Chromatography for Industrial Chemical

Different Types of chromatography techniques for Various Industries.

February 3, 2018


Different Types of chromatography techniques for Various Industries.

Application of Chromatography in Chemistry

In Industries pertaining to the synthesis of bio-molecules require a special technique which does not denature the characteristics of the bio-molecules during Separation and Purification. Column Chromatography is one such technique having different modes which are used on a large scale in Industrial level as it is bio-compatible and requires relatively short time for separation and purification. The Column Chromatography Principle is mainly based on size, Charge, and Hydrophobicity of the sample which in turn lead to various Chromatographic techniques such as-Size exclusion chromatography or Gel permeation chromatography, Adsorption chromatography, Ion exchange chromatography and Affinity Chromatography. Each technique is based on the biological interaction between the sample bio-molecules and the packing material such as Alumina or Silica gel.

 

Types of chromatography and their applications

Size exclusion chromatography:

is also known as molecular sieve chromatography separates analyses according to their size as they pass on through the stationary phase namely Silica which is porous in nature and acts as a hydrophilic surface.

Biochemical research applicationsIn Adsorption chromatography,

the stationary phase is considered as the Adsorbent which is used in separating Non-volatile mixtures. In this method, the Adsorbent used can be Silica, Alumina or Cellulose.

Ion exchange chromatography

is used in the separation of polar molecules and ions such as proteins, amino acids containing both positive and negative charged chemical groups. In this method, the stationary phase used is slurry of either Silica or Alumina.

Affinity chromatography:

Affinity chromatography is used for the separation of recombinant proteins and enzymes and here the stationary phase act as a Support medium.

Chromatography techniques are widely used-

 

  • In Biochemical research applications to separate and identify the compound of biological origin,
  • In Petroleum industries to control the production of saturated carbons,
  • In Chemical Industries to purify the chemicals,
  • In Sugar factories to purify sugar from Molasses.

Sugar Factories

For any technique, if the packing material employed is Silica gel than the Silica gel column chromatography Procedure mainly depends on packing the column without cracks and air bubbles as it leads to poor separation and purification. Whereas if the packing material used in these techniques is Alumina Column Methanol, it contains Methanol, which is considered as a volatile eluting solvent and therefore used in Alumina Column Chromatography. Apart, from methanol to improve the effectiveness of purification and to adjust the alkalinity it is treated with either dilute acid or base considering the components nature of the Column. After adjusting the alkalinity of Alumina it is characterized as Neutral, Acidic and Basic Alumina. Among this Neutral Alumina is used in chromatographic cleanup procedures and also for the separation of Aliphatic, Aromatic and Polar fractions. Purification of free base Porphyrins and Metalloporphyrins is also performed using Neutral Alumina. When it is used as a stationary phase in the Column, for the preparative separation of the mixture having basic properties, it does not require a mobile phase modifier.

 Uses of Chromatography in medicine

Isolation of natural products from plants

Lack of efficiency of a medicine results in lethal accidents and has become a major concern. Therefore Chromatography opts as a detection tool in identifying and separating the desired medicine from the contaminants. The following are some of the medicines that are developed using Chromatography techniques-

  • Synthesis of Potential Anti Pertussis medicine involves Chromatography technique.
  • Separation of enantiomers which are useful in drug discovery is progressed through different Chromatography techniques.
  • Chromatography is also applied in the batch Fermentation process to determine the improvement of the process during antibiotic production.
  • Separation often different H1-Antihistamines which were structurally related was achieved by Chromatography, impregnating Silica gel with transition metal ions.
  • Purification of Proteins and Peptide hormones is attained either by Ion-exchange Chromatography or Gel Permeation Chromatography.
  • By Size exclusion chromatography using Alumina nanoparticles size separation of DNA molecules was achieved.

Application of Chromatography in Chemistry

Application of Chromatography in Chemistry

In chemistry one of the major aspects is to analyze the residue of all major classes which include Food, water, and air. It is mainly essential to emphasize the quality and safety of these classes or else lead to adverse effect on Human health. Therefore, this analysis of residues is accomplished by different Chromatography Techniques. The following are some of the techniques used in chemistry to separate and analyze the components which are essential in day to day life.

