[Nhcoll-l] Freezer storage of EtoH tissue samples

Fri Jan 30 14:31:00 EST 2015

John

We too used to store ethanol tissue samples in regular -80 freezers (although all ethanol was drained off to prevent further degrading of DNA by the ethanol) until we switched to our liquid nitrogen dewar system recently.  However at higher temps (-20 freezers) we were instructed that we need to have explosion proof freezers.  We too were cited NFPA 45  10.5.2 and 12.2.2 and Article 501 of code 70 the National Electrical code by our EGS and fire marshal at the time and as far as I know there is no way around this.  It all depends on your local EHS and fire marshal as to whether they pick up on this and enforce it.

Andy

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Andy Bentley
Ichthyology Collection Manager
University of Kansas
Biodiversity Institute
Dyche Hall
1345 Jayhawk Boulevard
Lawrence, KS, 66045-7561
USA

Tel: (785) 864-3863
Fax: (785) 864-5335
Email: abentley at ku.edu<mailto:abentley at ku.edu>
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From: nhcoll-l-bounces at mailman.yale.edu [mailto:nhcoll-l-bounces at mailman.yale.edu] On Behalf Of John P. Sullivan
Sent: Friday, January 30, 2015 12:24 PM
To: nhcoll-l at mailman.yale.edu
Subject: [Nhcoll-l] Freezer storage of EtoH tissue samples

I am curious to know how people on this list deal with cold storage for DNA samples in ethanol.

We have thousands of tissue samples and fish fin clips preserved in 95% EtOH in 2ml Sarstedt screw cap vials. We typically store 16 boxes of 81 tubes per freezer rack and can fit 20 racks in a refrigerator/freezer.  This makes for a maximum of 52 liters or 13.7 gallons of EtOH stored in such a refrigerator/freezer.

We have been informed by Environmental Health and Safety at our institution that it is improper to store these in regular freezers (as opposed to prohibitively expensive, specialized “flammable material” freezers).

This person cited NFPA 45  10.5.2 and 12.2.2 and Article 501 of code 70 the National Electrical code.

Here are links to EH&S pages at several institutions that seem to indicate ethanol storage requires flammable material freezers:

https://urldefense.proofpoint.com/v2/url?u=http-3A__www.usd.edu_research_research-2Dand-2Dsponsored-2Dprograms_upload_Flammable-2DLiquids-2DStorage.pdf&d=AwIGaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=-zLzbwvkpBlNUW26hT9V0sPo63ARzXk_fcfds1MgwqE&s=Lu9FZHVSvj_N1fx3xGuLSSX226uY8R4BNBQUcFDg6G0&e= <https://urldefense.proofpoint.com/v2/url?u=http-3A__www.usd.edu_research_research-2Dand-2Dsponsored-2Dprograms_upload_Flammable-2DLiquids-2DStorage.pdf&d=AwMGaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=sfUtvnZ0J8NsTEIgQdXNNr7uxyEnp6bwuvR7T0JbELE&s=KF4QbWrSBgmNxBdy9sGpN5ZoayJnK3fFYHaiR9pdSK8&e=>
https://urldefense.proofpoint.com/v2/url?u=http-3A__www.ehs.washington.edu_fsofire_flamfrig.shtm&d=AwIGaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=-zLzbwvkpBlNUW26hT9V0sPo63ARzXk_fcfds1MgwqE&s=5wFoH3E2ageakQ8bwy3Os761cGV6EXuhbMXtTkVQrUM&e= <https://urldefense.proofpoint.com/v2/url?u=http-3A__www.ehs.washington.edu_fsofire_flamfrig.shtm&d=AwMGaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=sfUtvnZ0J8NsTEIgQdXNNr7uxyEnp6bwuvR7T0JbELE&s=qdXV0xQBR7I0tPSImqpMt4pk8mwMg_bQcEQlWWOdHC4&e=>
https://urldefense.proofpoint.com/v2/url?u=http-3A__blink.ucsd.edu_safety_research-2Dlab_chemical_liquids_storage_index.html-23Determine-2Dallowable-2Dcontainer-2C-2D&d=AwIGaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=-zLzbwvkpBlNUW26hT9V0sPo63ARzXk_fcfds1MgwqE&s=mWOcZ1zEohkBp5yk3T_uvDrZWGnPlgXpxCeAxBtjE2I&e= <https://urldefense.proofpoint.com/v2/url?u=http-3A__blink.ucsd.edu_safety_research-2Dlab_chemical_liquids_storage_index.html-23Determine-2Dallowable-2Dcontainer-2C-2D&d=AwMGaQ&c=-dg2m7zWuuDZ0MUcV7Sdqw&r=CLFZJ3fvGSmDp7xK1dNZfh6uGV_h-8NVlo3fXNoRNzI&m=sfUtvnZ0J8NsTEIgQdXNNr7uxyEnp6bwuvR7T0JbELE&s=39ybek4t0u3PswjDfD_nsH9j4KPo9k7Ourm56R-91s8&e=>

However, my informal polling of colleagues on Facebook suggests that many of them do in fact store screw-top tubes with tissue samples in ethanol in regular freezers (though perhaps without the approval/oversight of their EH&S departments). My view is that fire/explosion risk from storage of these vials must be negligible to non-existent.  The flashpoint of pure EtoH is 16.6°C.  This temp would only be reached if a freezer failed and no action was taken for a long period of time. The risk of EtOH vapors concentrating in any quantity near internal mechanisms that could spark are minimal since tubes are sealed.

My questions are:  Has anyone here received explicit clearance from their EH&S to store EtOH tissue samples in regular freezers?  If you got this, what was required to obtain it?  If you have ethanol-preserved tissue samples, how do you store them?

This could make for an interesting and useful conversation, but feel free to contact me off-list if you’re concerned about being “on the record.”

- John Sullivan

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John P. Sullivan, PhD
Collections Archivist & Curatorial Affiliate - Cornell University Museum of Vertebrates
Research Associate - The Academy of Natural Sciences of Drexel University

Cornell University Museum of Vertebrates
159 Sapsucker Woods Road
Ithaca, New York 14850-1923 USA

jpsullivan at cornell.edu<mailto:jpsullivan at cornell.edu> / office 607.254.6237, cell 607.342.2234, fax 607.254.2415

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