What Are the Most Common Applications of HPLC?
Introduction Ever wondered how scientists ensure your medicine is pure, or your drinking water is
C18 HPLC Column
C18 Column Professional OEM Manufacturer
uHPLCs focus on engineering C18 columns that are not just effective but exceptional. C18 Column Series Designed to meet a diverse array of analytical demands, our C18 columns are celebrated for their unparalleled retentivity among alkyl-bonded phases. Ideal for a multitude of applications, these columns are extensively employed in the analysis of pharmaceuticals, steroids, fatty acids, phthalates, environmental samples, and much more.
Each column is meticulously crafted to deliver outstanding performance, ensuring that no matter your specific needs—whether in research, quality control, or production environments—our C18 columns stand ready to exceed expectations. As a professional OEM manufacturer, we offer a variety of column characteristics tailored to optimize your applications, embodying our commitment to quality, consistency, and scientific excellence.
Choose uHPLCs for a C18 column that truly fits your application and elevates your analytical capabilities to new heights.
🔬 Key Features & Technology
- Unique Bonding Process: Delivers excellent chemical stability and high loading capacity.
- Double End-Capping Technique: Improves peak shape, minimizes tailing, and ensures consistent performance across different batches.
- Superior Batch-to-Batch Reproducibility: Designed for laboratories that demand accuracy and repeatability.
💡 Product Line Overview
- The uHPLCs C18 Series includes four major column types, each optimized for specific applications:
- 1. C18-Pro – The Versatile Performer
- High carbon content for strong retention and high selectivity
- Ideal for pharmaceuticals, traditional Chinese medicine, environmental samples, and food testing
- Recommended for general-purpose reversed-phase applications
- 2. C18-AQ – The Polar Compound Specialist
- Specially designed for polar and hydrophilic compounds
- Fully compatible with 100% aqueous mobile phases
- Ideal for the analysis of organic acids, metabolites, and water-soluble compounds
- 3. C18-AR – The Low pH Expert
- Uncapped with side chain steric protection, ensuring excellent selectivity
- Stable at pH as low as 0.5, making it ideal for acidic environments
- Best for isomer separation, peptides, and polar/non-polar mixed systems
- 4. C18-T – The Extreme Conditions Champion
- Based on ultra-pure organic-inorganic hybrid silica
- Stable from pH 1 to 11, with excellent resistance to strong acids, bases, high salt concentrations, and high temperatures
- Extended column life and robustness in harsh applications
- Ideal for alkaline substances, biological compounds, and samples with complex matrices
🎯 Applications Across Industries
- Pharmaceutical development and QC
- TCM (Traditional Chinese Medicine) and herbal product testing
- Food safety and additives detection
- Environmental pollutant and water quality analysis
- Metabolite profiling and bioanalytical research
✅ Why Choose uHPLCs C18 Columns?
- Wide range of specifications and dimensions to meet various flow rates and detection needs
- Excellent mechanical strength and packing uniformity
- Competitive pricing and OEM customization options available
- Trusted by research labs, academic institutions, and commercial testing centers worldwide
Reverse Phase C18 HPLC Column
Related HPLC Columns
C18 Column Specification & Option
| Item | USP | Pore Size Option | Surface area (m²/g) | Carbon Load (%) | PH Tolerance Range | Column Specifications |
|---|---|---|---|---|---|---|
| USHD C18-Pro | L1 | 5μm, 150A 3.5μm, 150A | 200 | 10 | 2-8 | 4.6X250mm 4.6X150mm 4.6X100mm 4.6X50mm 3.0X100mm 3.0X50mm 2.1X100mm 2.1X50mm |
| USHD C18 | L1 | 5μm, 120A 3μm, 120A 1.8μm, 120A | 320 | 17 | 2-8 | 4.6X250mm 4.6X150mm 4.6X100mm 4.6X50mm 3.0X100mm 3.0X50mm 2.1X100mm 2.1X50mm |
| USHD C18-AQ | L1 | 5μm, 120A 3μm, 120A 1.8μm, 120A | 320 | 12 | 2-8 | 4.6X250mm 4.6X150mm 4.6X100mm 4.6X50mm 3.0X100mm 3.0X50mm 2.1X100mm 2.1X50mm |
| USHD C18-AR | L1 | 5μm, 120A 3μm, 120A 1.8μm, 120A | 320 | 10 | 0.5-8 | 4.6X250mm 4.6X150mm 4.6X100mm 4.6X50mm 3.0X100mm 3.0X50mm 2.1X100mm 2.1X50mm |
| USHD C18-T | L1 | 5μm, 120A 3μm, 120A 1.8μm, 120A | 320 | 14 | 1-11 | 4.6X250mm 4.6X150mm 4.6X100mm 4.6X50mm 3.0X100mm 3.0X50mm 2.1X100mm 2.1X50mm |
| USHD C8-PRO | L7 | 5μm, 150A 3.5μm, 150A | 200 | 7 | 2-8 | 4.6X250mm 4.6X150mm 4.6X100mm 4.6X50mm 3.0X100mm 3.0X50mm 2.1X100mm 2.1X50mm |
| USHD C8 | L7 | 5μm, 150A 3.5μm, 120A 1.8μm, 120A | 320 | 12 | 2-8 | 4.6X250mm 4.6X150mm 4.6X100mm 4.6X50mm 3.0X100mm 3.0X50mm 2.1X100mm 2.1X50mm |
HPLC Column Product Features
1.Superior Reproducibility:
Advanced bonding techniques ensure consistent performance across different batches.