  • Separation of Proteins and small Oligonucleotides made easy by using Ion-exchange chromatography.
  • It is also used for the separation of Aldehydes, Ketones, Quinones, Esters, Lactones, and Glucosides.
  • Environmental agency as made Chromatography a method to test the quality of the Drinking water.
  • It is also used to monitor the air quality.
  • It is employed in the separation of organic as well as Inorganic compounds.
  • It is an indispensable method to study the heavy oils containing hydrocarbon group.
  • Analysis of pesticide residues on fruits and vegetables is also carried by this analytical technique.
Isolation of natural products from plants

Extraction of Bio-active compounds from medicinal plants.

February 3, 2018


Extraction of Bioactive compounds from medicinal plants.

In today’s arena, Herbal medicine or medicine extracted from medicinal plants have found their waisolation of bioactive compounds from plantsy in ailing many diseases when compared with Synthetic drugs. In recent years their utilization has increased rapidly and accounted for a significant percentage in the medicine market. Even the WHO (World Health Organization) supports the use of medicinal plants and introduced measures to ensure the quality of the Herbal medicines using modern techniques such as Chromatography and for manufacturing practices.

 

Isolation of Natural products from plants

Natural products are the components or substances that are produced from natural sources such as plants. These products which occur naturally have an impact on human life and therefore, used as Medicines, dietary supplements and in cosmetics.

Isolation of natural products from plants

The components of plants that have medicinal value and healing properties are called as Bioactive compounds and they undergo a series of the process called as Extraction, Isolation, and Characterization. Column Chromatography procedure for plant extracts is one such technique for isolating and identifying the Bioactive compound using a stationary phase such as Silica gel or Alumina. Before Isolation and extraction, identification of the bioactive compound is done based on a method called as Fingerprint which defines the character and gives complete information about a specific plant. Fingerprint analysis gives an accurate identification with similar peaks and is helpful in determining the intrinsic quality of the Bioactive compound.

Isolation of bioactive compounds from plants.

The Development of analytical techniques has to lead to a qualitative as well as quantitative analysis of Isolation of bioactive compounds from Plants which are herbs with a complicated system of mixtures formed through various pathways. Chromatography is one such analytical technique used for the authentication and identification of bioactive compound in a plant and is readily available.High-Performance Liquid Chromatography (HPLC) is a most extensively used application in Pharmaceutical industries for the isolation and purification of bioactive compounds from medicinal plants. This application requires larger stainless steel Columns packed with normal Phase Silica which not only isolates but also gives information about the sample containing new synthetic products. This is the very helpful technique for a pharmaceutical industry to introduce a new product into the market within a less timeframe.

Many more feasible techniques are emerging to control the quality of the bioactive compounds by using Solid phases such as Silica Gel with less particle size and shorter Column size. Ultra High-Performance Chromatography(UHPLC) is such a technique which is gaining momentum because of its enhanced selectivity and decreased analysis time, in isolating bioactive compounds from medicinal plants.

Some of the Bioactive Compounds and Medicinal Plants

  1. Atropine is a Bioactive compound isolated from Atropa Belladonna by HPLC technique for preparing analgesic and sleeping potions.
  2. Isolation of an extraction of Quinidine quinine, a bioactive compound from Cortex cinchona by HPLC method for treating Arthritis.
  3. Isolation and extraction of Ephedrine Norephedrine bioactive compound from Ephedra Sinica by Liquid Chromatography Technique for treating Asthma, Narcolepsy, and Obesity.
  4. Isolation and extraction of Quercetin Kaempferol bioactive compound from Ginko Biloba by Column Chromatography on Silica gel or HPLC for cancer treatment.
  5. Isolation and extraction of Rhein Emodin bioactive compound from Rheum Palmatum by HPLC analysis which is used for Inflammatory diseases and hepatitis.
  6. Chromatographic analyses of natural volatiles from Tasmanian blue gum and fennel for resolving breathing problems
  7. Through Ion exchange chromatography, Hydrophilic and Lipophilic extraction of bioactive compound from Conabiola for treating skin problems

 Isolation of secondary metabolites from plants

isolation of plant pigments by column chromatography

The metabolites which are essential for plant growth and development are considered as Primary metabolites, whereas the metabolites produced by the plant which are highly diverse in the structure are considered as Secondary Metabolites. These metabolites are bio-synthetic in origin and are classified into four different categories.

