，8，9色谱峰面积值图。 </li><li>获得的线性回归方程，R 2为校准曲线（ 例如， </em&gt; Y = AX + B，X:药物，Y的浓度:峰面积）1，8，9。 </li></ol></li></ol></li><li>稀从吸附研究样品用甲醇安定或5％葡萄糖为他克莫司酌情使得它们落在校准范围内，并直接注入稀释样品的10μL的入HPLC系统。 </li></ol> 5.计算药物浓度与吸附量的<ol><li>计算使用校准曲线（未知x和已知的y）的样本的药物浓度。 </li><li>计算使用下面的等式药物的吸附量: <img alt="式（1）" src="/files/ftp_upload/55086/55086eq1.jpg" /> 其中S:吸附量（％） 合:注射的稀释后的药物浓度（微克/毫升） CP:药物浓度通过管子传递（微克/毫升） </li></ol>

<ol>
	<li>Jin, S. E., You, S., Jeon, S., Hwang, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diazepam+sorption+to+PVC-+and+non-PVC-based+tubes+in+administration+sets+with+quantitative+determination+using+a+high-performance+liquid+chromatographic+method.">Diazepam sorption to PVC- and non-PVC-based tubes in administration sets with quantitative determination using a high-performance liquid chromatographic method.</a> Int. J. Pharm. 506, 414-419 (2016).</li><li>Treleano, A., Wolz, G., Brandsch, R., Welle, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+into+the+sorption+of+nitroglycerin+and+diazepam+into+PVC+tubes+and+alternative+tube+materials+during+application.">Investigation into the sorption of nitroglycerin and diazepam into PVC tubes and alternative tube materials during application.</a> Int. J. Pharm. 369, 30-37 (2009).</li><li>Shibata, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adsorption+and+pharmacokinetics+of+cyclosporin+A+in+relation+to+mode+of+infusion+in+bone+marrow+transplant+patients.">Adsorption and pharmacokinetics of cyclosporin A in relation to mode of infusion in bone marrow transplant patients.</a> Bone Marrow Transplant. 25, 633-638 (2000).</li><li>Kawano, K., Tsubouchi, R., Nakajima, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Effect+of+the+Drip+Condition+(Container+Materials,+Concentration,+Drip+Rate)+on+the+Sorption+of+Nicardipine+Hydrochloride+from+Injection+to+the+Intravenous+Infusion+Set.">The Effect of the Drip Condition (Container Materials, Concentration, Drip Rate) on the Sorption of Nicardipine Hydrochloride from Injection to the Intravenous Infusion Set.</a> Jap. J. Hosp. Pharm. 18, 491-495 (1992).</li><li>Hanawa, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+the+release+behavior+of+diethylhexyl+phthalate+from+the+polyvinyl-chloride+tubing+for+intravenous+administration.">Investigation of the release behavior of diethylhexyl phthalate from the polyvinyl-chloride tubing for intravenous administration.</a> Int. J. Pharm. 210, 109-115 (2000).</li><li>Mason, N. A., Cline, S., Hyneck, M. L., Berardi, R. R., Ho, N. F., Flynn, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+affecting+diazepam+infusion:+solubility,+administration-set+composition,+and+flow+rate.">Factors affecting diazepam infusion: solubility, administration-set composition, and flow rate.</a> Am. J. Hosp. Pharm. 38, 1449-1454 (1981).</li><li>Roberts, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+solute+sorption+into+plastic+tubing+during+organ+perfusion+and+intravenous+infusions.">Modeling solute sorption into plastic tubing during organ perfusion and intravenous infusions.</a> J. Pharm. Sci. 85, 655-665 (1996).</li><li>Patel, P., Panchal, S., Mehta, T., Solanki, S., Patel, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversed-Phase+High+Performance+Liquid+Chromatographic+(RP-HPLC)+Method+For+Determination+of+tacrolimus+in+bulk+and+pharmaceutical+formulation.">Reversed-Phase High Performance Liquid Chromatographic (RP-HPLC) Method For Determination of tacrolimus in bulk and pharmaceutical formulation.</a> Int. J. Pharm. Pharm. Sci. 3, 220-222 (2011).</li><li>Mercolini, L., Mandrioli, R., Iannello, C., Matrisciano, F., Nicoletti, F., Raggi, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+analysis+of+diazepam+and+its+metabolites+in+rat+plasma+and+brain+tissue+by+HPLC-UV+and+SPE.">Simultaneous analysis of diazepam and its metabolites in rat plasma and brain tissue by HPLC-UV and SPE.</a> Talanta. 80, 279-285 (2009).</li><li>Wang, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deposition+of+diazepam+and+its+metabolites+in+hair+following+a+single+dose+of+diazepam.">Deposition of diazepam and its metabolites in hair following a single dose of diazepam.</a> Int. J. Legal. Med. , (2016).</li><li>Wallemacq, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multi-site+Analytical+Evaluation+of+the+Abbott+ARCHITECT+Tacrolimus+Assay.">Multi-site Analytical Evaluation of the Abbott ARCHITECT Tacrolimus Assay.</a> Ther. Drug. Monit. 31 (2), 198-204 (2009).</li><li>Morar-Mitrica, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+stable+low-dose+aglycosylated+antibody+formulation+to+minimize+protein+loss+during+intravenous+administration.">Development of a stable low-dose aglycosylated antibody formulation to minimize protein loss during intravenous administration.</a> MAbs. 7 (4), 792-803 (2015).</li><li>Trissel, L. A., Xu, Q. A., Baker, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug+compatibility+with+new+polyolefin+infusion+solution+containers.">Drug compatibility with new polyolefin infusion solution containers.</a> Am. J. Health Syst. Pharm. 63, 2379-2382 (2006).</li><li>Cloyd, J. C., Vezeau, C., Miller, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Availability+of+diazepam+from+plastic+containers.">Availability of diazepam from plastic containers.</a> Am. J. Hosp. Pharm. 37, 492-496 (1980).</li><li>Ceccherini, G., Perondi, F., Lippi, I., Grazia, G., Marchetti, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravenous+lipid+emulsion+and+dexmedetomidine+for+treatment+of+feline+permethrin+intoxication:+a+report+from+4+cases.">Intravenous lipid emulsion and dexmedetomidine for treatment of feline permethrin intoxication: a report from 4 cases.</a> Open Veterinary Journal. 5 (2), 113-121 (2015).</li><li>Lei, W., Yu, C., Lin, H., Zhou, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+tacrolimus-loaded+transfersomes+for+deeper+skin+penetration+enhancement+and+therapeutic+effect+improvement+in+vivo.">Development of tacrolimus-loaded transfersomes for deeper skin penetration enhancement and therapeutic effect improvement in vivo.</a> Asian J. Pharm. Sci. 8, 336-345 (2013).</li><li>Maiguy-Foinard, A., Blanchemain, N., Barthelemy, C., Decaudin, B., Odou, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+a+double-lumen+extension+tube+on+drug+delivery:+examples+of+isosorbide+dinitrate+and+diazepam.">Influence of a double-lumen extension tube on drug delivery: examples of isosorbide dinitrate and diazepam.</a> PLoS One. 11 (5), 0154917 (2016).</li></ol>

