Preparation L-Glutamine Stock Solution

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Stock Solution Preparation Guide – L‑Glutamine (200 mM)

L‑Glutamine is an essential amino acid used as a nutrient and nitrogen source in cell culture media. It is soluble in water and saline but unstable in solution, degrading over time into ammonia, which can be toxic to cells. Therefore, concentrated stock solutions should be prepared, sterile‑filtered, aliquoted, and stored frozen until use.

Objective

Preparation of 100 mL of 200 mM L‑Glutamine solution in water or 0.85% saline.

Composition

• 200 mM L‑Glutamine (C₅H₁₀N₂O₃) solution in cell culture grade water or 0.85% saline

Required Materials

• L‑Glutamine powder (analytical or cell culture grade)
• Cell culture grade water (Milli‑Q) or 0.85% saline
• Beaker or 250 mL conical flask
• Measuring cylinder or volumetric flask
• Magnetic stirrer or glass rod
• Sterile 0.22 µm syringe filter and sterile container
• Personal protective equipment (gloves, goggles, lab coat)

Procedure

• Step 1
  Weigh out **2.92 g of L‑Glutamine** and transfer it to a 250 mL beaker or conical flask containing **~90 mL** of cell culture grade water or 0.85% saline.
• Step 2
  Stir the solution with a magnetic stirrer or glass rod until the L‑Glutamine is completely dissolved and the solution is clear.
• Step 3
  Transfer the dissolved solution to a volumetric flask or graduated cylinder and adjust the final volume to **100 mL** with the same solvent you used (water or 0.85% saline).
• Step 4
  Sterile‑filter the solution through a **0.22 µm** membrane into a sterile container to ensure sterility (do not autoclave).

Storage: Aliquot the sterile solution into small tubes (e.g., 5 mL) and store at **–20 °C**. The frozen stock solution is stable for several months.

Available Reagents