Validation of liquid chromatography mass of liquid chromatography mass spectrometry (LC-MS) methods (analytical chemistry) course This is a practice-oriented on-line course on validation of ... Table of contents Course introduction 1. Validation: General 1.1. The

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  • Validation of liquid chromatography mass spectrometry (LC-MS) methods(analytical chemistry) course

    This is a practice-oriented on-line course on validation of analytical methods, specifically using LC-MS as technique.

  • Table of contents

    Course introduction

    1. Validation: General

    1.1. The purpose of validation

    1.2 Carrying out validation

    1.3. Scope of validation

    2. Selectivity and identity conrmation

    2.1. Selectivity: LC selectivity

    2.2. Selectivity: detector-side (MS) selectivity

    2.3. Selectivity examples

    2.4. Introduction to identity conrmation

    2.5. Identity conrmation: retention time

    2.6. Identity conrmation by MS

    2.7. Identity conrmation examples

    3. Linearity of signal, linear range, sensitivity

    3.1. Linearity

    3.2. Experiment setup and evaluation of the data

    3.3. Estimating the linear range

    3.4. Sensitivity

    4. Precision

    4.1. Repeatability, intermediate precision and reproducibility

    4.2. Calculation of precision

    4.3. Acceptance criteria

    4.4. Examples

    5. Trueness

    5.1 Bias and its constituents

    5.2 Determining bias in practice

    5.3 Qualitative estimation of matrix eect

    5.4 Quantitative estimation of matrix eect, recovery and process

    5.4 Quantitative estimation of matrix eect, recovery and processeciency6. Precision and trueness: some additional aspects

    6.1. Choosing sample type, concentration levels, replicates and timerange for precision6.2. Obtaining reference value for bias calculation

    6.3 Avoiding/minimizing bias and correcting for bias

    7. Accuracy

    8. Stability

    8.1 Dierent types of stability

    8.2 Examples of unstable analytes

    8.3 Evaluation of stability

    8.4 Aspects to be considered if analytes are unstable

    9. LoD and LoQ

    9.1. Denitions and important aspects

    9.2. Decision limit and Detection capability

    9.3. Estimation of LoD

    9.4. Estimation of LoQ

    10. Ruggedness, robustness

    10.1 Robustness and ruggedness relation to LC-MS methoddevelopment10.2 Matrix robustness

    10.3 Dierent ways to evaluate robustness

    10.4 Experimental design

    VaLChrom

    Acknowledgements

    References

    Glossary

    What our participants say?

  • Course introduction

    Course introduction

    http://www.uttv.ee/naita?id=23245

    https://www.youtube.com/watch?v=jbdA8PnPdLY

    Short description of the courseThis course LC-MS Method Validation is a practice-oriented on-line course on validation of analytical methods, specifically using LC-MS astechnique. The course introduces the main concepts and mathematical apparatus of validation, covers the most important method performanceparameters and ways of estimating them. The course is largely based on the recently published two-part tutorial review:

    Tutorial review on validation of liquid chromatographymass spectrometry methods: Part I. A. Kruve, R. Rebane, K. Kipper, M.-L. Oldekop,H. Evard, K. Herodes, P. Ravio, I. Leito. Anal. Chim. Acta 2015, 870, 29-44

    Tutorial review on validation of liquid chromatographymass spectrometry methods: Part II. A. Kruve, R. Rebane, K. Kipper, M.-L.Oldekop, H. Evard, K. Herodes, P. Ravio, I. Leito. Anal. Chim. Acta 2015, 870, 8-28

    The course contains lectures, practical exercises and numerous tests for self-testing. In spite of being introductory, the course intends to offersufficient knowledge and skills for carrying out validation for most of the common LC-MS analyses in routine laboratory environment. The real-life analysis situations for which there are either examples or self-tests are for example pesticide analyses in fruits and vegetables,perfluoroalkyl acids in water, antibiotics in blood serum, glyphosate and AMPA in surface water, etc. It is important to stress, that for successfulvalidation experience (both in analytical chemistry as such and also specifically in validation) is crucial and this can be acquired only throughpractice.

    Required preliminary knowledgeIntroductory level knowledge of analytical chemistry, as well as liquid chromatography and mass spectrometry is required. More advancedknowledge of analytical chemistry and introductory knowledge of mathematical statistics is an advantage.

    Study outcomesThe participant who has successfully passed the course knows:

    the main performance parameters of analytical methods, what they show and which of them are particularly important in differentsituations;

    the main mathematical concepts and tools in method validation;

    the main approaches for evaluation of the performance parameters in the case of LC-MS analysis.

