크로마토그래피 소개: 분리 및 분석 장비 소개

크로마토그래피는 물질의 특성에 따라 분리하는 다목적 분석 장비로, 종이 및 컬럼 크로마토그래피와 같은 다양한 방법을 사용할 수 있습니다.

### 크로마토그래피에 대한 종합 가이드: 크로마토그래피의 원리, 기술 및 응용
* 🔍 "크로마토그래피는 주로 분리에 사용되며 **증류 및 결정화와 같은 기존 방법에 비해 많은 이점**을 제공합니다."
* 오늘날 컬럼 크로마토그래피는 **가장 효과적이고 편리한 방법**으로 간주되며, 일반적으로 분석 또는 제조 목적으로 분류됩니다.
* 📜 19세기에 시작된 가장 오래된 방법인 **종이 크로마토그래피**는 용매와 염료가 형성하는 다양한 패턴을 기반으로 염료를 분석하는 데 사용됩니다.
* 📏 "효과적인 분리를 달성하려면 입자가 작을수록 피크가 더 날카로워지므로 **컬럼 패킹 재료의 크기를 줄이는 것**이 필수적입니다."
* ⚙ 고성능 액체 크로마토그래피(HPLC)는 **컬럼의 작은 입자**를 사용하여 고압에서 고분해능으로 효율적인 분리를 위해 설계된 장비입니다.
* "HPLC는 액체와 기체의 고유한 차이로 인해 **미량 분석에는 우수**하지만, 생산 목적으로 사용하기에는 어려움이 있습니다."
* 기체 상태에서는 분자 간 상호작용이 거의 없기 때문에 **헬륨이나 수소와 같은 운반 기체**를 사용하여 순수한 물질을 분리할 수 있습니다.
* 🎓 크로마토그래피는 최종 성질이 없는 매우 다재다능한 분석 장비로, **이동상과 정지상의 차이에 따라 물질을 분리**합니다.

Hello, in this video, I will briefly introduce chromatography. In fact, compared to other spectrometers like NMR or XRD, there is a lot of Korean material for chromatography equipment. However, the content seems either too easy for high school students or overly specialized. Therefore, I have prepared a video suitable for undergraduate or graduate students. I will introduce chromatography in an easy-to-understand flow that I have organized. Chromatography is primarily used for separation. It has many advantages over traditional and old methods like distillation and crystallization. It can separate substances with very small differences in properties with high precision. Nowadays, column chromatography is the most effective and convenient method. Chromatography is usually classified for analysis or manufacturing purposes. I will show you some representative chromatographies while explaining.

Paper chromatography is the oldest method used for the analysis of dyes, dating back to the 19th century. It was empirically discovered that when dyes are stained on white cloth with a solution of dye in seawater or a solvent, the resulting patterns differ depending on the solvent used. In this process, the mobile phase that moves is called the mobile phase, while the stationary phase that remains still is called the stationary phase. The driving force, Paul's driving force, is the affinity between the solvent and the dye. Therefore, in the above results, the spots are interpreted as dyes A and B. When the solvent moves more than dye B in a solvent system of 2:1, dye A is considered to have moved more than dye B. Under the same conditions, if the dye moves more than the solvent, it means that the dye has a higher affinity with the solvent, and if the dye moves less, it is interpreted as having a higher polarity and saturation. In summary, the interpretation is quite simple when considering the detailed content.

If you look a little closer, the paper chromatography conducted in the Boto laboratory is cut into two for easier and more economical viewing. By directly comparing these, it is easy to identify the substances. If it is difficult to compare directly under the same conditions, you can also compare by calculating the substance's RF factor based on the distance traveled by the substance divided by the distance traveled by the solvent. It is a very easy way to separate substances. However, there is a basic material made of paper or cloth, which is a porous material made of polymers, so it has uniformity. Additionally, it is vulnerable to humidity and cannot be dried for 5 minutes, so it is good for uniformity, and it is a new type of paper using TS, which has many other advantages. When pulling the layer with TS, the introduction of gravity is also possible. Typically, glass or aluminum supports are used as substrates.