2.High Separation Efficiency:
Lower carbon content allows for more efficient separation, improving analysis speed and accuracy.
3.Enhanced Stability:
The double end-capping process provides robust stability, delivering reliable results even for challenging acidic and basic compounds.
4.Improved Anti-Contamination:
With a lower specific surface area and slightly larger pore size, these columns resist contamination better, extending column life significantly.
5.Longer Column Life:
Especially effective in traditional medicine analysis, offering a longer lifespan compared to standard HPLC C18 Columns.
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 C18 Column
uHPLCs understand that choosing the right HPLC column is crucial for achieving optimal results in your lab. That’s why we offer not just high-quality products, but also exceptional customer service.
Expert Guidance Every Step of the Way:
Our knowledgeable and experienced technical support team is always available to answer your questions and provide expert advice. They’ll guide you through the selection process, ensuring you find the perfect column for your specific application.
More Than Just Products:
uHPLCs is a professional and reliable HPLC column OEM factory located in China. We take pride in our commitment to quality, offering a wide range of standard products alongside the ability to customize columns to your exact needs. This flexibility allows us to cater to a diverse clientele within the HPLC field.
In short, choosing uHPLCs means:
- Expert technical support: Get the guidance you need to select the ideal column for your application.
- High-quality products: Trust in the reliability and performance of our HPLC columns.
- Customization options: Tailor columns to your specific requirements for a perfect fit.
- Extensive product range: Find the standard column you need, or work with us to create a custom solution.
Let uHPLCs be your partner in achieving superior HPLC results. Contact us today!
1. Hassle-Free
Before & After Servic
uHPLCs prides itself on providing excellent customer service, both before and after the sale.
2. Quality Management Test Before Mass Manufacture
Quality is a top priority. We understand that the products we manufacture will be used in critical applications, which is why we take great care to ensure that they are of the highest quality.
3. Rigorous Production Process
uHPLCs specializes in the production of HPLC columns, and we take great care to ensure that the columns we produce are of the highest quality.
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Customized C18 HPLC Columns
to Power Your Lab Success
Looking for a C18 HPLC column that perfectly fits your needs?
Our expert team offers a comprehensive OEM service for C18 HPLC columns, designed to meet your lab’s specific requirements.
Here’s what sets us apart:
- Tailored Solutions: We don’t offer a one-size-fits-all approach. We work closely with you to understand your unique needs and develop a custom C18 HPLC column that delivers optimal performance.
- Deep Expertise: Our team possesses extensive knowledge in HPLC column production and application. This ensures we create a column that aligns with your specific analytical goals.
- Seamless Process: We collaborate with you every step of the way, from initial concept to final delivery.
- Uncompromising Quality: We utilize strict quality control procedures and cutting-edge technology to guarantee consistent, reliable results.
- Flexibility: Our adaptable OEM service allows us to craft C18 HPLC columns that meet your exact specifications.
- Enhanced Analysis: Experience superior separation and sharper peaks for more accurate and efficient HPLC analysis.
Ready to elevate your lab’s capabilities?
Contact us today to discuss your C18 HPLC column requirements.
High Quality HPLC Column 316L, 316 Stainless Steel Tube Materials
Design New HPLC Columns 100% As Clients Request
Make HPLC Column Samples And Test Before Mass Products.
What
Clients Say
I am extremely impressed with the quality and consistency of uHPLCs' HPLC columns. They have greatly improved the efficiency and accuracy of our analysis, and their OEM service has allowed us to develop columns that perfectly meet our lab's specific needs.
