Solid Phase Extraction (SPE)

Solid Phase Extraction (SPE) is a separation and purification technique used in analytical
chemistry to separate and purify compounds from a liquid sample.

It involves passing the sample through a solid material (sorbent) to selectively retain the target
compounds while unwanted substances are washed away.

Solid Phase Extraction (SPE) is employed to concentrate, purify, or extract specific compounds
for further analysis.

Principle

In Solid Phase Extraction (SPE), the working principle involves

Passing a liquid sample through a solid sorbent material contained in a cartridge or a column.
The sorbent selectively retains certain compounds based on their chemical properties while
allowing other components to pass through. After loading the sample, unwanted substances
are washed away using various solvents, and then the retained compounds are eluted using
another solvent.

Fundamentals

Here are some key fundamentals of Solid Phase Extraction (SPE). These fundamentals are
essential for the successful application of Solid Phase Extraction in various analytical and
preparative processes.

1. Sorbent Selection: Choosing the right solid-phase material (sorbent) based on its
compatibility with the target compounds’ properties (polarity, size, etc.) is crucial for successful
extraction.
2. Sample Loading: The liquid sample containing the compounds of interest is passed through
the SPE column or cartridge containing the sorbent.
3. Washing Step: Unwanted substances are removed by washing the sorbent with appropriate
solvents, allowing only the target compounds to be retained.
4. Elution: The retained compounds are then eluted or released from the sorbent using another
solvent that breaks the interaction between the compounds and the sorbent, allowing the
compounds of interest to be collected.

5. Optimization: The process parameters, including the choice of sorbent, sample volume,
solvent selection, and elution conditions, need to be optimized to achieve maximum recovery
and purity of the target compounds.
Sorbent Selection

Sorbent selection in Solid Phase Extraction (SPE) is critical as it determines which compounds
will be retained and which will pass through. It involves choosing a solid-phase material
(sorbent) that possesses specific chemical and physical properties suitable for the target
compounds in the sample.

The necessity of sorbent selection lies in the fact that different sorbents have varying affinities
for different compounds based on factors such as polarity, charge, size, and functional groups.
This selectivity helps in isolating the desired compounds efficiently.
Selection is done based on Several Factors:

1. Chemical Compatibility: The sorbent should be chemically compatible with the solvents and
compounds in the sample to avoid unwanted interactions.

2. Target Compound Properties: Polarity, size, and functional groups of the compounds of
interest aids in choosing a sorbent with complementary properties for effective retention.

3. Matrix Interference: Some samples contain complex matrices. Choosing a sorbent that
minimizes interference from these matrices helps in selectively retaining the target compounds.

4. Optimization and Efficiency: Each sorbent has its strengths and limitations. Optimization
involves testing different sorbents to find the most efficient one for a particular application.

5. Sorbent Formats: SPE sorbents come in various formats like cartridges, disks, or plates.
The selection might also depend on the specific format needed for the extraction process.

6. Manufacturer’s Recommendations: Manufacturers often provide guidance on which

sorbent is suitable for certain applications based on their specifications.

Sorbent Formats
Sorbent formats refer to the different physical configurations in which solid-phase materials
are used in Solid Phase Extraction (SPE). These formats cater to various applications and
extraction requirements:

1. Cartridges: These are the most common format, consisting of a tube or cylinder filled with
the sorbent material. Cartridges come in different sizes and capacities and are compatible with
automated SPE systems.

2. Disks: Similar to cartridges but in a disk shape, they offer specific advantages such as
compatibility with lower sample volumes or integration into specific instrumentation.

3. Plates: These are designed for high-throughput applications, typically used in 96-well or
384-well plate formats, often in conjunction with automated liquid handlers.
4. Pipette Tips: Miniaturized SPE sorbents incorporated into pipette tips enable direct
extraction from the sample source, offering convenience and efficiency for smaller sample
volumes.