Silica gel or aluminum oxide is thinly and uniformly coated to manufacture it. Therefore, when using THD, the development results are cleaner than paper, slightly faster, and have advantages such as shorter development time, lower UV exposure, etc. It is a slightly more expensive but still economical and effective separation method compared to paper that can be used in the laboratory as a routine and essential method. However, after the TS, rinsing is required. TLC is fundamentally suitable for analyzing solubility, making it challenging to use for manufacturing purposes. Each spot on the tag plate contains substances with different Rf values, making it really challenging. Companies like Central Corporation can refine materials as desired on thousands or tens of thousands of gel plates, but unless there are unavoidable circumstances like the aforementioned company, that would be too ignorant of a method.

You can use a column chromatography using silica-based fillers that K used. If you use a column chromatography, you can pull enough and also use it for manufacturing chromatography. The principle is the same as with the tag, but the order may change, so you can set the basic conditions for the column by using TLC. By developing it, you can achieve the desired spots well separated according to the conditions in the test. Even in the column, you can achieve a good selective separation that you want. Of course, you have to manually collect and dry it. This process is cumbersome and takes about 9 hours, which is a really critical disadvantage. However, there is one fatal flaw in using it negatively; the separation is not sufficient. Simply put, in a chromatogram like this, chromatogram is usually expressed visually in terms of the time and intensity of the chromatogram signal.

I will explain this in more detail in another video. In simple terms, in a chromatogram like this, you can see that each substance has a different retention time, and it is also difficult to separate the peaks effectively in some cases. To solve this, you need to find the width of the peak. In terms of TLC, you need to make the bands as narrow as possible, almost like lines. When you look at the chromatogram, there are many methods and theories presented for this, but the best way is actually to reduce the size of the column packing. This is the most certain way. The smaller the packing material, the sharper the peaks become. Empirically, when you want to remove impurities from dirty water, it is better to do it in a sand-filled tube rather than a tube filled with pebbles. It is better to proceed in clear water or water with sand rather than in cloudy water. However, using small-sized packing material leads to one problem.

The fine sand does not easily enter the moving bed. If you think about trying to filter water in a situation where there are coarse, medium, and fine sands, when the fine sand covers the water inlet of the filter, it will clog the sand a lot and just pile up on top of the sand. When rain falls on the sandy ground, it will also clog a lot and water will slowly seep on top of it. If the particle size is small enough, it will seep very slowly. Therefore, it is not possible to filter with just sand. You need to apply external strong pressure to make it go down. So, using glass anymore is not recommended because glass can break under high pressure. That's why stainless steel is used. From now on, I will briefly explain about HPLC. It's difficult to apply strong pressure selectively only to the column. It's easy to isolate the column and apply pressure to the front.

Continuous infusion of this statue is necessary, so a device is needed to maintain pressure starting from mobile phase 4 being injected into the column. Otherwise, when only the column is hung, the statue cannot be inserted. Also, a switch is needed to connect the system when injecting samples. The key point of this video is HPLC. To provide strong pressure to the column, all parts from the mobile phase injection step to sample elution and separation are operated as a single system, isolated from the outside. Everything is designed with small particles in the column to achieve high pressure for high separation efficiency. In other words, it is equipment designed for high performance for high resolution at high pressure. Now that the separation efficiency is high enough, you can use a small column for analytical purposes or a large column for high separation efficiency. Also, the types of column packing materials are divided into head vs. tail phases.

By alternating, you can effectively separate various substances. It is a versatile equipment that is useful for various purposes. Lastly, if I briefly introduce the chromatography, it is an analysis equipment that is extremely versatile and has no final nature. In the previous section, we saw that the subject of the analysis is substances that interact in various additional ways. It was mentioned that substances are separated according to the differences between their mobile and stationary phases. However, the distances between the molecules making up this substance, which is basically in a liquid state, are too close. Since the distances are close, unwanted intermolecular interactions act significantly, and conversely, in the gas state, the distances between molecules are far, so there is almost no intermolecular interaction. In this state, using gases like helium, which has low reactivity due to the large number of carrier gas molecules, and hydrogen, you can perform pure substance separation with only mutual adaptation between the mobile and stationary phases.

