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1.样品制备<ol><li>根据反应器的设计和实验步骤在文献10,11发现生成的混合的水/有机产物经由藻类的连续流HTL流。 </li><li>使用重力分离器的产物流分离成水相和有机相。 </li><li>过滤10毫升使用在维持在4℃的GC×GC-TOF-MS分析的冰箱的0.45微米注射器式滤器并储存在HTL水相。 </li></ol> 2.仪器组件<ol><li>使用配备有用于这些实验中的四喷射双台基于冷却调制器和时间飞行（TOF）质谱仪（MS）的气相色谱仪（GC）。 </li><li>配置自动进样器注入1微升每个样品或标准到GC的。如文献13中所述使用样品和标准注射随机区组设计为自动取样序列。该randomi捷思块设计是在定量研究通常用于控制仪器的操作。我们的实验室采用了常规设计，即使在比较研究，以验证仪器操作。 </li><li>调制器之前使用压紧连接器连接主次列。确保主要和次要列的两个边缘连接到压紧接头，再直减无锋利的棱角。 </li><li>广场上的气相色谱柱套圈，然后主列连接到GC进样器，使5个mm色谱柱的是在喷射器内。 </li><li>确保玻璃内胆，不粘内胆O形圈和隔为GC进样都是新的，无污染的。 </li><li>使用1/16×0.5毫米内径传输线套圈连接次级柱和传输线。放置次级柱的0.2μm的部分中的传输线。 </li><li>确保0.1μm的部分次级柱的是在调制器。 </li><li>使用超高纯度氦气气体作为用于以1.5ml分钟-1的流速GC载气。 </li><li>确保有在其充当在调制器的冷却剂的杜瓦足够液氮中。在杜瓦液氮的水平可以使用连接到其出口的压力计来预测。压力计的69千帕读数表明杜瓦是满的，而0千帕表示它是空的。 </li></ol> 3.协议之前分析样品<ol><li>确保有仪器没有大的泄漏。如果TOF-MS的真空计读数高于2.7×10 -5 Pa下1.5毫升分钟-1 GC色谱柱流速，这表明在该系统的一个主要的泄漏。 </li><li>建立质量控制（QC）方法，并内置"采集系统调整"协议运行使用制造商的协议，以实现最大的信号响应。 </li><li>运行内置的QC方法的"仪器优化"的协议，在系列 - filam耳鼻喉科重点，离子光学对焦并使用制造商的协议质量校准测试。确保质量校正测试通过。这种质量控制方法确保该仪器的所有硬件参数在最佳水平。 </li><li>执行"泄漏检查"使用制造商的协议。分析自动生成泄漏检查报告。确保的相对浓度28（N 2），32（O 2）和18（水分）离子必须低于分别小于10％，3％和69离子内标质谱5％。 </li><li>使用调谐TOF-MS制造商的协议。 </li><li>运行质量控制方法以及TOF-MS调协议前，泄漏检查后，也同时分析样品和标准。 </li></ol> 4.协议来确定调制器的最佳调制周期<ol><li>任意选择长调制周期（ 如 10秒或13秒）。在2.2中所述注入样本。 </li><lI&gt;确定的滞留时间在等值线图之后，无峰洗脱的第二维。选择识别出的第二维保留时间为最佳调制周期。 图2清楚地阐明保留时间识别的等高线图的第二维。 </li><li>增加在步骤4.1中使用的调制周期，并执行再次，如果"环绕"观察18分析。如果在第二维的峰值洗脱第一尺寸的基线下发生回绕现象。例如，对于"环绕"等高线图显示在补充信息如图3所示 。 </li><li>直到最佳值确定重复步骤4.2和4.3。 </li></ol> 5.仪器设置参数实验<ol><li>安装一个极性（60米×0.25毫米×0.5微米膜厚度）毛细管柱作为主柱和非极性（2.3米×0.25毫米×0.5微米膜厚度）卡皮亚RY列作为次级柱。至少2小时烘烤主和次级柱以除去水分，空气和用新的GC柱相关的污染物的痕量。 </li><li>以1.5ml分钟-1的流速使用超高纯度氦气作为载气进行GC。 </li><li>在GC注射器设定为260℃的温度和1分流比:250。 </li><li>使用用于主列下列温度程序:40℃持续0.2分钟的恒定温度，随后的升温至260℃以5℃分钟-1，随后的260℃的恒定温度下保持5分钟。 </li><li>维持调制器温度5℃比次级柱中，在比该主塔的高5℃的次级柱温度高。 </li><li>使用4秒的最佳调制周期以0.8秒的热脉冲的冷脉冲1.2秒。此值的基础上确定在节描述的协议离子4。 </li><li>设置传输线温度至270℃。 </li><li>设置采集延时或溶剂延迟0秒。 </li><li>分别设置的较低和较高的范围为m / z为35和800。 </li><li>设置在400谱/秒的质谱检测器采集速率。 </li><li>在比最佳值150V时更高保持质谱仪的电压。 </li><li>保持在225℃下的MS离子源温度。 </li></ol> 6.数据分析<ol><li>使用由仪器制造商提供的软件进行数据处理。 </li><li>选择数据分析方法以下任务 - 计​​算基准，找到基线，库搜索以上的山峰和计算是/高度。 </li><li>跟踪通过数据文件的基线。输入偏移基线0.5。 输入在第一维15秒，在第二维0.15秒的预期峰宽。 </li><li>用于鉴定的&gt; 850设置信噪比为5000和相似度值化合物fication。 </li><li>选择市售的质谱库，以确定样品中存在的化合物，并设置库搜索模式来转发。 </li><li>处理使用使用制造商的协议此数据分析方法的数据文件。它至少需要1小时来处理的数据文件。 </li></ol>