5. Spin Columns: These are small columns filled with sorbent material and are often used in
manual SPE protocols. They are spun to facilitate sample passing through the sorbent material.

Sample Preparation

Sample preparation before Solid Phase Extraction (SPE) involves several critical steps to
ensure the successful extraction and purification of target compounds:
1. Selection of Sample and Pretreatment: Choose the appropriate sample matrix (e.g., water,
blood, soil, etc.) and pretreat it if necessary. Pretreatment may involve filtration, centrifugation,
dilution, or pH adjustment to remove particulates, debris, or unwanted components that could
interfere with the extraction process.

2. Selection of Extraction Solvents: Determine the suitable solvents for sample preparation.
These solvents should effectively dissolve the compounds of interest while being compatible
with the SPE sorbent and elution solvents. Proper solvent selection ensures efficient extraction.

3. Homogenization or Mixing: Homogenize or mix the sample thoroughly to ensure a

representative aliquot for extraction. Proper mixing helps distribute the target compounds
evenly throughout the sample, enhancing their accessibility during the extraction process.

4. Cleanup and Filtration: Perform additional cleanup steps if needed, such as solid-phase
microextraction, liquid-liquid extraction, or filtration, to eliminate larger particles, proteins,
lipids, or other interfering substances that could affect SPE performance.

5. Adjustment of Sample pH: Depending on the analytes of interest, adjust the pH of the
sample if their extraction efficiency is pH-dependent. This step can enhance the extraction
efficiency of certain compounds.

6. Sample Concentration: Concentrate the sample if the target compounds are present in low
concentrations. Techniques like solvent evaporation, solid-phase extraction, or freeze-drying
may be employed for this purpose.

7. Compatibility Testing: Ensure that the prepared sample is compatible with the chosen SPE
sorbent and elution solvents. Compatibility testing helps avoid interactions that could
compromise extraction efficiency or result in contamination.

Compatibility Testing in the context of Solid Phase Extraction (SPE) involves assessing
whether the sample, solvents, and materials used in the extraction process are harmonious and
won’t adversely affect the extraction efficiency or compromise the quality of the analysis.

Here’s how compatibility testing is typically conducted in SPE:

1. Sample Compatibility: Ensure that the components present in the sample matrix (e.g., salts,
proteins, lipids, etc.) do not interfere with the SPE process. Compatibility testing verifies
whether the sample composition is suitable for the chosen SPE sorbent and the subsequent
elution process.

2. Solvent Compatibility:

Assess the solvents used for conditioning, sample loading, washing, and elution to confirm
their compatibility with the SPE sorbent material. Solvents should not degrade or swell the
sorbent, affecting its ability to retain or release target compounds.
3. Sorbent Compatibility:

Verify that the sorbent material selected for SPE is appropriate for the target compounds and
is not adversely affected by the sample matrix or solvents used. The sorbent should effectively
retain the target compounds without excessive interference from sample components.

5. Elution Solvent Compatibility:

Ensure that the solvent chosen for elution effectively releases the target compounds from the
sorbent without causing interference or contamination. This solvent should not damage the
analytical instrument used for subsequent analysis.

Importance Of Sample Preparation

Here are several key reasons highlighting the importance of sample preparation:

1. Enhanced Sensitivity: Proper sample preparation helps concentrate target analytes,

increasing their concentration in the sample. This enhances the sensitivity of analytical
techniques, allowing for the detection of lower concentrations of compounds.

2. Improved Selectivity: Sample preparation methods selectively remove unwanted matrix

components, interferences, or contaminants, improving the selectivity of analytical methods
and reducing interference in the analysis.

3. Matrix Compatibility: Sample preparation ensures the sample is compatible with the
analytical instrumentation and methods used, preventing instrument damage and enhancing
the accuracy of measurements.

4. Elimination of Interferences: It removes impurities, matrix interferences, or compounds

that might mask or interfere with the detection or quantification of target analytes, leading to
more accurate results.

