这份手稿基于以前发表的文章5。转载自5，版权到 2015 年，约翰 · 威利父子公司的许可。
我们很感激我们管理，珍妮特戚浩霖博士、 弗朗西斯科 · 阿尔瓦雷斯博士、 博士 Arwinder 凪博士大卫谢明，为他们的支持、 兴趣和鼓励执行这一研究项目，以及敏慧马博士和罗伯特 · 芒格，提供与我们甲状旁腺激素盐酸，其配方的片剂和药物有关的信息。我们也要感谢博士迈克尔 · 伯恩斯坦、 曼努埃尔 · 佩雷斯博士和博士圣地亚哥 · 多明格斯的建设性支持和发展目前商业版本的 Mnova 软件进行讨论，如它适用于 TD 核磁共振数据定量。此外，特别感谢你向先生 Regnar L.Madarang F.N.P.为我们提供通用环丙沙星片来进行我们的研究。

注意: 请使用任何化学品前咨询所有相关的材料安全数据表 (MSDS)。药物的物质通常是有毒，应使用适当的个人防护装备 (PPE; 处理例如， 手套、 安全眼镜、 白大褂、 全长的裤子和封闭的鞋子），在权衡粉末材料时特别小心。这推荐使用放置在罩与良好的空气流动平衡尽量把粉末从固体材料散发出周围平衡地区。
 注意: 软件指令是特定的 RINMR、 ri 值校正和 Mnova。对于其他 TD 核磁共振供应商设备和软件包，指示将会发生变化。
 1.制备的样品事先到 19 F TD 核磁共振测量 <ol><li>重校准 （粉原料药或活性药物成分 (API)） 样品中适当的核磁共振管 （2.5-3-5-，10-18-和 25 毫米 OD 核磁共振管） 直到标线 （为核磁共振探头线圈的高度），或在高度在 30 毫米左右。通知，高度可能取决于探头。 
	注: 甲状旁腺激素盐酸是 API 用于校准来确定甲状旁腺激素商业片中的药物含量的一个例子。重量将取决于密度的粉末。在甲状旁腺激素盐酸，权重的 API 时，周围 7.2、 3.5、 1.0、 0.3、 0.1，0.07 g 分别填土高度为 25-18-10-，5-，3-、 2.5 毫米核磁共振管，线圈。</li>
	<li>重配方的药物产品样本 （药片或胶囊） 在适当的核磁共振管直到标线 （为核磁共振探头线圈的高度） 或约 30 毫米的高度，如上文所述。 
	注意: 25-18 毫米管是适合于大型片胶囊或足够配方的药物产品不可用时。对于小片/胶囊或当没有足够的配方的药物产品是可用，但信号噪声比可能仍有足够的测量，使用 10 毫米管。直接，测量平板电脑或粉碎如果需要。他们的测量结果的变化没有观察观察了在这里，讨论 中所述。
	<ol><li>时准备商业片的甲状旁腺激素，重量大约十五个 90 毫克片剂 25 毫米核磁共振管 (约 9.2 g) 中，在 25 毫米核磁共振管 (约 9.6 g)，二十二个 30 毫克片剂 18 毫米 NMR 中的五十 30 毫克片剂管 (约 4.2 g)和 25 毫米核磁共振管 (约 8.8 g) 二十三 60 毫克片剂。</li>
	</ol></li></ol> 2。核磁共振仪的测量制备 <ol><li>调谐核磁共振仪器到 19 F <ol><li>进行的永磁核磁共振在温度测量仪器。在这里，使用 40 ° C;温度控制是没有供 26 毫米 19 F 核磁共振探头，用在这里。进行所有测量在 19 F 23 兆赫 （为 1 H) 核磁共振仪 22.0 兆赫频率。</li>
		<li>在 25 或 26 毫米核磁共振管内的铁氟龙标准样品并将其放入磁铁内的探头。允许为 5-10 分钟左右的平衡和调整到 19 F.选择仪器 &#34; 自动 O1 &#34; 下 &#34; 命令 &#34; RINMR 软件中的菜单。重复测量，至少 3 次，并采取 O 1 参数 （频率偏移量） 的最后一个值。</li>
	</ol></li> <li>19 F 90 度脉冲标定 <ol><li>F-19 与使用标准的自动的校准序列可用在文书中的铁氟龙标样标定 90 度脉冲。选择 &#34; 自动 P90 &#34; 下 &#34; 命令 &#34; RINMR 软件中的菜单。 
		注意: 通常情况下，90 度脉冲并不需要经常测量，除非该文书不执行或探针不用于一些时间。获取与这里使用 26 毫米 19 F 核磁共振探头铁氟龙为 7.05 微秒的 90 度值。</li>
	</ol></li></ol> 3。纯 API 标样的 T 1 或纵向弛豫时间测量 <ol><li>为更好的信号噪声比，放置在 25 毫米核磁共振管内 F-19 探头安装在磁铁的 API 样品让它平衡至少 10 分钟</li>
	<li>测量 T 1 弛豫时间，仪器手册，每个 API 示例，该示例使用一个标准的实验为反转恢复从 RINMR 软件的说明。 
	注: 推荐的试验参数都是 16 扫描收集与 0.1 微秒停留时间;8 个数据点;90 度脉冲，如上所述;和 10 微秒 F-19 26 毫米探测器死时间。8 从 100 微秒延误的数组 s (例如， 100 微秒，500 微秒，10 ms，100 ms，300 ms，700 毫秒，1 s，2 s，5 s 和 8 s) 药物与 CF 3 和延迟从 100 微秒到 20 s (如 100 微秒，500 微秒，1 ms，100 ms，500 ms，1 s，5 s，10 s，15 s 和 20 s) 与附加到芳香环的一氟组药物的建议。
	<ol><li>在 RINMR 软件、 负载 &#34; invrec.exe &#34; 脉冲序列，通过选择 &#34; 加载 &#34; 下 &#34; 序列 &#34; 和单击 &#34; 开放。 &#34; 检查的基本参数有正确的价值观，通过单击 &#34; A &#34; 或选择 &#34;收购 &#34; 下 &#34; 参数 &#34; 菜单。</li>
		<li>选择 &#34; 脚本 &#34; 下 &#34; 工具 &#34; 菜单; 将会弹出一个窗口。选择 &#34; T1.ris &#34; 选项卡，然后单击绿色箭头;将弹出一个窗口。选择具有适当时滞的延迟列表文件或单击后创建的列表 &#34; 开放。 &#34; 一旦延迟列表是令人满意，请单击 &#34; OK。 &#34; 注: 一个窗口将显示一个提示创建的文件的所有光谱将在收集为了计算 T 1 值。位置和文件夹和文件的名称是由用户。</li>
		单击 <li>&#34; 保存 &#34; 仪器开始获取数据并自动选定文件夹中保存的文件延误。 
