Qualification of the ABC Transfer® Alpha-Beta Connection for Sterile Process Applications
1. Abstract
This report presents the qualifications of the ABC Transfer® Alpha-Beta Connection . The goal was to evaluate the system’s performance in maintaining sterility during material transfers. Multiple tests assessed the effects of cleaning and contamination under simulated production conditions inside a SKAN isolator. No contamination was observed inside the isolator or the Beta container, even when seals were not cleaned, demonstrating the system’s exceptional containment capabilities. These results validate the ABC Transfer® system as a very safe sterile process and aseptic transfer solution for GMP pharmaceutical operations.
2. Introduction
Transfer systems are key to preserving sterile conditions in aseptic pharmaceutical environments. ABC Transfer has developed an Alpha-Beta system that aims to optimize containment, user ergonomics, and compliance with GMP and Annex 1 standards. This report summarizes qualification results from a series of experimental tests evaluating microbial containment and decontamination across standard and challenged conditions. Emphasis is placed on real-world handling, efficacy of cleaning procedures, and the system’s capacity to protect critical sterile zones.
3. Materials and Methods
3.1 Sterile Process Equipment Used for Alpha-Beta Connection
Tests were conducted in a SKAN PSI/SARA isolator in Grade A conditions with +50 Pa overpressure. The Isolator was installed in a non-controlled environment. The tested equipment included a 270 mm Alpha port and two 270 mm Beta stainless steel containers. No filters were used on the containers, and doors remained sealed to simulate conditions of a controlled sterile process. Environmental conditions (temperature, humidity, pressure) were monitored using calibrated devices.
3.2 Microbiological Analysis
TSA agar and Bacillus stearothermophilus BIs were used. Samples were incubated at 30–35°C for 3–5 days. Surface and personnel monitoring was conducted using contact and settle plates. All microbiological assessments complied with USP <61> and EP <2.6.12>.
4. Results of the Alpha-Beta System in Sterile Process Conditions
4.1 Test 1: Environmental Control Baseline
The objective of this test was to establish the microbiological baseline of the testing environment. To do this, settle plates and contact plates were employed to detect ambient contamination. Plates were exposed for four hours on the Alpha port and Beta container doors. Additional contact plates were applied to the protective clothing of the operators working within the isolator setup. The results indicated measurable CFUs on settle plates located on both the Alpha and Beta doors. Furthermore, moderate contamination levels were found on the operator garments, which was consistent with expectations for a controlled Grade C cleanroom environment. These results confirmed that the microbiological background was stable and representative, making it suitable for the simulation testing that followed.
4.2 Test 2: VHP Decontamination
This test was designed to validate the effectiveness of vaporized hydrogen peroxide (VHP) in eliminating microbial bioburden on the Alpha and Beta port surfaces. Biological indicators (BIs) containing Bacillus stearothermophilus spores were strategically placed on these surfaces prior to the application of a full VHP sterilization cycle. Post-exposure analysis showed that all BIs were completely inactivated, indicating the high efficacy of VHP in achieving thorough decontamination—a cornerstone of any sterile process system.. These results confirm the reliability of VHP treatment for decontaminating the Alpha-Beta interface in routine sterile operations.
4.3 Test 3: Sterility Without Cleaning
The purpose of this test was to assess whether sterility could be maintained during multiple transfer operations without any intermediate cleaning of the Alpha and Beta seals. Five consecutive transfers were performed without the application of any disinfectant. While the first two cycles yielded no contamination, one CFU was detected on the seals starting from the third cycle. Importantly, no CFUs were detected inside the Beta container or within the isolator at any stage, demonstrating that the system effectively prevents contamination of critical sterile areas even under suboptimal cleaning conditions.
4.4 Test 4: IPA Disinfection Efficiency
This test evaluated the effectiveness of wiping the Alpha and Beta seals with sterile wipes soaked in 70% isopropyl alcohol (IPA) between each of five transfer cycles. Microbiological sampling confirmed that this practice successfully prevented the development of any microbial contamination, both on the seals and within the container and isolator. These findings support the integration of IPA wiping as a practical and effective disinfection method during routine aseptic transfers.
4.5 Test 5: Deliberate Seal Contamination (100 CFU)
To simulate a worst-case scenario, 100 colony forming units (CFUs) of Bacillus stearothermophilus were deliberately applied to the Alpha and Beta seals prior to five successive transfers. No cleaning was performed between transfers. While external contamination was observed on the ring of concern and peripheral Beta seal surfaces, no CFUs were detected inside the Beta container or inside the isolator. This outcome demonstrates that the ABC Alpha-Beta system contains microbial contamination externally and maintains a sterile process even in highly challenging conditions.
4.6 Test 6: Hydrogen Peroxide Surface Bioburden Reduction
This test was conducted to confirm the bioburden reduction capacity of a 6% hydrogen peroxide (H₂O₂) solution when applied manually as a surface disinfectant. Following deliberate contamination, all surfaces were wiped using sterile wipes soaked in the H₂O₂ solution. Subsequent microbiological sampling demonstrated a significant reduction in CFUs, with residual contamination consistently below 10 CFU per surface. This confirms the effectiveness of hydrogen peroxide in high-level disinfection of transfer interfaces.
4.7 Test 7: Container Sterility Over Time
This test assessed the ability of a Beta container to maintain sterility when stored outside the isolator for an extended period. The sealed container was disconnected from the isolator and left under controlled conditions for 96 hours. Upon re-connection, microbiological samples were taken from internal surfaces. No microbial growth was detected, demonstrating that the system preserves internal sterility for at least four days.
5. Discussion: Performance and Reliability in Sterile Process Qualification
The comprehensive testing program confirmed the robustness of the ABC Transfer Alpha-Beta system under a wide range of conditions. Tests 1 and 2 established a reliable environmental baseline and validated the effectiveness of VHP decontamination. Tests 3 through 5 demonstrated that the internal sterile volume of the Beta container and the isolator remained sterile even when the seals were not cleaned or deliberately contaminated. The results from Test 4 supported the use of 70% IPA as a practical and effective cleaning solution. Test 6 showed that hydrogen peroxide provided high-level bioburden reduction, while Test 7 confirmed that the Beta container retained sterility for up to four days of storage outside the isolator. Importantly, in no test scenario was contamination detected inside the container or the isolator, highlighting the superior containment performance of the system.
6. Conclusion
The qualification study demonstrated that the ABC Transfer Alpha-Beta connection is a reliable and safe solution for aseptic material transfer in pharmaceutical environments. Across all test scenarios, no microbial contamination was found inside the Beta container or the isolator, even under adverse conditions such as multiple uncleaned transfers or deliberate inoculation. The system effectively contains potential contaminants at the seal interface and maintains sterile boundaries. Its compatibility with IPA and hydrogen peroxide disinfection protocols, along with the demonstrated integrity of the Beta container during storage, confirms the ABC system’s suitability for GMP-compliant sterile process manufacturing operations. It is recommended as a best-in-class aseptic transfer solution.
