Research Peptide Reconstitution

Written by Johnathon Anderson, Ph.D., a research scientist specializing in regenerative medicine and serving as an Associate Professor at the University of California Davis School of Medicine

Peptide Reconstitution Protocol

Materials Needed

1. Peptide vial (eg 5mg, lyophilized powder)

2. Bacteriostatic water (10mL via, only need 2mL to 2.5mL)

3. Sterile 3mL syringe with 23-25 gauge needle for bacteriostatic water transfer

4. Alcohol swabs (70% isopropyl alcohol)

5. Clean hands and workspace

Note: While laminar flow is ideal for sterility, a sanitized countertop with restricted airflow can be used with added precautions.

Safety and Preparation

• Sterile Technique: Wear gloves and a face mask throughout the procedure. Disinfect the workspace and all equipment with alcohol before beginning.

• Environment: Perform reconstitution in a clean, controlled environment, minimizing airflow disruptions to maintain sterility.

• Hand Hygiene: Clean hands thoroughly before donning gloves, and avoid touching surfaces unnecessarily.

Protocol Steps

Step 1: Preparation of Materials

1. Inspect the Peptide Vial: Check that the peptide vial is intact, with no visible signs of damage or contamination. The lyophilized peptide should appear as a white, fluffy powder at the base of the vial.

2. Disinfect the Bacteriostatic Water Vial Stopper: Using an alcohol swab, wipe the rubber stopper of the bacteriostatic water vial to reduce contamination risks. Let it air dry for a few seconds.

Step 2: Drawing Bacteriostatic Water into the Syringe

1. Aspirate Air into the Syringe: Pull the syringe plunger back to draw in 2mL to 2.5mL of air. This step helps balance internal pressure when withdrawing the bacteriostatic water.

2. Insert the Needle into the Bacteriostatic Water Vial: Insert the needle through the stopper of the bacteriostatic water vial at a slight angle to minimize coring (small particles breaking off from the stopper).

3. Inject Air and Withdraw Bacteriostatic Water: Inject the 2mL to 2.5mL of air into the vial to equalize pressure, then invert the vial and slowly withdraw 2.5mL of bacteriostatic water into the syringe. Make sure no air bubbles remain in the syringe.

Step 3: Reconstituting Research Peptides

1. Disinfect the Peptide Vial Stopper: Wipe the rubber stopper of the peptide vial with a fresh alcohol swab and allow it to air dry.

2. Insert the Needle into the Peptide Vial: Insert the needle containing the bacteriostatic water into the peptide vial at a slight angle.

3. SLOWLY Add Bacteriostatic Water to the Vial:

   • Angle the Vial: Tilt the vial slightly to allow the bacteriostatic water to flow down the side of the vial, avoiding direct contact with the peptide powder. This reduces foaming and preserves the peptide’s structure and integrity.

   • Inject Slowly: Depress the plunger very slowly to gently introduce bacteriostatic water, ensuring minimal turbulence to prevent peptide degradation.

4. Agitate Gently: After adding the bacteriostatic water, swirl the vial gently—do not shake—to dissolve the peptide fully. Allow the solution to rest briefly if needed until the powder is fully dissolved. Avoid vigorous agitation to maintain peptide integrity.

Step 4: Label and Storage

1. Label the Vial: Mark the vial with the date of reconstitution and concentration if necessary.

2. Storage Conditions: Store the reconstituted peptide vial in a refrigerator at 2–8°C. Avoid freezing, as this will significantly degrade the peptide.

   • Stability: Reconstituted peptides are typically stable for up to 3–4 weeks under refrigeration. Discard any unused solution after this period.

Concentration and Dosage Calculation

• Final Concentration: An example-- With 5mg of BPC-157 dissolved in 2.5mL of bacteriostatic water, the resulting concentration is 2mg/mL.

• Dosage Calibration: Based on this concentration, draw the appropriate dosage into an insulin syringe for accurate dosing according to research needs.

Additional Peptide Reconstitution Notes

• Avoid Cross-Contamination: Use a fresh, sterile needle and syringe for each administration to preserve sterility.

• Reconstitution Technique: Ensure that bacteriostatic water is added gradually to prevent denaturation of the peptide.

• Check for Particulates: Inspect the solution before each use. If particulates or cloudiness is observed, discard the vial.

Peptide Reconstitution Troubleshooting

• Incomplete Dissolution: If the peptide does not dissolve completely when following the peptide reconstitution protocol, allow the vial to sit at room temperature briefly, then swirl gently again. Never apply heat.

• Visible Particles: If particles are present after reconstitution, do not use the solution, as this may indicate contamination or instability.