15张酷炫动图让你秒懂色谱原理

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                    <p><span><strong><span></span></strong></span></p><section><section powered-by="gulangu"><section><section><section powered-by="gulangu"><section><section></section><section><section><section powered-by="gulangu"><section><section><p>点击上方<strong>蓝字</strong>关注“<span><strong>公众号</strong></span>”</p></section></section></section></section></section></section></section></section><section><section powered-by="gulangu"><section><p><img src="image/20201014/af7fff12fe4b43cfc7c3fc5591ac0e75_1.gif" /></p></section></section></section></section></section></section><p><span><strong><span></span></strong></span><br  /></p><p><span><strong><span>色谱</span></strong></span><span>（chromatography）</span><span>是分析测试物质理化性质的常用的有效手段，多用于生物分子及高聚物的分离检测等。</span></p><p><br  /></p><p><span>色谱包含多种操作方式不同但原理相同的分离、分析技术。按其流动相和固定相状态不同分为气相色谱和液相色谱。今天，随测了么小编来看一下色谱仪具体是怎么工作的吧！</span></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/c613b65ef5f28319ff4a9c75af4bd896_2.gif" /></p></p><p><span></span></p><p><span>气相色谱仪流程</span><br  /></p><p><span><br  /></span></p><p><span>你一般看到的气相色谱仪是长这样的。</span><br  /></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/2704f15fca735fcebf33d83a3335bae9_3.gif" /></p></p><p><span>色谱仪</span><br  /><span></span></p><p><span><br  /></span></p><p><span>以气相色谱仪为例，主要由三大部分构成：<strong>载气、色谱柱、检测器。</strong>每一模块具体工作流程如下。</span><br  /></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/fc546e102150cbd2216696db21e4a313_4.gif" /></p></p><p><span>注射器<br  /></span></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/e407d752ef28e3afea42542a21578992_5.gif" /></p></p><p><span>色谱柱<br  /></span></p><p><span><br  /></span></p><p><span></span></p><section><section powered-by="gulangu"><section><section><section><p><strong>色谱柱分类</strong></p></section><section><section></section><section></section><section></section></section></section></section></section></section><p><br  /><span></span></p><p><span></span></p><section><section powered-by="gulangu"><section><section><span></span> <span></span> <span></span> </section><section><p><strong>1. 非极性</strong></p></section></section></section></section><p><span>100%Dimethylpolysiloxane，100%聚二甲基硅氧烷</span><br  /><span></span></p><p><span>产品牌号：AC1，OV-101，OV-1，DB-1，SE-30，HP-1，RTX-1，BP-1</span></p><p><span><br  /></span></p><p><span></span></p><section><section powered-by="gulangu"><section><section><span></span> <span></span> <span></span> </section><section><p><strong>2. 弱极性</strong></p></section></section></section></section><p><span>5%Phenyl dimethylpolysiloxane, 5%二苯基(95%)二甲基聚硅氧烷</span><br  /><span></span></p><p><span>产品牌号：AC5，SE-52，</span></p><p><span>5% Phenyl 1%vinyldimethyl polysiloxane，5%二苯基1%乙烯基(94%)二甲基聚硅氧烷</span></p><p><span>产品牌号：OV-5，DB-5，SE-54，HP-5，RTX-5，BP-5</span></p><p><br  /></p><section><section powered-by="gulangu"><section><section><span></span> <span></span> <span></span> </section><section><p><strong>3. 中等极性</strong></p></section></section></section></section><p><span>50%Phenyl dimethylpolysiloxane, 50%二苯基(50%)二甲基聚硅氧烷</span></p><p><span>产品牌号：OV-17，HP-50，RTX-50</span></p><p><span>14%Cyanopropylphenyl polysiloxane, 14%氰丙基苯基(其中7%氰丙基7%苯基)(86%)二甲基聚硅氧烷</span></p><p><span>产品牌号：AC10，OV-1701，DB-1701，RTX-1701</span></p><p><span>50% Cyanopropylphenyl polysiloxane，50%氰丙基苯基(其中25%氰丙基25%苯基)(50%)二甲基聚硅氧烷</span></p><p><span>产品牌号：AC225，OV-225，BP-225，DB-225，HP-225，RTX-225</span></p><p><br  /></p><section><section powered-by="gulangu"><section><section><span></span> <span></span> <span></span> </section><section><p><strong>4. 