Amanda LeeCEO 
I've been using the HPLC columns from uHPLCs for over a year, and I can honestly say they are the best I've ever used. The performance is consistently excellent, and I've never had any issues with them. I highly recommend them to anyone looking for high-quality HPLC columns.
Adam CheiseDesigner 
We have been using uHPLCs' HPLC columns for quality control of our pharmaceutical products and have found them reliable and efficient. The column lifetime is also longer than any other column that we have used before.
Floyd StallerCo-Founder 
I recently switched to uHPLCs' HPLC columns and have been extremely impressed with the results. The improved resolution and peak symmetry have been game changers for my research, and the customer service is top-notch.
Floyd StallerCo-Founder 
I've tried columns from several other companies, but uHPLCs' HPLC columns are by far the best in terms of quality and value. They have always delivered consistent, reproducible results, and the company's technical support has been invaluable.
Floyd StallerCo-Founder 
C18 HPLC Column FAQ
1.) Types of C18 HPLC Column
High-performance liquid chromatography (HPLC) is a technique used to separate components of a mixture based on their interaction with the stationary and mobile phases.
The stationary phase is packed inside a column, and the mobile phase is a solvent that flows through the column. C18 columns are one of the most common types of HPLC columns used for separating non-polar and moderately polar compounds. They are packed with octadecylsilane (ODS), a bonded stationary phase with 18 carbon atoms.
There are several types of C18 HPLC columns available, each with its own unique properties:
1. Ultra-high purity (UHPLC) C18 columns:
These columns are designed for use with UHPLC instruments, which can operate at higher pressures than traditional HPLC instruments. UHPLC C18 columns typically have smaller particle sizes (sub-2 µm) than traditional HPLC columns (3-5 µm), which results in higher resolution separations.
2.Wide-pore C18 columns:
- These columns have larger pores than traditional C18 columns. This allows them to be used for separating larger molecules that may not be able to fit into the pores of a traditional C18 column.
3.Endcapped C18 columns:
- These columns have a small endcapping molecule bonded to the silica surface.
- This endcapping reduces the activity of the silica surface, which can improve peak shape for some analytes.
4. Specialty C18 columns:
- There are a number of other specialty C18 columns available, such as superficially porous C18 columns and core-shell C18 columns. These columns offer unique advantages for specific applications.
The choice of C18 column will depend on the specific needs of the separation. Factors to consider include the size of the molecules to be separated, the polarity of the analytes, and the desired resolution.
C18 Column Dimensions
C18 columns for HPLC come in various standard dimensions, with the most common being:
Column length (L):
This ranges from 50 to 250 mm. Longer columns provide higher resolution but take more time to run the analysis.
Column inner diameter (ID):
This typically ranges from 2.1 to 4.6 mm.
Smaller diameter columns use less solvent but may provide lower sensitivity.
Here’s a table summarizing the common dimension ranges for C18 HPLC columns:
| Dimension | Typical Range |
|---|---|
| Length (L) | 50 mm – 250 mm |
| Inner Diameter (ID) | 2.1 mm – 4.6 mm |
It’s important to choose the appropriate column dimensions based on your specific needs. Here are some factors to consider:
* Resolution requirements: If you need to separate very similar compounds, a longer column may be necessary.
* Sample amount: If you have a limited amount of sample, you may want to use a smaller diameter column.
* Analysis time: Longer columns take more time to run the analysis.
* Instrument compatibility: Make sure the column dimensions are compatible with your HPLC instrument.
There are also preparative HPLC columns available that have larger diameters (e.g., 10 mm or more) for purifying larger quantities of material.
These columns are not typically used for analytical separations.
C18 Column Vs C8 HPLC Column
C18 and C8 columns are both widely used stationary phases in HPLC for separating non-polar and moderately polar compounds. However, they differ in terms of their bonded alkyl chain length, which significantly impacts their polarity and analyte interactions. Here’s a breakdown of their key differences:
Chain Length:
- C18: As the name suggests, C18 columns have a stationary phase bonded with octadecylsilane (ODS) molecules, containing 18 carbon atoms in the alkyl chain.
- C8: C8 columns, on the other hand, utilize octylsilane (ODS) molecules with a shorter alkyl chain of 8 carbon atoms.
Polarity:
- C18: The longer carbon chain in C18 columns results in a more hydrophobic (water-repelling) stationary phase. This translates to weaker interactions with polar analytes in the mobile phase.
