Detection Limit and Sensitivity of Staphylococcus aureus Detection in Pharmaceutical Products

  • Nur Aini National Quality Control Laboratory of Drug and Food, Indonesian Food and Drug Authority, Jakarta
  • Sitoresmi Triwibowo National Quality Control Laboratory of Drug and Food, Indonesian Food and Drug Authority, Jakarta


Stapylococcus aureus is a pathogen that should be absent in pharmaceutical products. Contamination of certain microorganisms can potentially reduce or inactivate therapeutic activity, affect the stability, efficacy and cause infection of the patient. Microbial detection methods must be valid and sensitive to detect the contamination of microorganisms at low concentrations. The standard method of S. aureus refers to the Indonesian Pharmacopoeia (FI). LOD and sensitivity of the method in general is not stated in FI. This study aims to determine LOD and sensitivity of S. aureus method on pharmaceutical products with route of administration in cutan, oromucosal, gingival, auricular, vaginal and oral based on Indonesian Pharmacopoeia. Experiments used nine pharmaceutical products representing six dosage forms contaminated with S. aureus ATCC 6538 with three level concentrations of ±1, ±3, and ±5 CFU per g atau mL sample. LOD and sensitivity were determined and analyzed descriptively. This study shows that the detection limit is 1-3 colonies per g or mL sample and a sensitivity of 100%. Results of this study can be used as reference for LOD value in the validation or verification process of the S. aureus detection method in the laboratory to ensure validity of the methods used.


1. Kementerian Kesehatan Republik Indonesia. Farmakope Indonesia edisi IV. Jakarta: Kementerian Kesehatan RI; 2020.

2. Ratajczak M, Kubicka MM, Kamińska D, Sawicka P, Długaszewska J. Microbiological quality of non-sterile pharmaceutical products. Saudi Pharm J 2015;23(3):303–7.

3. Eissa M, Mahm A. Evaluation of Microbial Recovery from Raw Materials for Pharmaceutical Use. J Food Pharm Sci 2016;4(1):6–11.

4. The United States Pharmacopeia. United State Pharmacopoeia 2020 USP 43-NF 38. In: United State Pharmacopoeia, editor. United State Pharmacopoeia. United State: United State Pharmacopoeia; 2020.

5. Turner NA, Sharma-Kuinkel BK, Maskarinec SA, Eichenberger EM, Shah PP, Carugati M, et al. Methicillin-resistant Staphylococcus aureus: an overview of basic and clinical research. Nat Rev Microbiol [Internet] 2019;17(4):203–18. Available from:

6. Wikananda IDARN, Hendrayana MA, Pinatih KJP. Efek Antibakteri EKstrak Ethanol Kulit Batang Tanaman Cempaka Kuning (M. champaca L.) Terhadap Pertumbuhan Staphylococcus aureus. J Med 2019;8(5):2597–8012.

7. Spratt HG, Levine D, Tillman L. Physical therapy clinic therapeutic ultrasound equipment as a source for bacterial contamination. Physiother Theory Pract 2014;30(7):507–11.

8. Spratt HG, Levine D, McDonald S, Drake S, Duke K, Kluttz C, et al. Survival of Staphylococcus aureus on therapeutic ultrasound heads. Am J Infect Control [Internet] 2019;47(9):1157–9. Available from:

9. Spratt HG, Levine D, Bage J, Giles DK, Collier AG. Topical lotions utilized in outpatient rehabilitation clinics as a potential source of bacterial contamination. Physiother Theory Pract [Internet] 2019;35(2):163–70. Available from:

10. Farajnia S, Hassan M, Nezhadi SH, Mohammadnejad L, Milani M, Lotfipour F. Determination of Indicator Bacteria in Pharmaceutical Samples by Multiplex PCR. 2008;98(411):328–38.

11. International Conference on Harmonization. Validation of analytical procedures: text and methodology Q2 (R1). In: International Conference on Harmonization. Geneva: IFPMA; 2005.