5. Increased Accuracy and Precision: Properly prepared samples lead to more accurate and
reproducible results, reducing variability and enhancing the precision of analytical
measurements.
6. Extended Instrument Lifespan: Removing contaminants or incompatible matrix
components helps prevent damage or fouling of instruments, prolonging their lifespan and
reducing maintenance requirements.

7. Reduced Analysis Time: Effective sample preparation streamlines the analysis by

minimizing unwanted compounds, leading to shorter analysis times and higher laboratory
throughput.

8. Optimized Method Performance: Sample preparation can optimize the performance of

analytical methods by improving the extraction efficiency, ensuring better recovery rates, and
minimizing sample loss.

9. Compliance with Regulations: In many fields like environmental analysis or

pharmaceuticals, proper sample preparation ensures compliance with stringent regulations by
accurately measuring compounds of interest while adhering to established standards.

10. Facilitation of Complex Analyses: For complex samples, sample preparation simplifies
the analytical process by isolating target compounds, making it easier to analyze and interpret
results.

11. Reproducibility: Consistent and well-defined sample preparation protocols ensure

reproducibility of results across multiple experiments or different laboratories.

12. Cost Efficiency:

Proper sample preparation can reduce the need for repeated analyses due to better accuracy
and reliability, thereby saving resources and costs in the long run.

SPE as Sample Preparation Tool

Yes, Solid Phase Extraction (SPE) is widely used as a sample preparation technique in
analytical chemistry. SPE serves as an effective method for preparing samples by selectively
extracting, purifying, and concentrating target compounds from complex mixtures before their
analysis.

Here’s why SPE is a valuable sample preparation technique:

1. Selective Extraction: SPE allows the selective retention of target compounds on a solid-
phase material while unwanted substances are removed, enhancing the purity of the analytes.

2. Concentration of Analytes: It concentrates target compounds from large sample volumes

into smaller elution volumes, increasing their concentration and improving sensitivity during
analysis.

3. Matrix Cleanup: SPE effectively removes matrix interferences, impurities, or contaminants

from the sample, reducing interference in subsequent analytical methods.
4. Preparation for Instrumental Analysis: By reducing sample complexity, SPE prepares
samples for various instrumental analyses like chromatography, mass spectrometry, or
spectroscopy, improving the accuracy of results.

5. Versatility: SPE can be tailored by selecting different sorbents and methodologies to suit
specific analytes, matrices, or analytical requirements, making it applicable across various
industries and sample types.

6. Enhanced Recovery and Precision: SPE methods optimize recovery rates, improving
precision and reproducibility of results in subsequent analyses.
7. Automation Compatibility: SPE methods can be automated, allowing for high-throughput
sample preparation in laboratories handling a large number of samples.

8. Compliance with Regulations: In fields like environmental monitoring or pharmaceutical

analysis, SPE ensures compliance by effectively extracting and isolating compounds of interest
from complex samples.

Sample Loading Conditions

The loading conditions in Solid Phase Extraction (SPE) refer to the parameters and procedures
employed when introducing the sample onto the SPE sorbent material. These conditions are
crucial as they determine how effectively the target compounds are retained and subsequently
eluted.

Loading conditions include:

1. Sample Volume: The volume of the sample introduced onto the SPE column or cartridge. It
should be within the sorbent’s capacity to avoid overloading or inefficient extraction.

2. Sample pH: Adjusting the pH of the sample, if necessary, to optimize the retention of target
compounds on the SPE sorbent. pH adjustments can enhance the extraction efficiency of
certain compounds.

3. Sample Flow Rate or Dwell Time: The rate at which the sample passes through the SPE
sorbent. Controlling the flow rate or allowing sufficient dwell time ensures adequate
interaction between the sample components and the sorbent, maximizing retention.
4. Preconditioning of the Sorbent: Ensuring the sorbent is properly conditioned and
equilibrated before sample loading to enhance its interaction with the sample components and
improve extraction efficiency.

