🧬 What Are Peptides?

🔬 HPLC Device – Components & Functions

🧪 **Technical Description:

This image shows mobile phase solvent bottles typically made of amber or clear glass, containing high-purity solvents such as:
• Water (H₂O)
• Methanol (MeOH)
• Acetonitrile (ACN)
• Buffer solutions (e.g., phosphate, acetate)

The bottles are connected via thin polymer tubing to the HPLC system’s pump unit, marking the starting point of the chromatography process.

🔍 Why This Step Is Critical:

Mobile phase preparation is the foundation of any reliable HPLC analysis. Its quality directly impacts:
• Retention times
• Peak resolution
• Detector signal stability
• Reproducibility and accuracy of results

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🧼 Best Practices in the Lab:
• Use only HPLC-grade solvents to prevent contamination.
• Always filter the mobile phase using 0.22 or 0.45 μm filters.
• Degas solvents (manually or with an inline degasser) to eliminate air bubbles.

🧪 **Technical Description:

This image captures the moment a lab technician places a sample vial into the HPLC autosampler tray, which may contain dozens—even hundreds—of sample vials. The autosampler is programmed to inject each sample precisely into the flow path of the mobile phase, allowing the system to perform multiple injections automatically without manual intervention.

🎯 Purpose and Advantages of the Autosampler:
• Automates routine analysis: Greatly increases throughput by handling large sample batches.
• Ensures precision: Injects accurate and reproducible sample volumes (often 5–100 μL).
• Reduces human error: Limits variability associated with manual injection.
• Improves consistency: Critical for method validation, quality control, and regulated environments.

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⚙ Key Features Typically Shown in the Image:
• Circular tray or linear rack holding vials (2 mL or 1.5 mL).
• Robotic needle or arm mechanism for precise aspiration and injection.
• Integration with software for sequencing and method control.

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🧼 Best Practices:
• Use clean, labeled, and properly sealed vials.
• Filter all samples to prevent column clogging.
• Store samples at controlled temperatures if the autosampler has cooling functions.
• Regularly calibrate and maintain the autosampler needle and syringe.

This image highlights the analytical core of the HPLC system — the high-pressure pump and the detector module. These components work together to deliver the mobile phase at a controlled flow rate and pressure, and to detect compounds as they elute from the column.

You can typically observe:
• A digital interface on the pump displaying real-time pressure, flow rate, and gradient settings.
• Tubing connected to the mobile phase reservoir and column inlet.
• The detector, positioned after the column, collecting data on compound concentration via UV, DAD, RI, or MS technologies.

💡 Pump Function – The Engine of HPLC:
• Delivers mobile phase at high precision (e.g. 0.001 mL/min up to 10 mL/min).
• Maintains consistent pressure (up to 400 bar or more).
• Supports isocratic or gradient elution, crucial for separating compounds with different polarities.

💡 Detector Function – The Eye of the System:
• Measures how much of each compound is present based on absorbance (UV/Vis) or refractive index, fluorescence, or mass-to-charge ratio (MS).
• Sends real-time data to the software, producing a chromatogram that shows the separated peaks.

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🎯 Importance of This Stage:
• Precise flow and pressure control are critical for repeatable separation.
• Accurate detection ensures quantitative and qualitative analysis of each compound.
• Detector sensitivity determines how low of a concentration can be reliably measured (LOD/LOQ).

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🧼 Best Practices:
• Regularly check and clean pump seals and pistons to avoid leaks or pulsation.
• Calibrate detectors and use proper wavelength settings for UV analysis.
• Monitor backpressure and maintain the system to extend column and detector life.

🧪 Technical Description:

This image shows the complete, fully integrated HPLC system operating in a real laboratory environment. All major components are connected and functioning in unison:
• Solvent reservoirs at the top feeding the system.
• Pump unit delivering the mobile phase.
• Autosampler injecting samples from multiple vials.
• Column compartment where separation occurs.
• Detector analyzing eluted compounds.
• Computer screen displaying the real-time chromatogram and data analysis.

The workspace is clean, well-lit, and organized — reflecting professional lab standards.

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🔬 What This Stage Represents:

This is the operational phase of the HPLC workflow, where real sample analysis occurs:
• Solvents flow continuously under pressure.
• The system runs pre-set analytical methods.
• The software logs retention times, peak areas, and identifies components.

At this stage, the method has already been optimized, and the system is ready to analyze single or batch samples for routine or research purposes.

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🧠 Significance of the Full Setup:
• Demonstrates automation and reliability of modern chromatography labs.
• Integrates all analytical steps in a continuous flow process.
• Ensures high-throughput capability—critical for research, pharmaceutical QC, and method validation.

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🧼 Best Practices at This Stage:
• Verify method parameters (flow rate, gradient, injection volume, detection wavelength).
• Monitor baseline stability and system pressure throughout the run.
• Back up data and regularly check for signal drift or noise.