The authors have nothing to disclose.

药物吸附到给药装置在静脉给药意想不到的药物损失的原因。期间吸附，药物通常分配在输液的早期阶段管的聚合物材料;在达到吸附平衡后，药物的递送量被稳定1。药物的吸附量应评估和最小化。药物吸附数的评价方法进行了研究，例如泵的方法和滴注法。相比于滴的方法，该泵的方法可以容易地没有偏压操纵。虽然给药设置与流量调节器（常规形式）在滴注法的使用，所希望的流率小于5毫升/小时是很难达到的。因此，我们建议在行政套管吸附评价泵的方法。 当使用泵的方法，影响药物吸收到管理组管主要因素是药物properties（ 例如，疏水性和电荷），吸附试验的条件（ 例如，药物浓度，流速，溶剂的相容性，管的长度和温度），用于药品的分析方法（ 例如，HPLC和MS）和该管的聚合物在给药组（ 例如，PVC，PU，和PO）2，3，4，5，6，7，8，9，10，11，12。首先，模型药物的选择是用于获得精确和准确的实验结果是至关重要的。尽管西泮管理是由精神药物管制法跟踪，我们选择地西泮（ 图1a）和他克莫司（ 图1b </s仲&gt;），因为他们的高吸附量给药聚合物管的模型药物设置1或容器13，14。在这种情况下，药物以高浓度显示出比那些在低浓度时在输液1,2的早期阶段的吸附少。这些药物具有较高的log P值（地西泮:2.82 15，他克莫司:3.96 16）和低溶解度，作为分类的生物制药分类系统（BCS第2类）。因为它们的疏水性的，这些药物可与施用套管相互作用，从而导致吸附。其他药物显示出较高的吸附量（如硝酸甘油2和环孢霉素A 3）可以作为替代车型药物吸附评价。此外，大分子药物，如生物制剂（抗体治疗剂，胰岛素， <em&gt;等。 ），可以应用于关于药物吸附12给药组的质量评价。 我们建立了一个简单的动力学研究的吸附，用泵轻松获得精确的结果，并尽量减少工件（ 图2）。在泵的方法，药物溶液（ 图2a）通过安装后的管切割从给药组（ 图2b）被传递到输注泵（ 图2c）。除了从管理集管，所有的设备（瓶，量筒，和取样瓶）分别组成耐化学腐蚀的高硼硅玻璃，以防止其他药物吸附到聚合物。在这项研究中，管而不会在1米的固定长度其它配件被用来简化药物吸附的因素的施用套管中。如果临床情况需要它，可用于管的长度的乘法因子。在SORP化试验中，将稀释的药物溶液作为起始浓度1,7。分娩后，将药物溶液（ 图2d）收集到在不同时间点（ 图2e）的小瓶中。为取样药物溶液在流速和取样时间点的预选条件通过管完全通过。吸附一般发生在输液的早期阶段，并且图案后跟一个对流界面电阻扩散模型7。地西泮的吸附结果是可比的双腔延伸管模型17时，初始药物浓度被输送之前考虑。因此，采样时间点可以被修改，使得吸附评价花费较少的时间。的测试条件的所有因素的基础上的药物的临床应用得到了证实。 在这个协议中，我们选择了次根据之前的报道1，8，9药物分析E含量的方法。简单和可再现性HPLC方法已被开发出来。 HPLC条件列于表1。各种其它技术，如MS和免疫测定，也被开发为药物浓度10,11的替代分析方法。 MS / MS和免疫测定是用于检测的药物及其代谢物高度敏感。具体地，可以很容易地而不需要大型且昂贵的设备用于药物分析来进行的免疫测定。 关于给药装置，药物吸附质量评价PVC-和在施用套管使用非PVC基材料进行了研究。吸附在管理组管的评价开始选药，并与considera结束的可接受的吸附水平的标准化，如图（ 图3）。 PVC基管显示出高的吸附水平对于许多药物，如地西泮，他克莫司（ 表2），硝酸甘油2和环孢菌素A 3。间的方法，以减少药物吸附到施用的管设置为小于10％，替代材料或聚合物的组合已被开发，如基于PO的材料和层-层设计2，13。在PE / PB / PP共混在本研究中使用的给药组的基于PO管呈低吸附量，作为基于非PVC管。在另一方面，基于PE管不用于给药装置，但它们在商业上的市场作为一个注射器的延伸管，由于其硬度使用。 此协议可应用到的给药装置的质量控制相对于药物吸附。以上药物CLAS通过吸附量sified（最高，下，和最低）应当在吸附评价可用于给药装置的质量保证。该协议也可以在科研用于新的替代的聚合物材料或用于在不导致药物吸附1,13施用套管新设计的开发。