5. Compatibility with Sample Matrix: Confirming that the sample matrix and components are
compatible with the chosen sorbent and loading conditions to avoid undesired interactions or
inefficiencies during extraction.
6. Optimization of Loading Solvents: Choosing the appropriate solvent or solvent mixture for
loading the sample onto the sorbent. The solvent should effectively dissolve the target
compounds without adversely affecting the sorbent or interfering with subsequent steps.

Properly optimizing these loading conditions is crucial to achieving efficient and selective
retention of the target compounds on the SPE sorbent.

Methods for Sample Loading

In Solid Phase Extraction (SPE), several methods are used for sample loading, which involve
introducing the sample containing the target compounds onto the solid-phase material
(sorbent). These methods differ based on the SPE format and the specific requirements of the
sample and analysis. Common methods of sample loading in SPE include:

1. Gravity Flow Loading: Simply passing the sample through the SPE cartridge or column
under the influence of gravity. This method is straightforward and commonly used for manual
SPE protocols.

2. Vacuum-Assisted Loading: Applying vacuum pressure to draw the sample through the
sorbent material, increasing the speed and efficiency of sample loading. This method is suitable
for enhancing the flow rate and ensuring better contact between the sample and the sorbent.

3. Pump-Assisted Loading: Using a pump system to drive the sample through the SPE
cartridge or column, providing better control over flow rates and allowing for automation in
high-throughput applications.

4. Solid-Phase Microextraction (SPME): Involves direct extraction of analytes from the

sample matrix onto a coated fiber or solid-phase microextraction device. The extracted
compounds are then desorbed for subsequent analysis.
5. Dip-and-Dry Loading: Immerse the sorbent material directly into the sample, allowing the
target compounds to adsorb onto the sorbent, followed by drying before further processing.

6. Direct Injection Loading: In some cases, especially in smaller-scale applications or when

using specialized formats like SPE pipette tips, samples can be directly aspirated or injected
onto the sorbent.

Elution Conditions

The elution conditions in Solid Phase Extraction (SPE) refer to the parameters and procedures
employed to release the target compounds from the sorbent material after the sample has been
loaded and processed. These conditions are crucial for obtaining high recovery rates and
purity of the desired analytes. Key elements of elution conditions include:

1. Elution Solvent: aSelection of an appropriate elution solvent that efficiently breaks the
interactions between the target compounds and the sorbent material. The solvent should have
a suitable polarity or pH to elute the compounds of interest effectively.
2. Solvent Volume and Flow Rate: Optimizing the volume of elution solvent and the flow rate
through the SPE cartridge or column to ensure maximum recovery of the target compounds
without losing efficiency.

3. Elution Fractionation: Sometimes, elution is performed in multiple steps or fractions using

different solvents or solvent gradients to elute different analytes selectively or to achieve a more
refined separation.

4. Elution Time: Determining the appropriate duration for the elution process to allow
sufficient contact time between the elution solvent and the sorbent material, ensuring efficient
release of the target compounds.

5. Elution Method: Different elution methods such as gravity flow, vacuum, or pressure-
assisted elution may be used depending on the SPE format (cartridge, disk, plate) and the
equipment available.

6. Post-Elution Treatments: Sometimes, additional steps like evaporation or concentration of

the eluate are performed to further concentrate the target compounds before subsequent
analyses.

Elution Methods

In Solid Phase Extraction (SPE), elution methods refer to the various techniques used to release
or extract the target compounds from the sorbent material after the sample has been processed.
These methods vary based on the type of SPE format used and the specific requirements of the
analysis. Some common elution methods include:

1. Liquid-Phase Elution:

Gravity Flow: This method involves passing the elution solvent through the SPE column or
cartridge by gravity.

Vacuum-Assisted: Applying vacuum pressure helps draw the elution solvent through the
sorbent material, enhancing the elution efficiency.
2. Pressure-Driven Elution: Using a pump system to apply controlled pressure to drive the
elution solvent through the SPE cartridge or column, allowing for faster and more precise
elution.

3. Solvent Gradient Elution: Gradient elution involves using a series of elution solvents with
varying polarities or compositions in a stepwise manner. This method helps elute compounds
selectively based on their affinity to different solvents.

4. Stepwise Elution: Employing multiple elution steps with different solvents or solvent
mixtures to sequentially extract analytes of varying polarities or affinities from the sorbent
material.
5. Dry Elution: Using gases (e.g., nitrogen) to aid in eluting compounds by passing the gas
through the sorbent, evaporating the analytes from the solid phase.

6. Temperature-Assisted Elution: Heating the SPE cartridge or column, or using a heated

elution solvent, to increase the mobility of the target compounds, aiding their release from the
sorbent.

The choice of elution method depends on factors such as the nature of the target analytes, the
sorbent material, the sample matrix, the desired purity of the eluted compounds, and the
subsequent analytical techniques used for detection and quantification.

Post-elution Treatments

Post-elution treatments in Solid Phase Extraction (SPE) refer to additional steps or procedures
performed after the elution of target compounds from the sorbent material. These treatments
aim to further process the eluate or enhance the concentration, purity, or compatibility of the
extracted analytes for downstream analyses. Some common post-elution treatments include:

1. Solvent Evaporation: Concentrating the eluate by evaporating the solvent, resulting in a

more concentrated sample suitable for analysis by techniques requiring higher analyte
concentrations.

2. Solvent Exchange: Changing the elution solvent to a different solvent that is more
compatible with the subsequent analytical method or better suited for the detection technique.

3. Reconstitution: Reconstituting the eluted sample by dissolving it in a specific solvent or

buffer to achieve the desired concentration or compatibility with the analysis method.

4. Sample Cleanup: Further cleaning or purification steps to remove remaining impurities,

residual matrix components, or interfering substances that might affect the accuracy or
sensitivity of the analysis.

5. Concentration: Using techniques like solvent evaporation, solid-phase concentration, or

freeze-drying to concentrate the eluate further, especially when the initial sample volume was
large or the analyte concentration was low.

6. Derivatization: Modifying the chemical structure of the extracted analytes to improve their
detectability, stability, or compatibility with the chosen analytical method.

7. pH Adjustment: Adjusting the pH of the eluate to achieve the desired pH range for the
subsequent analytical technique or to stabilize the analytes.

8. Sample Storage: Proper storage of the eluted sample under suitable conditions
(temperature, light exposure, etc.) to maintain the integrity of the analytes before analysis.
Packed Particle Beds And Disks

In Solid Phase Extraction (SPE), packed particle beds and disks refer to the physical formats
of the sorbent material used in the extraction process.

1. Packed Particle Beds:

- In SPE cartridges or columns, the sorbent material is often packed as small particles within
the column. These particles create a bed through which the sample is passed for extraction.

- These beds can be packed with various sorbents like silica, reversed-phase materials (such
as C18 or C8), ion-exchange resins, or other functionalized materials suitable for the target
compounds.
- The sample passes through this packed bed, allowing the target compounds to interact and
adhere to the sorbent while unwanted components flow through.

- Packed particle beds offer a high surface area for interaction, facilitating efficient
extraction and separation of compounds.

2. Disks:

- SPE disks are solid-phase materials in a disk-shaped format, similar to a thin wafer, often
used in specialized applications.

- These disks are typically used for lower sample volumes or specialized instrumentation.

- They can also be packed with various sorbents like silica, reversed-phase, ion-exchange,
or mixed-mode materials, depending on the specific requirements of the extraction.

Sorbent Capacity And Recovery

In Solid Phase Extraction (SPE), sorbent capacity and recovery are crucial factors that
determine the efficiency and effectiveness of the extraction process.

1. Sorbent Capacity:

- Sorbent capacity refers to the maximum amount of analyte that a particular sorbent
material can retain or adsorb under specified conditions.

- It’s important to ensure that the amount of sorbent used in the SPE process is sufficient to
effectively capture the target analytes present in the sample.

- The capacity depends on the specific characteristics of the sorbent, such as surface area,
pore size, and chemical composition, as well as the properties of the analytes, such as polarity,
size, and concentration.
2. Recovery:

- Recovery in SPE is the percentage of the target analytes that are successfully eluted or
extracted from the sorbent material relative to the total amount present in the sample.

- It represents the efficiency of the extraction process in capturing and releasing the target
compounds without significant losses.

- Factors affecting recovery include the sorbent selectivity, sample matrix complexity, elution
solvent choice, and elution conditions (solvent type, volume, flow rate, etc.).

Sorbent capacity and recovery optimizations

Here are optimization strategies for improving sorbent capacity and recovery in Solid Phase
Extraction (SPE) in point form:

1. Sorbent Selection: Choose suitable sorbents based on analyte properties.

2. Conditioning Optimization: Ensure effective activation of sorbent surfaces.

3. Sample pH Adjustment: Adjust pH if needed for better analyte-sorbent interaction.

4. Sample Loading Optimization: Adjust flow rate and volume for optimal loading.

5. Washing: Use selective washing for interference removal without losing target compounds.

6. Elution: Experiment with solvents, volumes, and techniques for maximum recovery.

7. Validation and Method Optimization: Validate SPE method and optimize based on results.

8. Matrix Interference Mitigation: Implement pretreatment steps to minimize interferences.

9. Quality Control Check: Monitoring sorbent performance for consistency and reliability.

Offline and Online approaches

Here are the differences between offline and online approaches in the context of sample
preparation and analysis:

Offline Approaches:

1. Separate Process: In offline approaches, sample preparation and analysis are distinct steps
performed separately.

2. Sample Preparation: Sample preparation steps, such as extraction, purification, and

concentration, are completed independently from the analytical instrument.

3. Time-consuming: It may be more time-consuming as it involves additional steps and

processes before analysis.
4. Flexibility: Offers flexibility in optimizing each step of sample preparation and analysis
independently.

5. Manual Intervention: Often requires manual handling and transfer of samples between
different preparation and analysis stages.

6. Sample Storage: Allows for storing prepared samples for a period before analysis.

Online Approaches

1. Integrated Process: In online approaches, sample preparation seamlessly integrates with

the analytical instrument.

2. Real-time Analysis: Sample preparation occurs just before analysis, allowing for immediate
and continuous analysis of the sample.

3. Efficiency: Typically faster as there is no delay between sample preparation and analysis.

4. Automation: Often involves automated systems or instrumentation, reducing manual

intervention and increasing throughput.

5. Limited Flexibility: May have limited flexibility in optimizing individual steps as they are
integrated into a single system.

6. No Sample Storage: Samples are processed immediately, eliminating the need for sample
storage between preparation and analysis.

Method development and protocols

In the context of Solid Phase Extraction (SPE), method development and protocols refer to the
systematic process of designing, optimizing, and establishing procedures for extracting,
purifying, and concentrating target analytes from sample matrices using SPE techniques. Here
are key aspects of method development and protocols

Here are details on method development and protocols in the context of Solid Phase Extraction
(SPE):

1. Objective Definition: Identify the specific objectives of the extraction: target analytes,
sample matrix, desired recovery, detection limits, and subsequent analysis method.

2. Sorbent Selection: Choose the most suitable sorbent(s) based on analyte properties, sample
matrix, and desired selectivity. Evaluate different sorbents to determine the best fit.

