TRIS HCl (Hydrochloride) – C4H12CINO3

TRIS HCl, & TRIS or TRIS BASE

A Tris Buffer solution has two formulations a Tris Base formulation and a Tris HCl formulation. The only difference between the two is hydrochloric acid added to Tris Base to create Tris HCl. Tris base (C₄H₁₁NO₃) is an effective buffer for biological processes, which is why it is also known as a Tris Buffer. Tris HCl (C₄H₁₂ClNO₃) is almost equally an important component and used extensively for molecular biology. Tris HCl is normally more expensive than Tris base but they are both known as Tris Buffer solutions.

INTRODUCTION

• Tris base and Tris or Tris Buffer are somehow closely similar compounds with the chemical formula C₄H₁₁NO₃. These two terms may be different slightly according to their associated compound. Tris represents the chemical formula C₄H₁₁NO₃. On the other hand, tris bases are the compounds having this formula in association with other compounds [3].
• Tris base or Tris Buffer contains tris (hydroxymethyl) aminomethane chemical group while Tris HCl contains similar group tris (hydroxymethyl) aminomethane along with the presence of attached Hydrochloric acid molecule (C₄H₁₂ClNO₃) [3].
• Tris base or Tris Buffer is available under the names of different labels in the market like 75376-20-4, 3-Amino-3-(2-hydroxyethyl)-1,5-pentanediol, DTXSID10332268. Tris HCl (C₄H₁₂ClNO₃) is obtainable under the names of various labels like Tris (hydroxymethyl) aminomethane hydrochloride, Tromethamine hydrochloride, TRIS HCL (C₄H₁₂ClNO₃), and Tris chloride [7].

STRUCTURE

• The structure of the Tris base (C₄H₁₁NO₃) or Tris Buffer is shown below..

 Figure 1: Structure of Tris-base [1]

• Only difference between the structures of Tris base (C₄H₁₁NO₃) or Tris HCl (C₄H₁₂ClNO₃) is the presence of hydrochloric acid. The structure of Tris HCl (C₄H₁₂ClNO₃) is also shown below.

Figure 2: Structure of Tris HCl [2]

PROCEDURE

The procedures for the preparation of tris buffer, both tris base and tris HCl are described below [9] [10].

Preparation method of Tris Buffer:

To prepare tris buffer in laboratory, following material and chemicals are needed.

Required Chemical:

The chemicals required for the preparation are:

• Distilled water
• Hydrochloric acid (for Tris HCl only)
• Tris (hydroxy methyl) aminomethane

Required Material:

The laboratory material needed for the preparation are:

• Suitable container
• Beakers
• pH Meter
• stirrer

      Preparation Steps:

• First of all, take distilled water (deionized) of about 800ml and pour it into the suitable beaker.
• Measure 121.14 grams of Tris Base (hydroxymethyl) aminomethane into the beaker containing distilled water.
• Stir it well by using the stirrer until the powder is fully dissolved in the deionized water.
• In order to achieve the desired pH of the tris buffer solution, add hydrochloric acid until the solution is adjusted according to the desired pH for Tris HCl only.
• Add the solution into the suitable container and check the pH of the buffer solution by using a pH meter (an instrument used to measure the concentration of hydrogen ions in the solution). The pH is checked after cooling the solution.
• The Tris Buffer is diluted by adding distilled water. After the preparation, the stock solution can be preserved at room temperature for future use.

SPECIFICATIONS

The specifications of tris buffer, both tris base and tris HCl are described below in the table [4] [5] [6] [8].

APPLICATIONS

The applications of tris base and tris HCl are described below [11] [12].

• Protein Electrophoresis:
  A tris buffer is important for protein electrophoresis. A tris buffer helps in keeping the proteins linear. An important amino acid is present in the structure of protein named glycine that plays a pivotal role in stacking gel.

• DNA Agarose Electrophoresis:
  DNA Agarose electrophoresis is a technique used to separate DNA pieces on the basis of their weight and charge. Smaller fragments separate rapidly than larger fragments of DNA.

• pH control:
  Tris and Tris Hydrochloride both are used in maintaining the pH of biological systems. It additionally controls pH for body fluids.

• Starting material for polymers:
  Tris base and tris HCl are used as a starting raw material in the production of polymers.

• Standardization:
  Tris is used in the direct standardization of a strong acid solution.

• DNA Extraction:
  Tris HCl is equally used for the extraction of DNA fragments.

• Surfactants:
  Tris bases are used in the production of surfactants.

• Protein crystal growth:
  Tris base is also used in enhancing the growth of protein crystals at balanced pH conditions.

SAFETY AND HAZARD:

The hazards and safety measures of a tris buffer is explained below [13] [14].

Health Effects: Following are the health effects that are caused by tris bases and tris HCl that are described below.

A) Eyes Exposure to Tris base and Tris HCl:
If eyes are exposed inadvertently to the powdered form or to the splashes of stock solution, it can be harmful to the eyes and cause itching, irritation, and redness. It can also cause temporary damage to the eyes.
First Aid Measure:
Carefully wash the eyes with excess water for 10 to 15 minutes. In case of severe irritation and pain, consult medical advice immediately.

B) Skin Exposure:
In case of skin exposure, skin rash, itching and irritation may occur on the contaminated body part.
First Aid Measure:
Promptly remove the contaminated clothes. Wash the clothes properly so that no trace of chemicals is left on them. Wash the skin with plenty of water for 15 to 20 minutes. Use neutral soap for washing purposes.

C) Accidentally Ingested:
Tris bases and Tris HCl on ingestion can cause disturbance in the digestive system and may be harmful.
First Aid Measure:
Don’t offer anything to eat or drink to the unconscious victim. Instantly assist the victim to the hospital so that he can be handled accordingly.

D) Inhalation:
On inhalation, it can cause respiratory tract infections which can lead to severe irritation or itching problems in the upper respiratory tract.
First Aid Measure:
Carefully move the victim to a spacious place where fresh air is available. Give artificial breathing, if the victim is not breathing.

REFERENCES

1. Santa Cruz Biotechnology – Tris base (CAS 77-86-1)
2. GoldBio – This vs That
3. Difference Between – Tris and Tris Base
4. Chemical Book – Tris Base
5. Chemical Book – Tris hydrochloride
6. PubChem NCBI NIH – Tris hydrochloride 3 Chemical and Physical Properties
7. PubChem NCBI NIH – Tris hydrochloride 2.4.1 Depositor-Supplied Synonyms
8. PubChem NCBI NIH – Tris hydrochloride 2.1.2 InChI
9. AAT Bioquest – Tris Buffer (1 M, pH 7.2) Preparation and Recipe
10. Thought Co. – How to Make Tris Buffer Solution for Medical or Lab Use
11. Bio Rad – Uses of Tris Buffer in Protein Electrophoresis and Western Blotting
12. Cosmo Bio – Tris and Tris hydrochloride
13. Sigma-Aldrich – Tris and Tris hydrochloride Safety Data Sheet
14. Cayman Chem – Tris HCl Safety Data Sheet