SFKKA9021U Principles and Practice of Bioanalysis

One part of the course will cover the separation techniques GC, HPLC and CE and their coupling with spectroscopic detection principles such as UV-VIS, fluorescence, MS og NMR. Focus will be on separation mechanisms and selective detection in order to obtain reliable quantitative bioanalytical data.
Another part of the course will cover sample preparation techniques including liquid-liquid extraction, solid phase extraction and protein precipitation. The advantages and limitations of each technique will be discussed in order to enable the student to make the best choice of analytical method taking into account the nature and concentration of the analyte, the matrix and the accuracy, precision and time of analysis.The course will cover the complete bioanalytical process starting with sampling, sample conditioning, sample treatment / preparation, instrumental analysis, derivatisation / structure modification, detection / identification, data handling, data interpretation /evaluation, decision-making and troubleshooting.
The most relevant course topics will be:

• Sampling of biological materials (e.g. representative samples, homogeneity, stability, traceability)
• Types of samples needed for different investigations
• Physicochemical composition of biological matrices (e.g., drug-protein binding, matrix interferences, enzymatic effects)
• Matrix effects during instrumental analysis (e.g., clogging/blocking effects, ion-suppression/enhancement effects, robustness /ruggedness)
• Physicochemical properties of analytes and matrices with respect to chemical analysis (e.g., acid-base properties, solubility, polarity)
• Sample handling (e.g. initial sample treatment, storage, stability)
• Sample clean-up and preparation (off-line, at-line, on-line, in-line approaches to improve selectivity and or sensitivity or to perform a phase transfer. In addition to the traditional sample preparation techniques, new developments with respect to selective sorbents, miniaturized approaches and high-throughput robotic devices will be included);
• Analytical separation methodologies, both traditional (e.g., LC, GC, CE, SFC) and advanced (e.g., UPLC, HPLC, assay-based techniques) separation techniques will be discussed)
• Instrumental identification and detection approaches, both traditional (e.g., absorbance, fluorescence, diode array, IR, MS) and advanced (e.g., MS/MS, MSⁿ, NMR, biochemical) detection/identification techniques will be discussed
• Bio-pharmaceutical analysis (e.g., amino acids, peptides/proteins, nucleotides)
• Industrial aspects (e.g., automation, robotics, high-throughput devices)
• Forensic application will be demonstrated
• Validation issues (e.g., bio-statistics, data-acquisition/evaluation, chemometrics)

Robustness/ruggedness testing (e.g. techniques and approaches to guarantee the quality of the developed methodologies), troubleshooting, matrix. The first half of the course will cover the separation techniques GC, HPLC og CE and their coupling with spectroscopic detection principles such as UV-VIS, fluorescence, MS and NMR. Focus will be on separation mechanisms and selective detection in order to obtain reliable quantitative bioanalytical data.

The second half of the course will cover sample preparation techniques including liquid-liquid extraction, solid phase extraction and protein precipitation. The advantages and limitations of each technique will be discussed in order to be able to enable the student to make the best choice of analysis method taking the nature of the analyte, the concentration af the analyte and the matrix as well as the accuracy, precision and the time of analysis.