Number
071-EN
Section
General Section
Use
Sector
Scientific research and development
Function
Solvent
Process
Manual activities involving hand contact
Product category
laboratory chemicals
Application
Seperating RNA from DNA and Proteins
Abstract
Chloroform is commonly used in a method to isolate RNA from DNA and proteins. Since chloroform is a neurotoxicant, an endocrine disruptor and possibly also a carcinogen, substitution of the substance is advantageous from a workers' safety perspective. It was found that 1-Bromo-3-chloropropane (BCP) could work as a fully functional substitute for chloroform without any changes in the experimental process.
Substituted substances
Chloroform
CAS No. 67-66-3 EC No. 200-663-8 Index No. 602-006-00-4
Chemical group
Chloroalkanes
Classification: hazard statements
H351 Suspected of causing cancer
H361d Suspected of damaging the unborn child
H331 Toxic if inhaled
H302 Harmful if swallowed
H372 Causes damage to organs through prolonged or repeated exposure
H315 Causes skin irritation
H319 Causes serious eye irritation
Other adverse effects
The substance is: 2B carcinogen (IARC), as listed in the Substance Database according to SUBSPORT Screening Criteria (SDSC).
Alternative Substances
1-Bromo-3-chloropropane
CAS No. 109-70-6 EC No. 203-697-1 Index No.
Chemical group
Halocarbons
Reliability of information
Evidence of implementation: there is evidence that the solution was implemented and in use at time of publication
Reason substitution
CMR
skin/respiratory sensitizing
other toxic effects
Hazard Assessment
Substance to be substituted: Chloroform is a Substance of very high concern and is included on the REACH restriction list, according to Article 73 of Regulation (EG) No. 1907/2006(REACH Regulation). Alternative substance: 1-Bromo-3-chloropropane is possibly carcinogenic to humans (Group 2B), according to the IARC Monographs. » Check the Substance Database according to SUBSPORTplus Screening Criteria (SDSC)
Description of Substitution
In phenol-chloroform extraction RNA is separated from DNA and proteins via a 2-step procedure. The same procedure can also be used to separate DNA or proteins from other compounds. Chloroform is used in the first step where a mixture of chloroform and phenol is mixed with an aqueous solution of the sample containing DNA, RNA and proteins. The combined mixture is then centrifuged, which results in one organic phase and one inorganic phase. The inorganic phase, containing both nucleic acids, is found on top of the vessel allowing it to be extracted. The organic phase, mainly consisting of chloroform will be at the bottom. This phase will contain the proteins of the mixture. DNA can then be separated from RNA in a further step using 2-propanol or ethanol. The substitution of chloroform with BCP does not include any changes in the experimental process. BCP is a less hazardous solvent than chloroform which results in safer working place conditions mainly for the laboratory assistant.
Case/substitution evaluation
The alternative is a less hazardous substance than chloroform, which is of advantage mainly for the laboratory personnel. However, the alternative should be handled carefully, because it is listed in the Substance Database according to SUBSPORT Screening Criteria (SDSC) as well. But in the future this substance should be substituted too because of it risks.
State of implementation
Full capacity
Date and place of implementation
01.09.2010 in Sweden
Enterprise using the alternative
Availability ofAlternative
Fully available.
Producer/Provider
http://www.appliedbiosystems.com/absite/us/en/home.html
Type of information supplier
Research
Date, reviewed
December 11, 2020