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Normal Phase HPLC Column

Normal Phase HPLC Column Supplier

uHPLCs, we specialize in providing top-tier normal phase HPLC columns designed to meet the diverse needs of your laboratory. Our columns are crafted with precision and high-quality materials, ensuring reliable and reproducible results for your chromatographic applications.

Why Choosing uHPLCs Normal Phase HPLC Columns

  • High Efficiency and Resolution: Our columns are engineered to deliver sharp, well-resolved peaks, enhancing the accuracy and reliability of your analyses.
  • Superior Reproducibility: Each column is manufactured to stringent quality standards, guaranteeing consistent performance across different batches.
  • Wide Range of Selectivities: We offer a variety of stationary phases, allowing you to choose the most suitable column for your specific separation needs.
  • Enhanced Durability: Constructed with robust materials, our columns withstand high pressures and extended use, providing excellent longevity and value for your investment.
  • Optimized Particle Size: Our columns feature optimized particle sizes that improve separation efficiency and reduce analysis time.
  • Excellent Chemical Compatibility: Compatible with a broad range of solvents and samples, our columns are versatile for various applications.
  • Custom Solutions Available: We provide tailored solutions to meet unique separation requirements, ensuring you achieve the best possible results.

Choose uHPLCs for your normal phase HPLC columns and experience superior performance, reliability, and expert support. Contact us today to learn more about our products and how we can assist with your chromatographic needs.

Normal Phase HPLC Column

3.0 Empty HPLC Column Hardware

USHD Sio2
Normal Phase HPLC Column

2.1mm Empty HPLC Columns Normal Pressure for Lab Equipment

USHD NH2
Normal Phase HPLC Column

2.1mm High Pressure Empty UHPLC Columns

USHD CN
Normal Phase HPLC Column

Normal Phase HPLC Column Specification

Item USP Pore Size Option Surface area (m²/g) Carbon Load (%) PH Tolerance Range Column Specifications Features and Application
USHD Sio2
L3
5μm, 120A
200
#
2-8
4.6X250mm; 4.6X150mm; 3.0X100mm;
1.) Ultra-pure spherical silica gel (metal impurities <10ppm), with high column efficiency and peak shape Good features.
2.) It has special selectivity for structurally isomeric compounds and is especially suitable for easy separation. Tailing highly polar compounds, basic organic compounds and fat-soluble compounds, Isomers, such as vitamins, steroids, and many other drug molecules.
USHD NH2
L8
5μm, 120A
320
4
2-8
4.6X250mm; 4.6X150mm; 3.0X100mm;
1.) Ultra-pure silica matrix, bonded aminopropyl stationary phase.
2.) Analyze polar compounds under normal phase elution conditions, which are soluble in weak compounds such as alkanes and alkenes. Polar compounds.
3.) Separate sugar substances such as fructose, glucose, and sucrose under reversed-phase conditions.
4.) In low pH buffer, the amino phase changes into weak anion exchange, which can separate Some negatively charged molecules.
USHD CN
L10
5μm, 120A
320
7
2-8
4.6X250mm; 4.6X150mm; 3.0X100mm;
1.) Ultra-pure silica gel matrix, bonded fluoropropyl stationary phase.
2.) The gas base of medium polarity allows it to be used in both forward and reverse phases.
3.) In reversed phase chromatography mode, hydrophobic compounds elute faster and due to The gas group with π-electron interaction shows different selectivity from C18. strong base The peak shape of the separation of chemical compounds is better, and it is also more compatible with working conditions with a high proportion of water.
4.) The most polar reversed-phase column, use standard C18 for very hydrophobic compounds. When reversed-phase columns such as C8 and C8 cannot elute, gas-based columns can be used.
HPLC Colun in the HPLC System Connect Diagram by uhplcs

In addition to custom columns, uHPLCs also offers a wide range of standard columns for various applications such as reversed-phase, normal-phase, ion exchange, size exclusion, and HILIC. We also offers prepacked columns and accessories, such as frits, end-fittings, and Guards Columns.

uHPLCs uses state-of-the-art manufacturing techniques to produce high-quality columns that are consistent, reliable, and provide excellent performance. we have a strict quality control program to ensure that all columns meet the highest CE , SGS and UL standards for performance and reproducibility.

Contact Us For Excellent HPLC Columns

Experience the Precision and Reliability of Our High-Quality C18 HPLC Columns Today

WHY uHPLCs Normal Phase HPLC Column ?

You might be interested in uHPLCs for normal phase HPLC columns for a couple of reasons:

  • Focus on normal phase: Unlike some other companies that offer a wider variety of HPLC columns, uHPLCs specialize in normal phase columns, This means you will have a wider selection and more expertise in this particular type of column.
  • Quality and reputation: uHPLCs emphasizes strict quality control and uses state-of-the-art manufacturing for their columns , We are also a supplier to major pharmaceutical companies, which suggests good reputation within the industry.
 