12. Cabicarová T, Kaclíková E, Mader A, Minarovičová J, Koreňová J, Kuchta T. Improvement of the Detection Sensitivity for Staphylococcus aureus in Spices and Herbs. Food Anal Methods 2016;9(7):1980–4.

13. Salihah NT, Hossain MM, Abdul Hamid MRW, Ahmed MU. A novel, rapid, and sensitive real-time PCR assay for cost-effective detection and quantification of Staphylococcus aureus in food samples with the ZENTM double-quenched probe chemistry. Int Food Res J 2019;26(1):193–201.

14. Marlowe EM, Bankowski MJ. Conventional and molecular methods for the detection of methicillin-resistant Staphylococcus aureus. J Clin Microbiol 2011;49(9 SUPPL.):53–6.

15. Alipour F, Ahmadi M, Javadi S. Evaluation of different methods to detect Methicillin Resistance in
Staphylococcus aureus (MRSA). J Infect Public Health [Internet] 2014;7(3):186–91. Available from:

16. Samadi N, Alvandi M, Fazeli MR, Azizi E, Mehrgan H, Naseri M. PCR-based detection of low levels of Staphylococcus aureus contamination in pharmaceutical preparations. J Biol Sci 2007;7(2):359–63.

17. Rao V, Haranatha K, Raju BVS, Chandra J, Vijayalakshmi PBKM. Detection of indicator pathogens from pharmaceutical W nished products and raw materials using multiplex PCR and comparison with conventional microbiological methods. 2008;1007–18.

18. Aini N, Berlina M, Triwibowo S, Z NS. Penetapan Limit of Detection ( LOD ) dan Sensitivitas Metode Deteksi Pseudomonas aeruginosa dalam Berbagai Matriks Sediaan Obat. Eruditio 2021;1(2):30–6.

19. Singapore Accreditation Councils. Method Validation of Microbiological Methods. Enterprise Singapore; 2019.

20. Sugiyono. Metode Penelitian Kuantitatif, Kualitatif, dan R&D. Bandung: Alfabeta; 2013.

21. Araújo de Assis P, Borba de Andrade S, Maria Carvalho de Oliveira C, Menezes de Araújo P, Grangeiro Júnior S, Verônica Vieira Ramos S. Development and validation of a microbial counting method for mebendazole oral suspension. 2011.

22. Nuryati. Farmakologi. Kementerian Kesehatan Republik Indonesia; 2017.

23. Kruszewska H, Zarȩba T, Tyski S. Examination of antibacterial and antifungal activity of selected non-antibiotic products. Acta Pol Pharm - Drug Res 2008;65(6):779–82.

24. Djajusman SK, Tedjosasongko U, Irmawati I. Daya Hambat Xylitol dan Nistation terhadap Pertumbuhan Candida albicans (in vitro) (Inhibition Effect of Xylitol and Nistatin Combination on Candida albicans Growth (in vitro)). Dent J (Majalah Kedokt Gigi) 2014;47(3):164.

25. ncbi. Benzydamine Hydrochloride (Code C1581) [Internet]. 2021 [cited 2021 Sep 13];Available from:

26. Eissa ME. Distribution of bacterial contamination in non-sterile pharmaceutical materials and assessment of its risk to the health of the final consumers quantitatively. Beni-Suef Univ J Basic Appl Sci 2016;5(3):217–30.

27. Guidance-Validation NGA. Validation and Verification of Quantitative and Qualitative Test Methods. 2018.
How to Cite
AINI, Nur; TRIWIBOWO, Sitoresmi. Detection Limit and Sensitivity of Staphylococcus aureus Detection in Pharmaceutical Products. JURNAL ILMU KEFARMASIAN INDONESIA, [S.l.], v. 21, n. 1, p. 49-56, apr. 2023. ISSN 2614-6495. Available at: <>. Date accessed: 10 june 2023. doi: