USP Bacteriostatic Water 30ML

Research Overview

USP Bacteriostatic Water 30mL serves as an essential research tool for laboratories conducting peptide research, protein studies, and various experimental protocols requiring sterile, preserved water. This pharmaceutical-grade solution combines ultrapure water meeting USP water for injection standards with 0.9% benzyl alcohol as an antimicrobial preservative, creating a bacteriostatic environment suitable for multi-dose research applications.

The 30mL volume represents a strategic choice for research laboratories with higher volume requirements, extended experimental protocols, or institutional research programs utilizing multiple peptide compounds. Compared to standard 10mL bacteriostatic water vials, the 30mL format provides three times the volume, reducing the frequency of vial replacement, minimizing waste from partially used vials, and offering cost efficiency for laboratories with consistent peptide reconstitution needs.

Bacteriostatic water’s primary research application involves reconstitution of lyophilized peptides and proteins. When research peptides are supplied as lyophilized powder, they require reconstitution with appropriate solvent before experimental use. Bacteriostatic water provides an ideal reconstitution medium that combines sterility with antimicrobial preservation, allowing researchers to draw multiple doses from a single reconstituted peptide vial over extended periods without contamination concerns.

The benzyl alcohol preservative functions by inhibiting bacterial growth through disruption of bacterial cell membrane integrity and interference with cellular metabolism. At 0.9% concentration, benzyl alcohol provides effective bacteriostatic action while maintaining compatibility with most peptides and proteins. This concentration represents the USP standard for bacteriostatic water and has been validated across decades of pharmaceutical use.

Molecular and Chemical Characteristics

Water Specifications:

• CAS Registry Number: 7732-18-5
• Molecular Weight: 18.02 Da
• Molecular Formula: H2O
• Chemical Name: Water, purified water, aqua
• Classification: USP Water for Injection quality

Benzyl Alcohol Preservative:

• CAS Number: 100-51-6
• Concentration: 0.9% w/v (weight/volume)
• Molecular Formula: C₇H₈O
• Molecular Weight: 108.14 Da
• Function: Antimicrobial preservative, bacteriostatic agent

Solution Specifications:

• Appearance: Clear, colorless solution
• pH Range: 4.5-7.0 (USP specification)
• Osmolality: Hypotonic solution
• Sterility: Sterile, non-pyrogenic
• Particulate Matter: Meets USP particulate requirements
• Container: Multi-dose vial with rubber stopper

The water component meets USP standards for Water for Injection (WFI), representing the highest pharmaceutical water quality. WFI standards require water produced through distillation or reverse osmosis with additional purification, ensuring removal of microorganisms, endotoxins, dissolved solids, and organic contaminants. This ultrapure water base provides an inert, neutral medium for peptide reconstitution.

Benzyl alcohol’s antimicrobial mechanism involves multiple cellular targets in bacteria. The alcohol disrupts bacterial cell membrane phospholipid bilayers, increasing membrane permeability and causing leakage of cellular contents. Additionally, benzyl alcohol interferes with bacterial enzymatic processes and protein synthesis. At 0.9% concentration, these effects prevent bacterial multiplication without causing immediate bacterial death, thus functioning as bacteriostatic rather than bactericidal.

Bacteriostatic Properties and Sterility

Antimicrobial Preservation:

Bacteriostatic water differs fundamentally from sterile water for injection. While both begin as sterile solutions, sterile water contains no preservative and must be used immediately after vial entry. Any remaining solution must be discarded. Bacteriostatic water’s benzyl alcohol preservative allows multiple entries into the vial over extended periods, provided proper aseptic technique is maintained.

Research validating bacteriostatic effectiveness demonstrates that 0.9% benzyl alcohol inhibits growth of common bacterial contaminants including:

• Staphylococcus aureus (gram-positive cocci)
• Escherichia coli (gram-negative rods)
• Pseudomonas aeruginosa (gram-negative rods)
• Bacillus species (gram-positive rods)

The bacteriostatic effect persists for 28 days after initial vial entry when stored properly and handled with aseptic technique. This multi-use capability provides significant practical advantages for research laboratories conducting extended experiments with reconstituted peptides.

