Quality of Non-Sterile Herbal Pharmaceutical Products – Detection of Fungi


  • Manuela-Claudia Curticapean George Emil Palade University of Medicine, Pharmacy, Sciences, and Technology of Targu Mures, Faculty of Pharmacy, Cell Biology and Microbiology Department, Gh. Marinescu Street, no.38, Targu Mures, 540139, Romania


identification of fungi, non-sterile herbal pharmaceutical products, quality control, qPCR


The aim of this paper was to use real-time quantitative PCR (qPCR) to perform microbiological purity tests, aiming to identify the presence/absence of fungal DNA and its quantification from non-sterile pharmaceuticals, with different manufacturers and various compositions. For identification a potential presence of fungi in non-sterile drug samples, fungal DNA has been isolated, amplified and quantified under real-time qPCR conditions. qPCR analysis of non-sterile pharmaceutical samples showed the presence of fungi in all investigated non-sterile herbal drugs. The highest amount of fungal DNA was recorded for S2 sample (0.0767 ng) and the lowest fungal DNA quantities have been registered for S1 sample (0.000032 ng). qPCR-based methods provided an earlier and more sensitive detection, identification, and quantification of fungal contamination compared to standard methods.


. G.F.M. Gad, R.A.I. Aly, M.S. El-din Ashour. “Microbial evaluation of some non-sterile pharmaceutical preparations commonly used in the Egyptian market.” Tropical Journal of Pharmaceutical Research., vol. 10(4), pp. 437-445, Aug. 2011. http://dx.doi.org/10.4314/tjpr.v10i4.9

. Ph.Eur. 5.1.4., “Microbiological quality of non-sterile pharmaceutical preparations and substances for pharmaceutical use”, in Ph. Eur., 10th edition, supplement 10.2., Council of Europe, Strasbourg, France, 2020, pp. 627-628.

. Ph.Eur. 5.1.8., “Microbiological quality of herbal medicinal products for oral use and extracts used in their preparation”, in Ph. Eur., 10th edition, supplement 10.2., Council of Europe, Strasbourg, France, 2020, pp. 638-807.

. N. Vu, J.R. Lou, T.C. Kupiec. “Quality Control: Microbial limit tests for nonsterile pharmaceuticals, Part 1”. Int J Pharm Compd., vol. 18(3), pp. 213-221, May-Jun. 2014. PMID:25306769

. N. Sahoo, P. Manchikanti, S. Dey. “Herbal drugs: Standards and regulation”. Fitoterapia, vol. 81(6), pp. 462- 471, Sep. 2010. doi:10.1016/j.fitote.2010.02.001

. B.H. Kroes. “The legal framework governing the quality of (traditional) herbal medicinal products in the European Union”. Journal of Ethnopharmacology, vol. 158(Part B), pp. 449-453, Dec. 2014. https://doi.org/10.1016/j.jep.2014.07.044

. M. Ratajczak, M.M. Kubicka, D. Kaminska, P. Sawicka, J. Długaszewska. “Microbiological quality of non-sterile pharmaceutical products”. Saudi Pharm J., vol. 23(3), pp. 303-307, Jul. 2015. https://doi.org/10.1016/j.jsps.2014.11.015

. S. Pan, A. Neeraj, K.S. Srivastava, P. Kishore, M.K. Danquah, I.P. Sarethy. “A Proposal for a Quality System for Herbal Products”. Journal of Pharmaceutical Sciences, vol. 102(12), pp. 4230-4241, Dec. 2013. https://doi.org/10.1002/jps.23732

. S.R. Aghili, A. Hossein nejad, M.R.J. Amiri, M. Abastabar. “Recovery and detection of fungal contaminants in some ointments and tablets after opening of the packages in hospitals”. Iranian Journal of Health Sciences, vol. 4(4), pp. 1-13, 2016. doi: 10.18869/acadpub.jhs.4.4.1

. J. Zhang, B. Wider, H. Shang, X. Li, E. Ernst. “Quality of herbal medicines: Challenges and solutions”. Complementary Therapies in Medicine, vol. 20(1-2), pp. 100-106, Feb.-Apr. 2012. https://doi.org/10.1016/j.ctim.2011.09.004

. M. Nemati, A. Hamidi, S.M. Dizaj, V. Javaherzadeh, F. Lotfipour. “An Overview on Novel Microbial Determination Methods in Pharmaceutical and Food Quality Control”. Adv Pharm Bull., vol. 6(3), pp. 301-308, Sep. 2016. doi: 10.15171/apb.2016.042

. R. Vijayakumar, T. Sandle, C. Manoharan. “A review on fungal contamination in pharmaceutical products and phenotypic identification of contaminants by conventional methods”. European Journal of Parenteral & Pharmaceutical Sciences, vol. 17(1), pp. 4-18, 2012.