- C8: Due to the shorter chain, C8 columns exhibit a slightly less hydrophobic character compared to C18. This allows for somewhat stronger interactions with polar analytes.
Analyte Retention:
- C18: The increased hydrophobicity of C18 leads to stronger retention of non-polar analytes. They tend to spend more time interacting with the stationary phase, resulting in longer elution times.
- C8: With a less hydrophobic surface, C8 columns exhibit weaker retention of non-polar analytes. This leads to faster elution times compared to C18.
Selectivity:
- C18: C18 columns are generally considered the more versatile option for separating a broad range of non-polar and moderately polar compounds due to their strong hydrophobic interactions.
- C8: C8 columns offer an alternative selectivity for compounds with a wider range of polarities. They can be particularly useful for separating analytes that might be too strongly retained on C18, leading to peak broadening or overlapping.
Choosing Between C18 and C8:
The selection between C18 and C8 columns depends on the specific characteristics of your analytes and desired separation:
- For non-polar analytes: C18 is a good starting point due to its strong retention and versatility.
- For moderately polar analytes: C8 can be a better choice if you encounter issues with excessive retention or peak broadening on C18.
- For analytes with a wider range of polarities: A gradient elution method with either C18 or C8 can be used, where the mobile phase composition is gradually changed to influence analyte elution.
Additional factors to consider:
- Starting with C18: Due to its broader applicability, C18 is often the preferred starting point for method development.
- Compatibility with mobile phases: Both C18 and C8 columns are compatible with a wide range of commonly used HPLC mobile phases.
Ultimately, the best way to determine the optimal column type is through experimentation. Running test separations with both C18 and C8 columns can help you achieve the desired resolution and separation efficiency for your specific analytes.
How HPLC Column Volume Calculator ?
Do You Know How to Calculator HPLC Column Volume ? Here We Give Some Idea, Hope It will be Helpful for You to Know Details.
Calculating the HPLC column volume is essential for determining the amount of solvent required for column equilibration and for estimating the elution times of compounds during chromatographic runs. The HPLC column volume can be calculated using the following formula:
Column Volume (V) = π * (Column Internal Diameter/2)^2 * Column Length
Here’s how to use the formula:
Gather Information:
- You will need to know the internal diameter (ID) of the HPLC column and its length. Both are typically provided by the column manufacturer or can be measured directly.
Convert Units (if necessary):
- Ensure that the units for both the internal diameter and length are consistent (e.g., millimeters or centimeters).
- If the dimensions are given in different units (e.g., inches or centimeters), convert them to the same unit before proceeding with the calculation.
Apply the Formula:
- Divide the internal diameter by 2 to get the radius (r).
- Square the radius (r^2).
- Multiply the squared radius by π (pi, approximately 3.14159).
- Multiply the result by the column length to obtain the column volume.
Here’s a step-by-step example using the formula:
Suppose we have an HPLC column with an internal diameter of 4.6 mm and a length of 150 mm.
Step 1: Calculate the radius (r):
- r = Internal Diameter / 2 = 4.6 mm / 2 = 2.3 mm
Step 2: Calculate the squared radius (r^2):
- r^2 = (2.3 mm)^2 ≈ 5.29 mm^2
Step 3: Calculate the column volume (V):
- V = π * r^2 * Column Length = 3.14159 * 5.29 mm^2 * 150 mm ≈ 1183.13 mm^3
Step 4: Convert the column volume to a more convenient unit (optional):
- In this example, 1 cm^3 (cubic centimeter) is equivalent to 1000 mm^3, so the column volume is approximately 1.18 cm^3.
Now you have calculated the HPLC column volume, which is useful for various aspects of HPLC method development and optimization.
How to Choose Right C18 HPLC Column ?
In Order to Choose Right HPLC Column for Your HPLC System or Project, You Should Care Some Factors As Following:
Choosing the right HPLC column is crucial for obtaining accurate and reliable chromatographic results. The selection process involves considering various factors based on the specific analytical requirements. Here are some essential steps to guide you in choosing the right HPLC column:
Analyte Characteristics:
- Consider the nature of the compounds you want to analyze. Are they polar or nonpolar? Ionic or neutral? Acidic or basic?
- For nonpolar compounds, a reverse-phase column like C18 may be suitable, while polar compounds may require a normal phase or HILIC column.
- For charged analytes, an ion-exchange column might be appropriate.
Sample Matrix:
- Evaluate the sample matrix, including potential contaminants or interfering compounds.
- Ensure the column can handle the sample matrix without clogging or irreversible adsorption.
Separation Requirements:
- Determine the resolution and separation efficiency needed for your analysis.
- Smaller particle sizes (sub-2 μm) or specialized columns may be necessary for high-resolution applications.
Column Length and Diameter:
- The column length affects the separation time, while the internal diameter (ID) influences sensitivity and flow rate.
- Longer columns offer higher resolution but might require longer analysis times.
- Smaller ID columns provide higher sensitivity but may generate higher backpressure.
Particle Size:
- Consider the particle size of the stationary phase.
- Smaller particle sizes offer higher efficiency and resolution but might require specialized equipment to handle the increased backpressure.
Column Chemistry and Stationary Phase:
- Choose the appropriate stationary phase chemistry based on analyte interactions.
- C18 columns are widely used for nonpolar compounds, while alternative phases cater to specific applications.
Column Compatibility:
- Ensure the chosen column is compatible with your HPLC system in terms of pressure and hardware requirements.
Sample Load and Throughput:
- If you need to analyze a large number of samples, consider a column with high throughput capabilities.
Budget Considerations:
- Take into account the cost of the column and its longevity.
- Balancing performance with cost is crucial for routine analyses.
Column Reputation and Support:
- Choose columns from reputable manufacturers with good technical support and resources.
Application Notes and Recommendations:
- Check if the column manufacturer provides application notes or recommendations for specific types of analyses.
Remember that column selection may involve some trial and error, particularly for complex separations. It is often beneficial to consult with experienced chromatographers, seek advice from column manufacturers, and conduct small-scale method development experiments to ensure optimal column selection for your specific HPLC analysis.
Analytical C18 Column or Preparative C18 Column, How to Choose ?
Think Some Newbie Don’t Know How to distinguish Between Analytical and Preparative Columns, Here we List some difference Between Analytical and Preparative Columns, Hope it will be helpful.
- Objective: Determine the main goal of your chromatographic analysis. If you need to separate, identify, and quantify compounds in a sample, an analytical column is appropriate. If your goal is to purify and collect larger amounts of specific compounds, a preparative column is the right choice.
- Sample Size: Consider the available sample size for analysis. Analytical columns are suitable for smaller samples, while preparative columns are necessary for larger quantities.
- Throughput: Assess the required throughput and purification capacity. Analytical columns are efficient for high-resolution analyses, while preparative columns allow for faster purification of larger quantities.
- Budget: Analytical columns are generally more budget-friendly, while preparative columns involve higher costs due to their larger size and specialized nature.
Ultimately, the decision comes down to the specific needs of your application. For routine analytical work and high-resolution separation, an analytical column is appropriate, while a preparative column is essential when purification and collection of larger quantities of target compounds are the primary objectives.
C18 HPLC Column， C8 HPLC Column，c4 Column HPLC，SIO₂ How to choose ?
Choosing between C18, C30, C4, and C8 HPLC columns involves understanding their different selectivity and interactions with analytes. Each type of column offers unique separation characteristics, making them suitable for specific applications. Here’s how to choose among them:
C18 HPLC Column:
- Selectivity: C18 columns are the most widely used and versatile HPLC columns.
- Chemistry: The stationary phase is made of octadecylsilane (C18), a long hydrocarbon chain.
- Application: C18 columns are best for separating nonpolar and hydrophobic compounds.
- Typical Uses: Analyzing drugs, lipids, natural products, and other hydrophobic analytes.
C30 HPLC Column:
- Selectivity: C30 columns provide extended hydrophobicity compared to C18 columns.
- Chemistry: The stationary phase consists of a longer hydrocarbon chain (30 carbons) than C18.
- Application: C30 columns are useful for separating highly hydrophobic compounds.
- Typical Uses: Analyzing complex natural products, lipids, and compounds with multiple hydrophobic moieties.
C4 HPLC Column:
- Selectivity: C4 columns have intermediate hydrophobicity between C18 and C8.
- Chemistry: The stationary phase is made of butyl groups (C4).
- Application: C4 columns are suitable for moderately hydrophobic compounds.
- Typical Uses: Analyzing small polar compounds, peptides, and moderately hydrophobic analytes.
C8 HPLC Column:
- Selectivity: C8 columns have lower hydrophobicity compared to C18.
- Chemistry: The stationary phase consists of octyl groups (C8).
- Application: C8 columns are used for separating polar and moderately hydrophobic compounds.
- Typical Uses: Analyzing peptides, small polar compounds, and moderately hydrophobic analytes.
