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
Summary
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.
Abstract
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。
Introduction
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。
Protocol
Representative Results
Discussion
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?…
Divulgaciones
The authors have nothing to disclose.
Acknowledgements
这份手稿已根据合同号DE-AC05-76RL01830撰写由巴特尔纪念研究所与美国能源部门。美国政府保留和发布，通过接受的文章发表，承认美国政府保留了非排他性的，缴足，不可撤销的，全球性的许可发布或复制本手稿的出版形式，还是让别人做所以，对于美国政府的目的。
Materials
| GC × GC – TOF/MS | Leco | PEG4D11DLN15 | Commercial Pegasus 4D |
| ChromaTOF version 4.50 | Leco | Data analysis software | |
| Rxi-5MS GC column | Restek | 13420 | 2.3 m column was used from this column. |
| Stabilwax GC column | Restek | 10626 | |
| HP-5 GC column | Agilent | 19091J-416 | |
| Stabilwax GC column | Restek | 15121 | |
| Presstight Connector | Restek | 20430 | |
| GC injector liner | Restek | 23305.5 | |
| GC Injector ferrules | Agilent | 5181-3323 | |
| Non-stick liner O-rings | Agilent | 5188-5365 | |
| Transfer line ferrules | Restek | 20212 | |
| Ethanol | Sigma-Aldrich | 459844 | Chromatography grade |
| Acetone | Sigma-Aldrich | 414689 | Chromatography grade |
| Acetic acid | Sigma-Aldrich | 320099 | Chromatography grade |
| 2-butanone | Sigma-Aldrich | 360473 | Chromatography grade |
| Propanoic acid | Sigma-Aldrich | 402907 | Chromatography grade |
| Butanoic acid | Sigma-Aldrich | 19215 | Chromatography grade |
| Pyridine | Sigma-Aldrich | 270970 | Chromatography grade |
| Pyrazine | Sigma-Aldrich | 65693 | Chromatography grade |
| Acetamide | Sigma-Aldrich | 695122 | Chromatography grade |
| 2,5-pyrrolididione | Sigma-Aldrich | S9381 | Chromatography grade |
| N-methylsuccinimide | Sigma-Aldrich | 325384 | Chromatography grade |
| N-(2-hydroxyethyl)succinimide | Sigma-Aldrich | 444073 | Chromatography grade |
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