How could I forget the message I got about twelve copies of?  It is tacked
onto the end for those interested in archiving the complete summary.

Noelle Patterson
Naval Medical Research Center
Bethesda, Md
pattersonn@NMRIPO.NMRI.NNMC.NAVY.MIL


----------
From:  Patterson, Noelle
Sent:  Monday, January 04, 1999 1:19 PM
To:  'Histonet'
Subject:  Summary:  Denatured alcohol + 1


This is a summary note of replies to whether I could use some denatured
ethanol I found lying around in great quantities.  I must admit, I am a bit
confused.  Naturally, I got opposing perspectives.  But, I think that I the
ethanol may be best saved for a non-histologic purpose.  I may try it
anyway, as there seem to be a few notes of support, and it will only go to
waste if I don't use it.  We are due to move/evacuate the building by
October to merge with either NIH or WRAIR (perhaps sooner), only reasonable
amounts of chemical will be moved. (Don't you just love military minds!
They figure it costs more to move it than to buy new stuff, I guess). What I
have is Fisher A407-500 Ethyl Alcohol (Denatured) Permit No. SDA-NJ-2517.
It has been denatured with SD-1 (a.k.a.  to every 100 gallons of ethyl
alcohol is added:  1 gallon ethyl acetate, 1 gallon methyl iso-butyi ketone
and 1 gallon hydrocarbon solvent.  Any more experienced users are free to
comment.

Here are the responses to my request:

Noelle:
I use denatured alcohol with no problems, but it all depends on how it was
denatured as there are different ways and I have had a bad experience with
one.
Janet Maass
We regularly sell denatured alcohol for histology use.   The one factor you
should keep in mind is the type of denaturant.  If the denaturants include
other alcohols such as methanol, Isopropanol, MIBK, etc. then you should
have no problems.  On the other hand, denaturants also include ethyl acetate
and other more volatile chemicals which could interfere with staining or
even the machinery ( ie processors, stainers, etc. )   So read the labels
before using your mystery solvents.

THIS WILL JUST GO INTO A TISSUE TEK MANUAL STAINING RACK FOR DEHYDRATING
AFTER H&E'S OR IMMUNOHISTOCHEMISTRY COUNTERSTAINS (WITH HEMATOXYLIN).  WHAT
DO YOU THINK?
Hope this is helpful.    Now go home and enjoy the New Year !!
I DID, THANKYOU.  I HOPE YOU ENJOYED YOURS AS WELL!!
Cliff Berge
President
Decal Chemical Corp

I've always used denatured alcohol for all histology procedures and found it
makes no difference at all. Using 100 percent ETOH is a waste of money.
HERE HERE, AND A WASTE OF TIME IN PAPERWORK ALSO, WHICH IS ONE OF THE
REASONS I AM LOOKING TO USE UP THESE LEFTOVER STOCKS!

Tim Morken, B.S., EMT(MSA), HTL(ASCP)
Infectious Disease Pathology
Centers for Disease Control

Alcohol labelled as "Reagent alcohol" is a denatured alcohol.  You may want
to dig around in some MSDS sheets for your "denatured alcohol" and ask for
or call technical services at Fisher to find out what their Reagent alcohol
contains.   If you get lucky, the container actually tells you what is in
these alcohols, often a mixture of methanol, ethyl alcohol, and maybe some
isopropyl, or maybe someone will be able to give the info via H'net so you
can compare and use if it is comparable to "reagent alcohol".
I know people who use Reagent alcohol in processing and staining.  I would
avoid using it for dissolving dyes that are listed as 1% eosin Y in alcohol,
learned years ago this meant ethyl alcohol, when the dye did not go into
solution.
Happy New Year
Gayle Callis

Hi!  I don't see any reason that you could not use this alcohol.  Hopefully,
the seals have not been broken.  Otherwise, I personnally don't see any
reason it could not be used.   H&E staining is usually very versatile as
far, as the alcohol that could be used.  Happy New Year!  Marianne Osborne,
Univ. of Nebraska Medical Center Eppley Cancer Institute,  Omaha, NE.  We
here in Omaha received about 4.3 inches of snow yesterday.  This is our
first measurable snow this year.  It is very cold outside also.  Happy New
Year!
M. Osborne
Replying directly, due to repeating.
Yes you can use it, BUT . . . are you certain of it's percentage now, if it
is old? Could it have broken down, evaporated, absorbed water, whatever,
while it was stored in the abandoned lab?  How old is it?
IT IS STILL SEALED.  NO NOTABLE EVAPORATION.  NOT TOO OLD, DOESN'T EVEN HAVE
DUST ON IT.
I might suggest trying it with an extra control slide that you don't mind
wasting, just to see what happens. If it is absolute (100%) denatured
alcohol, you might want to dilute it, and use it as 95% denatured alcohol,
just in case it has absorbed water, or is not as pure as it should be. Use
your "Good" absolute denatured as the final alcohols before the xylene.
THIS IS A GOOD IDEA!
If you have a hydrometer, you could always measure it's percent of water.
But most labs don't have one of these "floating thermometer-look-alikes"
laying around.
(It's been snowing on and off around here in Michigan. On Saturday, we
should be hit with about 6 inches around the Detroit area. Parts of the
Upper Peninsula of Michigan got almost 7 feet in the last week, so I'm not
really complaining about #189# foot.)