However, it’s important to note that normal phase UHPLC columns aren’t as common as reversed-phase ones.

The lower viscosity of normal phase mobile phases means the pressure limitations of standard HPLC systems are usually sufficient .

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Normal Phase HPLC Column FAQ

Main Features and Advantange of Normal Phase?

Normal phase HPLC columns offer a distinct separation approach compared to their widely used counterparts, reversed-phase columns. Here’s a breakdown of their key features and advantages:

Main Features:

  • Stationary phase: The heart of a normal phase column is a polar stationary phase. Most commonly, unmodified silica gel (SiO2) is used, with silanol (Si-OH) groups on its surface. However, bonded phases with functional groups like amine (NH2) or cyano (CN) can be employed for specific interactions.
  • Mobile phase: The mobile phase in normal phase chromatography is nonpolar. Common solvents include hexane, heptane, chloroform, and dichloromethane. Sometimes, small amounts of polar modifiers like methanol or ethanol are added for fine-tuning separations.
  • Separation principle: Separation relies on the relative polarities of the analytes and the stationary phase. Analytes with a greater degree of polarity interact more strongly with the polar stationary phase, leading to a longer retention time as they elute (flow through) the column. Conversely, less polar analytes interact minimally with the stationary phase and elute faster.
 

Advantages:

  • Effective for polar compounds: Normal phase chromatography excels at separating polar compounds that pose challenges in reversed-phase mode. This includes small organic molecules (alcohols, phenols), carbohydrates (sugars), and various natural products (lipids, pigments).
  • Isomer separation: Normal phase can be particularly useful for distinguishing positional isomers that might coelute (elute together) in reversed-phase chromatography. This can be crucial for applications where differentiating these specific structures is important.
  • Simpler mobile phases: Nonpolar mobile phases used in normal phase are often readily available and less expensive compared to the water-organic solvent mixtures required in reversed-phase chromatography.
 

Additional points to consider:

  • Challenges: Normal phase chromatography has its limitations. Unmodified silica can be deactivated by trace amounts of water in the mobile phase, requiring careful mobile phase preparation and storage. Additionally, silica gel dissolves at high and low pH extremes, limiting the usable pH range of the mobile phase. Some analytes might also be sensitive to the nonpolar solvents used.
  • Choosing between Normal and Reverse Phase: The choice depends on your analytes. If you’re dealing with polar compounds, normal phase is a good starting point. However, for nonpolar and moderately polar analytes, reversed phase might be more suitable.

Some Applications of Normal Phase

Normal phase HPLC columns, despite their limitations, find application in various analytical chemistry tasks due to their unique separation abilities. Here are some prominent examples:

1. Separation of Polar Natural Products:

  • Lipids: Normal phase is a preferred method for separating complex mixtures of lipids, including triglycerides, fatty acids, and phospholipids. The varying polarities of these molecules allow for their effective differentiation.
  • Carbohydrates: Monosaccharides, disaccharides, and polysaccharides (sugars and starches) can be separated based on their size and polarity using normal phase chromatography. This is crucial for analyzing food products, studying plant extracts, and characterizing carbohydrate-based drugs.
  • Pigments: Natural pigments like chlorophylls and carotenoids, found in plants and algae, can be separated and identified using normal phase HPLC. The technique helps analyze food colorants and study natural pigments in biological systems.
 

2. Analysis of Small Organic Molecules:

  • Alcohols and Phenols: Normal phase is well-suited for separating various alcohols and phenols, which are often polar and struggle with reversed-phase chromatography. This can be valuable for analyzing alcoholic beverages, studying plant metabolites, and identifying pharmaceutical impurities.
  • Organic Acids and Esters: Organic acids and esters with moderate polarity can be effectively separated using normal phase columns. Applications include analyzing organic acids in food and beverages, characterizing fermentation products, and identifying organic contaminants in environmental samples.
 

3. Separation of Functionalized Molecules:

  • Amino Acids and Peptides: While reversed-phase is dominant for protein analysis, normal phase can be used for separating smaller peptides and amino acids based on their polarity and functional groups (e.g., amine and carboxylic acid). This can be useful for studying peptide synthesis products or analyzing amino acid composition in biological samples.
  • Nucleotides and Nucleosides: The separation of nucleotides (building blocks of RNA and DNA) and nucleosides (nucleotides without phosphate groups) can be achieved using normal phase chromatography. This is valuable for analyzing nucleic acid samples, studying RNA modifications, and investigating nucleoside-based drugs.
 