The authors declare that they have no competing financial interests.

结果清楚地说明极性×非极性的列组合，以解决目前在藻类生物原油，恕不另行样品制备技术的水相极性化合物和光挥发物的能力。观察到的有机酸和在使用非极性×极性柱组合N化合物急剧峰拖尾。这拖尾没有为早期洗脱光有机物观察。此行为已重现的检验仪器是无泄漏（在TOF-MS中的真空度为低于2.7×10 -5 Pa下为1.5ml分钟-1 GC载气流量）时。可以预计，如果有与压紧连接器，或者如果死体积的问题冷喷流率将是过度的，该行为将整个色谱进行观​​察。然而，即使晚洗脱的化合物（没有确定的数字）没有尾巴。因此，我们的结论是，这是aque的结果OU的进样/列的配置组合。 分流比样品进入柱与输给分流量的体积。的分流比较高的小样品引入到色谱柱的量。一般来说，这会产生更有效的峰这将改善峰容量。确定适当的分流比样品可以防止与复合检测（分流比​​太高）从列超载的问题（分流比太低）或问题。因此，1:1的分流比:在GC×两个柱的组合，以防止柱加载的GC-TOF-MS的数据采集方法用于250并且还提高峰值容量。 用于鉴定化合物的相似值在850-999的范围内。这表明，化学化合物与超过85％的置信识别。这是通过在气相×GC使用400谱图/秒的MS采集率达到211; TOF-MS的数据采集方法。 A 400谱图/秒采集速率提高峰这增加鉴定的化学化合物17的相似性值的信噪比。较高的相似价值观使我们能够识别具有高可信度化合物。然而，这种高MS采集速率结果在相当长的数据分析时间。因此，建议使用200光谱/秒MS采集速率对这些样品这降低了数据分析时间的量化。 在GC×GC-TOF-MS的数据采集方法用于极性×非极性可通过增加辅助列的长度能够进一步提高表征水性藻类生物原油开发的。通过增加辅助列的长度，分辨率可以在第二维使存在于样品16,17中的异构体的分离得到改善。峰容量也随着增加而进一步提高次级柱的长度。 HTL藻类水域特征在本文中是稀11（含有碳的约3总重量％），并且可能不需要较长的次级柱。然而，这一建议可能是复杂的，浓缩的水样鉴定过程中有益的。 由于极性列的最大可编程温度是260℃，这种方法不能洗脱作为长链脂肪酸，单甘油酯，二甘油酯，三酸甘油酯和氨基酸低聚物以及糖16个这样的高沸点化合物。含有这些化合物的样品中，分析时，可能会污染GC进样和列。 GC进样和列的污染导致峰拖尾，在化学化合物的保留时间的变化，以及高噪声或低信噪比该MS检测器，其是不希望的定性和定量表征。因此，当utilizin克分析含有高沸点化合物分析师应采用适当的质量控制方法，水样此列组合。 在藻类生物粗的水性级分中确定的化学化合物具有广泛的应用。吡啶，吡嗪以及它们的烷基衍生物是生产农用化学品的中间化学品，药物36,37，并且在均相催化38,39被广泛地用作溶剂。同样，琥珀酰亚胺的衍生物也具有广泛的应用，包括聚合物中间体，去污剂40，临床药物41,42，燃料添加剂和润滑油添加剂40。存在于HTL藻类水中的有机酸可被用作在催化过程的原料，以产生酮或酯用于从水相43容易分离。 在GC×GC-TOF-MS方法开发的第极性×非极性本文电子柱组合也可以采用以从生物过程，食品加工，以及环境的废物分析水样。研究人员使用此列组合有机样品44-47表征。据报道，该列的组合是最好的不同类别的烃的有效分离-脂族，芳族化合物，烷基苯和双核芳烃44-46。因此，利用对分离和非极性分离的第二维的所述第一尺寸的极性的分离将是水性的表征合适的列配置以及生物粗生物质的热液液化产生的有机部分。

