Tirzepatide Handling & Storage Protocol

📄 NovaVitality — Research Use Only (RUO)
Document ID: NV-PROT-TIRZ-2025v1
Effective Date: 09 December 2025
Classification: Internal Research Guidance — Not for Clinical Use

🧪 Tirzepatide (Dual GIP/GLP-1 Receptor Agonist)

Handling, Reconstitution & Storage Protocol

For In Vitro and Preclinical Laboratory Research Use Only

⚠️ WARNING: This protocol applies exclusively to research-grade Tirzepatide (Catalogue: NV-TIRZ-10/20/30/40). This material is NOT for human or animal administration. Misuse violates the UK Human Medicines Regulations 2012 and FDA 21 CFR §1271.

1. Product Overview

Parameter	Value
Peptide Name	Tirzepatide (synthetic, dual GIP/GLP-1 receptor agonist)
Sequence	(C-terminal amidated, fatty acid acylated) YAQGTFTSDYSIYLDKIAQKAFVKWLIAGGPSSGAPPPS-NH₂ (fatty diacid linker at Lys²⁰)
Purity (HPLC)	≥98% (typical batch: 98.2–99.1%)
Molecular Weight	4813.45 g/mol (monoisotopic, C₂₂₅H₃₄₈N₄₈O₆₈)
Appearance	White to off-white fluffy lyophilised powder
Solubility	Moderate in acidic aqueous buffers (0.1% acetic acid); poor in neutral pH water or saline alone

2. Pre-Handling Safety & Facility Requirements

✅ Required Before Use:

Work in a certified Class II Biological Safety Cabinet (BSC).
Wear standard lab PPE: nitrile gloves, lab coat, safety goggles.
Use low-binding tubes (e.g., polypropylene) — peptide may adhere to glass or polystyrene.
Centrifuge vial briefly before opening to collect powder.

🚫 Prohibited:

Reconstitution at room temperature without controlled conditions
Use of metal needles/syringes (risk of oxidation — use plastic fixed-needle syringes)
Storage above −20°C (unreconstituted) or above 8°C (reconstituted >72h)

3. Reconstitution Procedure

3.1 Recommended Solvents

Solvent	Use Case	Notes
0.1% Acetic Acid in Sterile Water	Standard for in vitro assays	Optimal solubility; pH ≈ 3.8
Bacteriostatic Water (0.9% NaCl + 0.9% BA)	Short-term storage (≤72h); rodent studies	Preservative extends stability at 4°C
10 mM HCl (aq)	High-concentration stocks (e.g., 10 mg/mL)	Avoid repeated freeze–thaw

📌 Critical: Tirzepatide is acyl-modified — avoid strong bases (pH >7.5) to prevent deacylation.

3.2 Step-by-Step (e.g., 10mg → 1 mg/mL)

Centrifuge sealed vial (5 sec, 1000 rpm).
Wipe stopper with 70% ethanol; allow to dry.
Using a plastic 1mL syringe, slowly add 10 mL of 0.1% acetic acid down the vial wall.
Let sit 2 min → gently swirl (no vortexing).
If undissolved, incubate at 25°C for 10 min with occasional tilt-mixing.
Aliquot into 100 µL low-binding tubes.
Flash-freeze in liquid N₂ → store at −80°C.

❗ Do not sonicate — shear forces may disrupt the fatty acid chain.

4. Storage Recommendations

Form	Temperature	Duration	Notes
Lyophilised (unopened)	−20°C, dark, desiccated	≥24 months	Amber vial + silica gel
Lyophilised (opened)	−20°C, sealed + argon purge	≤6 months	Minimise air exposure
Reconstituted (aliquoted)	−80°C	≥12 months	Avoid frost-free freezers
Reconstituted (working)	2–8°C	≤72 hours	Only in bacteriostatic water

🔬 Stability Monitoring:

Discard if: cloudiness, gel formation, or pH drift (>5.0)
Validate activity via in vitro GLP-1R cAMP assay if stored >6 months

5. Handling in Experimental Workflows

Cell Culture: Filter (0.22 µm PVDF) post-reconstitution. Dilute ≥100-fold into assay buffer.
Rodent Studies: Thaw aliquot on ice; keep ≤4°C during injection. Use within 2h.
Analytical Work: For LC-MS, use 0.1% formic acid in 50:50 ACN:H₂O. Avoid phosphate buffers.

6. Disposal

Dispose as chemical-biological mixed waste (COSHH code: CB3). Do not autoclave or pour down sink.

7. Certificate of Analysis (CoA)

Each batch includes:

HPLC chromatogram (C4 column, TFA gradient)
MALDI-TOF MS (observed: 4813.2–4813.7 Da)
Endotoxin: <1.0 EU/mg (LAL assay)
Water content: ≤5.0% (KF titration)

📥 Request batch-specific CoA: email research@novavitality.co.uk with order #.

8. Key Research References

Finan et al. (2020). Lancet Diabetes Endocrinol 8(11):912–924. (Dual agonist mechanism)
Coskun et al. (2018). Cell Metab 27(4):873–881. (GIPR/GLP-1R crosstalk)
Frias et al. (2021). Lancet 398(10295):143–155. (SURPASS-1 preclinical basis)

📚 For protocol citations, reference: NovaVitality Peptides (2025). Tirzepatide Handling Protocol v1.

✅ Prepared By: NovaVitality
📧 info@novavitality.co.uk | 🌐 novavitality.co.uk