PDA Journal of Pharmaceutical Science and Technology最新文献

As described in USP <1207>, the deterministic leak test methods using laser-based gas headspace analysis and helium leakage are those with the highest sensitivities. As stated in the chapter, "no single package leak test or package seal quality test method is applicable to all product-package systems"; therefore, knowing the advantages and disadvantages of both of these techniques, and the extent to which they can be substituted for each other, is valuable. In an effort to begin addressing this issue, a systematic study using these two techniques has been performed. This study used the same well-defined positive controls prepared with microcapillaries for both measurement techniques. For the headspace gas analysis technique, the headspace carbon dioxide content was measured at multiple time points during three separate conditioning cycles using either a 0.5, 1.0, or 2.0 bar CO₂ overpressure; the observed change in headspace carbon dioxide was then used to determine an ingress rate for each positive control. For the helium leakage technique, the positive controls were measured with a standard helium leak detector with 100% helium atmosphere on the atmospheric pressure side of the artificial defects. The resulting leakage rates from both techniques were compared for ingress into both ISO 2 R and ISO 10 R vials. The obtained correlation between helium and carbon dioxide leakage rates resulted in a minimum R² coefficient of 0.98 across all 12 runs. Additionally, both setups met the acceptance criteria for accuracy with their respective calibrated standards.

An increasing number of pharmaceutical products require deep cold storage at cryogenic conditions, approximately -150°C to -190°C, to maintain stability and/or activity. Previous work has revealed that, at these extreme conditions, a typical pharmaceutical package configuration (vial, stopper, crimp cap) may lose container closure integrity (CCI) due to both the glass transition temperature (-55°C to -70°C) of the rubber stopper used to seal the vial and the different thermal expansion coefficients of the primary packaging components. Importantly, this type of temporary breach in CCI frequently reseals itself when the vial is brought back to ambient temperature. The following study illustrates a CCI test method using laser-based headspace oxygen analysis for identifying both temporary and permanent defects in vial assemblies exposed to cryogenic storage conditions. The method takes advantage of the nitrogen-enriched environment that exists in most cryogenic storage conditions. When vials are placed into cryogenic storage, the cold temperature creates a pressure gradient across the vial seal that, if a leak defect develops, drives nitrogen gas into the vial headspace and, thereby, decreases the headspace oxygen concentration for vials originally packaged with an air headspace. This decrease in headspace oxygen after the cryogenic storage period can be used to identify a breach in CCI, regardless of whether the defect is temporary or permanent. Experimental data obtained for three different vial types (a 2R glass vial, a 2mL plastic CZ vial, and a 2mL plastic Aseptic Technologies [AT] vial) demonstrated that this CCI test method can robustly and readily detect laser-drilled micron-sized positive controls down to 5 μm (the smallest defect size tested) in the body of the vial and type controls prepared using a Ø64 µm wire at the stopper-seal interface (effective defect size varied depending on the vial).

A system applied to clinical microbiology was adapted for the high throughput assessment of environmental monitoring plates collected from a parenteral manufacturing site. Proof of concept and industrialization of the instrument necessary to ensure a robust counting system where false negatives results could not be tolerated. Here we describe the side-by-side comparison of the system compared side by side to qualified microbiologists in routine use over multiple months.

Joint presentation - Ecolab and HiTech Health (CDMO for CGT)The purpose of this presentation is to review the impact cleaning and disinfectant residues can present to an ATMP cGMP manufacturing facility and discuss how to assess and manage these residues.The term "disinfectant residue" can illicit different responses. For some, they pose a risk potential product contamination, or can alternatively be seen as evidence that cleaning and disinfection has been performed. For others it could be a sign of lack of control. With the most recent revision to Annex 1 there has been a renewed focus on control and removal of disinfection residues. This presentation will discuss how to minimize disinfectant residues by regime design and investigate the methodology associated to quantifying residues beyond subjective visual assessment.

The typing of micro-organisms in pharmaceutical factories often relies on expensive and time-consuming molecular techniques. So, the implementation of cheap, fast and reliable typing methods in the routine would speed up the investigation procedures improving the contamination control strategy. The Fourier-transform infrared (FT-IR) spectroscopy is a method that generates spectra, that enables to micro-organisms typing within 3 h. This study aimed to evaluate the FT-IR for typing S. epidermidis strains isolated from an immunobiological pharmaceutical industry in Brazil. Fifty strains were evaluated by FT-IR using IR Biotyper®. A dendrogram was created with the raw data to cluster the separation spectrum and the cut-off value was automatically calculated. Forty-four FT-IR profiles were obtained, a ratio of 1.14 strain/profile. From the five clusters formed, Cluster 1, 2 and 3 (6 strains) were isolated from environmental monitoring of air and operators (EMO). Cluster 4 (3 strains) were isolated from EMO and bioburden assays, suggesting that the environment could be the main source of bacterial contamination in the product analyzed in bioburden assay. Cluster 5 (2 strains) were isolated from EMO and a cell culture lineage used in quality control assays, suggesting that the environment could also be the main source of cell contamination.

