Department of Antibiotics and Microbiology (MA)

Personnel and structure:

Head of the Department: Stefan Tyski, Professor
 e-mail: 

Secretary: Grażyna Stańczuk
phone: +48 022 841 36 83
phone/fax: +48 022 851 52 15
fax +48 022 841 06 52
 e-mail: 

Biology and Standardisation of Antibiotics Lab
Head: Wanda Grzybowska Ph.D; tel. 022 841 36 83 ex. 488
Ewa Bocian M.Sc.; tel. 022 841 36 83 ex. 399
Bożena Bukowska M. Sc. tel. 022 841 36 83 ex. 487

Physico-Chemical Properties of Antibiotics Lab
Head: Genowefa Pajchel Ph.D; tel. 022 841 36 83 ex. 476
Katarzyna Michalska M.Sc.; tel. 022 841 36 83 ex. 461
Rafał German M. Sc.; tel. 022 841 36 83 ex. 461
Bogumiła Andrzejak; tel. 022 841 36 83 ex. 461
Małgorzata Kaliszewska; tel. 022 841 36 83 ex. 462
Joanna Wajszczyk; tel. 022 841 36 83 ex. 461

General Microbiology Lab
Head: Tomasz Zaręba V.D.; tel. 022 841 36 83 ex. 253
Hanna Kruszewska M.Sc.; tel. 022 841 36 83 ex. 480
Agnieszka Mrówka tel. 022 841 36 83 ex. 174
Ewa Wasińska; tel. 022 841 36 83 ex. 174
Zofia Piekot; tel. 022 841 36 83 ex. 491
Krystyna Umięcka; tel. 022 841 36 83 ex. 492

Activities:

Control of medicinal products:

Tests performed in the department:
  • Total pharmaceutical analysis - chemical and microbiological assays of all pharmaceutical dosage forms of antibiotics, in mono or multi-antibiotic preparations, for human and veterinary use,
  • Sterility and microbial purity testing of all medicinal products supplied from other departments of the Institute,
  • Biocidal activity of antiseptics:
Collaborative trials for proficiency testing, e.g. the assignment of an assay value for antibiotics CRS (certified reference standard), organised by EDQM.

Physico-chemical analysis
Complex physico-chemical analysis of antibiotic drugs and antibiotic substances, including: identification, purity and content are performed by different methods (e.g. TLC, UV-VIS spectrophotometry, titrations and HPLC as well as microbiological assays). Recently capillary electrophoresis was introduced, mainly in beta-lactam antibiotics evaluation - several scientific papers (including Journal of Chromatography A and B editions and Journal of Pharmaceutical and Biomedical Analysis) were also published.
This lab is equipped with four HPLC systems (Shimadzu), CE apparatus (Waters) and spectrophotometer (Shimadzu).

Microbiological analysis
Pharmacopoeial turbidimetric and diffusion cylinder-plate methods are used for estimation of biological activity and content of antibiotics.
Dissolution test is performed with the usage of Hanson Research Dissolution Test system.
Released antibiotics are determined by spectrophotometrical, microbiological or HPLC methods.
Efficacy of antiseptics including bactericidal and fungicidal activity, according to EN standards are here tested, as well as efficacy of antimicrobial preservation of medicinal products.
Some scientific investigations of antibiotics interaction with the use of standard as well as clinical bacterial isolates by checkerboard, time-kill methods and E-tests have been performed. Antiseptics efficacy against clinical bacterial strains is also analysed.

Pharmaceutical microbiology assays
Sterility or microbial purity of all kinds of drugs and some medical devices supplied from other institute's departments are investigated by pharmacopoeial plate count or membrane filtration methods. If necessary, investigations according to EN standards are applied. Steritest system (Millipore) is used for analysis of liquids and ATB system (BioMerieux) is applied for isolate identification. Some conductivity analyses of preservatives efficacy were performed by the Bactometer system (BioMerieux) in comparison to pharmacopoeial assays, in order to speed up obtaining of the results.
Environmental microbiological monitoring of classified areas not only in the department but also in other clean rooms of the Institute is also performed by the lab staff. Microbial contamination of the air is analysed by MAS 100 apparatus (Merck).

Evaluation of analytical documentation of antibiotic medicinal products, which are submitted for registration on Polish market Quality Assurance

  • Participation in Proficiency Testing Studies organised by European Directorate for the Quality of Medicines and European Pharmacopoeia.
  • Creating of Standard Operation Procedures.
  • Microbiological monitoring of drug testing environment.

