From:

The Department of Pathology I and

the Department of Technology,
University of Umd, Sweden.
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CORROSION DURING AUTOCLAVE STERILIZATION

11. VOLATILE ORGANIC AMINES AS CORROSION INHIBITORS
bY
C-M. FAJERS
L. G. HOLMLUND
E. STENMAN
For personal use only.

INTRODUCTION

Autoclaving, i.e. sterilization in saturated steam under pressure, is generally

considered to be the most certain method known for destroying all forms
of microbial life. The humidity and temperature are the most important
factors in this process (Konrich, 1938; Walter, 1948; Berger, 1954; Perkins,
1960).
Instruments suffer, however, from corrosion during autoclaving due to
the effect of the hot steam medium. The corrosive attack on carbon steel
instruments is particularly great (Holmlund, 1963 f). Surgical and dental
cutting instruments must be both hard and flexible and these properties
depend to a considerable extent on the carbon content in such steels. Chro-
mium steel instruments are much more corrosion resistant but do not have
the hardness and flexibility of carbon steel instruments and are moreover
much more expensive. If the corrosion occurring on carbon steels during
autoclaving could be reduced, or completely eliminated, this material would
then be superior to chromium steels for the manufacture of dental and
surgical cutting instruments.
Considerable corrosion inhibition has been reported in power plants and
industrial steam-heating systems when volatile amines were mixed with the
steam (Dreyfus, 1942; Berk & Nigon, 194%;Kahler & Brown, 1954; Obrecht,
*) This work war supported by the Swedish Council for Applied Research.
 24 C-M. FAJERS, L. G. HOLMLUND AND E. STENMAN

1959). It is therefore of interest to investigate whether also the corrosion

during autoclaving can be reduced or completely eliminated by adding a
volatile organic amine to the corrosive medium. Several different types of
amine must of course be tested and their effectivity compared in order to
find out which ones are most suitable, with respect to effectivity, cost, tox -
icity, ease of handling etc., for practical clinical use.
The present study was thus carried out in order to examine the effecti-
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veness of different types of volatile organic amines as corrosion inhibitors

during the autoclave sterilization of carbon steel materials.

MATERIAL AND METHODS

Uniform carbon steel test samples (Sandvik 21T10P) which corresponded

as far as composition and pretreatment were concerned, to cutting instru-
ments (e.g. dental round burs, root canal files, scalpel blades etc.) were
tested in a high pressure autoclave at 1 4 ° C for 25 min.
The samples were cylindrical (length ca 45 mm, diameter 5 mm) and
were protected during storage by means of a thin layer of oil. The same
For personal use only.

autoclave - chamber volume 14 1 - was used for all of the experiments.

A temperature registration device was connected to a bimetallic strip inside
the autoclave. Deionized water (500 ml) with a resistance of 400.000-500.000
ohm. cm was used as autoclave nfeed water)). The test samples were enclosed
in envelopes in some of the experiments.
The following amines were tested:
Cyclohexylamine (CH),(CH.J4CHNH,, benzylamine (C,H,CH,NH,),
morpholine (OCH,CH,NHCH,CH,), decylamine (CH,(CH,),NH,) and
octadecylamine (CH,(CH,),,NH,).
Before each corrosion experiment the following precleaning procedure
(Holmlund, 1%3 b, d, e) was carried out. The randomly selected test samples
were first washed with ether and rinsed thoroughly in redistilled water and
finally cleaned by means of ultrasonic treatment. The air in the autoclave
was not evacuated, however, in order to avoid loss of the volatile amine
(Holmhnd, 1%3 d, e, f, 1%). In all of the tests, the autoclave feed water
had an initial temperature of 70°C (Holmbnd, 1963 d). During the experi-
ment, each sample was laid on a U-shaped :upport of pyrex glass
which was either enclosed in an envelope or placed, when the sample was
autoclaved freely exposed, in a low pyrex dish. The envelope was sealed
with autoclave tape after the air had been pressed out as completely as pos-
sible.
The experimental procedure is shown in Table I together with the com-
mentaries given below. The amine to be tested was added to the corrosive
 CORROSION DURING AUTOCLAVE STERILIZATION 25

medium in different ways. Except for the series 6, (11) and 15, the amount
of amine used was close to the lower boundary for maximum corrosion
inhibition under the present experimental conditions (Fajers C Holmlund,
1966,1967). Each series comprised 10 autoclave tests (8 in a few exceptional
cases).

Commentaries to the experimental procedure (Table I).

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Series 1-7: In each test, three freely exposed samples - which had just
been cleaned ultrasonically - were exposed simultaneously to one of the

Table I. Ezperirnental procedure

Type A . Samples exposed freely in the autoclave

Amine added directly to the Blank tests without amine

autoclave nfeed water!) N N
Series 1 Cyclohexylamine 0.5 ml 10
For personal use only.