强极性</strong></p></section></section></section></section><p><span>polyethylene glycol,聚乙二醇</span></p><p><span>产品牌号：AC20，PEG20M，HP-INNOWAX</span></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/e4952eef2a55383a2b78293767db2df0_6.gif" /></p></p><p><span>氢焰检测器</span><br  /></p><p><span><br  /></span></p><p><strong><span></span></strong></p><section><section powered-by="gulangu"><section><section><section><section><section></section><section></section></section><section><section powered-by="gulangu"><section><section><p><strong>Size exclusion chromatography</strong></p><p><strong>&nbsp;(SEC)</strong></p></section></section></section></section></section><section><section></section><section></section></section></section></section></section></section><p><br  /></p><p><span>体积排阻色谱法（SEC）是利用多孔凝胶固定相的独特特性，主要依据分子尺寸大小的差异来进行分离的方法，样品分子与固定相之间不存在相互作用。</span><br  /></p><p><br  /></p><p><span>根据所用凝胶的性质，可以分为使用水溶液的凝胶过滤色谱法（GFC）和使用有机溶剂的凝胶渗透色谱法（GPC）。</span></p><p><span><br  /></span></p><p><p><img src="image/20201014/a8343afe4c11a09840b0ed696d7d09bb_7.gif" /></p></p><p><span>只依据尺寸大小分离，大组分最先被洗提出</span></p><p><br  /></p><p><br  /></p><p><strong><span>原理：</span></strong><span>色谱固定相是多孔性凝胶，只有直径小于孔径的组分可以进入凝胶孔道。大组分不能进入凝胶孔洞而被排阻，只能沿着凝胶粒子之间的空隙通过，因而最大的组分最先被洗提出来。</span></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/ea558f802ae0deff82a6e89fa21a4a65_8.gif" /></p></p><p><span>直径小于孔径的组分进入凝胶孔道</span><br  /></p><p><br  /></p><p><br  /></p><p><span>小组分可进入大部分凝胶孔洞，在色谱柱中滞留时间长，会更慢被洗提出来。溶剂分子因体积最小，可进入所有凝胶孔洞，因而是最后从色谱柱中洗提出。这也是与其他色谱法最大的不同。</span></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/dea1db380f8adbb267afd4829229c609_9.gif" /></p></p><p><span>依据尺寸差异，样品组分分离</span><br  /></p><p><span><br  /></span></p><p><span>体积排阻色谱法适用于对未知样品的探索分离。凝胶过滤色谱适于分析水溶液中的多肽、蛋白质、生物酶等生物分子；凝胶渗透色谱主要用于高聚物（如聚乙烯、聚丙烯、聚苯乙烯、聚氯乙烯、聚甲基丙烯酸甲酯）的分子量测定。</span><br  /></p><p><span><br  /></span></p><p><span></span></p><section><section powered-by="gulangu"><section><section><section><section><section></section><section></section></section><section><section powered-by="gulangu"><section><section><p><strong>Ion exchange chromatography</strong></p><p><strong>(IEC)</strong><br  /></p></section></section></section></section></section><section><section></section><section></section></section></section></section></section></section><p><span></span><br  /></p><p><span>离子交换色谱是利用被分离组分与固定相之间发生离子交换的能力差异来实现组分分离。</span></p><p><span><br  /></span></p><p><span>离子交换色谱主要用于分离离子或可离解的化合物。</span><br  /></p><p><span><br  /></span></p><p><span>原理：固定相为离子交换树脂，树脂分子结构中存在许多可以电离的活性中心。流动相最常使用水缓冲溶液，也有有机溶剂如甲醇，或乙醇同水混合。</span><br  /></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/c00afa1b5f50213e8820cc1645a3ebaa_10.gif" /></p></p><p><span></span></p><p><span>初始平衡状态</span><br  /></p><p><span><br  /></span></p><p><span>流动相带着样品组分电离生成的离子通过固定相时，待分离组分中的离子与树脂分子结构中可电离的活性中心发生离子交换，形成离子交换平衡，从而在流动相与固定相之间形成分配。</span><br  /></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/dabf89ddaa301bcaede11b005dcccc26_11.gif" /></p></p><p><span>样品组分与活性中心发生离子交换</span><br  /></p><p><span><br  /></span></p><p><span>固定相的固有离子与待分离组分中的离子之间相互争夺固定相中的离子交换中心，并随着流动相的运动而运动，最终实现分离。</span><br  /></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/94d328ab5e6d0b2051f1035736d30d56_12.gif" /></p></p><p><span>洗提过程<br  /></span></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/d99e73d4add6377d558ea0f521ca9619_13.gif" /></p></p><p><span>洗提过程</span><br  /></p><p><span><br  /></span></p><p><span></span></p><section><section powered-by="gulangu"><section><section><section><section><section></section><section></section></section><section><section powered-by="gulangu"><section><section><p><strong>Affinity chromatography</strong></p><p><strong>(AFC)</strong></p><p><br  /></p></section></section></section></section></section><section><section></section><section></section></section></section></section></section></section><p><span></span><br  /></p><p><span>亲和色谱法是利用与固定相不同程度的可逆结合的亲和性，使成分与杂质分离的色谱法。