为液体燃料，有限的化石燃料资源，化石燃料供应的不确定性，并在大气中的温室气体浓度的增加，需求的担忧平稳增长增加了全球意识再生资源1。太阳能（光伏，包括太阳能热），风能，水能，地热能和生物质能是主要的可再生能源，它可能替代化石所产生的能量2。在这些中，生物质是用于生产液体运输燃料和高价值化学品3的仅基于碳的可替代能源资源。生物质包括任何有机材料，如森林资源，农业残余物，藻类，油籽，城市固体废物，和富含碳的产业废弃物（ 例如 ，从纸浆和造纸工业或食品加工）1。基于COM木质纤维和非木质原料:生物量分为两大类位置特性。木质纤维素生物质包括碳水化合物和木质素的，而非木质原料有蛋白质，碳水化合物和脂质/油4。木质纤维素原料，从陆生植物来源的，只能满足当前液体燃料（汽油，喷气燃料，柴油）需求的30％，如果持续培育和收获5,6。用于生产可再生液体燃料，因此，非木质水生微生物，如微藻和真菌，被认为是潜在的原料，以补充木质素纤维素资源。 微藻原料必须满足当前的液体运输燃料的需求7,8的潜力。藻类有许多优点:高面生产率8，在低质量的，微咸水或海水9和积累能量密度高的甘油三酯或烃7,8的能力中生长的能力。热液液化（HTL）是一个可行的和可扩展的共n版本的过程，利用自然与藻类或水生原料10,11相关的水。它是具有10-25兆帕其产生的液体产物，或生物原油，可升级到燃料调和油料的250-400℃，操作压力的工作温度的热化学处理。生物原油从藻类HTL产生具有区别的和容易分离有机和含水馏份。生物的粗的有机部分可以通过催化加氢处理过程11被有效地转换成一个炼油厂准备共混原料。生物的粗的水性馏分含有〜存在于藻类原料中的总碳的30％。虽然数以千计的化合物已在HTL水性流中被识别，主要的级分组成的低分子量含氧化合物（其中包括酸，醇，酮和醛类）由碳水化合物和脂质，和氮杂环（​​包括吡咯，吡啶的降解形成的，吡嗪ES，和咪唑）从蛋白质分解12而得。研究利用水相，以提高整体工艺的经济性以及可持续性正在进行中。合成气可从藻类生物原油经由催化热液气化10,13，14中的水相来制备。可替代地，在水级分的有机物也能催化转化为燃料添加剂和特殊化学品。研究在水性液相有机物转化优化催化气化热液和催化剂的筛选研究，目前是西北太平洋国家实验室（PNNL）正在进行中。对于这项工作，定性以及藻类生物粗的水性馏分定量表征是必需的。由于藻类生物粗的含水馏分被认为是废料流，也有已经分析的藻类生物粗13,15的含水馏分很少有研究。此外，最近的研究的结论是，这HTL藻类水转化成高价值的生物制品将改善的可持续性以及基于HTL-生物精炼厂11的经济性。因此，本研究集中于通过二维气相色谱加上时间飞行质谱（GC×GC-TOF-MS）开发用于从藻类HTL获得生物粗​​的水性馏分的定性表征的方法。 的GC×GC-TOF-MS是最有前途的色谱分析技术来提高（样品中或化学原料的分离）的分辨率，峰值容量（ 即分辨的峰的数目），信噪比（用于化学化合物的鉴定以高置信度），以及避免化学化合物16的共洗脱。为了最大限度地提高分辨率，峰值容量，和信号噪声比，具有不同固定相位的两个GC色谱柱使用压配合Ç串联onnector或微工会17（ 见图1，它是GC×在本研究中使用的GC-TOF-MS系统的框图）。调制器位于压入配合连接器和辅助列之间陷阱，重新调整，和从主塔重新注入流出物进入第二塔18。 如图1上，在本研究中的次级柱调制发生。然后将该次级柱经由传输线组件连接到TOF-MS。 的GC×GC-TOF-MS以前用于定性以及有机样品的定量分析如原油16,19，汽油，喷气燃料，柴油，生物柴油和生物燃料的有机部分20-从热化学以及热催化转化产生22处理23,24。对于GC这些有机样品的表征×GC-TOF-MS仪器，长非极性色谱柱W¯¯作为用作第一列中，而短极性柱用作第二列。这种传统的列配置解析基于波动性在第一个维度的差异，其次是极性第二个维度18化合物。从生物过程，食品加工，以及环境的废物水性或水样品也使用类似的初级/次级柱配置，其特征后的样品已经通过制备步骤17,25-30。样品制备技术，如衍生化，固相萃取，和有机溶剂萃取已全部之前利用GC×GC-TOF-MS分析17,27-29,31,32。这些技术的目的是使用常规的柱结构33减少样品分析中的化合物的极性。另一种策略在基于样品的性质该研究中采用的（在水中这里极性有机化合物）利用反向初级/次级柱用于GC×GC-TOF-MS分析的配置。因为生物原油从HTL产生的水性级分具有极性化合物13，一个主极性柱和一个第二非极性柱的柱组合在GC×GC-TOF-MS用于无需任何上游样品制备。这个初级/次级柱组合解析基于在第一维中的极性差异的化合物，接着在第二维的波动。限于分析方法在文献中存在使用二维气相色谱无需事先检测体处理15含水样品的表征。 本研究的目的是确定存在于藻类生物粗的含水馏分中的化合物。为了实现这一目标，一个GC×GC-TOF-MS的数据采集方法与极性列的列组合（开发普里姆进制）×非极性（二次）。 Klenn 等 。 （2015）建议，相对于增加主柱（尤其60米GC柱）和降低次级柱的偏移温度的长度与主塔将最大化峰值容量和分辨率16-18。因此，60米的主塔和5℃偏移二次柱的温度相对于该主列在该研究中使用。确定最佳调制周期遵循本研究中所描述的协议（见第4节）。 GC柱温度的最适斜坡率通过试错法确定，并且类似于在文献16-18所建议的值。为了探讨水样此列组合的优势，我们分析HTL藻水样与非极性×极性常规列的组合。在文献中建议的操作参数被用于分析所述水藻类生物原油用非极性×极性柱组合18的级分。