While it may be preferable to refer to it as HTC, it's actually better to call it HPLC. No matter how hard you try to pass the column with fine particles and high-pressure pumps, it's difficult to overcome the inherent difference between liquid and gas. This is why HPLC is superior for trace analysis. On the contrary, using it for production purposes is very challenging because the injection of the gas is minimal, and the smell of the solvent is too weak, leading to excessively low efficiency. Of course, using TLC in the same column conditions is impossible. Just accept that even without a large amount of solvent, TLC can have high separation efficiency. Please understand that the state of the substance being analyzed differs due to the nature of gas and liquid. So, this concludes a brief introduction to chromatography. From the perspective of analytical and manufacturing purposes.

I introduced some representative chromatographies in my own order and logic. I hope it was helpful in understanding chromatography a little bit. Since this is an introductory video, I omitted some details, but there is a lot of content. In the future, I will gather and explain one by one about chromatography. Then, I will try to provide more helpful videos. Thank you for watching.

[분석화학] Chromatography - 간단하게 살펴보는 크로마토그래피의 종류 및 원리

모든 영상은 자유롭게 퍼가세요. 수업, 과제, 발표등 비영리적인 목적이면 어디에서나 사용하셔도 됩니다!

화학 튜터

크로마토그래피 소개: 크로마토그래피 소개: 원리 및 방법

크로마토그래피는 작은 물성 차이를 바탕으로 물질을 분리하는 매우 정밀한 방법으로, 컬럼 크로마토그래피가 가장 효과적인 현대 기술입니다.

19세기에 시작된 이 공정은 이동상과 고정상을 사용하여 물질을 이동시키고 친화도와 극성을 기반으로 결과를 해석합니다.

종이 크로마토그래피는 습기에 취약하지만 물질 분리의 단순성과 균일성 때문에 널리 사용됩니다.

박층 크로마토그래피(TLC)와 컬럼 크로마토그래피의 장단점

박층 크로마토그래피(TLC)는 종이보다 더 깨끗하고 빠른 현상 결과를 제공하며, 현상 시간이 짧고 자외선 노출이 적다는 장점이 있습니다.

경제적이고 효과적인 분리 방법이지만 사용 후 헹굼이 필요하고 플레이트에 있는 물질의 Rf 값이 다르기 때문에 제조 목적으로 사용하기에는 어렵습니다.

반면에 컬럼 크로마토그래피는 선택적 분리가 가능하지만 약 9시간이 소요되는 번거로운 공정이며 충분한 분리를 제공하지 못할 수 있습니다.

크로마토그래피에서 피크 분리 개선

크로마토그램의 물질마다 머무르는 시간이 다르기 때문에 피크 분리가 어려울 수 있습니다.

컬럼 패킹의 크기를 줄이는 것이 피크를 더 선명하게 만드는 가장 효과적인 방법입니다.

그러나 작은 패킹 재료를 사용하면 막힘이 발생할 수 있으므로 일반적으로 스테인리스 스틸로 강한 압력을 가해야 합니다.

고성능 액체 크로마토그래피(HPLC)에서는 컬럼에 선택적으로 강한 압력을 가하는 것이 어렵기 때문에 일반적으로 분리를 위해 전면에 압력을 가합니다.

HPLC 시스템에서 연속 주입의 중요성

이동상 주입부터 HPLC 시스템에서 연속 주입을 위한 압력을 유지하기 위한 장치가 필요합니다.

HPLC 시스템은 단일 장치로 작동하여 분리 효율을 극대화하기 위해 높은 압력을 제공하므로 컬럼 패킹 재료를 다양하게 사용할 수 있습니다.

HPLC는 이동상과 고정상의 차이에 따라 물질을 분리하는 매우 다재다능한 분석 장비로 분자 간 상호 작용이 밀접한 물질을 분리할 수 있습니다.

미량 분석을 위한 HPLC의 우수성 및 생산에서의 과제

액체와 기체의 고유한 차이로 인해 미량 분석에는 HPLC가 HTC보다 우수합니다.

생산 목적으로 HPLC를 사용하면 가스 주입이 적고 용매 냄새가 약해 효율이 낮습니다.

반면 TLC는 소량의 용매로도 높은 분리 효율을 얻을 수 있지만 HPLC와 동일한 컬럼 조건에서 사용하는 것은 불가능합니다.