Choosing the Right Column:
- Analyte Hydrophobicity: If your analytes are highly hydrophobic, consider using a C30 or C18 column. For moderately hydrophobic analytes, C18 or C4 may be suitable.
- Sample Complexity: For complex mixtures with a wide range of compounds, C18 columns are a good starting point due to their versatility.
- Analyte Size: Larger molecules may require C18 or C30 columns with longer hydrophobic chains to accommodate their interactions.
- Method Development: If you have previous experience with specific columns and achieved good separations, it may be beneficial to stick with those columns.
In summary, C18 HPLC columns are a reliable choice for a wide range of applications, while C30, C4, and C8 columns offer specialized selectivity for particular analytes. Consider the hydrophobicity of your analytes and the complexity of your samples to make an informed decision. If possible, perform small-scale method development experiments using different columns to assess which one provides the best separation for your specific application.
What is Column Efficiency in HPLC ?
Column efficiency, also known as plate count or theoretical plates, is a critical parameter in High-Performance Liquid Chromatography (HPLC) that quantifies the separation capability of an HPLC column. It provides a measure of how well a column can separate individual components in a sample as they pass through the stationary phase.
In HPLC, analytes are separated based on their interactions with the stationary phase and the mobile phase. The more interactions an analyte undergoes with the stationary phase during its elution, the longer it takes to pass through the column. Efficient columns can provide sharper and narrower peaks, resulting in better resolution and sensitivity.
Column Efficiency is Quantified in Terms of Theoretical Plates:
The theoretical plate is a concept borrowed from distillation, where it represents a hypothetical equilibrium stage in the separation process. In HPLC, it refers to a hypothetical section of the column where an analyte gets evenly distributed between the mobile phase and the stationary phase. Higher theoretical plates indicate better separation efficiency.
Factors Affecting Column Efficiency:
Several factors influence column efficiency in HPLC:
Particle Size: Columns with smaller particle sizes generally offer higher efficiency because they provide a larger surface area for interactions.
Column Length: Longer columns tend to have more theoretical plates and better efficiency. However, longer columns may require longer analysis times.
Column Diameter: Smaller diameter columns offer higher efficiency due to reduced diffusion distances and a narrower band spreading.
Mobile Phase Flow Rate: Lower flow rates can increase column efficiency, but very low flow rates may increase the analysis time significantly.
Temperature: The column efficiency can be affected by temperature, with higher temperatures leading to reduced efficiency in some cases.
Assessing Column Efficiency:
Column efficiency is often expressed as the number of theoretical plates (N) or the plate count per unit length of the column (N/m):
N = 16 * (tR / w)^2
Where:
- N is the number of theoretical plates.
- tR is the retention time of the peak.
- w is the peak width at its base.
In practice, chromatographers aim to achieve a higher number of theoretical plates for better peak resolution and sensitivity. Efficient columns provide well-defined and sharp peaks, leading to accurate quantification and identification of analytes in complex mixtures. However, it’s essential to balance column efficiency with the desired analysis time and instrument capabilities to achieve optimal chromatographic performance.
How C18 HPLC Column Storage ?
Proper storage of High-Performance Liquid Chromatography (HPLC) columns is essential to maintain their performance and longevity. The following guidelines can help you store your HPLC columns correctly:
Cleanliness: Before storage, ensure that the column is clean and free from any residual analytes or contaminants. If necessary, perform a thorough cleaning according to the manufacturer’s instructions.
Mobile Phase: If the column will be stored for an extended period, remove the mobile phase from the system and the column. Empty the solvent reservoir and thoroughly flush the column with an appropriate solvent to remove any remaining mobile phase.
Protection from Contaminants: Install a column end cap or column stop on both ends to protect the column from airborne contaminants, dust, and moisture. This prevents particles from entering the column during storage.
Storage Conditions: Store the column in a clean and dry environment away from direct sunlight and extreme temperatures. Room temperature is generally suitable for short-term storage, but for long-term storage, consider refrigeration at a stable temperature between 4°C to 10°C.
Column Orientation: Store the column in an upright position to prevent settling of packing material and to ensure uniform distribution when the column is used again.
Transportation: If the column needs to be transported, ensure it is securely packed in a protective case or box to prevent damage during transit.
Avoid Aggressive Solvents: If the column is not in use for an extended period, avoid storing it in aggressive solvents, such as strong acids or bases, as they may deteriorate the stationary phase over time.
Follow Manufacturer’s Recommendations: Always follow the storage recommendations provided by the column manufacturer, as different types of columns may have specific storage requirements.