<table><tbody><tr><td>GC &amp;times; GC&amp;#8211;TOF/MS</td><td>Leco</td><td>PEG4D11DLN15</td><td>Commercial Pegasus 4D</td></tr><tr><td>ChromaTOF version 4.50&amp;nbsp;</td><td>Leco</td><td></td><td>Data analysis software</td></tr><tr><td>Rxi-5MS GC column</td><td>Restek</td><td>13420</td><td>2.3 m column was used from this column.</td></tr><tr><td>Stabilwax GC column</td><td>Restek</td><td>10626</td><td></td></tr><tr><td>HP-5 GC column</td><td>Agilent</td><td>19091J-416</td><td></td></tr><tr><td>Stabilwax GC column</td><td>Restek</td><td>15121</td><td></td></tr><tr><td>Presstight Connector</td><td>Restek</td><td>20430</td><td></td></tr><tr><td>GC injector liner</td><td>Restek</td><td>23305.5</td><td></td></tr><tr><td>GC Injector ferrules</td><td>Agilent</td><td>5181-3323</td><td></td></tr><tr><td>Non-stick liner O-rings</td><td>Agilent</td><td>5188-5365</td><td></td></tr><tr><td>Transfer line ferrules</td><td>Restek</td><td>20212</td><td></td></tr><tr><td>Ethanol</td><td>Sigma-Aldrich</td><td>459844</td><td>Chromatography grade</td></tr><tr><td>Acetone</td><td>Sigma-Aldrich</td><td>414689</td><td>Chromatography grade</td></tr><tr><td>Acetic acid</td><td>Sigma-Aldrich</td><td>320099</td><td>Chromatography grade</td></tr><tr><td>2-butanone</td><td>Sigma-Aldrich</td><td>360473</td><td>Chromatography grade</td></tr><tr><td>Propanoic acid</td><td>Sigma-Aldrich</td><td>402907</td><td>Chromatography grade</td></tr><tr><td>Butanoic acid</td><td>Sigma-Aldrich</td><td>19215</td><td>Chromatography grade</td></tr><tr><td>Pyridine</td><td>Sigma-Aldrich</td><td>270970</td><td>Chromatography grade</td></tr><tr><td>Pyrazine</td><td>Sigma-Aldrich</td><td>65693</td><td>Chromatography grade</td></tr><tr><td>Acetamide</td><td>Sigma-Aldrich</td><td>695122</td><td>Chromatography grade</td></tr><tr><td>2,5-pyrrolididione</td><td>Sigma-Aldrich</td><td>S9381</td><td>Chromatography grade</td></tr><tr><td>&lt;em&gt;N&lt;/em&gt;-methylsuccinimide</td><td>Sigma-Aldrich</td><td>325384</td><td>Chromatography grade</td></tr><tr><td>&lt;em&gt;N&lt;/em&gt;-(2-hydroxyethyl)succinimide</td><td>Sigma-Aldrich</td><td>444073</td><td>Chromatography grade</td></tr></tbody></table>

qualitative characterization of the aqueous fraction from hydrothermal liquefaction of algae using 2d gas chromatography with time-of-flight mass spectrometry

