This article gives a detailed overview on Disinfection and Antiseptic Practices to be included in the Indian Pharmacopoeia to be included in Chapter IP 2026. It will be published officially from July 2024. A comprehensive cleaning and sanitization program is crucial in pharmaceutical manufacturing to prevent microbial contamination of drug products. Contamination can occur through various sources, including ingredients, water, packaging, equipment, and personnel. Current Good Manufacturing Practices (cGMPs) stress the importance of facility design, construction, and material flow to facilitate cleaning and maintenance. When using disinfectants, it’s essential to avoid contaminating drug products with toxic chemicals. Aseptic processing requires cleanable surfaces, environmental controls, and strict cleaning procedures to maintain sterility. The program must meet cleanliness standards, control microbial contamination, and prevent chemical contamination of products and equipment. Antiseptics are used to decontaminate skin, while chemical sterilants sanitize surfaces and articles. UV irradiation can also sanitize surfaces. Proper selection, efficacy demonstration, application, and safety considerations of disinfectants, antiseptics, and sterilants are crucial in pharmaceutical manufacturing.
Classification of Disinfectants:
Chemical disinfectants are grouped based on their composition, including aldehydes, alcohols, halogens, peroxides, quaternary ammonium compounds, and phenolic compounds. Their effectiveness relies on factors like concentration, contact time, surface type, and the presence of organic matter. In India, disinfectants are regulated under the Drug and Cosmetic Act of 1940 and its rules from 1945, along with IS 1061:2017 standards. Manufacturers must provide usage information, and certain liquid chemical sterilizers are overseen by the Food and Drug Administration (FDA) for medical device applications, where CDSCO and SUGAM also play critical roles in overseeing registration and compliance.
| Chemical Entity | Classification | Example |
| Aldehydes | Sporicidal agent | 2% Glutaraldehyde |
| Alcohols | General purpose disinfectant, antiseptic, antiviral agent | 70% Isopropyl alcohol, 70% alcohol |
| Chlorine and sodium hypochlorite | Sporicidal agent | 0.5% Sodium hypochlorite |
| Phenolics | General purpose disinfectant | 500 µg/g Chlorocresol, 500 µg/g chloroxylenol |
| Ozone | Sporicidal agent | 8% Gas by weight |
| Hydrogen peroxide | Vapor phase sterilant, liquid sporicidal agent, antiseptic | 4 µg/g H2O2 vapor, 10%-25% solution, 3% solution |
| Substituted diguanides | Antiseptic agent | 0.5% Chlorhexidine gluconate |
| Peracetic acid | Liquid sterilant, vapor phase sterilant | 0.2% Peracetic acid, 1 µg/g peracetic acid |
| Ethylene oxide | Vapor-phase sterilant | 600 µg/g Ethylene oxide |
| Quaternary ammonium compounds | General-purpose disinfectant and antiseptic | Concentration varies by application, e.g., Benzalkonium chloride |
| β-Propiolactone | Sporicidal agent | 100 µg/g β-Propiolactone |
Selection of an Antiseptic for Hand and Surgical Site Disinfection:
In hospitals, effective hand and surgical site disinfection is crucial for reducing harmful bacteria associated with hospital-acquired infections. Antiseptics like 4% chlorhexidine, 10% povidone-iodine, and others are preferred for their ability to reduce bacterial counts on the skin, promoting hygiene and infection prevention.
Selection of a Disinfectant for Use in Pharmaceutical Manufacturing:
Choosing the right disinfectant for pharmaceutical environments requires considering factors such as
- Microorganism diversity
- Efficacy,
- Endorsement by standards/regulatory authorities
- Application methods
- Surface compatibility
- Presence of Organic material
- Lasting effectiveness
- Safety
- Compatibility with cleaning agents
- Rotation strategies
- Product contamination prevention
Addressing the above ensures the selection of a suitable disinfectant supporting safety and quality in manufacturing processes.
Theoretical Discussion of Disinfectant Activity:
Disinfectant effectiveness follows a sigmoid curve, starting slowly and accelerating over time, governed by first-order kinetics. The rate constant (K) is determined by initial (N0) and final (N) microbial counts. Temperature positively influences disinfection rates, with the Q10 coefficient indicating acceleration with each 10° rise. Disinfectant concentration, described by a concentration exponent (n), significantly impacts microbial reduction, guiding appropriate dilution and neutralization procedures. pH also affects efficacy, determined by the disinfectant’s ionization state relative to its pKa and environmental pH, crucial for selecting disinfectants in pharmaceutical manufacturing.
The rate constant, K, for the disinfection process can be calculated by the formula: (1/t) (log N0/N)
Where, t = time, in minutes, for the microbial count to be reduced from N0 to N; N0 = initial number of microorganisms in cfu per ml and N = final number of microorganisms in cfu per ml.