给药装置是由一个尖峰，一个滴注室，管，鲁尔接头（连接器），以及用于保护针头盖。其他配件，如气道单向阀，一调节夹具，一个在线过滤器，一个Y型管帽（喷射口），和一个针，也可附连到给药装置。药物吸附到管是在注射药物1的递送的一个关键问题。吸附描述了一种药物到聚合物表面和药物的吸收入聚合物基质2的吸附。药物吸附到管在给药装置会导致不可预测的药物损失，使得难以控制递送的药物的浓度。因此，药物吸附到聚合物管是一大障碍的注射药物的精确传递到体内。然而，存在用于药物吸附的评价在施用套管没有标准的方法或法规准则。 <html>药物吸附到管在施用套水平已使用各种评价方法1，2，3，4，5的报道。试验方法和模型药物在吸附评价的关键因素。泵方法1,3和滴注法4,5已被广泛地用于吸附试验。在一般情况下，泵的方法应该在药物与低浓度和低流率作为输注条件的情况下使用。使用各种评价方法，药物吸附的许多研究已经报道了聚氯乙烯（PVC） -和在施用非PVC基管套1，2，3，&gt; 5。许多吸附药物可以选择调查施用套管是否有药物吸附潜力或没有。地西泮（ 图1a）1,2，他克莫司（ 图1b）5，硝酸甘油2和环孢菌素A 3是代表性的药物，在PVC-高吸附和非PVC基管中。 药物吸附在管子的评价，试验条件如流速和药物浓度是基于临床使用所选药物1，6,7。在地西泮的情况下，以1毫升/分钟的流速，使用100微克/毫升的高浓度，以模仿为癫痫持续状态1的治疗的初始剂量。对于他克莫司，10微克/ mL的浓度以10毫升/小时的流速被交付。葡萄糖溶液（5％）用于药物注射的稀释，并且管长为固定在1微米。玻璃瓶和小瓶应当用来防止在实验和贮存过程中附加的吸附。 在这项研究中，我们进行了动力学吸附研究与模型药物，地西泮和他克莫司，用泵方法。该协议的具体细节，从管准备吸附评价，以前描述。药物吸附的评价方法已用于确认注射药物性质和建议的临床使用给药组注射上的情况下，逐案1，2，3，4，5，6，7。这个协议可以被用作标准技术的给药装置吸附评价。药物吸附的评价管国际标准可能是必要的，以确保药物输送的安全性和有效性。