固定相为相互间具有高度特异亲和性的二种物质之一。</span><br  /></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/da9f541e75391a01bf78fc9c2cdef12e_14.gif" /></p></p><p><span>初始平衡状态<br  /></span></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/31288e051f48941eff4ff26b3c2c147b_15.gif" /></p><span></span></p><p><span>亲和吸附</span><br  /></p><p><span><br  /></span></p><p><span>不同分子与固定相间亲和力大小有差异，选择适当的洗脱条件便可将混合物中的组分逐个洗脱下来，达到分离纯化的目的。</span><br  /></p><p><span><br  /></span></p><p><span></span></p><p><p><img src="image/20201014/db257fae91ab57e567ac1aa6cbe9d823_16.gif" /></p></p><p><span>解吸附</span><br  /></p><p><span><br  /></span></p><p><span>主要用于分离活体高分子物质、过滤性病毒及细胞。优点是操作简便、效率高、条件温和，缺点是使用局限性大。</span><br  /></p><p><span><br  /></span></p><p><br  /></p><section><section powered-by="gulangu"><section><section><section><section><section></section><section></section></section><section><section powered-by="gulangu"><section><section><p><strong>Hydrophobicinteraction chromatography</strong></p><p><strong>&nbsp;(HIC)</strong><br  /></p></section></section></section></section></section><section><section></section><section></section></section></section></section></section></section><p><span></span><br  /></p><p><br  /></p><p><span>Hydrophilicinteraction chromatography (HILIC),similar to normal phase chromatography (NPC)</span><br  /></p><p><span><br  /></span></p><p><span>Reversed phasechromatography (RPC)</span><br  /></p><p><span><br  /></span></p><p><span>Mixed modechromatography</span><br  /></p><p><span><br  /></span></p><p><strong><span>色谱仪的选用依据：</span></strong></p><p><strong><span><br  /></span></strong></p><section><section powered-by="gulangu"><section><section><span></span> <span></span> <span></span> </section><section><p><strong>1. 相对分子质量</strong></p></section></section></section></section><p><span>相对分子质量小于200、挥发性比较好的化合物，可用气相色谱仪剖析。</span><br  /></p><p><span>相对分子质量在200～2000的化合物，可用液-液分配色谱仪、液-固吸附色谱仪和离子交换色谱仪剖析。</span></p><p><span>相对分子质量大于2000的化合物，可用凝胶色谱仪剖析。</span></p><p><br  /></p><section><section powered-by="gulangu"><section><section><span></span> <span></span> <span></span> </section><section><p><strong>2. 溶解性</strong></p></section></section></section></section><p><span>水溶性样品选用离子交换色谱仪和液-液分配色谱仪剖析。</span></p><p><span>微溶于水，但用酸或碱能很好电离的化合物，可用离子交换色谱仪剖析。</span></p><p><span>油溶性和非极性的样品，可选用液固吸附色谱仪。</span></p><p><span><br  /></span></p><p><span></span></p><section><section powered-by="gulangu"><section><section><span></span> <span></span> <span></span> </section><section><p><strong>3. 化学构造</strong></p></section></section></section></section><p><span>含有离子型或可离子化的化合物，或者能与离子型化合物相互作用的化合物（如配位体和有机螯合剂），可首要考虑用离子交换色谱仪，或液-液分配色谱仪和凝胶色谱仪。</span></p><p><span>具有不同官能团的化合物、同系物，可用液-液分配色谱仪剖析。</span></p><p><span>异构体可用液固吸附色谱仪剖析。</span></p><p><span>高分子聚合物可用凝胶色谱仪。</span></p><p><br  /></p><p><span><strong><span>- The End -</span></strong></span></p><p><br  /></p><p><br  /></p><section><section powered-by="gulangu"><section><section><span><section><span>推荐阅读：</span></section></span></section><section><section powered-by="gulangu"><section><section><p><span><strong><span>1.&nbsp;</span></strong></span><strong><span>又来！20张酷炫动图,秒懂四大名谱（IR、MS、NMR、UV）原理！</span></strong></p><p><span><strong><span>2.&nbsp;</span></strong></span><strong><span>叒来了！20张动图秒懂十大3D打印原理之高分子篇</span></strong></p><p><span><strong><span>3.&nbsp;</span></strong></span><strong><span>超逼真20张动图，秒懂四大显微分析工作原理（SEM, TEM, AFM, 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