加上时间飞行质谱二维气相色谱法是用于识别与在复杂的混合物定量化学成分的有力工具。它经常被用于分析汽油，喷气燃料，柴油，生物柴油和生物原油/生物油的有机部分。在大多数这些分析的，分离的第一维是非极性的，其次是极性的分离。生物原油和其他水样的生物燃料生产中的水部分已审查了类似的列组合。然而，样品制备技术，如衍生化，溶剂萃取，固相萃取是必要 分析前。在这项研究中，来自藻类的水热液化得到的水级分特征在于二维气相色谱加上时间飞行质谱不使用在第一维极性分离，随后前样品制备技术通过在第二非极性的分离。从这个分析二维曲线与来自更传统的柱结构中所得到的那些进行比较。从藻类生物粗的水性级分的定性表征结果进行了详细的讨论。使用极性分离后跟一个非极性分离为含水样品中的有机物的表征二维气相色谱加上时间飞行质谱的优点突出。

Watch this Scientific Journal Video about Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

A two-dimensional gas chromatography-time-of-flight mass spectrometry method is described for characterization of the aqueous fraction of bio-crude produced from hydrothermal liquefaction of algae. This protocol can also be employed to analyze the aqueous fraction of liquid products from fast pyrolysis, catalytic fast pyrolysis, catalytic deoxygenation and hydro-treating.

从藻类水热液化使用二维气相色谱时间飞行质谱法水提取部分的定性鉴定

Two-dimensional gas chromatography coupled with time-of-flight mass spectrometry is a powerful tool for identifying and quantifying chemical components in ...

qualitative-characterization-aqueous-fraction-from-hydrothermal

Universidad San Sebastian

Research

JoVE Journal

Bioengineering

9.3K 次观看.  Pacific Northwest National Laboratory. A two-dimensional gas chromatography-time-of-flight mass spectrometry method is described for characterization of the aqueous fraction of bio-crude produced from hydrothermal liquefaction of algae. This protocol can also be employed to analyze the aqueous fraction of liquid products from fast pyrolysis, catalytic fast pyrolysis, catalytic deoxygenation and hydro-treating. 

视频: Qualitative Characterization of the Aqueous Fraction from Hydrothermal Liquefaction of Algae Using 2D Gas Chromatography with Time-of-flight Mass Spectrometry

Analysis of Fatty Acid Content and Composition in Microalgae

A method to determine the content and composition of total fatty acids present in microalgae is described. Fatty acids are a major constituent of microalgal biomass. These fatty acids can be present in different acyl-lipid classes. Especially the fatty acids present in triacylglycerol (TAG) are of commercial interest, because they can be used for production of transportation fuels, bulk chemicals, nutraceuticals (&omega;-3 fatty acids), and food commodities. To develop commercial applications, reliable analytical methods for quantification of fatty acid content and composition are needed. Microalgae are single cells surrounded by a rigid cell wall. A fatty acid analysis method should provide sufficient cell disruption to liberate all acyl lipids and the extraction procedure used should be able to extract all acyl lipid classes.
With the method presented here all fatty acids present in microalgae can be accurately and reproducibly identified and quantified using small amounts of sample (5 mg) independent of their chain length, degree of unsaturation, or the lipid class they are part of.
This method does not provide information about the relative abundance of different lipid classes, but can be extended to separate lipid classes from each other.
The method is based on a sequence of mechanical cell disruption, solvent based lipid extraction, transesterification of fatty acids to fatty acid methyl esters (FAMEs), and quantification and identification of FAMEs using gas chromatography (GC-FID). A TAG internal standard (tripentadecanoin) is added prior to the analytical procedure to correct for losses during extraction and incomplete transesterification.

A method for the determination of fatty acid content and composition in microalgae based on mechanical cell disruption, solvent based lipid extraction, transesterification, and quantification and identification of fatty acids using gas chromatography is described. A tripentadecanoin internal standard is used to compensate for the possible losses during extraction and incomplete transesterification.

