Dionex Ultimate 3000 UHPLC Manual

The Dionex Ultimate 3000 is a high-performance ultra-high performance liquid chromatography system designed for analytical laboratories. It features advanced pump technology, precise temperature control, multiple detection options, and intuitive software control. Below are key sections covering system overview, installation, operation, maintenance, and troubleshooting.

1. System Overview 2. Installation and Setup 3. System Specifications 4. Operating Principles 5. Control and Software 6. Method Development 7. Maintenance Procedures 8. Troubleshooting Guide 9. Regulatory Compliance 10. Technical Support

System Overview

Key components: Binary pump, autosampler, column compartment, multiple detectors (DAD, FLD, RID), and Chromeleon software.

Component	Description
Binary Pump	High-pressure mixing with flow rates from 0.001 to 10 mL/min
Autosampler	Temperature-controlled with 96-well plate capacity
Column Compartment	Temperature range 5-110°C with active preheating
DAD Detector	Wavelength range 190-800 nm
FLD Detector	Programmable excitation and emission wavelengths
RID Detector	Refractive index detection
Software	Chromeleon Chromatography Data System
Solvent Cabinet	4-channel degasser with 2L bottle capacity
Waste Container	2L capacity with level sensor
Power Supply	100-240V AC, 50/60Hz

Installation and Setup

Unpack and inspect all components. Ensure proper laboratory environment.

Place system on stable bench: Allow 15cm clearance on all sides for ventilation.
Connect solvent lines: Prime pump with appropriate solvents.
Install columns: Follow manufacturer's instructions for column installation.
Connect to power: Use provided power cables and ensure proper grounding.
Install Chromeleon software: Follow installation wizard instructions.

WARNING! System contains high-pressure components. Wear appropriate PPE during installation.

System Specifications

Pressure Range: Up to 1000 bar (14,500 psi)
Flow Rate Range: 0.001 to 10 mL/min
Injection Volume: 0.1 to 100 μL
Temperature Control: 5-110°C (±0.5°C)
Detection Wavelengths: 190-800 nm (DAD)
Carryover: < 0.05%
Data Rate: Up to 200 Hz
Dimensions: 65 cm W × 55 cm D × 50 cm H
Weight: 45 kg
Power Requirements: 100-240V AC, 50/60Hz, 500W

Operating Principles

The Ultimate 3000 operates on UHPLC principles with sub-2μm particle columns for high resolution and fast separations.

Pump Operation: Dual piston design with active inlet valve and electronic pressure control.

Autosampler: Uses needle-in-loop design for minimal carryover and precise injections.

Detection: Multiple detection options including diode array, fluorescence, and refractive index.

Control and Software

System controlled via Chromeleon Chromatography Data System.

Method Development: Create and optimize separation methods
Sequence Setup: Program sample sequences with injection parameters
Data Acquisition: Real-time monitoring and data collection
Data Analysis: Peak integration, quantification, and reporting
System Monitoring: Pressure, temperature, and flow monitoring

Interface: Touch-screen display and computer software control.

Method Development

Develop and optimize UHPLC methods for various applications.

Column Selection: Choose appropriate column chemistry and dimensions
Mobile Phase: Optimize solvent composition and gradient
Flow Rate: Adjust for resolution and analysis time balance
Temperature: Optimize column temperature for separation
Detection: Select appropriate detection wavelength or mode
Validation: Validate method performance parameters

Tip: Start with generic gradients and optimize for specific analytes.

Maintenance Procedures

Regular maintenance ensures optimal performance and extends system lifetime.

Daily: Check pressure readings and system leaks
Weekly: Clean autosampler needle and wash ports
Monthly: Replace pump seals and check detector lamps
Quarterly: Perform system performance qualification
Annually: Complete system calibration and certification

CAUTION! Always depressurize system before performing maintenance.

Troubleshooting Guide

Symptom	Possible Cause	Corrective Action
High pressure	Column blockage	Replace column; flush system
Pressure fluctuations	Air bubbles	Purge system; degas solvents
Peak tailing	Column degradation	Replace column; check pH
Retention time shift	Temperature variation	Check column thermostat
No injection	Autosampler error	Check needle position; clean ports
Baseline noise	Detector lamp aging	Replace lamp; check mobile phase
Low sensitivity	Flow cell contamination	Clean flow cell; check detector settings

Regulatory Compliance

CE marked; complies with IVDR 2017/746 for in-vitro diagnostics. FDA 21 CFR Part 11 compliant software options available. RoHS and WEEE compliant.

Quality Standards: ISO 9001, ISO 13485 certified manufacturing.

Technical Support

For technical assistance, service, or training:

Phone Support: 1-800-723-1161
Email Support: [email protected]
Online Resources: www.thermofisher.com/ultimate3000
Service Contracts: Available for preventive maintenance and repairs
Training: On-site and online training courses available

Emergency Service: Available 24/7 for critical system failures.