Sterility Assurance:

Each batch undergoes comprehensive sterility testing according to USP standards. Sterility testing involves inoculation of culture media with test samples and incubation under conditions supporting growth of aerobic bacteria, anaerobic bacteria, fungi, and yeasts. Absence of visible growth after incubation confirms sterility.

Manufacturing processes include terminal sterilization through steam sterilization (autoclaving) or sterile filtration through 0.22-micron filters validated to remove bacteria and mycoplasma. Fill processes occur in ISO Class 5 cleanroom environments with appropriate environmental monitoring. Each batch receives unique lot identification allowing complete traceability.

Large Volume Research Applications

30mL Volume Advantages:

The 30mL bacteriostatic water vial provides specific advantages for research laboratories:

1. Extended Multi-Use Capacity: Three times the volume of standard 10mL vials supports extended experimental protocols without vial replacement. A single 30mL vial can reconstitute multiple peptide vials or provide dilution solution for numerous experiments.

2. Cost Efficiency: Per-milliliter cost decreases with larger volumes, providing budget advantages for institutional research programs and laboratories with consistent bacteriostatic water requirements.

3. Reduced Waste: Laboratories frequently discard partially used 10mL vials when experiments conclude or when the 28-day multi-use period expires. The 30mL volume better matches research consumption patterns, reducing waste.

4. Inventory Management: Fewer vials require less storage space and reduce reordering frequency. Research programs can maintain adequate supply with reduced inventory investment.

5. Batch Consistency: Using solution from a single 30mL vial for multiple experiments ensures batch-to-batch consistency within experimental series.

Institutional Research Applications:

Research institutions, university laboratories, and pharmaceutical research facilities conducting extensive peptide research benefit significantly from 30mL bacteriostatic water vials. Typical institutional applications include:

• Peptide Screening Programs: Research programs evaluating multiple peptide compounds require consistent reconstitution solution for comparative studies.

• Dose-Response Studies: Experiments examining peptide concentration effects require multiple dilutions from reconstituted stock solutions.

• Temporal Studies: Research protocols examining effects over time require multiple peptide administrations prepared from the same reconstituted solution.

• Multi-Investigator Facilities: Shared research facilities with multiple investigators using peptides benefit from larger volume vials serving multiple users.

• Teaching Laboratories: Academic laboratories conducting peptide research demonstrations or training require larger volumes for multiple student groups.

Peptide Reconstitution Protocols

General Reconstitution Procedure:

Proper peptide reconstitution technique ensures accurate concentration, maintains peptide stability, and prevents contamination. Follow these research-validated protocols:

Preparation Steps:
1. Remove lyophilized peptide vial and bacteriostatic water from storage
2. Allow both to reach room temperature (15-20 minutes)
3. Clean work surface with 70% isopropanol
4. Gather sterile supplies: alcohol swabs, appropriate volume syringe, sterile needle
5. Perform reconstitution in biosafety cabinet when available

Reconstitution Steps:
1. Calculate required bacteriostatic water volume based on desired final peptide concentration
2. Clean rubber stopper of bacteriostatic water vial with alcohol swab, allow to dry
3. Draw calculated volume of bacteriostatic water using sterile syringe and needle
4. Clean rubber stopper of peptide vial with alcohol swab, allow to dry
5. Add bacteriostatic water slowly down the vial side (avoid direct stream onto lyophilized powder)
6. Gentle swirling motion to dissolve (avoid vigorous shaking that may denature peptides)
7. Allow complete dissolution (typically 1-3 minutes, some peptides require longer)
8. Inspect solution for clarity and complete dissolution
9. Label reconstituted vial with peptide name, concentration, reconstitution date, and lot number

Concentration Calculations:

Accurate concentration determination requires consideration of peptide content, not just labeled amount. Research-grade peptides include certificate of analysis documenting actual peptide content by weight, typically 80-95% depending on the peptide.