脂肪酸组分和含量微藻分析

A method to determine the content and composition of total fatty acids present in microalgae is described. Fatty acids are a major constituent of ...

科研

环境科学

On-line Analysis of Nitrogen Containing Compounds in Complex Hydrocarbon Matrixes

The shift to heavy crude oils and the use of alternative fossil resources such as shale oil are a challenge for the petrochemical industry. The composition of heavy crude oils and shale oils varies substantially depending on the origin of the mixture. In particular they contain an increased amount of nitrogen containing compounds compared to the conventionally used sweet crude oils. As nitrogen compounds have an influence on the operation of thermal processes occurring in coker units and steam crackers, and as some species are considered as environmentally hazardous, a detailed analysis of the reactions involving nitrogen containing compounds under pyrolysis conditions provides valuable information. Therefore a novel method has been developed and validated with a feedstock containing a high nitrogen content, i.e., a shale oil. First, the feed was characterized offline by comprehensive two-dimensional gas chromatography (GC &#215; GC) coupled with a nitrogen chemiluminescence detector (NCD). In a second step the on-line analysis method was developed and tested on a steam cracking pilot plant by feeding pyridine dissolved in heptane. The former being a representative compound for one of the most abundant classes of compounds present in shale oil. The composition of the reactor effluent was determined via an in-house developed automated sampling system followed by immediate injection of the sample on a GC &#215; GC coupled with a time-of-flight mass spectrometer (TOF-MS), flame ionization detector (FID) and NCD. A novel method for quantitative analysis of nitrogen containing compounds using NCD and 2-chloropyridine as an internal standard has been developed and demonstrated.

A method combining comprehensive two-dimensional gas chromatography with nitrogen chemiluminescence detection has been developed and applied to on-line analysis of nitrogen containing compounds in a complex hydrocarbon matrix.

在线含硫化合物在复杂油气矩阵氮分析

The shift to heavy crude oils and the use of alternative fossil resources such as shale oil are a challenge for the petrochemical industry. The ...

化学

Single-throughput Complementary High-resolution Analytical Techniques for Characterizing Complex Natural Organic Matter Mixtures

Natural organic matter (NOM) is composed of a highly complex mixture of thousands of organic compounds which, historically, proved difficult to characterize. However, to understand the thermodynamic and kinetic controls on greenhouse gas (carbon dioxide [CO2] and methane [CH4]) production resulting from the decomposition of NOM, a molecular-level characterization coupled with microbial proteome analyses is necessary. Further, climate and environmental changes are expected to perturb natural ecosystems, potentially upsetting complex interactions that influence both the supply of organic matter substrates and the microorganisms performing the transformations. A detailed molecular characterization of the organic matter, microbial proteomics, and the pathways and transformations by which organic matter is decomposed will be necessary to predict the direction and magnitude of the effects of environmental changes. This article describes a methodological throughput for comprehensive metabolite characterization in a single sample by direct injection Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS), gas chromatography mass spectrometry (GC-MS), nuclear magnetic resonance (NMR) spectroscopy, liquid chromatography mass spectrometry (LC-MS), and proteomics analysis. This approach results in a fully-paired dataset which improves statistical confidence for inferring pathways of organic matter decomposition, the resulting CO2 and CH4 production rates, and their responses to environmental perturbation. Herein we present results of applying this method to NOM samples collected from peatlands; however, the protocol is applicable to any NOM sample (e.g., peat, forested soils, marine sediments, etc.).

This protocol describes a single throughput for complementary analytical and omics techniques culminating in a fully-paired characterization of natural organic matter and microbial proteomics in different ecosystems. This approach permits robust comparisons for identifying metabolic pathways and transformations important for describing greenhouse gas production and predicting responses to environmental change.

用于表征复杂天然有机物混合物的单吞吐量互补高分辨率分析技术

Natural organic matter (NOM) is composed of a highly complex mixture of thousands of organic compounds which, historically, proved difficult to ...