		注: RIMNR 软件会创建 T 1 松弛曲线和计算 T 1 弛豫时间为每个案件。与 T 1 松弛曲线和时间常数或 T 1 值 . 会弹出一个窗口在这里，甲状旁腺激素 HCl T1 的值是 2.4 s.</li>
	</ol></li></ol> 4。测量校准样品与纯 API <ol><li>测量 API 样品建立校准曲线 <ol><li>地方一校准样品在磁铁为一个特定的 API。平衡为最小的小管 5 分钟，10 分钟大管样品。</li>
		<li>获得自由感应衰减 (FID) 或固体 (90 x 90y FID) 实验与仪器手册中的说明适当的参数。在 RINMR 软件中，加载 &#34; solid.exe &#34; (或 &#34; fid.exe &#34;) 脉冲序列，通过选择 &#34; 加载 &#34; 下 &#34; 序列 &#34; 并单击 &#34; 开放。 &#34; 注: 推荐的试验是固体固体材料的快速放松与短 T 2 s （横向弛豫时间）。推荐的实验参数是 128 扫描收集，与 0.1 微秒停留时间，1,024 的数据点，90 度脉冲 （在本例中，calibr 7.05 微秒使用铁氟龙不等式），和 10 微秒 19 F 26 毫米探测器死时间。7.5 循环延迟药物与 CF 3 组和 20 s s 药物与附加到芳香 （由于长 T 1 弛豫时间） 的 F 组有用于案件在这里，提出了一种基于其 T 1测量。</li>
		<li>测量所有样品的每一个 API 编写后他们都到了磁铁为 5-10 分钟来生成每个 API 示例的校准曲线的温度平衡。将数据保存在文件夹中，给每个实验运行一个不同的名称。</li>
		<li>计算量在每个管和考虑纯度的大管中的 API 示例是基于样品的重量百分比。</li>
	</ol></li></ol> 5。药物产品测量样品作为平板电脑 <ol><li>早已准备好的地方制定药物产品样本 （药片或胶囊） 的磁铁来衡量他们一次。平衡 5 分钟小核磁共振管或样品的更大的核磁共振管 10 分钟样本。</li>
	<li>获得相同核磁共振实验 （即 固体） 与校准样品相同的条件为每个特定 API。</li>
</ol> 6。一代的校准曲线与纯 API 示例添加数据从药物产品样本作为片剂的校准曲线 <ol><li>在这些情况下，选择该区域的固体实验从 5 到 300 点来创建校准曲线。如下所述，可以在三种不同的方式，分析了的数据。</li>
	<li>打开 RI 校准软件。通过单击要打开的 FID 或固体的文件从一个特定的 API 的剪辑符号撤回所有固体的数据。对于样本测的情况下，输入下的重量百分比 &#8220; 浓 &#8221; 列和下的重量 &#8220; 大众 &#8221; 列。选择相应的区域 （5-300 分） 使用平均方法来建立校准曲线 (打开的图标 &#8220; 显示核磁共振数据 &#8221; 选择范围的计算点)。保存的校准曲线。 
	注: Cal 列将提供药物测定样品中的百分比。</li>
	<li>在 RI 校准软件相同的选定区域 （5-300 点），与使用拟合的方法来建立校准曲线。保存的校准曲线。按照相同的说明与步骤 4.2.2。</li>
	<li>打开软件 （例如，Mnova）。从一个特定的 API 拖出固体的所有数据并输入重量百分比的标准样品;可以通过单击找到手动说明 &#8220; 内容 &#8221; 下 &#8220; 帮助 &#8221; 菜单。在 &#8220; 先进 &#8221; 菜单上，选择 &#8220; 时间域 &#8221;，然后 &#8220; 定量; &#8221; 将会弹出一个窗口。单击蓝色的加号来选择一体化区域。整合，相应的区域 （5-300 分） 用震级模式打造的校准曲线。</li>
	<li>保存校准曲线。在表中，输入每个样品的药物的百分比只根据标准 &#8220; 浓度 (x) &#8221; 列和下全部样品重量 &#8220; 质量 (m) &#8221; 列;校准样品应在红色 （检查）。请确保 &#8220; 信号函数 &#8221; 是 &#8220; y = s/m &#8221; （以正常化到信号/质谱）。</li>
	<li>为重复性、 再现，测量每个样品在不同的日子 3 倍和建立在这两个软件程序包的校准曲线。保存的校准曲线文件。</li>
</ol> 7。API 含氟药物计算重量在制定药物产品使用校准曲线 <ol><li>计算每片/胶囊的含氟 API 的重量百分比和平均剂量药物配方药物产品 <ol><li>使用的文件从 RI 校准软件和平均校准曲线法。从测得的片 （或胶囊） 读取核磁共振数据并输入他们的权重，以确定氟 API 在配方的药片或胶囊从校准曲线的重量百分比。 
		注: &#8220; Cal &#8221; 列将提供药物 （校准和商业片或未知） 所测样品中的百分比。请注意，O1 参数必须相同校准样品和片剂或胶囊为 RI 校准软件。</li>
		<li>使用该文件从 RI 校准软件校准曲线，拟合的方法。读片 （或胶囊） 的核磁共振数据测量和输入他们的权重，以确定氟 API 在配方的药片或胶囊从校准曲线的重量百分比。按照步骤 5.2.1 相同的说明。请注意，O 1 参数必须相同校准样品和片剂或胶囊为 RI 校准软件。</li>
		<li>使用 Mnova 文件和拖动所获得的 NMR 数据为制定 API （或测量它们都在同一天）。请按照步骤 4.2.4 中的说明。输入的权重确定氟 API 在配方的药片或胶囊从校准曲线的重量百分比。 
		注: 在表中，校准样品将红色 （检查） 和未知或商业片/胶囊的蓝色 （未选中）。&#8220; 浓度 (x) &#8221; 列会显示未知样品商业片的计算的值在基于值的校准样品在红色的蓝色。该软件不需要使用相同的 O1 参数，计算重量百分比的通知。</li>
		<li>作为平均值从下面公式计算每药片或胶囊含氟药物的量 (D = API 在毫克，W 核磁共振 剂量 = 重量百分比的 API 获得从校准曲线，w = 总重量制定产品从配方药物来自测量的片剂或胶囊和 N 的重量 = 一些配方的药片或胶囊用于测量) 与基于软件计算值比较校准曲线: <img alt="Equation 1"src="/files/ftp_upload/55850/55850eq1.jpg"/></li></ol></li></ol>