Column Care: Regularly inspect the column for any signs of damage or leakage. If any issues are observed, contact the manufacturer or technical support for guidance.
Record Keeping: Keep a record of the column usage, storage duration, and any maintenance or cleaning performed, as this information can help track the column’s performance and aid in troubleshooting potential issues.
Proper storage practices help preserve the performance and integrity of HPLC columns, ensuring reliable and consistent results during analyses. Before putting a stored column back into operation, it is advisable to recondition it by flushing with the appropriate solvent and following the manufacturer’s recommended procedures for equilibration.
Some Applications of C18 HPLC Column
High-Performance Liquid Chromatography (HPLC) is a versatile analytical technique with a wide range of applications in various industries. The choice of HPLC column depends on the specific analytes and separation requirements of each application. Here are some common applications and the recommended HPLC column types for each:
Pharmaceutical Analysis:
- Application: Analyzing drugs, active pharmaceutical ingredients (APIs), impurities, and formulation components.
- Recommended HPLC Column: C18 column (Reverse Phase). C18 is the most commonly used column for pharmaceutical analysis due to its broad applicability for separating a wide range of compounds.
Environmental Analysis:
- Application: Monitoring environmental pollutants, pesticides, herbicides, and other contaminants in water, soil, and air samples.
- Recommended HPLC Column: C18 column (Reverse Phase) or HILIC column. The choice depends on the hydrophobicity/polarity of the target compounds. HILIC columns are suitable for polar analytes.
Food and Beverage Analysis:
- Application: Determining food additives, preservatives, vitamins, and other components in food and beverage samples.
- Recommended HPLC Column: C18 column (Reverse Phase). C18 is widely used in food analysis due to its ability to separate a variety of food-related compounds.
Biochemical Analysis and Proteomics:
- Application: Identifying and quantifying biomolecules, such as peptides, proteins, amino acids, nucleic acids, and metabolites.
- Recommended HPLC Column: C18 column (Reverse Phase) or C4 column. C18 is commonly used for peptides and proteins, while C4 is preferred for small polar compounds.
Clinical and Forensic Toxicology:
- Application: Detecting and quantifying drugs, metabolites, and toxins in biological samples for medical and forensic purposes.
- Recommended HPLC Column: C18 column (Reverse Phase). C18 is suitable for analyzing a wide range of drugs and metabolites.
Natural Product Analysis:
- Application: Characterizing and identifying compounds from plant extracts and natural products.
- Recommended HPLC Column: Depending on the complexity of the sample, C18 column (Reverse Phase) or C30 column. C30 provides additional hydrophobic interactions for complex mixtures.
Chiral Separations:
- Application: Separating enantiomers (optical isomers) of chiral compounds.
- Recommended HPLC Column: Chiral column. Chiral columns have immobilized chiral selectors that differentiate enantiomers based on their stereoisomeric configurations.
Pharmaceutical Preparative Purification:
- Application: Isolating and purifying larger quantities of drugs and intermediates for drug development and manufacturing.
- Recommended HPLC Column: Preparative C18 column (Reverse Phase) or other preparative columns with larger dimensions for increased loading capacity.
For each application, it is essential to consider the sample characteristics, the nature of the analytes, and the separation requirements to choose the most suitable HPLC column. Method development and optimization may also be necessary to achieve the desired results.
Why Buy C18 HPLC Column From uHPLCs
When you choose HPLC Columns, You should care some factors, Here we list some factors you should care, why choose uHPLCs to buy HPLC Columns. Hope it will be helpful for your choose.
Quality Assurance: Buy HPLC Columns from Reputable suppliers or manufacturers often adhere to strict quality control standards. Buying from such sources can ensure that you receive high-quality, reliable, and consistent products.
Wide Range of Selection: Established suppliers typically offer a wide range of HPLC columns with various chemistries, particle sizes, and dimensions. This allows you to find the most suitable column for your specific analytical needs.
Technical Support: Reputable suppliers often provide excellent technical support to help you select the right column, troubleshoot issues, and optimize your chromatographic methods.
Application Expertise: Suppliers with expertise in different industries or applications (pharmaceuticals, environmental analysis, biochemistry, etc.) can offer tailored column recommendations based on your specific field of interest.
Batch-to-Batch Consistency: Buying from a trusted source ensures consistent quality across different batches, leading to reproducible results and fewer method adjustments.
Compatibility with Instruments: Established suppliers ensure their HPLC columns are compatible with a wide range of HPLC instruments and systems, reducing the risk of compatibility issues.