Concentration Exponents of Common Antiseptics, Disinfectants and Sterilant
| Disinfectant | Concentration Exponent |
| Hydrogen peroxide | 0.5 |
| Sodium hypochlorite | 0.5 |
| Mercuric chloride | 1 |
| Chlorhexidine | 2 |
| Formaldehyde | 1 |
| Alcohol | 9 |
| Phenol | 6 |
| Quaternary ammonium compounds | 0.8 to 2.5 |
| Aliphatic alcohols | 6.0 to 12.7 |
| Phenolic compounds | 4 to 9.9 |
Mechanism of Disinfectant Activity Against Microbial Cell
| Target | Disinfectants |
| Cell wall | Formaldehyde, hypochlorite, glutaraldehyde |
| Cytoplasmic membrane, action on membrane potential | Anilides, hexachlorophene |
| Membrane enzymes, action on electron-transport chain | Hexachlorophene |
| Action on ATP | Chlorhexidine, ethylene oxide |
| Action on enzymes with -SH groups | Ethylene oxide, glutaraldehyde, hydrogen peroxide, hypochlorite, iodine |
| Action on general membrane permeability | Alcohols, chlorhexidine, quaternary ammonium compounds |
| Cell contents, general coagulation | Chlorhexidine, aldehydes, quaternary ammonium compounds |
| Ribosomes | Hydrogen peroxide |
| Nucleic acids | Hypochlorites |
| Thiol groups | Ethylene oxide, glutaraldehyde, hydrogen peroxide, hypochlorite |
| Amino groups | Ethylene oxide, glutaraldehyde, hypochlorite |
| General oxidation | Hypochlorite |
Microbial Resistance to Disinfectants
Microbial resistance to disinfectants is less likely than to antibiotics due to their high potency against low microorganism populations and reduced selective pressure. Periodic testing may be needed to confirm susceptibility of frequently isolated microorganisms to specific disinfectants due to variability in resistance levels between species.
The Resistance of Some Clinically Important Microorganisms to Chemical Disinfectants (Listed in Order of Decreasing Resistance)
| Type of Microorganisms | Examples |
| Bacterial spores | Bacillus subtilis, Clostridium sporogenes |
| Mycobacteria | Mycobacterium tuberculosis |
| Nonlipid-coated viruses | Poliovirus, rhinovirus |
| Fungal spores and vegetative molds and yeast | Trichophyton, Cryptococcus, Candida spp. |
| Vegetative bacteria | Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella spp. |
| Lipid-coated viruses | Herpes simplex virus, hepatitis B virus, human immunodeficiency virus |
Disinfectant Challenge Testing
Under the Drug and Cosmetic Act of 1940 and its related rules, Indian regulatory bodies require thorough testing for safety and effectiveness of antimicrobial agents like disinfectants and antiseptics. Companies must provide detailed information on their products’ chemical composition and demonstrate their safety and effectiveness through testing. Official testing methods, such as the Phenol-Coefficient Test, ensure compliance. In pharmaceutical manufacturing, disinfectant efficacy is validated through tests like use-dilution and surface challenge tests to meet regulations and ensure cleanliness. Acceptance criteria for these tests must be realistic, considering factors like microbial growth and physical removal during application.
Typical Challenge Organisms:
| Microorganisms | Typical Environmental Isolates | |
| Bactericide | E. coli, ATCC 11229; S. aureus, ATCC 6538; P. aeruginosa, ATCC 15442 | M. luteus, S. epidermidis, Corynebacterium jeikeium, P. vesicularis |
| Fungicide | C. albicans, ATCC 10231 or 2091; Penicillium chrysogenum, ATCC 11709; A. brasiliensis, ATCC 16404 | P. chrysogenum, A. brasiliensis |
| Sporicide | B. subtilis, ATCC 19659 | B. sphaericus, B. thuringiensis |
Typical Surfaces to be Decontaminated by Disinfectants in a Pharmaceutical Manufacturing Area
| Material | Application |
| Stainless steel 304L and 316L | Work surfaces, Filling Equipment and Tanks |
| Glass | Windows and Vessels |
| Plastic, Vinyl | Curtains |
| Plastic, Polycarbonate | Insulation Coating |
| Lexan® (plexiglass) | Shields |
| Epoxyl-coated Gypsum | Walls and Ceilings |
| Fiberglass-reinforced Plastic | Wall Paneling |
| Tyvek® | Equipment Wraps |
| Terrazzo Tiles | Floors |
Determination of Bactericidal, Fungicidal and Yeasticidal Activity of Antiseptic Medicinal Products
This section outlines a test to evaluate the antimicrobial effectiveness of water-miscible antiseptic medicinal products for skin or mucous membrane contact. The test assesses bactericidal, fungicidal, or yeasticidal activity against specific standards, depending on the product’s claimed properties.Explore this essential aspect of disinfection and antiseptic practices to ensure the efficacy and safety of your pharmaceutical products.