    The participant who has successfully passed the course is able to:

    decide what data are needed for evaluating the different method performance parameters, understand the meaning of the available dataand decide whether the available data are suitable and sufficient;

    select the validation approach and design the experiments for obtaining suitable data;

    quantify the relevant performance parameters using the available data and assess whether the obtained values are realistic;

    assess the fitness of the method for the intended purpose based on the values of the evaluated performance parameters.

    Organization of the course materialThe course is organized in 11 thematic sections, of which most are in turn split into smaller subsections. The following parts are found in thesections:

    1. The sections (and also many subsections) start with a brief introduction stating the main topic(s) and study outcomes of the section.2. The main topic of the respective section is explained in one or several short video lectures. The videos are by default streamed in high

    quality from the UT Video server, which needs quite good Internet connection. If you have slow Internet connection we recommendwatching the videos from YouTube, using the links below video screens.

    3. The lecture(s) is(are) complemented by a textual part. The textual part is in most cases meant to complement, not substitute the lecture(although in some cases the contents of the lecture are also repeated in some extent). It rather gives additional explanations andaddresses some additional topics that were not covered by the lecture.

    4. The learners are introduced to the software, which helps to carry out validation of chromatographic methods. Learners areencouraged to use VaLChrom throughout the whole course.

    http://www.uttv.ee/naita?id=23245https://www.youtube.com/watch?v=jbdA8PnPdLYhttp://dx.doi.org/10.1016/j.aca.2015.02.017http://dx.doi.org/10.1016/j.aca.2015.02.016https://sisu.ut.ee/lcms_method_validation/11-valchromhttps://sisu.ut.ee/lcms_method_validation/11-valchrom

  • 5. Most sections end with a self-test, which enables to assess the acquired knowledge and skills. The self-tests contain questions, as well ascalculation problems. The self-tests are on one hand meant for the participants to monitor their progress. On the other hand, however,they also promote thinking and provide (by the feedback of the questions) additional knowledge about validation in different practicalsituations. So, the self-tests are an intrinsic component of the course and participants are strongly recommended to take all of them.

    Self-testingThroughout the course there are numerous self-tests for enabling the participant to test his/her knowledge and skills in specific topics. Eachtest is graded as a percentage (100% corresponding to correctly answering all questions and correctly solving all problems).

    Feedback is given as:

    Correct answer, correctly recognised and marked by the student.

    Correct answer, not recognised and not marked by the student.

    Incorrect answer, however, considered correct by the student.

    Explanatory feedback is displayed when wrong answer is selected. All self-tests can be taken as many times as needed and the success oftaking these tests will not influence the final grade. We recommend that you take all the tests and work with them until you achieve score100% and only then move to next topic.

    Terminology and denitionsAt the end of the thematic section there is Glossary, which gives definitions and/or explanations of the terms used in the course. Whereverpossible, the used terminology adheres to the one used in the Tutorial review on validation of liquid chromatographymass spectrometrymethods (Literature sources 1 and 2).

    Literature sourcesThe main literature sources of the course are here:

    1. Tutorial review on validation of liquid chromatographymass spectrometry methods: Part I. A. Kruve, R. Rebane, K. Kipper, M.-L. Oldekop,H. Evard, K. Herodes, P. Ravio, I. Leito. Anal. Chim. Acta 2015, 870, 29-44

    2. Tutorial review on validation of liquid chromatographymass spectrometry methods: Part II. A. Kruve, R. Rebane, K. Kipper, M.-L.Oldekop, H. Evard, K. Herodes, P. Ravio, I. Leito. Anal. Chim. Acta 2015, 870, 8-28

    3. B. Magnusson and U. rnemark (eds.) Eurachem Guide: The Fitness for Purpose of Analytical Methods - A Laboratory Guide to MethodValidation and Related Topics, (2nd ed. 2014).

    4. SANTE/11945/2015 (Until 01.01.2016: SANCO/12571/2013), Guidance document on analytical quality control and method validationprocedures for pesticides residues analysis in food and feed. European Commission, 2015.

    5. European Commission Decision 2002/657/EC implementing Council Directive 96/23/EC concerning the performance of analytical methodsand the interpretation of results, Off. J. Eur. Commun. L221 (2002) 8-36.

    6. JCGM 200:2008, International vocabulary of metrology Basic and general concepts and associated terms (VIM), 3rd edition. BIPM, IEC,IFCC, ILAC, ISO, IUPAC, IUPAP and OIML, 2008.

    This list of mai