Example calculation for 5mg BPC-157 with 85% peptide content:

• Labeled amount: 5mg
• Actual peptide content: 5mg × 0.85 = 4.25mg
• For 2mg/mL solution: 4.25mg ÷ 2mg/mL = 2.125mL bacteriostatic water required
• For 1mg/mL solution: 4.25mg ÷ 1mg/mL = 4.25mL bacteriostatic water required

This calculation method ensures accurate final concentrations for experimental protocols requiring precise dosing.

Peptide-Specific Considerations:

Different peptides may have specific reconstitution requirements:

• Hydrophobic Peptides: Some peptides with hydrophobic character may require addition of small amounts of acetic acid or adjustment to slightly acidic pH for solubility enhancement.

• Aggregation-Prone Peptides: Peptides with tendency to aggregate benefit from slow reconstitution with gentle swirling and extended dissolution time.

• pH-Sensitive Peptides: Some peptides demonstrate optimal stability at specific pH ranges. Bacteriostatic water typically produces pH 5.5-6.5 when peptides are dissolved, suitable for most peptides.

• Multiple Reconstitution: For peptides requiring larger volumes than practical in original vial, consider partial reconstitution in original vial, transfer to sterile larger vial, then add remaining bacteriostatic water.

Reconstituted Peptide Storage and Stability

Storage Conditions:

After reconstitution, peptide solution stability depends on peptide-specific characteristics, concentration, pH, temperature, and storage duration. General guidelines include:

Short-Term Storage (up to 14 days):

• Temperature: 4°C (refrigerator)
• Container: Original vial with rubber stopper, or sterile vial
• Light protection: Store in darkness or amber vial
• Multiple use: Draw doses using aseptic technique
• Stability: Most peptides stable 7-14 days at 4°C

Long-Term Storage (beyond 14 days):

• Temperature: -20°C to -80°C (freezer)
• Container: Single-use aliquots in sterile cryovials
• Freeze-thaw cycles: Minimize to maximum 2-3 cycles
• Aliquot preparation: Divide reconstituted peptide into volumes matching single experimental use
• Thawing: Slow thawing at 4°C preferred over room temperature

Peptide-Specific Stability:

Certain peptides demonstrate exceptional stability allowing extended storage:

• BPC-157: Unusual stability across pH ranges and temperatures allows extended storage. Studies document activity retention at room temperature for weeks.

• TB-500: Relatively stable in solution, typically stable 14+ days at 4°C.

• Growth Hormone Secretagogues: Peptides like Ipamorelin, CJC-1295 generally stable 7-14 days at 4°C.

• Copper Peptides (GHK-Cu): Copper coordination may affect stability; typically use within 7 days.

Consult peptide-specific literature and certificate of analysis for optimal storage recommendations.

Research Applications Beyond Reconstitution

Dilution and Solution Preparation:

Bacteriostatic water serves numerous research applications beyond initial peptide reconstitution:

1. Serial Dilutions: Dose-response studies require preparation of multiple peptide concentrations. Bacteriostatic water provides sterile preserved diluent for creating concentration series from stock solution.

2. Working Solution Preparation: Concentrated peptide stocks can be diluted to working concentrations appropriate for specific experimental models.

3. Vehicle Control Preparation: Experimental controls require vehicle solution matching the peptide delivery vehicle. Bacteriostatic water serves as appropriate control for peptide solutions.

4. Cell Culture Applications: Some cell culture protocols require addition of peptides or growth factors. Bacteriostatic water-reconstituted compounds can be added to culture media at appropriate dilutions.

5. Assay Preparation: Biochemical assays, ELISA protocols, and analytical procedures may require dilution of samples or standards. Bacteriostatic water provides preserved diluent.

Multi-Compound Research:

Laboratories investigating multiple peptide compounds benefit from bacteriostatic water’s standardization. Using consistent reconstitution solution across different peptides eliminates variables related to solvent differences, enabling better experimental comparisons.