Coupling Carbon Capture from a Power Plant with Semi-automated Open Raceway Ponds for Microalgae Cultivation

In the United States, 35% of the total carbon dioxide (CO2) emissions come from the electrical power industry, of which 30% represent natural gas electricity generation. Microalgae can biofix CO2 10 to 15 times faster than plants and convert algal biomass to products of interest, such as biofuels. Thus, this study presents a protocol that demonstrates the potential synergies of microalgae cultivation with a natural gas power plant situated in the southwestern United States in a hot semi-arid climate. State-of-the-art technologies are used to enhance carbon capture and utilization via the green algal species Chlorella sorokiniana, which can be further processed into biofuel. We describe a protocol involving a semi-automated open raceway pond and discuss the results of its performance when it was tested at the Tucson Electric Power plant, in Tucson, Arizona. Flue gas was used as the main carbon source to control pH, and Chlorella sorokiniana was cultivated. An optimized medium was used to grow the algae. The amount of CO2 added to the system as a function of time was closely monitored. Additionally, other physicochemical factors affecting algal growth rate, biomass productivity, and carbon fixation were monitored, including optical density, dissolved oxygen (DO), electroconductivity (EC), and air and pond temperatures. The results indicate that a microalgae yield of up to 0.385 g/L ash-free dry weight is attainable, with a lipid content of 24%. Leveraging synergistic opportunities between CO2 emitters and algal farmers can provide the resources required to increase carbon capture while supporting the sustainable production of algal biofuels and bioproducts.

A protocol is described to utilize the carbon dioxide in natural gas power plant flue gas to cultivate microalgae in open raceway ponds. Flue gas injection is controlled with a pH sensor, and microalgae growth is monitored with real time measurements of optical density.

将发电厂的碳捕获与半自动开放式管道池相结合，用于微藻养殖

In the United States, 35% of the total carbon dioxide (CO2) emissions come from the electrical power industry, of which 30% represent natural gas ...

Medium Preparation for the Cultivation of Microorganisms under Strictly Anaerobic/Anoxic Conditions

In contrast to aerobic organisms, strictly anaerobic microorganisms require the absence of oxygen and usually a low redox potential to initiate growth. As oxygen is ubiquitous in air, retaining O2-free conditions during all steps of cultivation is challenging but a prerequisite for anaerobic culturing. The protocol presented here demonstrates the successful cultivation of an anaerobic mixed culture derived from a biogas plant using a simple and inexpensive method. A precise description of the entire anoxic culturing process is given including media preparation, filling of cultivation flasks, supplementation with redox indicator and reducing agents to provide low redox potentials as well as exchanging the headspace to keep media free from oxygen. Furthermore, a detailed overview of aseptically inoculating gas tight serum flasks (by using sterile syringes and needles) and suitable incubation conditions is provided. The present protocol further deals with gas and liquid sampling for subsequent analyses regarding gas composition and volatile fatty acid concentrations using gas chromatography (GC) and high performance liquid chromatography (HPLC), respectively, and the calculation of biogas and methane yield considering the ideal gas law.

As obligate anaerobic organisms are unable to grow upon oxygen exposure, the use of anaerobic culturing techniques is indispensable. Here, we demonstrate a simple and effective method to cultivate a mixed culture derived from a biogas plant from media preparation to gas and volatile fatty acid quantification.

严格厌氧/缺氧条件下微生物培养的介质制备

In contrast to aerobic organisms, strictly anaerobic microorganisms require the absence of oxygen and usually a low redox potential to initiate growth. As ...

生物化学

Quantitative Analysis by Thermogravimetry-Mass Spectrum Analysis for Reactions with Evolved Gases

During energy conversion, material production, and metallurgy processes, reactions often have the features of unsteadiness, multistep, and multi-intermediates. Thermogravimetry-mass spectrum (TG-MS) is seen as a powerful tool to study reaction features. However, reaction details and reaction mechanics have not been effectively obtained directly from the ion current of TG-MS. Here, we provide a method of an equivalent characteristic spectrum analysis (ECSA) for analyzing the mass spectrum and giving the mass flow rate of reaction gases as precise as possible. The ECSA can effectively separate overlapping ion peaks and then eliminate the mass discrimination and temperature-dependent effect. Two example experiments are presented: (1) the decomposition of CaCO3 with evolved gas of CO2 and the decomposition of hydromagnesite with evolved gas of CO2 and H2O, to evaluate the ECSA on single-component system measurement and (2) the thermal pyrolysis of Zhundong coal with evolved gases of inorganic gases CO, H2, and CO2, and organic gases C2H4, C2H6, C3H8, C6H14, etc., to evaluate the ECSA on multi-component system measurement. Based on the successful calibration of the characteristic spectrum and relative sensitivity of specific gas and the ECSA on mass spectrum, we demonstrate that the ECSA accurately gives the mass flow rates of each evolved gas, including organic or inorganic gases, for not only single but multi-component reactions, which cannot be implemented by the traditional measurements.

Precise determination of the evolved gases&#39; flow rate is key to study the details of reactions. We provide a novel quantitative analysis method of equivalent characteristic spectrum analysis for thermogravimetry-mass spectrum analysis by establishing the calibration system of the characteristic spectrum and relative sensitivity, for obtaining the flow rate.