4. Specialized Applications:

  • Chiral Separations: Normal phase chromatography, with the help of chiral stationary phases, can be used to separate enantiomers (mirror-image molecules) of certain polar compounds. This is crucial in the pharmaceutical industry for ensuring the purity and efficacy of chiral drugs.
  • Explosives Analysis: Normal phase columns can be employed for the separation and identification of explosives and their degradation products. This application is relevant in forensic science and environmental analysis.
 

It’s important to remember that normal phase chromatography isn’t a one-size-fits-all solution. While it excels at separating polar compounds, reversed-phase remains the dominant choice for many analytes. However, for specific applications where normal phase offers a distinct advantage, it remains a valuable tool in the analytical chemist’s arsenal.

Frequently Asked Questions

Normal phase HPLC columns excel at separating polar compounds. These include:

  • Small organic molecules: Alcohols, phenols, organic acids, and esters.
  • Carbohydrates: Monosaccharides, disaccharides, and polysaccharides.
  • Natural products: Lipids, alkaloids, and pigments.

They are particularly useful when reversed-phase HPLC columns struggle. This can happen when analytes are:

  • Too strongly retained on reversed-phase columns, leading to peak tailing and slow elution.
  • Insoluble in typical aqueous mobile phases used in reversed-phase chromatography.

 

The separation principle in normal phase HPLC relies on the polarity of the stationary phase and the mobile phase. Here’s a breakdown:

  • Stationary phase: In normal-phase chromatography, the stationary phase is typically polar. The most common material is unmodified silica gel, which has silanol (Si-OH) groups on its surface. Additionally, bonded phases with functional groups like amino (NH2) or cyano (CN) can be used for alternative selectivities.
  • Mobile phase: The mobile phase is nonpolar. Common solvents include hexane, heptane, chloroform, and dichloromethane. Sometimes, small amounts of polar modifiers like methanol or ethanol are added to fine-tune the separation.

Separation occurs based on the relative polarities of the analytes and the stationary phase. More polar analytes interact more strongly with the polar stationary phase, leading to a longer retention time. Conversely, less polar analytes interact less and elute faster.

Advantages:

  • Excellent for polar compounds: Normal phase is the go-to method for separating many polar analytes that are difficult to resolve using reversed-phase chromatography.
  • Separation of isomers: Normal phase can effectively separate positional isomers that might coelute in reversed-phase mode.
  • Simple mobile phases: Nonpolar mobile phases are often readily available and inexpensive.

Disadvantages:

  • Silica deactivation: Unmodified silica can be deactivated by trace amounts of water in the mobile phase, affecting column performance. Careful mobile phase preparation and storage are crucial.
  • Limited pH stability: Silica gel dissolves at high and low pH extremes. This restricts the usable pH range of the mobile phase.
  • Compatibility issues: Some analytes might be sensitive to nonpolar solvents used in the mobile phase.

Several factors influence the choice of a normal phase HPLC column:

  • The polarity of your analytes: Match the column’s polarity to the analytes. Unmodified silica is suitable for moderately polar analytes, while amino or cyano phases can be used for more polar analytes.
  • The desired selectivity: Different stationary phases offer different selectivities. Consider if you need to separate based on polarity, size, or other factors.
  • Sample solubility: Ensure your analytes are soluble in the chosen mobile phase.
  • Resolution requirements: Consider the column particle size and length for optimal peak resolution. Smaller particles and longer columns generally provide better resolution but require higher pressures.

Here are some tips for maximizing the performance of your normal phase HPLC column:

  • Mobile phase preparation: Use high-purity solvents and freshly prepared mobile phases to minimize silica deactivation. Consider using mobile phase filtering systems.
  • pH control: Maintain the mobile phase pH within the recommended range for your specific column.
  • Sample preparation: Ensure your samples are free of contaminants that could deactivate the column.
  • Column storage: Store your column in a cool, dry place with the endcaps on to prevent moisture ingress.
  • Regular maintenance: Follow the manufacturer’s recommendations for column cleaning and regeneration to maintain optimal performance.

By following these tips, you can get the most out of your normal phase HPLC column and achieve successful separations of your polar analytes.

Here’s the key difference between normal phase and reverse phase chromatography:

  • Normal phase: The stationary phase is polar (e.g., silica gel), and the mobile phase is nonpolar (e.g., hexane).
  • Reverse phase: The stationary phase is nonpolar (e.g., C18 chains bonded to silica), and the mobile phase is polar (e.g., water-acetonitrile mixture).

In a C18 column, the long hydrocarbon chains (C18 refers to 18 carbons) create a nonpolar environment on the silica particle surface. This attracts analytes based on their hydrophobic interactions, making it a reversed-phase mode.