<table>
	<tbody>
		<tr>
			<td>PVC IV sets</td>
			<td>Becton Dickinson 
			(BD) Co. Ltd. (Franklin Lakes, NJ, USA)</td>
			<td></td>
			<td>Internal diameter: 2.54 mm</td>
		</tr>
		<tr>
			<td>PU IV sets</td>
			<td>Tianjin Hanaco Medical (THM) Co.Ltd. 
			(Tianjin, China)</td>
			<td></td>
			<td>Non-PVC 
			Internal diameter: 2.54 mm</td>
		</tr>
		<tr>
			<td>Non-PVC Polyolefin IV sets</td>
			<td>Polyscientech, Co. Ltd (Anseong, Korea)</td>
			<td></td>
			<td>Non-PVC [PE elastomer/PP elastomer/PB elastomer(25/50/25, weight ratio) blend] 
			Internal diameter: 2.54 mm</td>
		</tr>
		<tr>
			<td>Syringe</td>
			<td>Korea Vaccine Co. Ltd. (Seoul, Korea)</td>
			<td>KOVAX-SYRINGE 1 mL 26G 1/2&#34;</td>
			<td></td>
		</tr>
		<tr>
			<td>Daewon diazepam injection</td>
			<td>Daewon Pharma. Co. Ltd. (Hwaseong, Gyunggi, Korea)</td>
			<td></td>
			<td>1 mg/mL, total 2 mL 
			Batch No.: P003 
			Composition: diazepam, propylene glycol, ethanol, benzyl alcohol, sodium benzoate, bezoic acid, water for injection</td>
		</tr>
		<tr>
			<td>Tacrobel injection</td>
			<td>Chong Keun Dang, Co. Ltd. (Seoul, Korea)</td>
			<td></td>
			<td>5 mg/mL, total 1 mL 
			Batch No.: AG002 
			Composition: tacrolimus hydrate, polyoxyl 60 hydrogenated castor oil (HCO-60), dehydrated alcohol</td>
		</tr>
		<tr>
			<td>5% Dextrose</td>
			<td>JW Pharmaceutical (Seoul, Korea)</td>
			<td>500 mL</td>
			<td></td>
		</tr>
		<tr>
			<td>5% Dextrose</td>
			<td>Daehan Pharmaceutical (Seoul, Korea)</td>
			<td>200 mL</td>
			<td>Bottle (glass)</td>
		</tr>
		<tr>
			<td>Amber vials</td>
			<td></td>
			<td>20 mL</td>
			<td>Glass</td>
		</tr>
		<tr>
			<td>Terumo infusion pump</td>
			<td>Terumo (Medical Corp., USA)</td>
			<td>TE-135</td>
			<td></td>
		</tr>
		<tr>
			<td>HPLC system with UV detector</td>
			<td>Agilent (Santa Clara, CA, USA)</td>
			<td>Agilent 1260</td>
			<td></td>
		</tr>
		<tr>
			<td>CAPCELL PAK C18 column&#160;</td>
			<td>Shiseido (Japan)</td>
			<td>90404</td>
			<td>1.5 mm x 250 mm, 5 &#956;m</td>
		</tr>
		<tr>
			<td>Diazepam</td>
			<td></td>
			<td></td>
			<td>From Daewon Pharma. Co., Ltd.</td>
		</tr>
		<tr>
			<td>Tacrolimus</td>
			<td>Teva Czech industries&#160; (Czech Republic)</td>
			<td></td>
			<td>From Chong Keun Dang, Co. Ltd.</td>
		</tr>
		<tr>
			<td>Acetonitrile</td>
			<td>Burdick and Jackson Co., Ltd. (MI, USA)</td>
			<td>3/1/9017</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Burdick and Jackson Co. Ltd. (MI, USA)</td>
			<td>AH230-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Water</td>
			<td>Burdick and Jackson Co. Ltd. (MI, USA)</td>
			<td>3/1/4218</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium dihydrogen phosphate</td>
			<td>Sigma (St. Louis, MO, USA)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphoric acid</td>
			<td>Sigma (St. Louis, MO, USA)</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

evaluation of drug sorption to pvc- and non-pvc-based tubes in administration sets using a pump

管理组是直接应用的药物进入体内交付工具和由一个尖峰，点滴室，管，鲁尔适配器（连接器），用于保护针头帽，配件等。药物吸附到施用套管是在安全性和有效性方面的关键问题。虽然药物吸附是在一个管理集的质量的一个重要因素，存在用于药物吸附在管子的调节没有标准的评价方法。在这里，我们描述了药物吸收的评估协议的管理组管。制成的聚氯乙烯（PVC）管 - 和非PVC基聚合物材料被切成1米的长度。地西泮和他克莫司被用作模型药物。在动力学吸附研究中，我们选择了基于这些药物的临床使用的药物浓度和流速。在玻璃瓶中每种药物的稀释后，将稀释的药物溶液通过管输送施用的设置使用的泵。样品收集在琥珀色小瓶在适当的时间点，并使用高效液相色谱法进行分析的药物。药物浓度和吸附各级行政套管进行了计算。建议上可接受的标准，以确保的给药装置的质量。