<ol>
	<li>El-Yazigi, A., Wahab, F. A., Afrane, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stability+Study+and+Content+Uniformity+of+Prochloroperazine+in+Pharmaceutical+Preparations+by+Liquid+Chromatography.">Stability Study and Content Uniformity of Prochloroperazine in Pharmaceutical Preparations by Liquid Chromatography.</a> J Chromatogr A. 690, 71-76 (1995).</li><li>Takeuchi, Y., Yoshida, M., Ito, A., Sunada, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uniformity+of+Drug+Content+During+Pharmaceutical+Dry+Granulation+by+Roller+Compaction+and+Tableting+Processes.">Uniformity of Drug Content During Pharmaceutical Dry Granulation by Roller Compaction and Tableting Processes.</a> J Drug Del Sci Tech. 19 (2), 119-124 (2009).</li><li>Toro, I., Dulsat, J. F., F&#225;bregas, J. L., Claramunt, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+Validation+of+a+Fully+Automated+method+of+the+Chromatographic+Determination+of+Content+Uniformity+of+Drug+Tablets.">Development and Validation of a Fully Automated method of the Chromatographic Determination of Content Uniformity of Drug Tablets.</a> J Pharm Biomed Anal. 36, 57-63 (2004).</li><li>Shi, Z., Hermiller, J. G., Gunter, T. Z., Zhang, X., Reed, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Novel+Sample+Selection+Strategy+by+Near-Infrared+Spectroscopy-Based+High+Throughput+Tablet+Tester+for+Content+Uniformity+in+Early-Phae+Pharmaceutical+Product+Development.">A Novel Sample Selection Strategy by Near-Infrared Spectroscopy-Based High Throughput Tablet Tester for Content Uniformity in Early-Phae Pharmaceutical Product Development.</a> J Pharm Sci. 101 (7), 2502-2511 (2012).</li><li>Silva Elipe, M. V., 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=Applications+of+19F+Time-Domain+NMR+to+Measure+Content+in+Fluorine-Containing+Drug+Products.">Applications of 19F Time-Domain NMR to Measure Content in Fluorine-Containing Drug Products.</a> Magn Reson Chem. 54 (6), 531-538 (2015).</li><li>Wang, J., 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=Fluorine+in+Pharmaceutical+Industry:+Fluorine-Containing+Drugs+Introduced+to+the+Market+in+the+Last+Decade+(2001-2011).">Fluorine in Pharmaceutical Industry: Fluorine-Containing Drugs Introduced to the Market in the Last Decade (2001-2011).</a> Chem Rev. 114, 2432-2506 (2014).</li><li>Dolbier, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide to Fluorine NMR for Organic Chemists. , (2009).</li><li>Barman, J. A., Scott, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cinacalcet+Hydrochloride.">Cinacalcet Hydrochloride.</a> Drugs. 65 (2), 271-282 (2005).</li><li>Horn, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Proton-Pump+Inhibitors:+Similarities+and+Differences.">The Proton-Pump Inhibitors: Similarities and Differences.</a> Clin Ther. 22 (3), 266-280 (2000).</li><li>LeBel, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ciprofloxacin:+Chemistry,+Mechanism+of+Action,+Resistance,+Antimicrobial+Spectrum,+Pharmacokinetics,+Clinical+Trials,+and+Adverse+Reactions.">Ciprofloxacin: Chemistry, Mechanism of Action, Resistance, Antimicrobial Spectrum, Pharmacokinetics, Clinical Trials, and Adverse Reactions.</a> Pharmacotherapy. 8 (1), 3-30 (1988).</li></ol>