Educational activity

  • Lectures and individual training concerning: pharmaceutical microbiology, analytical techniques, antiseptics and antisepsis;
  • Summer training for students from Warsaw University, Warsaw Agricultural University and Medical University of Warsaw;
  • Supervision over M.Sc. and Ph.D. thesis.

Research

  • Application of new, alternative techniques for pharmaceutical products testing;
  • Susceptibility testing of clinical bacterial isolates to antimicrobial agents;
  • Antimicrobial activity testing of antiseptics;
  • Antimicrobial activity testing of "non-antibiotics";
  • Microbiological monitoring of medicinal products and medical devices (sterility or purity)

Research projects:

Scientific grants of the Committee for Scientific Research - from 2000:
  • Investigation and characterisation of N. meningitidis, H. influenzae and S. pneumoniae carrier strains isolated from nasopharynx of healthy children and adults.
  • Phenotypic and genotypic characteristics of Neisseria meningitidis strains isolated in Poland.
  • Determination of beta-lactam antibiotics in selected medicinal products by capillary electrophoresis in comparison with HPLC.
  • Analysis of fungicidal activity of disinfectants and antiseptics - essential tools in eradication of fungal hospital infections.
  • Optymalizaction of capillary electrophoresis assays applied for analyzis of inmedicinal produkt containing antibiotics. 2007-2008.

Cooperation

  • European Directorate for the Quality of Medicines (EDQM) - PTS studies.
  • European Pharmacopoeia - establishing of strength of CRS (Certified Reference Standards) for antibiotics.
  • Polish Pharmacopoeia - the head of the department leads the Antibiotics and Biological Methods Commission of the Pharmcopoeia and there are 3 members of the Commission among the scientists of the department.
    Universities:
    • Faculty of Biology - Warsaw University,
    • Faculty of Biology, Faculty of Biotechnology - Warsaw Agricultural University,
    • Faculty of Pharmacy, Faculty of Medicine - Medical University of Warsaw,
    • Faculty of Biotechnology - Warsaw Technical University,
      (Students training: summer schools, preparing of M.Sc. thesis)
  • National Institute of Hygiene,
  • Military Institute of Hygiene and Epidemiology,
  • Institute of Oncology,
  • Polish Standardization Committee,
  • Polish Pharmacopoeia,
  • Soderborg Sygehus, Soderborg, Denmark.

Services:

  • Testing of medicinal products and medical devices concerning sterility or microbial purity.
  • Determination of microbial bioburden level.
  • Testing of antimicrobial activity of raw materials and final products of natural origin.
  • Microbiological control of environment.
  • Physico-chemical analysis of medicinal products.
  • Standardization of antibiotics by biological methods.
  • Determination of antibiotics content in food.
  • Testing of antimicrobial activity of antiseptics, according to PN, EN, ISO standards.
  • Efficacy of antimicrobial preservation testing
  • Training concerning pharmaceutical microbiology,
  • Validation of selected analytical methods

Equipment

  • HPLC systems (Shimadzu) with detectors: UV, spectrofluorometirc, amperometric, conductometric.
  • Capillary electrophoresis (Waters) with UV detector,
  • UV-Vis spectrophotometer (Shimadzu),
  • Dissolution system - detection: spectrophotometer, HPLC, EC, microbiological,
  • Automated system for microorganisms identification - ATB (BioMerieux),
  • Automated system for conductivity measurement - Bactometer (BioMerieux)
  • Systems for membrane filtration
  • Steritest (Millipore) - system for liquid sterility testing
  • General equipment for genetic research of microorganisms: PCR, RFLP, GE, PFGE (BioRad)
  • System for visual documentation of electrophoresis and TLC (BioRad),
  • Incubators, refrigerators, freezers, shakers, colony counters, pH-meters, centrifuges, balances, microscope, 5 autoclaves, 2 water purification systems,
  • Laminar flow cabinets located in air conditioned rooms with monitored microbial purity of environment (air and surfaces).

Main publications since 2004

  • Michalska K., Pajchel G., Tyski S.:
    Capillary electrophoresis method for simultaneous determination of penicillin G, procaine and dihydrostreptomycin in veterinary drugs.
    Journal of Chromatography B, 2004, 800 (1-2), 203-209.
  • Pajchel G., Michalska K., Tyski S.:
    Application of capillary electrophoresis to the determination of various benzylpenicillin salts.
    Journal of Chromatography A, 2004 1032, 265-272.
  • Michalska K, Pajchel G., Tyski S.:
    Determination of ciprofloxacin and its impurities by capillary zone electrophoresis.
    Journal of Chromatography A, 1051 (1-2), 267-272 (2004).
  • Kruszewska H. Zaręba T., Tyski S.:
    Examination of antimicrobial activity of selected non-antibiotic drugs.
    Acta Poloniae Pharmaceutica - Drug Research 2004, 61 (supl.) 18-21.
  • G. Pajchel, K. Michalska, S. Tyski:
    Analysis of phnoxymethylpenicillin potassium by capillary electrophoresis.
    Journal of Chromatography A, 1087, 197-202 (2005).
  • K. Dzierżanowska-Fangrat, K. Semczuk, P. Górska, S. Giedrys-Kalemba, M. Kochman, A. Samet, S. Tyski, D. Dzierżanowska, K. Trzciński:
    Evidence for tetracycline resistance determinant tet(M) allele replacement in a Streptococcus pneumoniae population of limitem geographical origin.
    International Journal of Antimicrobial Agents 27, 159-164 (2006).
  • H. Kruszewska, T. Zaręba, S. Tyski:
    Estimation of antimicrobial activity of selected non-antibiotic products.
    Acta Poloniae Pharmaceutica - Drug Research 63 (5) 457- 460 (2006).
  • G. Młynarczyk, W. Grzybowska, A. Młynarczyk, S. Tyski, D. Kawecki, M. Łuczak, M. Durlik, L. P±czek, A. Chmura, W. Rowiński:
    Occurrence of glycopeptide-resistant enterococci in transplant medicine internal wards in 2001-2005.
    Transplant. Proc., 39 (9), 2886-89 (2007).
  • K. Michalska, G. Pajchel, S. Tyski:
    Detemination of enantiomeric impurity of linezolid by capillary electrophoresis using heptakis-(2,3-diacetyl-6-sulfato)-Β-cyclodextrin.
    J. Chromatogr. A , 1180, 179-186, (2008)
  • K.. Michalska, G. Pajchel, S. Tyski:
    Determination of linezolid and its achiral impurities using sweeping preconcentration by micellar capillary electrophoresis.
    Journal of Pharmaceutical and Biomedical Analysis (2008).
Examination objects / groups of objects Examination character
Research method 		Norms &/or documented research procedures
1 2 3
Therapeutic, medicinal and biocidal products
  1. Sterile objects
  2. Non-sterile objects
 Microbiological contamination:
  1. Sterility test
  2. Microbiological examination of non-sterile product
  3. Efficacy of antimicrobial preservation
for each above methods:
Variant - pour-plate method
Variant - membrane filtration method
Ph. Eur. 5, 2005: 2.6.1; 2.6.12; 2.6.13; 5.1.3

FP. VII, 2006: 2.6.1; 2.6.12; 2.6.13; 5.1.3

FP VI, 2002, str. 114-125; 143-146
  1. Disinfectional and antiseptical objects
Efficacy of disinfectants / antiseptical:
  1. Examination of biocidal effect for microorganisms
 PN-EN 1040:2006;
PN-EN 1275:2006;
PN-EN 14347: 2005;
PN-EN 13727:2006;
prEN 13727:2005;
PN-EN 1276:2000/Ap 1:2001
PN-EN 1650:2002;
PN-EN 1656:2001;
PN-EN 1657:2006;
Therapeutic products
  1. Antibiotics and chemotherapeutics
Antimicrobial activity:
  1. Diffusion method
  2. Turbidimetric method
 Ph. Eur. 5, 2005: 2.7.2
FP. VII, 2006: 2.7.2
FP VI, 2002, str. 131-141; 185-189; 91-93
USP 29, 2006: 81
Dissolution for oral formulas:
  1. Dissolution method (microbiological or UV detection)
 Ph. Eur. 5, 2005: 2.9.3; 2.2.25
FP. VII, 2006: 2.9.3; 2.2.25
USP 29, 2006: 711; 851
Environ-ment
  1. Air
  2. Surface
 Microbiological contamination:
  1. Impaction method
  2. Contact-plate method
 Nr: MA/IT 5.3/060
ed. 2 from 27.03.2006