Substitution of methanol with ethanol for use in fixation of polyacrylamide gels in analysis of proteins and DNA/RNA.

Number

048-EN

Section

General Section

Use

Sector

Health services

Function

Other

Process

Other

Product category

laboratory chemicals

Application

Analysis of protein, DNA and RNA

Abstract

Methanol is used in a fixation process required for analysis of proteins and DNA/RNA on polyacrylamide gels. Fixation is necessary for transforming the proteins or nucleic acids into large, insoluble aggregates which are unable to react or degrade. Ethanol is a good substitute for methanol since it´s less toxic and therefore safer to work with.

The substitution does not work well with Western blot analysis's and processing of membranes. It should also not be used on PVDF-membranes.

Substituted substances

Methanol

CAS No. 67-56-1 EC No. 200-659-6 Index No. 603-001-00-X

Chemical group

Alcohols

Classification: hazard statements

H225 Highly flammable liquid and vapour
H331 Toxic if inhaled
H311 Toxic in contact with skin
H301 Toxic if swallowed
H370 Causes damage to organs

Other adverse effects

The substance is: neurotoxic cat. 4 (Vela et al.) as listed in the Substance Database according to SUBSPORT Screening Criteria (SDSC).

Alternative Substances

Ethanol

CAS No. 64-17-5 EC No. 200-578-6 Index No. 603-002-00-5

Chemical group

Alcohol

Classification: hazard statements

H225 Highly flammable liquid and vapour

Other adverse effects

The substance is: 1 carcinogen (IARC), as listed in the Substance Database according to SUBSPORT Screening Criteria (SDSC).

Reliability of information

Evidence of implementation: there is evidence that the solution was implemented and in use at time of publication

Reason substitution

other toxic effects

Hazard Assessment

Ethanol is highly flammable (H225). Methanol is on the  Subsport Database Screening Criteria (SDSC) and holds the risk phrases H225, 23/24/25 and 39/24/25.

Description of Substitution

Methanol is used in the fixation of polyacrylamide gels in analysis of proteins and DNA/RNA. Fixation is necessary to transform the proteins or nucleic acids into large insoluble aggregates which are unable to react or degrade. Proper fixation is also required to get sharp protein band in electrophoresis. Ethanol is a good substitute for methanol in all applications except western blot analysis and processing of membranes. In these cases the polarity of ethanol disrupts the process.
Since methanol is far more toxic by inhalation and in contact with skin, ethanol works well as a substituent since it improves working place safety and makes the analysis easier to carry out.

Ethanol should also not be used on PVDF-membranes.

Methanol is toxic by inhalation and by skin contact; ethanol isn't and is therefore easier and safer to work with.

Ethanol is a polar compound and this can disrupt some analyses including western blot and membrane processing. The substituent does also not work well on PVDF-membranes.

The substitution does not require any changes in the process and the results of the analysis are not affected by the change of fixing agent. Note however that the substitution does not work well with western blot analyses and membrane processing due to the polar nature of ethanol. Also take care not to use ethanol together with PVDF-membranes.

Case/substitution evaluation

The use of ethanol instead instead of methanol is advantageous from a health and environmental point of view.

State of implementation

Full capacity

Date and place of implementation

Sweden

Enterprise using the alternative

Public Health Agency of Sweden https://www.folkhalsomyndigheten.se/the-public-health-agency-of-sweden/

Availability of Alternative

Available on the market.

Type of information supplier

User

Contact

https://www.folkhalsomyndigheten.se/the-public-health-agency-of-sweden/

Date, reviewed

February 27, 2020