  • Alkaloids: These metabolites contain Nitrogen group and are extracted and isolated by HPLC Column Chromatography.
  • Phenylpropanoids: These metabolites contain aromatic compounds and are isolated by various Chromatographic techniques.
  • Polyketides: These metabolites are biosynthetic in origin, and their isolation and detection are carried by Liquid Chromatography or HPLC.
  • Terpenoids: These metabolites are the largest class of natural products in plants, and is extracted and isolated by Silica gel Column Chromatography.

Pigment extraction from plants

pigment extraction from plants

The pigment is a molecule which has an ability to absorb the color and reflect the color which is abundantly found in Plant Tissues. Pigments obtained from plants are of utmost importance in day to day life as they are used -for coloring foods, in medicines, in plastics, in fabrics, in cosmetics. Many types of research have also proved that consumption of diet which is rich in plant pigment slows the cellular aging.They have features such as resistant to high heat, chemical agents and different weather conditions that made them popular and lead to extensive usage. Therefore Pigment extraction from plants can be accomplished through Analytical techniques among which Chromatography is considered as the best technique and the pigments which are of interest in Pharmaceutical and Nutritional research are said to be –Anthocyanins, Beta-carotene, Curcumin, Lutein, Lycopene, Zeaxanthin.

Application of column chromatography in pharmacy

APPLICATION OF COLUMN CHROMATOGRAPHY IN PHARMACY

January 5, 2018


APPLICATION OF COLUMN CHROMATOGRAPHY IN PHARMACY

Analytical Separation and isolation of chemical compound from a mixture is called as Chromatography and purification of such compounds in larger quantities is achieved prominently using Column Chromatography. Column Chromatography is a separation Technique which is applied in various fields among which Pharmacy is the one, where preparation and dispensing of drugs is performed based on this technique, to ensure the effectiveness of the drug.In Column Chromatography, there are different types of columns such as Gravity Columns, Flash Columns, Low and Medium pressure Columns, Vacuum Columns and High-Pressure Columns. But the similar point in all these columns is that all require an Absorbent, which acts as a Stationary Phase through which sample containing different compounds flow down at differing rates. On the Other hand, the Adsorbent is applied to the Column in 2 ways namely-Slurry packing method and Dry packing Method. Slurry packing method is often used for macro scale separations and Dry packing method for microscale separations.

silica gel for column chromatography mesh size

Silica gel for Column Chromatography

Silica gel is often used for macro scale separations using Slurry pack method where the adsorbent is mixed with a small amount of Non-polar solvent until a consistent paste is formed and then poured in the column.  In dry pack method, a steady stream of the absorbent is poured through a funnel into the column by tapping the sides of the column so that packing remains even in the column. Silica gel for Column Chromatography is considered as the best absorbent in column packing for both dry pack and Slurry pack method, as it gets eventually distributed and forms a packed Stationary phase. The sample is then mixed with a polar solvent and added to the column where the component molecules which are to be separated are either adsorbed on the particle surface or adsorbed into the particle pores. Thus, the different components of a sample get split into separate bands in the column and get eluted at different rates.

Application of column chromatography in pharmacy

The elution is at different rates because of the particle size and pores of Silica gel absorbent which are available in different sizes called as Silica Gel mesh with small size and more pores which act as a Sieve. These Silica gel mesh sizes chromatography is typically used in Column as it retains the compounds for a longer time thereby enhancing the purification of the product. Silica gel for Column chromatography mesh size refers to the number of holes present in the mesh, per unit area of the absorbent to be used in the Column. Silica Gel 100, 200 mesh Merck are employed in Gravity Columns, whereas higher mesh Merck is used in Flash Columns.