作者没有透露。

随着在市场上更多的氟化药物可用，我们开发了一个具体和简单的方法，测量平均药物含量的含氟药物产品采用低场19F TD-核磁共振5。我们测试这个方法对三种商业药物产品: 含盐酸，甲状旁腺激素含有兰索拉唑，胶囊和片剂含环丙沙星盐酸盐单水合物。这种方法的优点是辅料和它的简单性氟缺乏特异性。我们测试的测量功能完整的配方 （胶囊或片剂） 甲状旁腺激素和?...

在制药行业，药物物质在他们单位的剂量形式 （例如，片剂和胶囊） 的含量必须在标签声称要满足质量控制和性能准则的范围内。高性能液相色谱法 (HPLC) 是用来衡量在他们制定的形式的药物的含量均匀度的常规分析技术。然而，该方法是冗长和破坏性，需要使用适当的溶剂分析前其配制成药物的溶出。甚至与自动化，过程可能需要 1-3 h 每个样品，根据适当的分析方法1，，23的发展。高效液相色谱法，除了近红外 (NIR) 被用作为同一目的和 PAT 的工具，有的需要的数据，使其执行长和更复杂4的化学计量学分析警告。这两种技术都具有破坏性，分析后丢弃的配方药物。
在这份手稿，我们描述了以前公布的5就我们组来测量含氟药物剂量形式 （例如，片剂和胶囊） 的平均含量方法的分步协议使用低场19F核磁共振。多年来，含氟药物通过世界各地的监管机构，作为制订的药物产品市场上的比例增至 2 %1970 年 2013年625%。因此，我们相信有需求，开发更简单，但更具体的方法，比当前可用的含氟药物在他们制定的形式内容的质量控制的方法。
药物含有氟-19 (19F) 在其结构中有优势的很容易被检测到19F 核磁共振由于相对于质子7其 100%的丰度和 83%的敏感性。1H TD-NMR 直接测量不是有用的因为有质子，赋形剂和 API 和总质子信号来自的药物产品，使得无法衡量药物产品使用1中的 API 内容的所有组件H TD-核磁共振。因此，测定药物含量的含氟药物产品采用19F TD-核磁共振的优点的赋形剂氟缺乏无干扰。为了展示我们的含氟药物产品平均药物含量测定的方法，我们选择了含氟的不同剂量范围内的三个商业配方的药物产品。图 1描述了药物选择，在两个结构的他们-甲状旁腺激素盐酸8和兰索拉唑9-三氟甲基 (CF3) 组在它们的结构，有剂量从 15 到 90 毫克和第三个其中一个环丙沙星-盐酸盐一水合物10 -包含一个附加到一个苯环，500 毫克剂量的氟原子。
在这里，我们表明我们有制定5量化使用低场台式 TD 核磁共振仪器 （23.4 兆赫为1H 和 22.0 兆赫为19F） 含氟药物产品的平均药物含量的方法。我们也比较两个软件包 （RI 校准和 Mnova 软件），可以用来报告结果。

<table><tbody><tr><td>MQC-23</td><td>Oxford Instruments</td><td>52-AM4044</td><td>23.4 MHz for &lt;sup&gt;1&lt;/sup&gt;H and 22.0 MHz for &lt;sup&gt;19&lt;/sup&gt;F</td></tr><tr><td>26 mm Probe (&lt;sup&gt;19&lt;/sup&gt;F)</td><td>Oxford Instruments</td><td>52-AM4061</td><td>&lt;sup&gt;19&lt;/sup&gt;F NMR probe</td></tr><tr><td>Cinacalcet HCl</td><td>Amgen</td><td>Lot 005002 M</td><td>Purity 99.8%</td></tr><tr><td>Cinacalcet commercial tablets</td><td>Amgen</td><td>Lot 0010021308</td><td>30 mg tablets</td></tr><tr><td>Cinacalcet commercial tablets</td><td>Amgen</td><td>Lot D1026396</td><td>30 mg tablets</td></tr><tr><td>Cinacalcet commercial tablets</td><td>Amgen</td><td>Lot D118714</td><td>30 mg tablets</td></tr><tr><td>Cinacalcet commercial tablets</td><td>Amgen</td><td>Lot D061829</td><td>60 mg tablets</td></tr><tr><td>Cinacalcet commercial tablets</td><td>Amgen</td><td>Lot 00100213</td><td>90 mg tablets</td></tr><tr><td>Cinacalcet commercial tablets</td><td>Amgen</td><td>Lot D064074</td><td>90 mg tablets</td></tr><tr><td>Cinacalcet commercial tablets</td><td>Amgen</td><td>Lot 1026356</td><td>90 mg tablets</td></tr><tr><td>Lansoprazole</td><td>Fluka</td><td>Lot LRAA1897</td><td>Purity 99.7%</td></tr><tr><td>Brand name Lansoprazole</td><td>Novartis</td><td>Lot DV1891</td><td>15 mg capsules</td></tr><tr><td>Lansoprazole generic</td><td>SUPERVALUE INC</td><td>Lot 2GE2027</td><td>15 mg capsules</td></tr><tr><td>Ciprofloxacin free base</td><td>Fluka</td><td>Lot BCBM7969V</td><td>99.9% purity</td></tr><tr><td>Ciprofloxacin HCl monohydrate</td><td>Fluka</td><td>Lot P500044</td><td>94% purity as HCl salt</td></tr><tr><td>Cipro generic</td><td>Pack Pharmaceuticals</td><td>Lot PUB3033</td><td>500 mg tablets</td></tr><tr><td>25 mm NMR tube</td><td>New Era Enterprises</td><td>NE-25TD-200-FB</td><td></td></tr><tr><td>18 mm NMR tube</td><td>New Era Enterprises</td><td>NE-18TD-200-FB</td><td></td></tr><tr><td>10 mm NMR tube</td><td>New Era Enterprises</td><td>NE-10TD-200-FB</td><td></td></tr><tr><td>5 mm NMR tube</td><td>New Era Enterprises</td><td>NE-HL5-7</td><td></td></tr><tr><td>3 mm NMR tube</td><td>New Era Enterprises</td><td>NE-H3-7</td><td></td></tr><tr><td>2.5 mm NMR tube</td><td>New Era Enterprises</td><td>NE-H5/2.5</td><td></td></tr></tbody></table>