Research programs commonly examining multiple peptides include:

• Tissue Repair Research: Studies comparing BPC-157, TB-500, GHK-Cu, and other repair peptides
• Growth Hormone Research: Investigations utilizing Ipamorelin, CJC-1295, GHRP-2, and related compounds
• Metabolic Research: Studies examining metabolic peptides like AOD9604, Fragment 176-191, and related analogs
• Neuropeptide Research: Investigations of Semax, Selank, Cerebrolysin, and cognitive peptides

Quality Assurance and Analytical Testing

USP Compliance:

USP Bacteriostatic Water must meet comprehensive United States Pharmacopeia specifications:

Water Quality Testing:

• Conductivity: Measures ionic impurities, must meet WFI specifications (<1.3 μS/cm at 25°C)
• Total Organic Carbon (TOC): Verifies organic purity (<500 ppb)
• Bacterial Endotoxins: LAL test confirms <0.25 EU/mL
• Sterility: USP sterility testing confirms absence of viable organisms
• Heavy Metals: Atomic absorption or ICP-MS verifies absence of heavy metal contaminants
• pH Testing: Confirms pH within 4.5-7.0 specification range

Benzyl Alcohol Content:

• HPLC Analysis: Quantifies benzyl alcohol concentration
• Specification: 0.9% w/v (9mg/mL)
• Acceptance Range: Typically 0.85-0.95% (±5%)
• Method Validation: HPLC method validated for accuracy, precision, linearity, and specificity

Particulate Matter Testing:

• Light Obscuration: Automated particle counting for particles ≥10μm and ≥25μm
• Microscopic Examination: Visual inspection under magnification
• USP Specifications: Particulate limits defined in USP

Container and Closure Testing:

• Container Integrity: Verifies seal integrity and absence of leaks
• Stopper Compatibility: Confirms rubber stopper does not leach contaminants
• Extractables Testing: Identifies compounds extracted from container closure system

Documentation:

Each batch includes comprehensive Certificate of Analysis documenting:

• Batch/lot number with manufacturing and expiration dates
• Water quality test results (conductivity, TOC, endotoxins, pH)
• Benzyl alcohol content verification
• Sterility test results
• Particulate matter testing
• Authorized quality control signature and date

Third-party analytical verification available upon request for institutional research applications requiring independent validation.

Proper Handling and Aseptic Technique

Aseptic Technique Principles:

Maintaining bacteriostatic water and reconstituted peptide sterility requires consistent aseptic technique. Proper procedures prevent introduction of microbial contamination during vial access:

1. Work Surface Preparation: Clean work area with 70% isopropanol before procedures. Use biosafety cabinet when available.

2. Hand Hygiene: Wash hands thoroughly and consider sterile gloves for critical procedures.

3. Stopper Disinfection: Clean rubber stopper with 70% isopropanol using sterile alcohol swab. Allow complete drying before needle insertion (15-30 seconds).

4. Needle Technique: Use sterile needles and syringes. Insert needle through stopper center. Avoid multiple needle insertions in the same location.

5. Air Pressure Management: When drawing solution, inject air volume equal to liquid volume to be withdrawn, preventing vacuum formation in vial.

6. No-Touch Technique: Avoid touching sterile needle tip, syringe plunger tip, or any surfaces contacting sterile solution.

7. Single-Use Supplies: Use sterile needles and syringes once only. Never reuse or attempt to resterilize.

Storage and Handling:

Proper storage maintains bacteriostatic water quality:

• Temperature: Store at controlled room temperature (15-30°C)
• Light Protection: Protect from direct sunlight and intense artificial light
• Sealed Storage: Keep vial sealed except during solution withdrawal
• Expiration Dating: Observe manufacturer expiration date for unopened vials
• Multi-Use Period: After initial vial entry, use within 28 days

Contamination Prevention:

Visual inspection before each use helps detect contamination:

• Cloudiness or turbidity indicates possible bacterial growth
• Particulate matter may indicate container degradation or contamination
• Color changes suggest chemical degradation or contamination
• Any visible abnormalities require vial disposal

Research Considerations and Best Practices

Experimental Design Factors:

Researchers should consider several factors when utilizing bacteriostatic water:

1. Benzyl Alcohol Compatibility: While 0.9% benzyl alcohol is compatible with most peptides and proteins, certain sensitive compounds may show reduced activity. Consider sterile water for particularly sensitive compounds.