Gravity Columns are mainly preferred in Gel Permeation Chromatography and are followed by Adsorption chromatography technique containing Silica gel 100-200 mesh adsorbent gives the higher efficiency of separations, improved reproducibility and require low solvent consumption. Whereas, Flash Columns containing Silica gel (250-400 mesh)  with small particle size, restricts the flow of solvent, therefore pressurized gas is applied to drive the solvent resulting in high resolution.The amount of Silica gel to be used for these Columns mainly depends on the amount of the sample. It means nearly 30 to 100 grams of Silica gel is required for easy separations but for difficult separations more than 30:1 ratio of silica gel is required. With the increase in the quantity of Silica gel, the time consumption for the separation also increases.

 Column chromatography applications

Purification of reaction mixture in chemical synthesis such as

  • Through Flash column chromatography using Silica gel 60(220-240 mesh), β-Ketoester and Desired alcohol were purified,
  • Purification of Polychlorinated biphenyls (PCB) from traces of sulphur using Silica gel 60(70-230 mesh) and Silver nitrate impregnated silica gel.
  • Silica gel (100-200 mesh) for Organ chlorine Pesticides, Phenols and Polynuclear aromatic compounds

Purification of biomolecules such as proteins for pharmaceutical research

  • Synthesis of Pramlintide which is an analogue of Amylin, a peptide hormone, for treating type 1 and type 2 Diabetics is also purified using Silica gel.
  • Purification of bioactive glycolipids, showing antiviral activity towards HSV-1(Herpes Virus)is also performed by Column chromatography using Silica gel as absorbent.
  • Purification of Nucleic acids in vivo and vitro cultures is also done using Silica gel as it absorbs the Nucleic acid.

Analysis of environmental samples

  • Purification of target chemical from co-extracted, non-chemical samples from the environment is done using Gas chromatography, in which the environmental samples are passed through an inert gas where the target chemical gets desorbed from aqueous phase to gas phase and gets separated from a stream of gas by absorption on the Silica gel mesh. On heating it the trapped chemical get released and can be analyzed
  • Extraction of pesticides from solid food samples of animal origin containing lipids, waxes and pigments as contaminants are performed using Silica gel adsorption chromatography in atmospheric condition.

 

RANGE OF SILICA GEL MESH FOR CHROMOTOGRAPHY

September 11, 2017


Discovery of Chromatography technique has made a revolution in the field of analytical laboratories for the separation of the chemicals after synthesis. Since the separation process for each compound is different based on their physical interactions with the solute and the absorbent, different methods were discovered for an efficient and reliable analysis. Since for any chromatographic technique, the absorbent remains the same i.e. Silica Gel adsorbent, because it is low acidic and available in definite particle size ranging from 60-800 mesh size. There are many Chromatography techniques theoretically, but only few can be operated practically in the laboratories. Among these techniques Column chromatographic techniques are mostly preferred as it is low in cost as well as requires minimum instrumentation. In turn based on the flow of the solvent down the column through the absorbent, Column chromatography is classified into Gravity Column Chromatography and Flash Chromatography. The flow of the solvent in the column is based on the definite pore size of the Silica Gel, which allows smaller molecules into its matrix and thereby excludes the larger molecules which flow down and get eluted faster.

 

Silica Gel Mesh for Chromatography

If the solvent flows down the column passing the Silica gel absorbent of definite particle size by gravity or fissure, it is referred to as Gravity Column Chromatography. But if the solvent flows down the column passing the Silica gel absorbent of definite particle size by positive air pressure, it is referred to as Flash Chromatography. Therefore the absorbent used in both these techniques is the same, the Silica Gel Mesh. There are many products of Silica Gel mesh available depending on the mesh size such as Silica Gel 60-120 mesh, Silica Gel 60-200 mesh, Silica Gel 70-230 mesh, Silica Gel 200-400 mesh and Silica Gel 400-800 mesh. Among this Silica Gel 70-230 mesh is used for Gravity Column Chromatography and Silica Gel 200-400 mesh is used in Flash Chromatography. These all are available in a thin white free flowing powder form with a density of 0.75 gm per ml. Best separation is achieved in Ordinary compounds by Silica Gel Absorbent. Apart from Gravity Column Chromatography and Flash Chromatography, there is another technique in Chromatography named as Reversed-phase Chromatography, which is employed for the solvent which is more polar than the Absorbent. Silica gel was the first polymer used formerly in Reversed-phase Chromatography for the purification of small organic molecules but later for the purification of synthesized Peptides. The separation is achieved by the partitioning mechanism between the Stationary phase and the mobile phase in Reversed-phase Chromatography, in which the stationary phase is Silica Gel.

 

Overall, in each and every Chromatographic techniques the main absorbent is the Silica gel and is considered as the best absorbent in the analytical field, as it is porous, insoluble and its hydrophobic interaction with the solvent makes the elution process progress at a faster rate as the polar components get eluted first. These Silica gels exist in different particle sizes, because of which they are used in many Column Chromatography Techniques and Sorbead being the best Silica gel 200-400 supplier and Silica gel 70-230 mesh supplier.

Use of Silica Gel in Flash Chromatography

Use of Silica Gel in Flash Chromatography

January 28, 2017


Flash Chromatography – An Introduction

Flash chromatography is also known as “medium pressure chromatography”. This form of chromatography works by air pressure driving the mixture, which is to be separated down the vertical glass column. This type of chromatography is ideal for separations that need to take place in a fast paced manner.

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thin layer chromatography

Silica Gel Desiccants in Thin Layer Chromatography

September 27, 2016


Understanding Thin layer Chromatography

Thin layer chromatography is a widely used form of chromatography. This type of chromatography mainly finds use in the separation process of non-volatile mixtures. Thin-layer chromatography consists of both a stationary phase as well as a mobile phase. The stationary phase is usually an adsorbent like Silica Gel, Aluminium Oxide or Cellulose. The thin-layer chromatography process works by the molecular adsorbent like Silica Gel absorbing the different components of the mixture, which has to be separated, at different levels. The adsorbents for thin-layer chromatography are always decided in such a manner that they will quicken as well as improve the separation as well as the subsequent purification process.

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Different types of Silica Gel Chromatography

May 6, 2016


Chromatography – A Brief Introduction

Chromatography is a method widely used in the separation of different compounds, solid and liquid. This method is very helpful in segregating and purifying various components of mixtures, which when separated can be analyzed individually. The chromatography process is a very simple method of separating compounds using a solid as well as liquid phase. Silica gel, alumina oxide is mainly used as the solid phase, which adsorbs all the unwanted particles as well as all kinds of impurities. When the mixture passes through the mobile phase, then only the required compounds move forward while the unwanted components of the main mixture are left behind.

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Thin Layer Chromatography

Thin Layer Chromatography and Its Many Uses

December 22, 2015


What is Thin Layer Chromatography?

 

Similar to Silica Gel Chromatography, Thin Layer Chromatography is another procedure used to separate individual components from a mixture. Thin Layer Chromatography also consists of a solid or stationary phase like a silica gel plate and a mobile phase. The Silica Gel acts like an adsorbent for thin layer chromatography. The mobile portion can be a solvent or a solvent mixture, which moves upwards using capillary action. The components that are to be separated are adsorbed at different levels and finally collected at the lower end of the Thin Layer Chromatography or TLC plate.

 
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Silica Gel Column Chromatography

The Benefits of Silica Gel Chromatography

January 27, 2015


Silica gel adsorbents are widely accepted as one of the best adsorbents that are used in column chromatography. One of the main benefits is that it has a huge affinity for absorption; besides, it is very easily available commercially in many different forms and sizes and there is a lot of research and information that is provided by the manufacturers on its numerous uses, especially in chromatography.

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Why Flash Chromatography method is inexpensive and quicker?

August 11, 2014


 

After the process of chemical synthesis, the compounds of interest need to be purified in a proper manner. Some of the popular purification techniques are Crystallization, Separation, Filtration, Liquid-Liquid clean up and Distillation. All these techniques are vastly used in the laboratories. But there is another purification method that is used for separating two almost same nature substances. This process known as Flash Chromatography.

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