Normal-phase and reversed-phase are two main approaches to column chromatography, a technique for separating mixtures based on the interaction between the components and the column. The key difference lies in the polarity of the stationary phase (the packing material in the column) and the mobile phase (the solvent that flows through the column).

Normal Phase Chromatography

  • Stationary phase: Polar. Most commonly, unmodified silica gel (SiO2) with silanol (Si-OH) groups on its surface. Other options include bonded phases with functional groups like amine (NH2) or cyano (CN) for specific interactions.
  • Mobile phase: Nonpolar. Common solvents include hexane, heptane, chloroform, and dichloromethane. Sometimes, small amounts of polar modifiers like methanol or ethanol are added for fine-tuning.

Separation principle: Analytes with greater polarity tend to interact more with the polar stationary phase, causing them to elute (flow through the column) slower. Conversely, less polar analytes elute faster with minimal interaction with the stationary phase.

Suitable for: Separating polar compounds such as:

  • Small organic molecules (alcohols, phenols, organic acids, esters)
  • Carbohydrates (monosaccharides, disaccharides, polysaccharides)
  • Natural products (lipids, alkaloids, pigments)

Advantages:

  • Effective for polar compounds: Ideal for separating analytes that are difficult to resolve in reversed-phase mode.
  • Isomer separation: Can effectively distinguish positional isomers that might coelute (elute together) in reversed-phase.
  • Simpler mobile phases: Nonpolar mobile phases are often readily available and inexpensive.

Disadvantages:

  • Silica deactivation: Unmodified silica can be deactivated by trace amounts of water in the mobile phase, affecting column performance. Careful mobile phase preparation is crucial.
  • Limited pH stability: Silica gel dissolves at high and low pH extremes, restricting the usable pH range of the mobile phase.
  • Compatibility issues: Some analytes might be sensitive to nonpolar solvents used in the mobile phase.

Reverse Phase Chromatography

  • Stationary phase: Nonpolar. Typically, silica particles chemically modified with long hydrocarbon chains (e.g., C18 reversed phase columns). This creates a hydrophobic environment.
  • Mobile phase: Polar. A mixture of water and organic solvents (e.g., acetonitrile, methanol) is commonly used. The ratio of organic solvent to water can be adjusted to influence separation.

Separation principle: Analytes with more hydrophobic character (greater affinity for nonpolar environments) interact more strongly with the nonpolar stationary phase, leading to a longer retention time. Conversely, more polar analytes elute faster with weaker interactions.

Suitable for: Separating nonpolar and moderately polar compounds, including:

  • Pharmaceuticals
  • Proteins and peptides
  • Environmental contaminants
  • Organic extracts

Advantages:

  • Wider applicability: Can handle a broader range of analytes compared to normal phase.
  • More flexible mobile phases: The ability to adjust the polarity of the mobile phase with water/organic solvent mixtures offers greater control over separations.
  • Generally more stable: Reversed-phase columns are less susceptible to deactivation by water compared to normal phase silica.

Disadvantages:

  • Potentially less effective for highly polar compounds: Highly polar analytes might interact too strongly with the mobile phase, leading to poor retention and peak broadening.
  • More complex mobile phases: Mixing water and organic solvents requires more considerations for proper preparation and disposal.

Choosing Between Normal and Reverse Phase

The choice between normal and reversed-phase chromatography depends on the properties of your analytes. Here’s a general guideline:

  • For polar analytes: Start with normal phase chromatography.
  • For nonpolar and moderately polar analytes: Consider reversed phase chromatography.

Additionally, factors like analyte solubility in the mobile phase and desired selectivity (separation based on specific properties) also influence the decision.

Normal Phase vs. Reverse Phase Chromatography

FeatureNormal PhaseReverse Phase
Stationary Phase PolarityPolar (e.g., silica gel)Nonpolar (e.g., C18 chains)
Mobile Phase PolarityNonpolar (e.g., hexane)Polar (e.g., water-acetonitrile mix)
Separation PrincipleMore polar analytes interact more with the stationary phase, eluting slower.More hydrophobic analytes interact more with the stationary phase, eluting slower.
Suitable forPolar compounds (alcohols, carbohydrates, natural products)Nonpolar & moderately polar compounds (pharmaceuticals, proteins, environmental contaminants)
Advantages* Effective for polar compounds * Separates some isomers * Simpler mobile phases* Wider applicability * More flexible mobile phases * Generally more stable
Disadvantages* Silica deactivation by water * Limited pH stability * Compatibility issues with some solvents* Less effective for highly polar compounds * More complex mobile phases

Need Help ?

Contact uHPLCs Today for Any Questions for HPLC / UHPLC 

+(86) 0755-28502380

sales@uhplcs.com

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