WOW, WE ONLY GOT HIT WITH 1 INCH OF SNOW, AND A WEEK LATER WITH AN ICE/SLEET
STORM THAT WAS MELTED BY THE NEXT MORNING.  MADE GETTING HOME AFTER A PARTY
RATHER TRICKY THOUGH!
Peggy A. Wenk, HTL (ASCP)
Anatomic Pathology
Wm. Beaumont Hospital
3601 W. 13 Mile Rd.
Royal Oak, MI 48073-6769

Dear Noelle,
I apologise for the lateness of my reply, I took a week off to go to the
beach and instead watched numerous planes and helicopters scouring the area
offshore for missing competitors in the Sydney/Hobart race.
SOUNDS INTERESTING, BUT NOT RELAXING!  AT LEAST YOU DIDN'T HAVE TO CANCEL
YOUR TRIP, LIKE MY MOTHER-IN-LAW, WHO COULDN'T GET A FLIGHT OUT THIS
WEEKEND...AT LEAST NOT IN A REASONABLE TIME.
we have used denatured anhydrous alcohol for nearly twenty years and have
substituted it in all procedures and recipes including processing without
any ill effects. Having said this, a word of caution, the denaturant in the
formula we use is MEK or Methyl Ethyl Ketone with about 2% Methanol, it also
contains a proprietery known as "Bitrene" which is an additive that tastes
incredibly bitter so you can't drink it. (Also used in toilet blocks etc. to
stop children from eating them); there are many form s of denatured alcohol
and a list of these may be found in the Merck Index and form part of the
regs. in the USA.

SO DENATURED WITH METHYL ISO-BUTYI KETONE WOULD NOT BE THE SAME, BUT IS IT
HARMFUL TO USE?
One of our net colleagues made mention of not using Methanol denatured
alcohol, and they are quite correct for absolute Methanol, when less than
10% Methanol is present there appears to be no problem for eosin binding
(you may recall that Methanol is in fact added to PAP E.A.50 and keratinised
cells have no problem taking up the Eosin)
Here in Australia, the only advantage for using denatured alcohol (or
100IMS) is that the user is not required to hold a Customs Excise
Permit/Licence, and no fully documented records have to be kept with respect
to useage or by whom and what for. So if record keeping is not your forte
and you have no wish to go through the hassle of Customs Officials going
through and auditing you then use denatured. There is no price advantage
whatsoever. All other forms of regulation that apply to Ethanol such as
storage, quantities, vessels etc. are the same.
From, Mick Rentsch
YOUR REPLY WAS NOT AT ALL UNTIMELY.  THANKS!

Denatured alcohol for histologic use:
Pure ethyl alcohol (ethanol) is the standard alcohol for histologic use,
but, because it is possible to drink it (anecdotes on request), using it
requires a lot of administrative hassle. Some histology services are able to
delegate this hassle to the hospital pharmacy, which however has to have
adequate locked storage space.
Ethyl alcohol is rendered (legally) undrinkable by naturing it. The Bureau
of Alcohol, Tobacco, and Firearms (BATF, under the Treasury department of
the US Gummint) has a large number of official denaturing formulas.
Alcohol S3DA is denatured with 5% methanol. For histologic use, it also has
5% isopropanol added (I don't know why), and this is S3DA Modified. This
mixture is the usual one called Reagent Alcohol in histology. I believe that
this is a generic usage, and not a trade name.
Another denaturant for histologic use is methyl isobutyl ketone (MIBK). The
major problem with this denaturant is that it smells bad.
IF SMELL IS THE ONLY PROBLEM, CAN I STILL USE IT FOR DEHYDRATING IHC'S?
Ethanol denatured with acetone is sometimes available in laboratory
catalogs.
It strips out eosin very rapidly.
As far as I know, reagent alcohol can be used for any histologic purpose for
which ethyl alcohol is specified. I don't know what happens to it, either
chemically or governmentally, if you distill it with a spinning band still
and try to re-use it.
Bob Richmond
Samurai Pathologist
Knoxville TN

AS USUAL, A VERY ENLIGHTENING AND USEFUL RESPONSE.  THANKS FOR SHARING YOUR
KNOWLEDGE.



Well, after reviewing the notes, I guess I will let it just gather dust on
the shelf.  MAYBE NOT, WHAT DO YOU THINK SAMURAI?

Thanks to all who responded,
Noelle