Isolators play a critical role in protecting both the product and the environment, as well as the personnel involved in pharmaceutical manufacturing, analytical procedures, and sterility testing. Gloves attached to the windows and doors of the isolator are designed to facilitate intervention, testing, and safety. However, due to their inherent characteristics and vulnerability to puncture or loss of integrity, they are recognized as a significant potential source of contamination. In addition to the possible pathways of contamination transfer, the size of glove holes plays a critical role in determining the risk of contamination. In this study, chlorosulphonated polyethylene (CSM) gloves were exposed to an aerosol containing Bacillus subtilis or Staphylococcus aureus This assessment aimed to ascertain the integrity of the gloves' seal. It was postulated that, below a certain aperture size, gloves used in isolator systems could establish an effective seal even if the external surface of the gloves exhibited modifications. Calibrated holes of different diameters (0.3, 0.5, 1, and 1.5 mm) were created using a femtosecond laser drilling technology. The holes were located on the tip of the middle finger. Based on the context of our study, passage of microorganisms through glove holes of a certain size does occur. Under the experimental conditions chosen, the cutoff for passage was determined to be a 0.5-mm hole, regardless of the microorganism evaluated. Although this study has some limitations, including the lack of a panel of microorganisms evaluated and the investigation of a single glove type called CSM, the high level of "worst case" challenge conditions provides compelling data to support our results. It would now be interesting to carry out studies at different production sites to assess their risk of contamination and relate this to their glove failure.

In pharmaceutical manufacturing, benefit is conferred in detection of specified microorganism (i.e., Burkholderia cepacia complex (BCC), E. coli, Pseudomonas aeruginosa, Salmonella enterica) not readily identified by culture-dependent methods. It's logical to test for the presence of "specified microorganism" using metagenomic analysis before culturing a "specified organism", especially when the organism isn't easy to culture. We developed a metagenomic analysis during enrichment to identify specified organisms. The enriched bacterial community consisted predominantly of Bacillus spp. and Stenotrophomonas spp., each contributing about 97-99% to total taxon abundance in TSB and 1/10× TSB. The specified microorganisms that were observed were Clostridium spp., Burkholderia spp., and Staphylococcus spp. (0.04 - 0.07%) in TSB, otherwise Burkholderia spp., Pseudomonas spp., Salmonella spp., Staphylococcus spp. and Escherichia spp. (0.01 - 1.73%) in 1/10× TSB. PreQ0 biosynthesis (PWY-6703) and guanosine ribonucleotides de novo biosynthesis (PWY-7221) were the most abundant pathways in 1/10× TSB-24 h. BCC chiefly contributed to the toluene degradation (PWY-5180 and PWY-5182) pathways. Initial results demonstrate the potential of the metagenomic approach during enrichment in water-based environments. These results indicate that a metagenomic enrichment approach to evaluating water samples can be useful to monitor specified organisms over time, including oligotrophs such as BCC in 1/10× TSB.

A challenge facing the biomanufacturing industry is the lengthy timeline for quality control testing that delays critical go/no-go decision making for the rapid release of drug products. The recommended USP methods for bioburden and sterility testing are surface-spread plating method and direct inoculation method, respectively. These compendial methods are reliable; however, results take approximately 5-7 days for bioburden testing (USP< 61>) and no less than 14 days for sterility testing (USP < 71>). ATP-bioluminescence detection is a rapid microbial method (RMM) that can reduce the time to result for both bioburden and sterility testing, taking 18-24 hours and 6 days, respectively. This study aims to evaluate the performance of ATP-bioluminescence in comparison to compendial methods to understand the implications of using this technology in bioburden and sterility testing processes. The research conducted consisted of simulating contamination to assess the detection capabilities of the ATP-bioluminescence assay. Results from compendia and ATP-bioluminescence detection were analyzed for comparability. Findings from this study will provide insight on the use of this platform as an alternative tool for in-process testing.

The Milliflex^® Rapid System is a proven automated ATP-based solution used for rapid bioburden and sterility testing. Combining membrane filtration, ATP bioluminescence and imaging, it provides microbial enumeration in significantly less time than traditional methods. Detecting microbial contamination in cell-based products remains a challenge due to filterability issues and interference from mammalian ATP. To address this, a sample processing method has been developed using a selective cell lysis solution and a free ATP removal enzyme effective at both low and high mammalian cells densities. The processing of Chinese Hamster Ovary (CHO) cells ranging from 106 to 107 cells/mL with lysis solution eliminates the mammalian ATP background in 15 minutes. This requires a ratio of 4 mL of lysis solution per mL of sample at 106 cells/mL or 9 mL of lysis solution per mL of sample at 107 cells/mL. CHO cells were inoculated separately with 6 microorganisms including Cutibacterium acnes ATCC 6919 and then processed with the lysis solution before enumeration with the Milliflex^® Rapid System. Results showed a rapid enumeration of the 6 microorganisms with good recovery according to the control.