吸附在给药组管使用模型药物，地西泮（ 图1a）和他克莫司被动力学监测（ 图1b），和泵的方法（ 图2）。稀释药物（ 图2a）的混合物在使用输液泵（ 图2c）的固定流动速率（ 图2b图 ）通过PVC-和非基于PVC的管子通过。玻璃瓶被稍微打开以允许施用集管的插入。该药物是通过管（ 图2d）递送后，将样品收集到琥珀色小瓶（ 图2e）。所有样品，包括在出发点的样品，用UV检测（ 图3a）的HPLC法进行分析。分析条件列于表1。对于药品标准的编制，地西泮和他克莫司溶解在满足因为它们在5％葡萄糖不溶性的hanol和乙腈。在吸附研究的起点药物浓度从样品中的药物稀释后的分析来计算。在PVC-和给药组的非PVC基管吸附水平通过通过从校准曲线（ 图3b），从100％中减去这些值（ 图3c）的管子流路后计算剩余药物含量的百分比来确定。药物吸附百分数的建议可接受的范围是小于10％，基于注射从药典12的内容。此外，特定的药物（ 如，抗癌药）应与临床准则来确认。我们确定了样品的药物吸附量是否适当（ 图3d）。 图4和图5显示在低和高的C代表色谱图的药物分别标准和样品溶液，oncentrations。每种药物的保留时间是8.2安定分钟，6.8分钟他克莫司。有从基体没有干扰峰。具体地，在地西泮吸附研究的样品中，干扰峰不与在色谱药物峰重叠，虽然不同的溶剂（5％右旋糖）中比在标准中使用。 表2示出地西泮和他克莫司的代表吸附量的计算。每种药物的吸附量是基于PVC的管中的最高的，在基于PU-管低，最低在基于PO管在动力学吸附试验的初始阶段。 <img alt="图1" src="/files/ftp_upload/55086/55086fig1.jpg" /> 图1的模型药物的化学结构:（1）地西泮和（b）他克莫司。地西泮是苯二氮衍生物和他克莫司是23元大环内酯类内酯。这个数字已被修改1。 <img alt="图2" src="/files/ftp_upload/55086/55086fig2.jpg" /> 图2.测试组使用泵的吸附动力学研究。 （ 一 ）药物在一个瓶子用5％葡萄糖稀释，（ 二 ）给药组（长度为1米）的管中，通过管子（C）输注泵，（D）药物，和（e）琥珀色小瓶储存。为了尽量减少额外药物吸附，制备并分别储存在玻璃瓶中和琥珀小瓶对于注射，药物溶液和样品。这个数字已被修改1。 <img alt="图3" src="/files/ftp_upload/55086/55086fig3.jpg" /> 图3.药物吸附量计算到的管理组管的关键步骤。 （ 一 ）分析使用带有UV检测，（b）由校正曲线的药物浓度和（c）吸附量（％），并进行药物吸附可接受准则（ 四 ）建议的计算的HPLC方法的药物。在吸附评价几个步骤，从药物选择的吸附试验，如药物浓度的分析和吸附量的计算，进行说明。 <img alt="图4" src="/files/ftp_upload/55086/55086fig4.jpg" /> 图4.安定的代表性色谱图。 （ 一 ）空白（甲醇）中，在标准（ 二 ）0.3125微克/毫升和（c）20微克/毫升，以及（d）样品（5％右旋糖）。在8.2分钟内检测到地西泮的峰，和在保留时间波动的色谱1分钟内发生。从溶剂中峰呈现befo重新在空白和标准色谱图的地西泮高峰。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55086/55086fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/55086/55086fig5.jpg" /> 图5.他克莫司的代表色谱图。 （ 一 ）空白（乙腈，用5％右旋糖），在标准（B）2.5微克/毫升和（c）20微克/毫升，和（d）的样品。在6.8分钟的他克莫司峰色谱成功分离。没有峰，与他克莫司的峰重叠。峰值为2.5微克/毫升检测，具有信噪比大于10 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55086/55086fig5large.jpg" target="_blank">，请点击此处查看该图的放大版本。</a> <table bo刻申="1"FO:保together.within页="1"FO:保持与 - next.within页="总是"&gt; <tbody><tr><td rowspan="2"> HPLC条件</td><td align="center" colspan="2">药物</td></tr><tr><td>地西泮</td><td>他克莫司</td></tr><tr><td>流动相</td><td>乙腈，甲醇，和磷酸钠缓冲液*中的混合物（29:47:24，体积/体积/体积，pH调至3.1，用磷酸） *磷酸二氢钠为1.2g / L的蒸馏水</td><td>乙腈（100％） </td></tr><tr><td>流量</td><td> 0.1毫升/分钟</td><td> 0.1毫升/分钟</td></tr><tr><td> UV波长</td><td> 232纳米</td><td> 213纳米</td></tr><tr><td>运行</td><td> 10分钟</td><td> 10分钟</td></tr><tr><td>保留时间</td><td> 8.2分</td><td> 6.8分</td></tr></tbody></table> 表1. HPLC条件。 <table border="1" fo:keep-together.wi薄页="1"FO:保持与 - next.within页="总是"&gt; <tbody><tr><td rowspan="2">药物</td><td colspan="3">吸附（％） </td></tr><tr><td> PVC </td><td> PU </td><td> PO </td></tr><tr><td>地西泮（0.05高） </td><td> 27.6±3.0 </td><td> 21.9±12.6 </td><td> 11.3±4.6 </td></tr><tr><td>他克莫司（1.05高） </td><td> 15.1±3.3 </td><td> 10.3±6.3 </td><td> 0.6±0.9 </td></tr></tbody></table> 表2代表性的吸附结果为在PVC-地西泮和他克莫司和非PVC基管（N = 3）1。

Watch this Scientific Journal Video about Evaluation of Drug Sorption to PVC- and Non-PVC-based Tubes in Administration Sets Using a Pump at JoVE.com

Evaluation of Drug Sorption to PVC- and Non-PVC-based Tubes in Administration Sets Using a Pump

我们报告使用泵和模型药物（地西泮和他克莫司），用于药物吸附的评价方法。后用高效液相色谱法，药物浓度和吸附水平的施用套管药物分析计算。

药物吸附的评价PVC-和非PVC为基础的管理管设置在使用泵

Administration sets are delivery tools for the direct application of drugs into the body and are composed of a spike, a drip chamber, tubes, Luer adapters ...

evaluation-drug-sorption-to-pvc-non-pvc-based-tubes-administration

Nanyang Technological University

Research

JoVE Journal

Medicine

10.5K 次观看.  Yonsei University. 我们报告使用泵和模型药物（地西泮和他克莫司），用于药物吸附的评价方法。后用高效液相色谱法，药物浓度和吸附水平的施用套管药物分析计算。 

视频: Evaluation of Drug Sorption to PVC- and Non-PVC-based Tubes in Administration Sets Using a Pump

Evaluation of Cancer Stem Cell Migration Using Compartmentalizing Microfluidic Devices and Live Cell Imaging

In the last 40 years, the United States invested over 200 billion dollars on cancer research, resulting in only a 5% decrease in death rate. A major obstacle for improving patient outcomes is the poor understanding of mechanisms underlying cellular migration associated with aggressive cancer cell invasion, metastasis and therapeutic resistance1. Glioblastoma Multiforme (GBM), the most prevalent primary malignant adult brain tumor2, exemplifies this difficulty. Despite standard surgery, radiation and chemotherapies, patient median survival is only fifteen months, due to aggressive GBM infiltration into adjacent brain and rapid cancer recurrence2. The interactions of aberrant cell migratory mechanisms and the tumor microenvironment likely differentiate cancer from normal cells3. Therefore, improving therapeutic approaches for GBM require a better understanding of cancer cell migration mechanisms. Recent work suggests that a small subpopulation of cells within GBM, the brain tumor stem cell (BTSC), may be responsible for therapeutic resistance and recurrence. Mechanisms underlying BTSC migratory capacity are only starting to be characterized1,4.
Due to a limitation in visual inspection and geometrical manipulation, conventional migration assays5 are restricted to quantifying overall cell populations. In contrast, microfluidic devices permit single cell analysis because of compatibility with modern microscopy and control over micro-environment6-9.
We present a method for detailed characterization of BTSC migration using compartmentalizing microfluidic devices. These PDMS-made devices cast the tissue culture environment into three connected compartments: seeding chamber, receiving chamber and bridging microchannels. We tailored the device such that both chambers hold sufficient media to support viable BTSC for 4-5 days without media exchange. Highly mobile BTSCs initially introduced into the seeding chamber are isolated after migration though bridging microchannels to the parallel receiving chamber. This migration simulates cancer cellular spread through the interstitial spaces of the brain. The phase live images of cell morphology during migration are recorded over several days. Highly migratory BTSC can therefore be isolated, recultured, and analyzed further. 
Compartmentalizing microfluidics can be a versatile platform to study the migratory behavior of BTSCs and other cancer stem cells. By combining gradient generators, fluid handling, micro-electrodes and other microfluidic modules, these devices can also be used for drug screening and disease diagnosis6. Isolation of an aggressive subpopulation of migratory cells will enable studies of underlying molecular mechanisms.

A compartmentalizing microfluidic device for investigating cancer stem cell migration is described. This novel platform creates a viable cellular microenvironment and enables microscopic visualization of live cell locomotion. Highly motile cancer cells are isolated to study molecular mechanisms of aggressive infiltration, potentially leading to more effective future therapies.

癌症干细胞迁移的使用进行划分的微流控芯片和活细胞成像的评价

In the last 40 years, the United States invested over 200 billion dollars on cancer research, resulting in only a 5% decrease in death rate. A major ...

科研

医学

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

股动脉和静脉导管采血，药品监督管理局和意识的血压和心率测量

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

测量和制止在弱视治疗

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

Intranasal Administration of CNS Therapeutics to Awake Mice

Intranasal administration is a method of delivering therapeutic agents to the central nervous system (CNS). It is non-invasive and allows large molecules that do not cross the blood-brain barrier access to the CNS. Drugs are directly targeted to the CNS with intranasal delivery, reducing systemic exposure and thus unwanted systemic side effects1. Delivery from the nose to the CNS occurs within minutes along both the olfactory and trigeminal neural pathways via an extracellular route and does not require drug to bind to any receptor or axonal transport2. Intranasal delivery is a widely publicized method and is currently being used in human clinical trials3.
Intranasal delivery of drugs in animal models allows for initial evaluation of pharmacokinetic distribution and efficacy. With mice, it is possible to administer drugs to awake (non-anesthetized) animals on a regular basis using a specialized intranasal grip. Awake delivery is beneficial because it allows for long-term chronic dosing without anesthesia, it takes less time than with anesthesia, and can be learned and done by many people so that teams of technicians can dose large numbers of mice in short periods. Efficacy of therapeutics administered intranasally in this way to mice has been demonstrated in a number of studies including insulin in diabetic mouse models 4-6 and deferoxamine in Alzheimer's mouse models. 7,8
The intranasal grip for mice can be learned, but is not easy and requires practice, skill, and a precise grip to effectively deliver drug to the brain and avoid drainage to the lung and stomach. Mice are restrained by hand using a modified scruff in the non-dominant hand with the neck held parallel to the floor, while drug is delivered with a pipettor using the dominant hand. It usually takes 3-4 weeks of acclimating to handling before mice can be held with this grip without a stress response. We have prepared this JoVE video to make this intranasal delivery technique more accessible.

A method to intranasally administer drugs to awake mice for the purpose of targeting the brain is described. This method allows for repeat dosing over long periods using intranasal administration of drug without anesthesia, and nose-to-brain delivery with minimal systemic exposure.

清醒小鼠滴鼻的CNS药物治疗管理

Intranasal administration is a method of delivering  therapeutic agents to the central nervous system (CNS). It is  non-invasive and allows large ...

Evaluation of a Novel Laser-assisted Coronary Anastomotic Connector - the Trinity Clip - in a Porcine Off-pump Bypass Model

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the Trinity Clip, is developed based on the excimer laser-assisted nonocclusive anastomosis technique. The Trinity Clip connector enables simplified, sutureless, and nonocclusive connection of the graft to the coronary artery, and an excimer laser catheter laser-punches the opening of the anastomosis. Consequently, owing to the complete nonocclusive anastomosis construction, coronary conditioning (i.e., occluding or shunting) is not necessary, in contrast to the conventional anastomotic technique, hence simplifying the off-pump bypass procedure. Prior to clinical application in coronary artery bypass grafting, the safety and quality of this novel connector will be evaluated in a long-term experimental porcine off-pump coronary artery bypass (OPCAB) study. In this paper, we describe how to evaluate the coronary anastomosis in the porcine OPCAB model using various techniques to assess its quality. Representative results are summarized and visually demonstrated.

This paper describes a novel nonocclusive coronary anastomotic connector in a porcine off-pump coronary artery bypass (OPCAB) model. This easy-to-use coronary connector has intrinsic potential to facilitate minimally invasive OPCAB surgery.

评价一种新型激光辅助冠状动脉吻合口连接器 - 三位一体夹 - 在猪非体外循环旁路模式

To simplify and facilitate beating heart (i.e., off-pump), minimally invasive coronary artery bypass surgery, a new coronary anastomotic connector, the ...

Quantification of the Immunosuppressant Tacrolimus on Dried Blood Spots Using LC-MS/MS

The calcineurin inhibitor tacrolimus is the cornerstone of most immunosuppressive treatment protocols after solid organ transplantation in the United States. Tacrolimus is a narrow therapeutic index drug and as such requires therapeutic drug monitoring and dose adjustment based on its whole blood trough concentrations. To facilitate home therapeutic drug and adherence monitoring, the collection of dried blood spots is an attractive concept. After a finger stick, the patient collects a blood drop on filter paper at home. After the blood is dried, it is mailed to the analytical laboratory where tacrolimus is quantified using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in combination with a simple manual protein precipitation step and online column extraction.
For tacrolimus analysis, a 6-mm disc is punched from the saturated center of the blood spot. The blood spot is homogenized using a bullet blender and then proteins are precipitated with methanol/0.2 M ZnSO4 containing the internal standard D2,13C-tacrolimus. After vortexing and centrifugation, 100 &#181;l of supernatant is injected into an online extraction column and washed with 5 ml/min of 0.1 formic acid/acetonitrile (7:3, v:v) for 1 min. Hereafter, the switching valve is activated and the analytes are back-flushed onto the analytical column (and separated using a 0.1% formic acid/acetonitrile gradient). Tacrolimus is quantified in the positive multi reaction mode (MRM) using a tandem mass spectrometer.
The assay is linear from 1 to 50 ng/ml. Inter-assay variability (3.6%-6.1%) and accuracy (91.7%-101.6%) as assessed over 20 days meet acceptance criteria. Average extraction recovery is 95.5%. There are no relevant carry-over, matrix interferences and matrix effects. Tacrolimus is stable in dried blood spots at RT and at +4 &#176;C for 1 week. Extracted samples in the autosampler are stable at +4 &#176;C for at least 72 hr.

Here we describe a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay to quantify the immunosuppressant tacrolimus in dried blood spots using a simple manual protein precipitation step and online column extraction.

在免疫抑制剂他克莫司对干血斑使用LC-MS / MS定量

The calcineurin inhibitor tacrolimus is the cornerstone of most immunosuppressive treatment protocols after solid organ transplantation in the United ...

Using Multi-fluorinated Bile Acids and In Vivo Magnetic Resonance Imaging to Measure Bile Acid Transport

Along with their traditional role as detergents that facilitate fat absorption, emerging literature indicates that bile acids are potent signaling molecules that affect multiple organs; they modulate gut motility and hormone production, and alter vascular tone, glucose metabolism, lipid metabolism, and energy utilization. Changes in fecal bile acids may alter the gut microbiome and promote colon pathology including cholerrheic diarrhea and colon cancer. Key regulators of fecal bile acid composition are the small intestinal Apical Sodium-dependent Bile Acid Transporter (ASBT) and fibroblast growth factor-19 (FGF19). Reduced expression and function of ASBT decreases intestinal bile acid up-take. Moreover, in vitro data suggest that some FDA-approved drugs inhibit ASBT function. Deficient FGF19 release increases hepatic bile acid synthesis and release into the intestines to levels that overwhelm ASBT. Either ASBT dysfunction or FGF19 deficiency increases fecal bile acids and may cause chronic diarrhea and promote colon neoplasia. Regrettably, tools to measure bile acid malabsorption and the actions of drugs on bile acid transport in vivo are limited. To understand the complex actions of bile acids, techniques are required that permit simultaneous monitoring of bile acids in the gut and metabolic tissues. This led us to conceive an innovative method to measure bile acid transport in live animals using a combination of proton (1H) and fluorine (19F) magnetic resonance imaging (MRI). Novel tracers for fluorine (19F)-based live animal MRI were created and tested, both in vitro and in vivo. Strengths of this approach include the lack of exposure to ionizing radiation and translational potential for clinical research and practice.

Using Multi-fluorinated Bile Acids and In Vivo Magnetic Resonance Imaging to Measure Bile Acid Transport

Tools to diagnose bile acid malabsorption and measure bile acid transport in vivo are limited. An innovative approach in live animals is described that utilizes combined proton (1H) plus fluorine (19F) magnetic resonance imaging; this novel methodology has translational potential to screen for bile acid malabsorption in clinical practice.

使用多氟化胆汁酸和<em&gt;在体内</em&gt;磁共振成像来衡量胆汁酸运输

Along with their traditional role as detergents that facilitate fat absorption, emerging literature indicates that bile acids are potent signaling ...