application and methodology of the non-destructive 19f time-domain nmr technique to measure the content in fluorine-containing drug products

在这里，我们描述了使用低场氟-19 (19F) 时间域 (TD) 核磁共振 (NMR) 在其制定的药品产品形态测量平均含量的含氟药物我们集团开发的一项议定书: 片剂或胶囊。此方法是特定于含氟药物，因为它检测到只有含量为氟、 避免从缺乏氟辅料的干扰。测量含氟药物使用低场19F TD-核磁共振与高场19F 固体 (SS) 核磁共振的活动内容的优点是方法的简单;低成本;和技术，可恢复完整形式 （例如，粉末，片剂和胶囊），所有样品的非破坏性使这项技术对于任何实验室负担得起。
我们已经测试的方法与三氟化药物可用市场上的产品-甲状旁腺激素、 兰索拉唑和环丙沙星-从 15 至 500 毫克的剂量。分析的结果，测定低场19F TD-核磁、 支持平均药物含量的报告的标签索赔。
我们简单和重现性分析的基础，设想这种方法在任何实验室里，作为一种过程分析技术 (PAT) 工具在制药行业包括制造工厂，正在实施。

我们已经测试低场19F TD-核磁共振 （甲状旁腺激素、 兰索拉唑和环丙沙星） 测量平均药物含量的含氟药物产品的三个商业药物产品。图 1显示了测试药物，结构显示氟原子的位置。
我们测试了几个很多和甲状旁腺激素片剂型 （即， 30、 60、 90 毫克），大量的兰索拉唑胶囊的品牌名称和泛?...

Watch this Scientific Journal Video about Application and Methodology of the Non-destructive 19F Time-domain NMR Technique to Measure the Content in Fluorine-containing Drug Products at JoVE.com

Application and Methodology of the Non-destructive 19F Time-domain NMR Technique to Measure the Content in Fluorine-containing Drug Products

本文提出了一种简单和非破坏性的技术措施制定的药物产品使用低场氟-19 (19F) 时间域 (TD) 核磁共振 (NMR) 含氟药物的平均含量是。可以应用这项技术制造的药物在制药行业的发展。

应用程序和非破坏性19F 时间域核磁共振技术的方法来衡量中含氟药物产品的内容

Here, we describe a protocol developed by our group that uses low-field fluorine-19 (19F) time-domain (TD) nuclear magnetic resonance (NMR) to measure the ...

application-methodology-non-destructive-19f-time-domain-nmr-technique

Research

JoVE Journal

Chemistry

Application and Methodology of the Non-destructive 19F Time-domain NMR Technique to Measure the Content in Fluorine-containing Drug Products

8.3K 次观看.  Amgen, Inc.. 本文提出了一种简单和非破坏性的技术措施制定的药物产品使用低场氟-19 (19F) 时间域 (TD) 核磁共振 (NMR) 含氟药物的平均含量是。可以应用这项技术制造的药物在制药行业的发展。

视频: Application and Methodology of the Non-destructive 19F Time-domain NMR Technique to Measure the Content in Fluorine-containing Drug Products

NMR Spectroscopy as a Robust Tool for the Rapid Evaluation of the Lipid Profile of Fish Oil Supplements

The western diet is poor in n-3 fatty acids, therefore the consumption of fish oil supplements is recommended to increase the intake of these essential nutrients. The objective of this work is to demonstrate the qualitative and quantitative analysis of encapsulated fish oil supplements using high-resolution 1H and 13C NMR spectroscopy utilizing two different NMR instruments; a 500 MHz and an 850 MHz instrument. Both proton (1H) and carbon (13C) NMR spectra can be used for the quantitative determination of the major constituents of fish oil supplements. Quantification of the lipids in fish oil supplements is achieved through integration of the appropriate NMR signals in the relevant 1D spectra. Results obtained by 1H and 13C NMR are in good agreement with each other, despite the difference in resolution and sensitivity between the two nuclei and the two instruments. 1H NMR offers a more rapid analysis compared to 13C NMR, as the spectrum can be recorded in less than 1 min, in contrast to 13C NMR analysis, which lasts from 10 min to one hour. The 13C NMR spectrum, however, is much more informative. It can provide quantitative data for a greater number of individual fatty acids and can be used for determining the positional distribution of fatty acids on the glycerol backbone. Both nuclei can provide quantitative information in just one experiment without the need of purification or separation steps. The strength of the magnetic field mostly affects the 1H NMR spectra due to its lower resolution with respect to 13C NMR, however, even lower cost NMR instruments can be efficiently applied as a standard method by the food industry and quality control laboratories.

