how to develop a method with
ChromSword® Auto?
The analytical scientist’s burden of profiling main compounds,
impurities and/or degradation products can be quite
challenging. Consequently, developing chromatographic methods to separate
impurities requires substantial time and effort. Thus, there is a tremendous
need to employ a comprehensive method development strategy to meet the
challenges of high throughput and rapid turn-round time.
key steps in chromatographic method
development
screening - selection of initial conditions
The first step of chromatographic methods development is identifying
the most promising column, buffer, and organic solvent that will be used to
separate impurities and/or degradation products in a particular sample.
Selecting the initial conditions based on experience, intuition and availability
of materials such as columns, solvent and buffers relies heavily on the users
experience.
run of experiments and evaluation of results
Once initial starting conditions have been identified, the next step is
to run a sample which contains representative impurities and/or degradation
products to be separated.
documentation of the method
Once a method is identified that satisfies the goals intended, the
scientist then documents the method in either an SOP or in a business specified
document that contains details about the method including data acquisition and
analysis.
Currently, the aforementioned steps in chromatographic method development are done separately using separate islands of technology.
With ChromSword®Auto it is possible to apply a streamlined
strategy for method development in which all the
screening, optimization, and documentation steps in the method
development process are automated and performed under one software platform.
Selecting the most appropriate starting conditions (with ChromSword RAPID
screening or USER defined screening) for method optimization leading to enhanced
methods that exhibit maximum resolution of components, symmetrical peak shape,
and minimum run time is of great importance during the development process (with
ChromSword FINE optimization).
ChromSword®Auto options
The ChromSword® Auto software package consists of on-line and off-line modes of optimization.
The off-line mode prompts the user for compound structures to be modeled for separation. Based on the functional groups of the structure(s) input by the user, the software can model chromatographic retention behavior with changes in organic modifier composition. Additionally, in the off-line mode, the software can use chromatographic data obtained from at least two initial experiments to predict optimum separation condition.
ChromSword® Auto performs method development in
an automated mode. ChromSword®
Auto takes control of the chromatographic data system and HPLC hardware.
The software serves as a real-time expert system. ChromSword®
Auto simulates the action of a human chromatographer in a method
development procedure such as evaluating results, solving problems (subtasks)
and making decisions. The user can specify method
goals such as attaining certain resolution between peaks of interest or
maximum possible resolution for all peaks in a mixture. After specifying
desired separation goals, the system conducts method development
independent of user involvement.
ChromSword® Auto offers two types of automated method development:
1) SCREENING of initial conditions (ChromSword® Auto supports column and solvent switching valves)
- RAPID (intelligent) method development or USER defined method development are usually used for SCREENING of initial conditions (e.g. column, organic solvent, aqueous solvent).
- The RAPID and USER defined method development options offer a crude method that may still be acceptable (sample profiling).
- The RAPID method development performs three gradient attempts at separating all the components in the given sample using a ChromSword® algorithm. The advantage of the rapid method development mode is that it is conducted in a 2 or 3h timeframe.
- The chromatographer determines specific screening conditions in the USER defined mode depending on experience. The user sets up a sequence of actions for the software to complete. The software controls the hardware to execute simple defined instructions.
- Selection of the best column/mobile phase system for a sample is subjective, but straightforward. The best column/mobile phase combination is based on the number of peaks generated, their peak shape, and total run time.
2) FINE (FULL) method optimization
- The best column, buffer, and organic solvent combination derived from the initial screening can be used in the FINE optimization, and the software takes over the rest of the experiment. The full optimization mode provides the best method of separation for a sample.
- The fully optimized method development option studies the elution of sample components in increments of percent organic and ascertains the total number of component peaks. The system either elects the best percent organic for an isocratic separation; or it performs various gradient runs before finding one or possibly more separation solutions to choose from.
- The final optimized method is documented in a MSWord® document containing all of the method development history performed by ChromSword®-Auto.
- Additionally, the software automatically
equilibrates columns prior to sample injection and
washes columns after use.
what sets ChromSword® Auto apart?
- Ability to optimize the separation of a sample mixture with
no user information provided regarding the number of components or structure of components in the sample. - Mixtures with unknown amounts of impurities and/or degradation products can be fully separated with little prior work done by the chromatographer. ChromSword®Auto is a software package of choice for conducting automated HPLC-methods development particularly with the burden of the quite challenging impurity identification and quantitation.
- ChromSword®Auto, paired with the column/solvent screen, offers a consistently faster and more efficient method development option than traditional manual method development.
- ChromSword®Auto allows for step-by-step documentation of the method development process
that is reliable, thorough, and critical for regulatory review.