Novel Methods for Intranasal Administration Under Inhalation Anesthesia to Evaluate Nose-to-Brain Drug Delivery

Intranasal administration has been reported to be a potential pathway for nose-to-brain delivery of therapeutic agents that circumvents the blood-brain barrier. However, there have been few reports regarding not only the quantitative analysis but also optimal administration conditions and dosing regimens for investigations of nose-to-brain delivery. The limited progress in research on nose-to-brain pathway mechanisms using rodents represents a significant impediment in terms of designing nose-to-brain delivery systems for candidate drugs.
To gain some headway in this regard, we developed and evaluated two novel methods of stable intranasal administration under inhalation anesthesia for experimental animals. We also describe a method for the evaluation of drug distribution levels in the brain via the nose-to-brain pathway using radio-labeled [14C]-inulin (molecular weight: 5,000) as a model substrate of water-soluble macromolecules.
Initially, we developed a pipette-based intranasal administration protocol using temporarily openable masks, which enabled us to perform reliable administration to animals under stable anesthesia. Using this system, [14C]-inulin could be delivered to the brain with little experimental error.
We subsequently developed an intranasal administration protocol entailing reverse cannulation from the airway side through the esophagus, which was developed to minimize the effects of mucociliary clearance (MC). This technique led to significantly higher levels of [14C]-inulin, which was quantitatively detected in the olfactory bulb, cerebrum, and medulla oblongata, than the pipette method. This appears to be because retention of the drug solution in the nasal cavity was substantially increased by active administration using a syringe pump in a direction opposite to the MC into the nasal cavity.
In conclusion, the two methods of intranasal administration developed in this study can be expected to be extremely useful techniques for evaluating pharmacokinetics in rodents. The reverse cannulation method, in particular, could be useful for evaluating the full potential of nose-to-brain delivery of drug candidates.

Here, we describe two novel methods of stable intranasal administration under inhalation anesthesia with minimal physical stress for experimental animals. We also describe a method for quantitative evaluation of drug distribution levels in the brain via the nose-to-brain pathway using radiolabeled [14C]-inulin as a model substrate of water-soluble macromolecules.

吸入麻醉下鼻内给药评价鼻腔给脑药物的新方法

Intranasal administration has been reported to be a potential pathway for nose-to-brain delivery of therapeutic agents that circumvents the blood-brain ...

Implantation of an Isoproterenol Mini-Pump to Induce Heart Failure in Mice

Isoproterenol (ISO), is a non-selective beta-adrenergic agonist, that is used widely to induce cardiac injury in mice. While the acute model mimics stress-induced cardiomyopathy, the chronic model, administered through an osmotic pump, mimics advanced heart failure in humans. The purpose of the described protocol is to create the chronic ISO-induced heart failure model in mice using an implanted mini-pump. This protocol has been used to induce heart failure in 100+ strains of inbred mice. Techniques on surgical pump implantation are described in detail and may be relevant to anyone interested in creating a heart failure model in mice. In addition, the weekly cardiac remodeling changes based on echocardiographic parameters for each strain and expected time to model development are presented. In summary, the method is simple and reproducible. Continuous ISO administered via the implanted mini-pump over 3 to 4 weeks is sufficient to induce cardiac remodeling. Finally, the success for ISO model creation may be assessed in vivo by serial echocardiography demonstrating hypertrophy, ventricular dilation, and dysfunction.

The chronic administration of isoproterenol via an implanted osmotic pump has been used widely to mimic advanced heart failure in mice. Here, we describe detailed methods in surgical mini-pump implantation for the continuous isoproterenol administration over 3 weeks, as well as, echocardiographic assessment for the successful model creation.

植入异丙酮醇微型泵，以诱导小鼠心力衰竭

Isoproterenol (ISO), is a non-selective beta-adrenergic agonist, that is used widely to induce cardiac injury in mice. While the acute model mimics ...

Evaluation of Cerebral Blood Flow Autoregulation in the Rat Using Laser Doppler Flowmetry

When investigating the body&#39;s mechanisms for regulating cerebral blood flow, a relative measurement of microcirculatory blood flow can be obtained using laser Doppler flowmetry (LDF). This paper demonstrates a closed skull preparation that allows cerebral blood flow to be assessed without penetrating the skull or installing a chamber or cerebral window. To evaluate autoregulatory mechanisms, a model of controlled blood pressure reduction via graded hemorrhage can be utilized while simultaneously employing LDF. This enables the real time tracking of the relative changes in the blood flow in response to reductions in arterial blood pressure produced by the withdrawal of circulating blood volume. This paradigm is a valuable approach to study cerebral blood flow autoregulation during reductions in arterial blood pressure and, with minor modifications in the protocol, is also valuable as an experimental model of hemorrhagic shock. In addition to evaluating autoregulatory responses, LDF can be used to monitor the cortical blood flow when investigating metabolic, myogenic, endothelial, humoral, or neural mechanisms that regulate cerebral blood flow and the impact of various experimental interventions and pathological conditions on cerebral blood flow.

This article demonstrates the use of laser Doppler flowmetry to evaluate the ability of the cerebral circulation to autoregulate its blood flow during reductions in arterial blood pressure.

使用激光多普勒流测量测量大鼠脑血流自动调节的评估

When investigating the body&#39;s mechanisms for regulating cerebral blood flow, a relative measurement of microcirculatory blood flow can be obtained ...