3. Optimization of Conditions: Optimize parameters such as sample pH, conditioning solvents,

sample loading volume, flow rate, washing steps, and elution conditions (solvents, volumes,
gradients) to maximize analyte recovery and purity.
4. Validation: Validate the developed SPE method by assessing parameters like recovery,
reproducibility, linearity, matrix effects, and limits of detection and quantification.

5. Protocol Development: Establish a detailed protocol documenting step-by-step procedures,

including sample preparation, sorbent conditioning, loading, washing, elution, and any post-
elution treatments.

6. Robustness Testing: Assess the robustness of the method by testing variations in parameters
(e.g., pH, flow rate, solvent composition) to ensure consistent results under different conditions.

7. Sample Matrix Compatibility: Evaluate the method’s applicability to different sample

matrices and assess any matrix effects that might interfere with the extraction process or
subsequent analysis.

8. Automation and High Throughput: Consider automating the SPE process for higher
throughput and consistency, especially in laboratories dealing with a large number of samples.

9. Quality Assurance and Control: Implement quality control measures to ensure the
reliability and reproducibility of the SPE method over time. This includes regular calibration
checks, monitoring of sorbent performance, and adherence to established protocols.

10. Documentation and Reporting: Maintain comprehensive documentation of method

development, optimization, validation results, and protocols for future reference and reporting.

Microchips & Nanochips

In the realm of Solid Phase Extraction (SPE), the terms “microchips” and “nanochips” don’t
directly relate to electronic devices but rather signify the scale of materials or structures used
within the SPE process.

Microchips in SPE: Microchips, in the context of SPE, can refer to the scale of the sorbent
materials utilized in the extraction process. These materials are often composed of particles or
beads that generally range in size from a few micrometers to several hundred micrometers.
These particles provide the surface area necessary for adsorption and interaction with the
analytes present in the sample solution. The packing of these particles within SPE columns or
cartridges influences the efficiency, selectivity, and capacity of the extraction process. The
larger surface area and proper packing increase the contact between the sample solution and
the sorbent, enhancing the extraction efficiency.

Nanochips in SPE: Nanochips, although not a conventional term in SPE, could theoretically
refer to nanoscale materials or structures integrated into the sorbent materials used in
specialized SPE applications. These materials might include nanoparticles, nanofibers, or
other nanoscale structures embedded within the sorbent matrix. The implementation of such
nanoscale materials aims to exploit their unique properties, such as high surface area-to-
volume ratio and tailored surface functionalities, to enhance the selectivity and efficiency of
the extraction process. Their small size allows for increased interaction sites and precise
capture of specific analytes from complex sample matrices.
Micro-LC & Nano-LC Systems

Certainly! In the context of Solid Phase Extraction (SPE), “Micro-LC” and “Nano-LC” refer
to liquid chromatography systems used for analyzing small sample volumes with varying
degrees of sensitivity and resolution:

1. Micro-LC:

Column Diameter: Micro-LC systems typically employ columns with larger internal diameters,
commonly in the range of 1-2 mm.
Flow Rates: Operates at higher flow rates than conventional LC systems, allowing faster
separations.

Sample Volume: Analyzes sample volumes in the microliter range.

Applications in SPE: After extracting target compounds using SPE, Micro-LC is employed for
subsequent analysis when dealing with limited sample volumes. It offers increased sensitivity
and resolution compared to standard LC, enabling efficient analysis of extracted compounds.

2. Nano-LC:

Column Diameter: Nano-LC systems use columns with significantly smaller internal
diameters, often below 100 µm.

Flow Rates: Operates at lower flow rates than Micro-LC and standard LC systems, providing
increased resolution and sensitivity.

Sample Volume: Analyzes very small sample volumes in the nanoliter range.

Applications in SPE: Post-extraction, Nano-LC systems are useful for highly concentrated and
limited sample volumes. They offer exceptional sensitivity and resolution, crucial for analyzing
minute quantities of extracted compounds.