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Course, academic year 2023/2024
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Instrumental Methods - GAF301
Title: Instrumental Methods
Guaranteed by: Department of Analytical Chemistry (16-16140)
Faculty: Faculty of Pharmacy in Hradec Králové
Actual: from 2018
Semester: summer
Points: 0
E-Credits: 6
Examination process: summer s.:combined
Hours per week, examination: summer s.:14/56, C+Ex [HS]
Capacity: unlimited / unlimited (unknown)
Min. number of students: unlimited
4EU+: no
Virtual mobility / capacity: no
Key competences:  
State of the course: taught
Language: English
Teaching methods: full-time
Teaching methods: full-time
Note: deregister from the exam date if a requisite was not fulfilled
course can be enrolled in outside the study plan
enabled for web enrollment
Guarantor: doc. RNDr. Miroslav Polášek, CSc.
Comes under: 2.ročník 2023/24 Pharmacy (EN19)
Co-requisite : GAF300
Interchangeability : GAF274
Is co-requisite for: GAF314, GAF302, GAF313
Is pre-requisite for: GAF385, GAF306
Annotation -
Last update: doc. RNDr. Miroslav Polášek, CSc. (21.09.2023)
The aim of the subject is to provide the students with firm and sufficiently wide theoretical and methodological fundamentals for pursuing (in a rational way) instrumental analysis of inorganic and organic substances. The teaching is focused on the explanation of theoretical aspects of instrumental methods and putting the accent on problem-solving tuition. In practical training individual approach of students to solving analytical problems is required.
Course completion requirements -
Last update: doc. RNDr. Miroslav Polášek, CSc. (21.09.2023)

Requirements for credit and exam are based on:

Practical course

  1. Participation in all practical classes – (missing practical classes must be revisited or it can be substituted if permitted by the teacher)
  2. Submission of all reports of practical exercises

  3. Minimum 66% of points for correct results in practical tasks and for correct answers in the monitoring and credit tests



  1. The exam from Instrumental methods is organized in terms presented in the Study information system, the time of exam is always specified. Simple calculator is needed for written exam. Evaluation is carried out directly after evaluation of the test.

  2. The exam is organized together with the subject Analytical chemistry.

  3. For the exam, credits from both practical classes are needed.

  4. Exam organization:

The first term – written test + possibility of oral examination

The second term - written test + oral examination

The third term – oral examination

  1. Exam scope is specified in the Study materials – Study KACH – Instrumental methods

Literature -
Last update: doc. RNDr. Miroslav Polášek, CSc. (21.09.2023)


  • . 1079. In Robinson, James W., Frame, Eileen M. Skelly Frame, George M.. Undergraduate instrumental analysis . New York: M. Dekker, 2005, s. -. ISBN 0-8247-5359-3..

Syllabus -
Last update: doc. RNDr. Miroslav Polášek, CSc. (21.09.2023)

Syllabus – Instrumental methods

Introduction to instrumental methods

  • Instrumental methods, their advantages, drawbacks, classification of instrumental methods, their application in qualitative and quantitative analysis.

Optical methods

  • Basic optical methods, interaction of radiation with particulate matter, classification of optical methods, their principles and scope, fundamentals of luminescence analysis, practical use of optical methods.

  • Spectrophotometry in UV spectral region, types of electron transfer, light absorption of organic compounds, absorption spectrum, absorption band, absorption coefficient, double-beam spectrophotometers, diode-array detector, analytical applications – identification, purity of substances, quantitative assay, calibration curve, analysis of multi-component systems.

  • Infra-red spectrophotometry, types of vibrations, characteristic vibrations of functional groups, absorption spectrum, absorption bands, instrumentation, measuring techniques, analytical applications – interpretation of IR spectra, identification, purity, structure of organic compounds.

  • Flame photometry, refractometry, polarimetry, principles, and analytical applications.

Electroanalytical methods

  • Basic electroanalytical methods, their classification and principles. Potentiometry, principles, instrumentation, selection of electrode system, ion-selective electrodes, potentiometric titrations and their evaluation, analytical applications. Conductimetry, principles, conductimetric titrations and other applications.

  • Voltammetry and polarography, principles, instrumentation, qualitative and quantitative analysis.

Radioanalytical methods

  • Basic radioanalytical methods, their classification, principles, applications.

Separation methods

  • Basic separation methods, classification according to the principle of separation. Chromatographic methods and their classification.

  • Gas chromatography, instrumentation, analytical applications.

  • Liquid chromatography in planar and packed-bed arrangement. High-performance liquid chromatography (HPLC). Thin-layer chromatography (TLC, HPTLC). Paper chromatography. Instrumentation and analytical applications.



    1)    Acidimetric titration with potentiometric end-point indication. Calibration of the electrode system. Determination of sodium tetraborate in solid sample. Graphical evaluation and end-point estimation according to Hahn method. Graphical evaluation and end-point estimation according to Hahn method.

    2)    Dichromatometric titration with potentiometric end-point indication. Determination of % content of ferrous salt in a solid sample.

    3)    Argentometric titration with potentiometric end-point indication. Determination of Cl-, Br- and I- in a mixture.

    4)    Spectrophotometric assay of sulphosalicylic acid in a solution. Finding absorption maximum, optimum amount of reagent. Mesurement of calibration curve, graphical and statistical evaluation.

    5)    Spectrophotometric assay of cupric salts in the form of complex with EDTA. Preparation of calibration solutions, evaluation of calibration curve. Determination of Cu2+ content in a solid sample.

    6)    Extraction - spectrophotometric assay of ferric salts. Formation of the complex with 8-quinolinol and its extraction into chloroform. Preparation of calibration solutions, evaluation of calibration curve. Determination of Fe(III) content in a test solution.

    7)    Identification of organic compound by the measurement of the melting point of derivative.

    8)    Thin-layer chromatography (TLC). Derivatization for separation purposes, detection, evaluation of chromatograms. Identification of an unknown organic compound.

    9)    UV spectrophotometry. Measurement of UV spectra by an UV spectrophotometer with a diode array detector and their evaluation for identification purposes. Identification of an unknown drug.

    10)  Raman spectroscopy. Measurement and evaluation of  Raman spectra for identification purposes. Identification of unknown drug.

    11)  Quantitative elemental analysis (EA), UV and IR spectrometry. Identification of an unknown organic compound with use of EA, UV and IR data.

    12)  Gas chromatography. Measurement and evaluation of chromatograms. GLC identification of organic compounds. Determination of pesticides permethrin and fenoxycarb in a veterinary preparation.

    13)  Polarimetric analysis. Quantitative polarimetric analysis of a sample containing unknown amount of glucose, sucrose or lactose.

    14)  High performance liquid chromatography (HPLC) I. Measurement and evaluation of chromatograms. Determination of sodium diclofenac in pharmaceutical preparations. ). Determination of hydrocortisone acetate in a test solution.

    15)  High performance liquid chromatography (HPLC) II. Optimization of the mobile phase for the UHPLC separation of a mixture of acidic, neutral and basic compounds.

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