2. Neonatal Research Exclusion: Benzyl alcohol has been associated with toxicity in neonatal animals. Neonatal research models should use preservative-free sterile water rather than bacteriostatic water.

3. Large Volume Administration: For research protocols requiring large volume administration (>10mL in rodent models), benzyl alcohol concentration becomes relevant. Consider dilution or use of sterile water for large volume preparations.

4. Cell Culture Direct Addition: Some cell culture applications involve direct addition of bacteriostatic water-reconstituted peptides to culture media. Final benzyl alcohol concentration in culture should remain below 0.5% to avoid cellular toxicity. Most research protocols involve sufficient dilution that this is not problematic.

5. Control Consistency: When using bacteriostatic water-reconstituted peptides, vehicle controls should also use bacteriostatic water at equivalent dilution to maintain experimental consistency.

Quality Considerations:

Selecting bacteriostatic water source requires quality consideration:

• USP Compliance: Verify USP certification and compliance documentation
• Certificate of Analysis: Request and review COA for each batch
• Sterility Validation: Confirm sterility testing methodology and results
• Supplier Reputation: Choose established suppliers with pharmaceutical manufacturing standards
• Traceability: Ensure lot number traceability and complete documentation

Cost-Benefit Analysis for 30mL Volume:

Laboratories should evaluate 30mL versus 10mL bacteriostatic water based on utilization patterns:

30mL Advantages:

• Lower per-mL cost (typically 20-40% reduction)
• Reduced ordering frequency and shipping costs
• Better volume utilization for high-usage laboratories
• Fewer vials to manage and store
• Reduced waste from partially used vials

10mL Considerations:

• Better for low-usage laboratories
• Lower absolute cost per vial
• Reduced waste if usage doesn’t justify larger volume
• More portable for small-scale field research

Laboratories reconstituting 5+ peptide vials monthly typically benefit economically from 30mL bacteriostatic water vials. Lower usage laboratories may find 10mL vials more appropriate.

Compliance and Safety Information

Regulatory Status:

USP Bacteriostatic Water 30mL is provided as a research solution for in-vitro laboratory studies and preclinical research only. This product has not been approved by the FDA for human therapeutic use, direct human administration, or medical applications.

Intended Research Use:

• Reconstitution of research-grade peptides and proteins
• Dilution preparation for experimental protocols
• In-vitro cell culture applications
• In-vivo preclinical research in approved animal models
• Laboratory investigation requiring sterile preserved water
• Academic and institutional research applications

NOT Intended For:

• Human consumption or administration
• Therapeutic treatment or diagnosis
• Direct injection without appropriate peptide reconstitution
• Neonatal research applications (use preservative-free sterile water)
• Dietary supplementation
• Veterinary therapeutic applications without appropriate oversight

Benzyl Alcohol Safety Information:

Benzyl alcohol serves as the preservative in bacteriostatic water. Safety considerations include:

• Antimicrobial Agent: 0.9% concentration provides bacteriostatic effect
• Generally Recognized as Safe: Long pharmaceutical history at this concentration
• Neonatal Precaution: Associated with gasping syndrome in neonates; avoid in neonatal research
• Toxicity Threshold: Toxicity observed at concentrations far exceeding 0.9% after dilution in experimental use
• Allergen Potential: Rare allergic reactions documented; monitor experimental animals for adverse reactions

Laboratory Safety Protocols:

Researchers should follow standard laboratory safety practices:

• Use appropriate personal protective equipment (lab coat, gloves, safety glasses)
• Handle in well-ventilated areas
• Follow institutional biosafety and chemical safety guidelines
• Dispose of used vials, needles, and syringes according to biohazard/sharps protocols
• Clean spills immediately with appropriate disinfectant
• Consult Safety Data Sheet (SDS) for additional safety information

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