Principle: The method involves exposing test microorganism suspensions to the antiseptic at 33°C for specified times, followed by immediate neutralization using validated methods.
Test Organisms and Conditions:
- Use stable suspensions of specific strains, with microbial counts reported in CFU/ml.
- Employ designated strains and include others relevant to the product’s use.
| Strains for Bactericidal activity testing | |
| Microorganisms | Growth Conditions |
| Staphylococcus aureus ATCC 6538 Enterococcus hirae ATCC 10541 Escherichia coli NCTC 10538 Pseudomonas aeruginosa ATCC 15442 | Grow each of the bacterial species separately in Casein soyabean digest agar or Casein soyabean digest broth.After incubation, harvest the growth and obtain a microbial count of 1-5×108 CFU per ml. For Preparation of test strains incubate at 30° to35° for 18 to 24 hours and subculture at least twice.For testing of product and validation of test incubate at 30° to 35° for ≤3 days. |
| Strains for Yeasticidal activity testing | |
| Candida albicans ATCC 10231 | Grow in Sabouraud dextrose agar or Sabouraud Dextrose broth. After incubation, harvest the growth and obtain a microbial count of 1-5×107 CFU per ml.For Preparation of test strains incubate at 20° to25° for 48 to 72 hours and subculture at least twice.For testing of product and validation of test incubate at 20° to 25° for ≤5 days. |
| Strains for Fungicidal activity testing | |
| Candida albicans ATCC 10231Aspergillus brasiliensis ATCC 16404 | Grow in Sabouraud dextrose agar or Sabouraud Dextrose broth. After incubation, harvest the growth and obtain a microbial count of 1-5×107 CFU per ml.For Preparation of test suspension of C.albicans incubate at 20° to 25° for 48 to 72 hours.For Preparation of test suspension of A.brasiliensis spores incubate at 20° to 25° for 5 to 7 days or until good sporulation.For testing of product and validation of test withC. albicans and A. brasiliensis incubate at 20° to 25° for ≤5 days. |
Preparation Steps:
- Prepare test strains in saline solution.
- Concentrate the antiseptic solution to 1.25 times the in-use concentration, considering dilution during testing.
- Use neutralizing agents to stop antimicrobial activity.
Methods:
- Ensure reagent temperatures are at 33°C.
- Apply the dilution neutralization or membrane filtration method to determine viable microorganism counts.
Controls and Validation:
- Validate testing conditions, including neutralizing agent efficacy and method controls.
- Ensure at least a 0.5 × 10^(-1) recovery rate of CFU from the validation suspension.
Acceptance Criteria:
- A product is considered bactericidal if it achieves a ≥5 log10 reduction in CFU, and fungicidal/yeasticidal if it reaches a ≥4 log10 reduction, unless different criteria are justified.
This framework ensures antimicrobial efficacy before market release, though it does not replace clinical efficacy assessments.
Disinfectants in a Cleaning and Sanitization Program:
- Selection and Validation: Choose suitable disinfectants and confirm their effectiveness through surface challenge tests.
- Program Establishment: Develop detailed procedures, train staff, decide on disinfectant rotation, and ensure correct application methods and contact times.
- cGMP Guidelines: Follow documented procedures for cleaning, maintenance, and sanitization of pharmaceutical equipment, including responsibilities, schedules, and record-keeping.
- Staff Training: Train staff on microbiology, safe handling and disposal of disinfectants, and proper application techniques.
- Dilution and Preparation: Prepare disinfectants at correct dilutions, especially in aseptic areas, and establish the shelf life of diluted solutions.
- Disinfectant Rotation: Rotate disinfectants, particularly those with sporicidal agents, based on environmental monitoring data.
- Safety Measures: Handle concentrated disinfectants safely, avoid mixing incompatible agents, and provide access to safety gear, Material Safety Data Sheets, and safety showers.
- these essential elements within the realm of disinfection and antiseptic practices to uphold stringent hygiene standards and ensure the safety of pharmaceutical products and medical devices.
These essential elements of disinfection and antiseptic practices uphold stringent hygiene standards and ensure the safety of pharmaceutical products and medical devices.
Conclusion:
In conclusion, establishing an effective cleaning and sanitization program is essential in pharmaceutical manufacturing to ensure product safety and regulatory compliance. Choosing appropriate disinfectants, validating their efficacy, and implementing procedures are criticalsteps in this process. Morulaa HealthTech is committed to assisting clients in navigating these complexities by providing guidance on disinfectant selection, training staff on proper usage, and ensuring compliance with regulatory standards. Additionally, incorporating MD-14 forms and the SUGAM portal can streamline the registration and compliance processes, making it easier for companies to adhere to CDSCO guidelines and maintain high standards in the manufacturing of medical devices and pharmaceutical products. Contact us to know more on [email protected] or Click here.