A CE-based analytical system configured optimally for the methods development process, where a focus is placed on spectral analysis and the automation of separation strategies. The system includes a P/ACE MDQ configured with both a photo diode array and selectable-wavelength UV/Vis (200, 214, 254 and 280 nm filters included) detector, UV source optics, temperature-controlled sample storage module and 32 Karat™ Software configured on an IBM personal computer. Installation Qualification, Operation Qualification 1 (OQ1) and documentation to aid in software validation is also included.
Multi-format Sample Introduction
Automated Sample Introduction directly from 96 well plates, 2ml autosampler vials, PCR vials and 0.5 ml tubes. Automation is certainly a means by which many laboratory bottle necks may be reduced. One of the major bottle necks is in sample handling. The use of 96 well plates as a sampling vessel not only increases system capacity but allows compatibility with automated sample preparation devices which utilize the 96 well plate format. Additionally, 2 ml vials (which are autosampler industry standards) may be used; or sampling can be achieved directly from PCR vials and microfuge tubes.
Methods Development with 36 Position
Methods Development through a 36 position buffer pair array independent from sampling vials. When optimizing a methods development strategy or scouting for successful separation conditions; the buffer pH, concentration and ion type plays a dramatic role in the process. An array of 36 buffer pairs independent from the samples allows your methods development strategies to be automated while maintaining system sample capacity. To ensure proper separation one must match both the inlet and outlet buffers requiring the necessity of buffer pairs.
Multiple Separation Modes
Modes of separation include: Voltage, Current, Power, Pressure and Vacuum. All electrophoretic separations allow the programming of both step and linear gradients along with the simultaneous application of pressure or vacuum on both ends of the capillary. Voltage gradient programming is beneficial for gel-sieving techniques improving separations over a wide range of fragment sizes. This is especially critical for the separation of nucleic acids. The application of simultaneous voltage and pressure is beneficial for the high efficiency mobilization of proteins during iso-electric focusing as well as detecting material not migrated off the capillary. Also, several CEC methods have specified the use of simultaneous pressure applied to both inlet and outlet reservoir. This system is compatible with those methods.
High Sensitivity UV Detector
Modular High Sensitivity Selectable Wavelength UV Detector utilizing up to seven filters in an assembly which allows wavelength changes during a run. Detector sensitivity in capillary electrophoresis is critical since small mass loads are introduced and small detection pathlengths are used (governed by the capillary dimensions i.e. 25 - 200 um). Fixed wavelength detection in a filter wheel assembly maximizes detector sensitivity yet still allows wavelength changes during a run. In a Q.C. or regulated environment it is beneficial to "lock-down" a method once it is developed in order to retain the integrity of the data. The P/ACE™ high sensitivity UV detector may be configured with a "single" wavelength from a filter traceable to NIST making both the instrument certification and method validation much simpler. Additionally the system employs complete fiber-optic technology bypassing the necessity of an optical bench. This lowers the noise on the detection system (by removing reflective surfaces where energy loss can occur) and simplifies the system making it more rugged for routine use. Diode array is an essential detection tool for methods development work, but will reduce the lifetime of a "coated" capillary. Therefore, once a method is developed and is in routine use it is beneficial to switch to a UV detector where the capillaries will last longer.
Dual Wavelength LIF Option
Optional - Dual Wavelength Laser-Induced Fluorescence (LIF) Detection System. High sensitivity laser-induced fluorescence detection extends the capabilities of capillary electrophoresis for the analysis of carbohydrates and nucleic acids and other compounds which either naturally fluorescence or that can be made to fluoresce through sample derivitization. LIF provides both selectivity and sensitivity to any analysis and typically yields 500 x better sensitivity than UV for compounds which fluoresce. The P/ACE™ MDQ LIF detector is modular and will allow simultaneous excitation using two laser sources. And with the second channel option installed, it will collect simultaneously emissions from two different wavelengths. This is important where you wish to run your sample and standard together in the same run, using two "colors" to discriminate between the two species.
GLP/CGMP Compliance Features
32 Karat Software includes the following GLP/CGMP compliance features: 1) Original method is stored with sample data. 2) Raw data cannot be overwritten. 3) The instrument logs all major events 4) Software security includes password protection with multiple levels of operator access. Many laboratories are mandated by law to follow current good manufacturing practices (CGMP) and good laboratory practices (GLP) the 32 Karat Software has many features which are geared towards the maintenance of data integrity. With the GLP flag turned on, sample data and methods parameters cannot be overwritten or overridden, ensuring the data that was generated was as described. To track instrument operation, all events including errors are logged onto the system. To provide security of operation users may be coded to different security levels with selected access to: system configuration, methods programming, data reprocessing, specific instrument operation and data comparison. All of these features can be used to "lock down" the system for regulated use or logged in as a "toggled off" state in laboratories where flexibility of operation is the critical issue.
Specialized CE Calculations
32 Karat Software provides specialized CE calculations including: Apparent Mobility, Mobility, Corrected Peak Area, Molecular weight and Iso-electric Point. The software also includes system suitability which will use these calculations in actuating suitability decisions. The suitability algorithms are capable of triggering re-injections, methods pausing, methods aborting or running shutdown methodology upon failure. Capillary electrophoresis requires specialized data handling. Mobility calculations establish the identity of a compound independent from chang