Nano LC-MS Columns (C18,75µm×15cm)

With the continuous advancement of proteomics, metabolomics, and related life science research, there is an increasing demand for high-resolution and high-sensitivity separation and analysis tools. Nano-scale liquid chromatography technology (Nano-LC), known for achieving highly efficient separation and ultra-sensitive detection at extremely low flow rates, has gradually become a common approach for protein and peptide separation analysis. For many complex samples that require high-precision identification, Nano-LC not only reduces sample and reagent consumption but also enhances signal response and coverage in subsequent mass spectrometry detection. Therefore, a high-quality Nano LC column plays a critical role in proteomics, scientific testing, pharmaceutical R&D, and other fields. Based on years of technical accumulation and industry experience, MtoZ Biolabs, a supplier specializing in protein sample preparation consumables, has launched a Nano LC-MS column (C18, 75 µm × 15 cm), aimed at helping researchers and technical personnel achieve more stable, accurate, and high-throughput analysis results in micro-scale protein/peptide separation.

 

Figure 1. The Nano LC Column Provided by MtoZ Biolabs.

  

Product Overview

The Nano LC-MS column (C18, 75 µm × 15 cm) from MtoZ Biolabs features a narrow inner diameter, uniform packing, appropriate particle size, and stable bonded phase. The column is built upon a high-purity silica matrix and undergoes a rigorous C18 bonding process to ensure reliable separation efficiency and durability under low flow rate conditions. The small inner diameter (75 µm) maximizes peak resolution, which is especially beneficial for capturing signals from low-abundance peptide segments in subsequent mass spectrometry analysis. Meanwhile, the 15 cm length strikes a balance between separation depth and resolution, enabling high peak capacity while reducing analysis time and solvent consumption.

 

In proteomics and related scientific analyses, samples often exhibit a wide molecular weight distribution and large differences in concentration. MtoZ Biolabs fully considered these characteristics in the design of the Nano LC-MS column (C18, 75 µm × 15 cm), selecting packing materials with high stability and inertness to maintain excellent separation performance in complex matrix backgrounds. Thanks to its superior surface modification process, the column effectively reduces peak tailing and nonspecific adsorption of target molecules, thereby achieving more accurate quantitative analysis while maintaining a high signal-to-noise ratio.

 

Protocol

1. Recommended Mobile Phase

Phase A: 0.1% FA in water; Phase B: 0.1% FA in 80% ACN.

 

2. New Column Conditioning

When using a new column, flush it with 95% organic phase at a flow rate of 200 nL/min for 30 minutes. Then increase the flow rate to 400 nL/min and continue flushing with 95% organic phase for another 30 minutes. Adjust the organic phase to 50% and flush the column for 30 minutes. Finally, adjust the organic phase to 8% and equilibrate for 30 minutes.

 

3. Pre-Run Preparation

Before starting the experiment, flush the Nano LC column with the appropriate initial mobile phase for several minutes to ensure a stable baseline and match the solvent environment within the column to that of the sample.

 

4. Sample Loading

Dilute the peptide solution obtained from proteolysis or other pretreatment steps to an appropriate concentration (typically within nanoliter volumes). Use a micro-injection device or autosampler to introduce the sample at low flow rates to the Nano LC column inlet, avoiding high backpressure or sample overloading.

 

5. Gradient Elution

Set the elution gradient according to experimental requirements. As the proportion of organic solvent increases programmatically, peptides with different hydrophobicities elute sequentially, achieving good chromatographic separation. A recommended gradient duration is 60-120 minutes, with an effective gradient range of 8%-40% phase B.

 

6. Detection and Data Acquisition

Nano LC columns are commonly coupled with mass spectrometry for highly sensitive peptide detection. During gradient elution, the mass spectrometer continuously scans the eluate and performs qualitative and quantitative analysis of the separated components.

 

7. Post-Run Maintenance

After analysis, flush the column with a high organic phase (e.g., >90% acetonitrile or methanol) to remove residual hydrophobic molecules, and then restore column equilibrium with the equilibration solvent. For long-term storage, use a preservation solution containing a suitable amount of organic phase (e.g., 50% acetonitrile + 0.1% formic acid) to extend the column's service life and maintain stable performance.

 

Features and Benefits

1. High Resolution and Sensitivity

The 75 µm inner diameter combined with high-purity C18 packing facilitates efficient separation of target peptides under micro-flow conditions. The sharp peak shapes and enhanced signal intensity provide more accurate data for subsequent mass spectrometry analysis.

 

2. Excellent Peak Capacity

The 15 cm column length ensures sufficient separation depth while also reducing overall analysis time, playing a key role in high-throughput experiments.

 

3. Stable and Durable

Meticulous manufacturing processes ensure the stability of the bonded phase, reducing the risk of performance degradation with frequent use and enabling consistent separation performance over extended periods.

 

4. Reduced Nonspecific Adsorption

The optimized inertness of the packing surface effectively minimizes nonspecific interactions between the sample and the column, lowers background noise, and improves the reliability of detection results.

 

5. Broad Compatibility

The Nano LC-MS column (C18, 75 µm × 15 cm) is suitable for various gradient elution modes and commonly used liquid chromatography systems. It is compatible with nano-LC-MS platforms from multiple brands, meeting the analytical needs of different laboratories.

 

Applications

1. Proteomics Research

In the analysis of complex protein samples such as cell lysates and tissue homogenates, high-resolution peptide separation combined with mass spectrometry enables rapid screening and identification of various proteins using the Nano LC-MS column (C18, 75 µm × 15 cm), providing strong data support for elucidating biological mechanisms.

 

2. Biomarker Discovery

In disease diagnosis and drug mechanism studies, accurate separation and detection of low-abundance peptides are crucial for identifying potential biomarkers and developing novel diagnostic reagents.

 

3. Protein Modification Detection

Using Nano-LC-MS/MS technology, post-translational modifications (such as phosphorylation and glycosylation) can be precisely identified, contributing to a deeper understanding of signaling pathway regulation and cellular processes.

 

4. New Drug Development and Quality Control

In the biopharmaceutical field, high-resolution separation is essential for the characterization and quantification of biomacromolecules and their degradation products. It supports impurity identification and drug consistency evaluation, providing reliable data for R&D and quality control.

 

5. Trace Sample Analysis

MtoZ Biolabs' Nano LC-MS column (C18, 75 µm × 15 cm) is suitable for analyzing valuable or limited biological materials (such as serum and microbial culture supernatants), helping researchers obtain reliable detection results even when sample availability is constrained.