热重分析-质谱分析对演化气体反应的定量分析

During energy conversion, material production, and metallurgy processes, reactions often have the features of unsteadiness, multistep, and ...

工程学

用于从猪废沼气中获取生物甲烷的微藻培养

Biogas Purification through the use of a Microalgae-Bacterial System in Semi-Industrial High Rate Algal Ponds

In recent years, a number of technologies have emerged to purify biogas into biomethane. This purification entails a reduction in the concentration of polluting gases such as carbon dioxide and hydrogen sulfide to increase the content of methane. In this study, we used a microalgal cultivation technology to treat and purify biogas produced from organic waste from the swine industry to obtain ready-to-use biomethane. For cultivation and purification, two 22.2 m3 open-pond photobioreactors coupled with an absorption-desorption column system were set up in San Juan de los Lagos, Mexico. Several recirculation liquid/biogas ratios (L/G) were tested to obtain the highest removal efficiencies; other parameters, such as pH, dissolved oxygen (DO), temperature, and biomass growth, were measured. The most efficient L/Gs were 1.6 and 2.5, resulting in a treated biogas effluent with a composition of 6.8%vol and 6.6%vol in CO2, respectively, and removal efficiencies for H2S up to 98.9%, as well as maintaining O2 contamination values of less than 2%vol. We found that pH greatly determines CO2 removal, more so than L/G, during cultivation because of its participation in the photosynthetic process of microalgae and its ability to vary pH when solubilized due to its acidic nature. DO, and temperature oscillated as expected from the light-dark natural cycles of photosynthesis and the time of day, respectively. Biomass growth varied with CO2 and nutrient feeding as well as reactor harvesting; however, the trend remained primed for growth.

作者聚焦:扩展微藻生物技术以提高生物甲烷产量 

Air pollution impacts the quality of life of all organisms. Here, we describe the use of microalgae biotechnology for the treatment of biogas (simultaneous removal of carbon dioxide and hydrogen sulfide) and the production of biomethane through semi-industrial open high-rate algal ponds and subsequent analysis of treatment efficiency, pH, dissolved oxygen, and microalgae growth.

在半工业化高速率藻类池塘中使用微藻细菌系统进行沼气净化

In recent years, a number of technologies have emerged to purify biogas into biomethane. This purification entails a reduction in the concentration of ...

Derivatization and Separation of Barley Flour Metabolites into Four Fractions

Metabolomic Analysis of Barley by Gas Chromatography/Mass Spectrometry

Climate change increases drought risk to agriculture and impacts both food nutrient content and overall food security. Metabolomics is one way to observe and quantify the impacts of drought on grain and other agricultural products. The identified metabolites may allow for the identification of the biochemical response that allows the plant to tolerate stressful environments. The methodology presented herein allowed for the total metabolomic analysis of barley flour using gas chromatography/mass spectrometry (GC/MS). 
Barley flour metabolite extracts were fractionated into four fractions based on polarity. To allow for analysis by GC/MS, metabolites were derivatized to increase volatility and metabolite separation: fatty acids esters were derivatized into fatty acid methyl esters; sugars were oximated into their straight chain form; and metabolites with hydroxyl groups were converted to their corresponding silyl ethers. The derivatized samples were injected into the GC/MS and the generated mass spectra were used for metabolite identification by comparing the generated spectra to the National Institute of Standards and Technology (NIST) Tandem Mass Spectra library. The method described here can also be used to examine the total metabolome for other plants, furthering our understanding of the biochemical responses of stressed plants.

A method for metabolomic analysis of barley is presented. The method entails fractionation and derivatization of metabolites and analysis thereof by gas chromatography/mass spectrometry (GC/MS). Metabolomic analysis can be used to determine the effect of intercropping and drought stress on the grain.

Climate change increases drought risk to agriculture and impacts both food nutrient content and overall food security. Metabolomics is one way to observe ...

从藻类水热液化使用二维气相色谱时间飞行质谱法水提取部分的定性鉴定

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用于表征复杂天然有机物混合物的单吞吐量互补高分辨率分析技术

将发电厂的碳捕获与半自动开放式管道池相结合，用于微藻养殖

严格厌氧/缺氧条件下微生物培养的介质制备

热重分析-质谱分析对演化气体反应的定量分析

在半工业化高速率藻类池塘中使用微藻细菌系统进行沼气净化

在半工业化高速率藻类池塘中使用微藻细菌系统进行沼气净化

在半工业化高速率藻类池塘中使用微藻细菌系统进行沼气净化

Metabolomic Analysis of Barley by Gas Chromatography/Mass Spectrometry