. I. Altyn, M. Twaruzek. “Profile of fungi in dietary supplement, based on plant raw material”. Reference Module in Life Sciences, Jan. 2020. https://doi.org/10.1016/B978-0-12-809633-8.21047-9

. L.X. Yu, M. Kopcha. “The future of pharmaceutical quality and the path to get there”. International Journal of Pharmaceutics, vol. 528, pp. 354-359, Jun. 2017. http://dx.doi.org/10.1016/j.ijpharm.2017.06.039

. R.M. Haleem, M.Y. Salem, F.A. Fatahallah, L.E. Abdelfattah. “Quality in the pharmaceutical industry – A literature review”. Saudi Pharm J., vol. 23(5), pp. 463–469, Oct. 2015. http://dx.doi.org/10.1016/j.jsps.2013.11.004

. N. Vu, J.R. Lou, T.C. Kupiec. “Quality Control: Microbial limit tests for nonsterile pharmaceuticals, Part 2”. Int J Pharm Compd., vol. 18(4), pp. 305-310, Jul.-Aug. 2014. PMID:25474859

. L. Jimenez, R. Ignar, S. Smalls, P. Grech, J. Hamilton, Y. Bosko, D. English. “Molecular detection of bacterial indicators in cosmetic/pharmaceuticals and raw materials”. J. Ind. Microbiol. Biotechnol, vol. 22, pp. 93-95, Feb. 1999. https://doi.org/10.1038/sj.jim.2900611

. R.M. Baird. “Monitoring microbiological quality: conventional testing methods”, in Guide to microbiological control in pharmaceuticals and medical devices, 2nd ed., S.P. Denyer, R.M. Baird, Ed. SUA: CRC, Taylor & Francis Group, 2007, pp 155-182.

. S.P. Denyer. “Monitoring microbiological quality: application of rapid microbiological methods to pharmaceuticals”, in Guide to microbiological control in pharmaceuticals and medical devices, 2nd ed., S.P. Denyer, R.M. Baird, Ed. SUA: CRC, Taylor & Francis Group, 2007, pp 183-195.

. J.D. Oliver. “Recent findings on the viable but nonculturable state in pathogenic bacteria”. FEMS Microbiol Rev., vol. 34(4), pp. 415-425, Jul. 2010. doi: 10.1111/j.1574-6976.2009.00200.x

. H. Shintani. “Methods of Rapid Microbiological Assay and Their Application to Pharmaceutical and Medical Device Fabrication”. Biocontrol Sci., vol. 21(4), pp. 193-201, 2016. doi:10.4265/bio.21.193

. H. Shintani. “Validation study of rapid assays of bioburden, endotoxins and other contamination”, Biocontrol Sci., vol. 21(2), pp. 63-72, 2016. doi:10.4265/bio.21.63

. J. Peris-Vicente, S. Carda-Broch, J. Esteve-Romero. “Validation of rapid microbiological methods”, J Lab Autom., vol. 20(3), pp. 259-264, Oct. 2015. doi: 10.1177/2211068214554612

. T. Sandle. “Rapid microbiological methods”. Chapter 17, in Pharmaceutical microbiology: essentials for quality assurance and quality control, No. 80, T. Sandle, Ed. USA: WP Woodhead Publising, Elsevier, 2016, pp 219-231. http://dx.doi.org/10.1016/B978-0-08-100022-9.00017-7

. R. Noor, N. Zerin, K.K. Das. “Microbiological quality of pharmaceutical products in Bangladesh: current research perspective”. Asian Pacific Journal of Tropical Disease, vol. 5(4), pp. 264-270, Apr. 2015. https://doi.org/10.1016/S2222-1808(14)60781-7

. J. Dlugaszewska, M. Ratajczak, D. Kaminska, M. Gajecka. “Are dietary supplements containing plant-derived ingredients safe microbiologically?” Saudi Pharmaceutical Journal, vol. 27(2), pp. 240-245, Feb. 2019. https://doi.org/10.1016/j.jsps.2018.11.005

. I. Rizzo, G. Vedoya, S. Maurutto, M. Haidukowski, E. Varsavsky. “Assessment of toxigenic fungi on Argentinean medicinal herbs”. Microbiological Research, vol. 159(2), pp. 113-120, Jun. 2004. https://doi.org/10.1016/j.micres.2004.01.013

. H.K. Chourasia. “Mycobiota and mycotoxins in herbal drugs of Indian pharmaceutical industries”. Mycological Research, vol. 99(6), pp. 697-703, Jun. 1995. https://doi.org/10.1016/S0953-7562(09)80531-5

. V. Mugoyela, K.D. Mwambete. “Microbial contamination of nonsterile pharmaceuticals in public hospital settings”, Ther Clin Risk Manag., vol. 6, pp. 443-448, Oct. 2010. doi: 10.2147/TCRM.S12253

. C. Sornchaithawatwong, S. Tadtong, M. Tangkiatkumjai. “The prevalence of acceptable quality herbal products in Thailand”. Journal of Herbal Medicine, vol. 24, 100391, 2020. https://doi.org/10.1016/j.hermed.2020.100391

. L. Jimenez, S. Smalls, R. Ignar. “Use of PCR analysis for detecting low levels of bacteria and mold contamination in pharmaceutical samples”. Journal of Microbiological Methods, vol. 41(3), pp. 259-265, Aug. 2000. https://doi.org/10.1016/S0167-7012(00)00164-0

. S. Govindaraghavan, N.J. Sucher. “Quality assessment of medicinal herbs and their extracts: Criteria and prerequisites for consistent safety and efficacy of herbal medicines”. Epilepsy & Behavior, vol. 52 (Part B), pp. 363-371, Nov. 2015. http://dx.doi.org/10.1016/j.yebeh.2015.03.004

. Y.T. Lo, P.C. Shaw. “Application of next-generation sequencing for the identification of herbal products”. Biotechnology Advances, vol. 37(8), 107450, 2019. https://doi.org/10.1016/j.biotechadv.2019.107450




How to Cite

Curticapean, M.-C. . (2021). Quality of Non-Sterile Herbal Pharmaceutical Products – Detection of Fungi. International Journal of Sciences: Basic and Applied Research (IJSBAR), 57(2), 57–64. Retrieved from https://gssrr.org/index.php/JournalOfBasicAndApplied/article/view/12516