Here, high-resolution 1H and 13C Nuclear Magnetic Resonance (NMR) spectroscopy was used as a rapid and reliable tool for quantitative and qualitative analysis of encapsulated fish oil supplements.

核磁共振作为鱼油补充剂的血脂的快速评价一个强大的工具

The western diet is poor in n-3 fatty acids, therefore the consumption of fish oil supplements is recommended to increase the intake of these essential ...

科研

化学

In vivo 19F MRI for Cell Tracking

In vivo 19F MRI allows quantitative cell tracking without the use of ionizing radiation. It is a noninvasive technique that can be applied to humans. Here, we describe a general protocol for cell labeling, imaging, and image processing. The technique is applicable to various cell types and animal models, although here we focus on a typical mouse model for tracking murine immune cells. The most important issues for cell labeling are described, as these are relevant to all models. Similarly, key imaging parameters are listed, although the details will vary depending on the MRI system and the individual setup. Finally, we include an image processing protocol for quantification. Variations for this, and other parts of the protocol, are assessed in the Discussion section. Based on the detailed procedure described here, the user will need to adapt the protocol for each specific cell type, cell label, animal model, and imaging setup. Note that the protocol can also be adapted for human use, as long as clinical restrictions are met.

In vivo 19F MRI for Cell Tracking

We describe a general protocol for in vivo cell tracking using MRI in a mouse model with ex vivo labeled cells. A typical protocol for cell labeling, image acquisition processing and quantification is included.

在体内 19女性 MRI对细胞跟踪

In vivo 19F MRI allows quantitative cell tracking without the use of ionizing radiation. It is a noninvasive technique that can be applied to humans. ...

医学

Automation of a Positron-emission Tomography (PET) Radiotracer Synthesis Protocol for Clinical Production

The development of new positron-emission tomography (PET) tracers is enabling researchers and clinicians to image an increasingly wide array of biological targets and processes. However, the increasing number of different tracers creates challenges for their production at radiopharmacies. While historically it has been practical to dedicate a custom-configured radiosynthesizer and hot cell for the repeated production of each individual tracer, it is becoming necessary to change this workflow. Recent commercial radiosynthesizers based on disposable cassettes/kits for each tracer simplify the production of multiple tracers with one set of equipment by eliminating the need for custom tracer-specific modifications. Furthermore, some of these radiosynthesizers enable the operator to develop and optimize their own synthesis protocols in addition to purchasing commercially-available kits. In this protocol, we describe the general procedure for how the manual synthesis of a new PET tracer can be automated on one of these radiosynthesizers and validated for the production of clinical-grade tracers. As an example, we use the ELIXYS radiosynthesizer, a flexible cassette-based radiochemistry tool that can support both PET tracer development efforts, as well as routine clinical probe manufacturing on the same system, to produce [18F]Clofarabine ([18F]CFA), a PET tracer to measure in vivo deoxycytidine kinase (dCK) enzyme activity. Translating a manual synthesis involves breaking down the synthetic protocol into basic radiochemistry processes that are then translated into intuitive chemistry &#34;unit operations&#34; supported by the synthesizer software. These operations can then rapidly be converted into an automated synthesis program by assembling them using the drag-and-drop interface. After basic testing, the synthesis and purification procedure may require optimization to achieve the desired yield and purity. Once the desired performance is achieved, a validation of the synthesis is carried out to determine its suitability for the production of the radiotracer for clinical use.

Automation of a Positron-emission Tomography PET Radiotracer Synthesis Protocol for Clinical Production

Positron-emission tomography (PET) imaging sites that are involved in multiple early clinical research trials need robust and versatile radiotracer manufacturing capabilities. Using the radiotracer [18F]Clofarabine as an example, we illustrate how to automate the synthesis of a radiotracer using a flexible, cassette-based radiosynthesizer and validate the synthesis for clinical use.

用于临床生产的正电子发射层析成像 (PET) 放射性同位素示踪合成协议的自动化

The development of new positron-emission tomography (PET) tracers is enabling researchers and clinicians to image an increasingly wide array of biological ...

A New Straightforward Method for Lipophilicity (logP) Measurement using 19F NMR Spectroscopy

Fluorination has become an effective tool to optimize physicochemical properties of bioactive compounds. One of the applications of fluorine introduction is to modulate the lipophilicity of the compound. In our group, we are interested in the study of the impact of fluorination on lipophilicity of aliphatic fluorohydrins and fluorinated carbohydrates. These are not UV-active, resulting in a challenging lipophilicity determination. Here, we present a straightforward method for the measurement of lipophilicity of fluorinated compounds by 19F NMR spectroscopy. This method requires no UV-activity. Accurate solute mass, solvent and aliquot volume are also not required to be measured. Using this method, we measured the lipophilicities of a large number of fluorinated alkanols and carbohydrates.

A New Straightforward Method for Lipophilicity logP Measurement using 19F NMR Spectroscopy

A novel and straightforward variation of the shake-flask method was developed for accurate lipophilicity measurement of fluorinated compounds by 19F NMR spectroscopy.

一种利用19f 核磁共振光谱测定脂质性 (日志 p) 的新方法

Fluorination has become an effective tool to optimize physicochemical properties of bioactive compounds. One of the applications of fluorine introduction ...

Radiotracer Administration for High Temporal Resolution Positron Emission Tomography of the Human Brain: Application to FDG-fPET

Functional positron emission tomography (fPET) provides a method to track molecular targets in the human brain. With a radioactively-labelled glucose analogue, 18F-fluordeoxyglucose (FDG-fPET), it is now possible to measure the dynamics of glucose metabolism with temporal resolutions approaching those of functional magnetic resonance imaging (fMRI). This direct measure of glucose uptake has enormous potential for understanding normal and abnormal brain function and probing the effects of metabolic and neurodegenerative diseases. Further, new advances in hybrid MR-PET hardware make it possible to capture fluctuations in glucose and blood oxygenation simultaneously using fMRI and FDG-fPET.
The temporal resolution and signal-to-noise of the FDG-fPET images is critically dependent upon the administration of the radiotracer. This work presents two alternative continuous infusion protocols and compares them to a traditional bolus approach. It presents a method for acquiring blood samples, time-locking PET, MRI, experimental stimulus, and administering the non-traditional tracer delivery. Using a visual stimulus, the protocol results show cortical maps of the glucose-response to external stimuli on an individual level with a temporal resolution of 16 s.

This manuscript describes two radiotracer administration protocols for FDG-PET (constant infusion and bolus plus infusion) and compares them to bolus administration. Temporal resolutions of 16 s are achievable using these protocols.

人脑高时态分辨率正态发射断层扫描的放射跟踪器管理:在FDG-fPET中的应用

Functional positron emission tomography (fPET) provides a method to track molecular targets in the human brain. With a radioactively-labelled glucose ...

行为学

NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode

Fragment-based screening (FBS) is a well-validated and accepted concept within the drug discovery process both in academia and industry. The greatest advantage of NMR-based fragment screening is its ability not only to detect binders over 7-8 orders of magnitude of affinity but also to monitor purity and chemical quality of the fragments and thus to produce high quality hits and minimal false positives or false negatives. A prerequisite within the FBS is to perform initial and periodic quality control of the fragment library, determining solubility and chemical integrity of the fragments in relevant buffers, and establishing multiple libraries to cover diverse scaffolds to accommodate various macromolecule target classes (proteins/RNA/DNA). Further, an extensive NMR-based screening protocol optimization with respect to sample quantities, speed of acquisition and analysis at the level of biological construct/fragment-space, in condition-space (buffer, additives, ions, pH, and temperature) and in ligand-space (ligand analogues, ligand concentration) is required. At least in academia, these screening efforts have so far been undertaken manually in a very limited fashion, leading to limited availability of screening infrastructure not only in the drug development process but also in the context of chemical probe development. In order to meet the requirements economically, advanced workflows are presented. They take advantage of the latest state-of-the-art advanced hardware, with which the liquid sample collection can be filled in a temperature-controlled fashion into the NMR-tubes in an automated manner. 1H/19F NMR ligand-based spectra are then collected at a given temperature. High-throughput sample changer (HT sample changer) can handle more than 500 samples in temperature-controlled blocks. This together with advanced software tools speeds up data acquisition and analysis. Further, application of screening routines on protein and RNA samples are described to make aware of the established protocols for a broad user base in biomacromolecular research.

Fragment-based screening by NMR is a robust method to rapidly identify small molecule binders to biomacromolecules (DNA, RNA, or proteins). Protocols describing automation-based sample preparation, NMR experiments &#38; acquisition conditions, and analysis workflows are presented. The technique allows for optimal exploitation of both 1H and 19F NMR-active nuclei for detection.

基于核磁共振的片段筛选，样品最少，自动化程度最高

Fragment-based screening (FBS) is a well-validated and accepted concept within the drug discovery process both in academia and industry. The greatest ...

生物化学

18F-Labeling of Radiotracers Functionalized with a Silicon Fluoride Acceptor (SiFA) for Positron Emission Tomography

The para-substituted di-tert-butylfluorosilylbenzene structural motif known as the silicon-fluoride acceptor (SiFA) is a useful tag in the radiochemist&#39;s toolkit for incorporating radioactive [18F]fluoride into tracers for use in positron emission tomography. In comparison to conventional radiolabeling strategies, isotopic exchange of fluorine-19 from SiFA with [18F]fluoride is carried out at room temperature and requires minimal reaction participants. The formation of by-products is thus negligible, and purification is greatly simplified. However, while the precursor molecule used for labeling and the final radiolabeled product are isotopically discrete, they are chemically identical and are thus inseparable during purification procedures. The SiFA tag is also susceptible to degradation under the basic conditions arising from the processing and drying of [18F]fluoride. The &#39;4 drop method&#39;, wherein only the first 4 drops of eluted [18F]fluoride are used from the solid-phase extraction, reduces the amount of base in the reaction, facilitates lower molar amounts of precursor, and reduces degradation.

18F-Labeling of Radiotracers Functionalized with a Silicon Fluoride Acceptor SiFA for Positron Emission Tomography

The synthesis of fluorine-18 (18F) labeled radiopharmaceuticals for positron emission tomography typically requires months of experience. When incorporated into a radiotracer, the silicon-fluoride acceptor (SiFA) motif enables a simple 18F-labeling protocol that is independent of costly equipment and preparatory training, while reducing precursor quantity needed and utilizing milder reaction conditions.

18放射性跟踪器的F-标签，其功能与硅氟化物接受器（SiFA）进行正电子发射断层扫描

The para-substituted di-tert-butylfluorosilylbenzene structural motif known as the silicon-fluoride acceptor (SiFA) is a useful tag in the ...

Radiosynthesis of 1-(2-[18F]Fluoroethyl)-L-Tryptophan using a One-pot, Two-step Protocol

The kynurenine pathway (KP) is a primary route for tryptophan metabolism. Evidence strongly suggests that metabolites of the KP play a vital role in tumor proliferation, epilepsy, neurodegenerative diseases, and psychiatric illnesses due to their immune-modulatory, neuro-modulatory, and neurotoxic effects. The most extensively used positron emission tomography (PET) agent for mapping tryptophan metabolism, &#945;-[11C]methyl-L-tryptophan ([11C]AMT), has a short half-life of 20 min with laborious radiosynthesis procedures. An onsite cyclotron is required to radiosynthesize [11C]AMT. Only a limited number of centers produce [11C]AMT for preclinical studies and clinical investigations. Hence, the development of an alternative imaging agent that has a longer half-life, favorable in vivo kinetics, and is easy to automate is urgently needed. The utility and value of 1-(2-[18F]fluoroethyl)-L-tryptophan, a fluorine-18-labeled tryptophan analog, has been reported in preclinical applications in cell line-derived xenografts, patient-derived xenografts, and transgenic tumor models.
This paper presents a protocol for the radiosynthesis of 1-(2-[18F]fluoroethyl)-L-tryptophan using a one-pot, two-step strategy. Using this protocol, the radiotracer can be produced in a 20 &#177; 5% (decay corrected at the end of synthesis, n &#62; 20) radiochemical yield, with both radiochemical purity and enantiomeric excess of over 95%. The protocol features a small precursor amount with no more than 0.5 mL of reaction solvent in each step, low loading of potentially toxic 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo[8.8.8]hexacosane (K222), and an environmentally benign and injectable mobile phase for purification. The protocol can be easily configured to produce 1-(2-[18F]fluoroethyl)-L-tryptophan for clinical investigation in a commercially available module.

Radiosynthesis of 1-2-[18F]Fluoroethyl-L-Tryptophan using a One-pot, Two-step Protocol

Here, we describe the radiosynthesis of 1-(2-[18F]Fluoroethyl)-L-tryptophan, a positron emission tomography imaging agent for studying tryptophan metabolism, using a one-pot, two-step strategy in a radiochemistry synthesis system with good radiochemical yields, high enantiomeric excess, and high reliability.

1-（2-[18F]氟乙基）-L-色氨酸的放射性合成，采用一锅两步实验方案

The kynurenine pathway (KP) is a primary route for tryptophan metabolism. Evidence strongly suggests that metabolites of the KP play a vital role in tumor ...

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 ...

Automated Radiochemical Synthesis of [18F]3F4AP: A Novel PET Tracer for Imaging Demyelinating Diseases

3-[18F]fluoro-4-aminopyridine, [18F]3F4AP, is a radiofluorinated analog of the FDA-approved drug for multiple sclerosis 4-aminopyridine (4AP). This compound is currently under investigation as a PET tracer for demyelination. We recently described a novel chemical reaction to produce metafluorinated pyridines consisting of direct fluorination of a pyridine N-oxide and the utilization of this reaction for the radiochemical synthesis of [18F]3F4AP. In this article, we demonstrate how to produce this tracer using an automated synthesizer and an in-house made flow hydrogenation reactor. We also show the standard quality control procedures performed before releasing the radiotracer for preclinical animal imaging studies. This semi-automated procedure may serve as the basis for future production of [18F]3F4AP for clinical studies.

Automated Radiochemical Synthesis of [18F]3F4AP: A Novel PET Tracer for Imaging Demyelinating Diseases

We demonstrate the semi-automated radiochemical synthesis of [18F]3F4AP and quality control procedures.

自动放射化学合成[<sup&gt; 18</sup&gt; F] 3F4AP:一种用于成像脱髓鞘疾病的新型PET示踪剂

3-[18F]fluoro-4-aminopyridine, [18F]3F4AP, is a radiofluorinated analog of the FDA-approved drug for multiple sclerosis 4-aminopyridine (4AP). This ...

应用程序和非破坏性¹⁹F 时间域核磁共振技术的方法来衡量中含氟药物产品的内容

摘要

探索更多视频

此视频中的章节

核磁共振作为鱼油补充剂的血脂的快速评价一个强大的工具

在体内 ^19女性 MRI对细胞跟踪

用于临床生产的正电子发射层析成像 (PET) 放射性同位素示踪合成协议的自动化

一种利用¹⁹f 核磁共振光谱测定脂质性 (日志 p) 的新方法

人脑高时态分辨率正态发射断层扫描的放射跟踪器管理:在FDG-fPET中的应用

基于核磁共振的片段筛选，样品最少，自动化程度最高

¹⁸放射性跟踪器的F-标签，其功能与硅氟化物接受器（SiFA）进行正电子发射断层扫描

1-（2-[^18F]氟乙基）-L-色氨酸的放射性合成，采用一锅两步实验方案

使用多氟化胆汁酸和

自动放射化学合成[