Warranty and Returns: Reputable suppliers often offer warranties and return policies, giving you peace of mind in case of any unexpected issues.
Value for Money: While quality products may come at a higher price, the investment in high-quality HPLC columns often translates into better performance, increased efficiency, and reduced downtime, ultimately offering value for money.
Before purchasing HPLC columns or any other scientific equipment, it’s essential to do thorough research and read reviews or testimonials from other customers. Additionally, consider factors such as the supplier’s reputation, customer service, and overall track record in the industry. Choose a supplier that aligns with your specific analytical needs and offers the support and expertise required for successful chromatographic analysis.
Frequently Asked Questions
As Follow is Some Frequently Questions People Asked
C18 HPLC columns are a type of reversed-phase HPLC column packed with stationary particles coated with octadecyl (18-carbon) alkyl chains. These hydrophobic chains interact with the sample components through van der Waals forces. More hydrophobic molecules spend more time interacting with the stationary phase, leading to longer retention times. The mobile phase (a solvent or solvent mixture) carries the sample through the column, separating the components based on their hydrophobicity.
- Versatility: C18 columns offer broad applicability for separating a wide range of non-polar, moderately polar, and even some ionic compounds.
- High resolution: The hydrophobic interactions allow for good separation of closely related compounds.
- Reproducibility: Standardized manufacturing processes ensure consistent performance across different C18 columns.
- Wide availability: C18 is the most common type of HPLC column, making them readily available from various manufacturers.
- Particle size: Smaller particles (e.g., 2-5 µm) provide higher resolution but require higher pressure. Larger particles (e.g., 10 µm) offer faster analysis times but lower resolution.
- Pore size: Larger pore sizes (e.g., 100 Å) allow for better separation of larger molecules. Smaller pore sizes (e.g., 30 Å) are better suited for smaller molecules.
- Carbon chain coverage: Higher coverage leads to stronger hydrophobic interactions and longer retention times. Lower coverage allows for better separation of polar compounds.
- Endcapped vs. non-endcapped: Endcapped columns minimize unwanted secondary interactions with silanol groups on the silica surface, improving peak shapes for acidic or basic compounds.
- Proper storage: Store the column in the appropriate solvent (usually acetonitrile or methanol) to prevent drying out.
- Mobile phase selection: Use mobile phases compatible with the pH range of your analytes to avoid column degradation.
- Sample filtration: Filter your samples to remove particulates that can clog the column.
- Regular cleaning: Periodically clean the column with appropriate solvents to remove residual sample components.
- Decreased peak resolution: Broadened peaks indicate potential column contamination or degradation.
- Increased backpressure: A significant rise in pressure suggests clogged frits or channeling within the column.
- Changes in retention times: Unexpected shifts in retention times could be due to column deterioration.
- Ghost peaks: The appearance of extraneous peaks may indicate irreversible sample adsorption on the column.
Reusing mobile phase can be economical, but it’s crucial to consider potential drawbacks:
- Contamination buildup: Reused mobile phase may accumulate sample components, affecting future separations.
- Composition changes: Evaporation of solvents can alter the mobile phase composition, impacting retention times.
- Increased baseline noise: Contaminants can contribute to higher baseline noise in chromatograms.
For critical separations, it’s generally recommended to use fresh mobile phase.
Both are reversed-phase columns, but C18 has a longer alkyl chain (18 carbons) compared to C8 (8 carbons). This makes C18 more hydrophobic, leading to stronger interactions with analytes and longer retention times. C8 columns are preferred for separating more polar compounds that wouldn’t retain well on C18.
C18 vs. C8 HPLC Columns
| Feature | C18 HPLC Column | C8 HPLC Column |
|---|---|---|
| Alkyl Chain Length | 18 carbons | 8 carbons |
| Hydrophobicity | More hydrophobic | Less hydrophobic |
| Analyte Interaction | Stronger interaction with non-polar and moderately polar analytes | Weaker interaction with non-polar and moderately polar analytes |
| Retention Time | Longer retention times for analytes | Shorter retention times for analytes |
| Analyte Suitability | Ideal for non-polar and moderately polar compounds | Better for polar compounds and those that wouldn’t retain well on C18 |
Absolutely! Gradient elution, where the mobile phase composition changes over time, is a common technique used with C18 columns. It allows for controlled elution of a wider range of compounds with varying polarities.
Used HPLC columns may contain residual hazardous solvents or sample components. Consult your local regulations for proper disposal procedures. Generally, specialized waste disposal companies can handle the safe and compliant disposal of HPLC columns.
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