How to Reconstitute Semaglutide: Step-by-Step Visual Guide

Materials Needed

Required materials: lyophilized semaglutide vial (commonly 2 mg or 5 mg), bacteriostatic water (30 mL vial), U-100 insulin syringes (1 mL, 29–31 gauge), alcohol prep pads, exam gloves, and a sharps container. Semaglutide is a GLP-1 receptor agonist used in preclinical metabolic research. FDA-approved formulations include Ozempic (subcutaneous, diabetes) and Wegovy (subcutaneous, obesity). This guide covers research-grade lyophilized semaglutide only.

Calculating BAC Water Volume

For a 5 mg vial targeting 1 mg/mL: 5 mg ÷ 1 mg/mL = 5 mL BAC water. This is a practical concentration for research doses ranging from 0.25 mg (25 units) to 2.4 mg (240 units, i.e., 2.4 mL across multiple syringe fills).

For a 2 mg vial targeting 0.5 mg/mL: 2 mg ÷ 0.5 mg/mL = 4 mL BAC water. At 0.5 mg/mL, each 100 units on the syringe delivers 0.5 mg.

Reconstitution Steps

Step 1 — Swab both vial stoppers with alcohol pads. Allow 30 seconds to air-dry.

Step 2 — Draw calculated BAC water volume into syringe. For 5 mL total, fill the 1 mL syringe 5 times.

Step 3 — Direct the needle tip toward the inner wall of the semaglutide vial. Inject BAC water slowly along the glass surface, not onto the powder cake.

Step 4 — Swirl the vial gently in slow circles until the powder fully dissolves. The solution should be clear and colorless. Do not shake.

Step 5 — Inspect the solution: clear, colorless, free of visible particles. Discard if cloudy or discolored.

Calculating Units on Syringe per Dose

Using 5 mg in 5 mL (1 mg/mL):

0.25 mg dose: 0.25 mg ÷ 1 mg/mL = 0.25 mL = 25 units on a U-100 syringe.

0.5 mg dose: 0.5 mg ÷ 1 mg/mL = 0.5 mL = 50 units.

1.0 mg dose: 1.0 mg ÷ 1 mg/mL = 1.0 mL = 100 units (one full 1 mL syringe).

2.4 mg dose: 2.4 mg ÷ 1 mg/mL = 2.4 mL = two full syringes (200 units) + 40 additional units.

Storage After Reconstitution

Store at 2–8°C. Per FDA prescribing information for Ozempic, the approved subcutaneous semaglutide formulation is stored refrigerated and stable for 56 days after first use. Research-grade reconstituted semaglutide in BAC water is generally considered stable 4–6 weeks at refrigerator temperature. Do not freeze. Label with date and concentration.

Safety Notes and Research Framing

Semaglutide is a GLP-1 receptor agonist approved for type 2 diabetes (Ozempic) and chronic weight management (Wegovy). Research-grade lyophilized semaglutide is used in preclinical metabolic, appetite regulation, and neuroprotection studies. This guide is for research reference only. Any use in human subjects requires IND authorization and IRB approval.