Series 2 Decylamine 0.5 ml 10 Series 3 (blank for 1 and 2) 10

Series 4 Octadecylamine 0.5 g 8 Series 5 (insufficient cleaning) 6
(blank for 4) 6
Series 6 Morpholine 1.0 ml 10 Series 7 (blank for 6) 10
Type B. Samples sealed in envelopes

Amine added directly to the Blank tests without amine

autoclave nfeed waterw N N
Series 8 Cyclohexylamine 0.5 ml 10 Series 9 (blank for 8 )
~
10
Type C. Samples exposed to direct or indirect amine action
~ ~~

I. Amine placed inside one or several of the envelopes

Series 10 Cyclohexylamine 0.010 ml per envelope 10
Series 11 Cyclohexylamine ca 0.035 g per envelope 10
Series 12 Morpholine 0.005 ml per envelope 10
Series 13 Morpholine ca 0.016 g per envelope 10
Series 14 Benzylamine 0.010 ml per envelope 10
Series 15 Octadecylamine 0.25 g per envelope 10
Series 16 Octadecylamine 0.01 g per envelope 10
Series 17 Decylamine 0.05 ml per envelope 10
11. Blank tests without amine in the envelopes N
Series 18 envelope material of same type as in series
10,12,14, 15-17 10
Series 19 envelope material of same type as in series
11,13 10
 26 C-11. FAJERS, L. G. H O L M L U N D A N D E. STENMAN

corrosion inhibitors. In the tests in series 4 and 5, difficulties were encoun-

tered with respect to the cleaning of the autoclave (see washing procedure)
and therefore a number of intermediate tests without amine were per-
formed.
Series 8,9, 11, 16, 17, 19: In each test, the corrosion occurring on a sample
enclosed in an envelope was determined.
Series 10, 12, 13, 14: Each test comprised three test samples as follows:
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a. Test sample enclosed in an envelope together with amine.

h. Test sample in a shallow pyrex dish.
c . Test sample enclosed in an envelope.
Series 18: As for the series immediately above but without amine in a.
Series 15: Each test included three samples as follows:
a. Test sample enclosed in an envelope.
b. Test sample enclosed in an envelope together with amine.
c. Glass support alone (blank).
d. Test sample.
e. Glass support enclosed in an envelope together with amine (blank).
For personal use only.

After each experiment, the corrosion products formed were collected from:
1) the test sample by subjecting it to the same ultrasonic treatment as that
employed before the autoclave test, i.e. post-cleaning,
2) the U-shaped support on which the sample was placed during the test,
3) the shallow pyrex dish and
4) the envelope material by wet-combusting it (Holmlund, 1963 d, e, f;
Fajers & Holmlund, 1967).
The Fe in the corrosion products collected was ionized by acidification
and taken as a quantitative measure of the corrosion. Fe in the envelope
material itself was also determined (blank). From previous investigations,
it is known that the Fe amount dissolved from the autoclave chamber is SO
small that it can be neglected (Holmlund, 1963 d, e). This was verified once
again in series 15 (c). Fe was determined spectrophotometrically and in
duplicate according to a modified orthophenanthroline method (Holmlund,
1963a).
The quantitative measure of the corrosion, expressed as pg Fe dissolved
out/cm2 sample surface area, was obtained for each sample by adding the
Fe amounts in the post-cleaning liquids (see Holmlund 1963 C) to the Fe
amounts from the support, dish and envelope. Correction was made for
the Fe in the envelope blank.

R a s h i n g procedure
The gla>a\\are and plastic-covered pincers used for transferring the test
 CORROSION DURING AUTOCLAVE STERILIZATION 27

Table 11.

Amount of Fe dissolved (pg/cma sample surface area) from each sample during autoclaving

Type A . Samples exposed freely in the autoclave on a U-shaped support in a low pyrex dish

Amine added directly to autoclave Blank test without

feed water N Mean SD amine N Mean SD
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Series 1 Cyclohexylamine 10 0.46 0.16 Series 3 Steam alone 10 119.4 48.2

Series 2 Decylamine 10 0.99 0.59 Series 6 Steam alone 6 355.87*) 82.25
Series 4 Octadecylamine 8 19.76 18.19 Steamalone 6 83.53 17.35
Series 6 Morpholine 10 3.% 1.82 Series 7 Steam alone 10 220.21 87.43
Type B. Samples enclosed in envelopes

Amine added directly to autoclave Blank test without

feed water N Mean SD amine N Mean SD
Series 8 Cyclohexylamine 10 15.20 8.14 Series 9 Steam alone 10 246.5 44.6

Type C. Samples either enclosed in envelopes or freely exposed in the autoclave

For personal use only.

~ ~

I. Direct and indirect amine action

~ ~

Sample freely Sample enclosed in Sample enclosed in

Amine used N exposed envelope without amine envelope with amine
Mean SD Mean SD Mean SD
~ ~~~~

Series 10 Cyclohexylamine 10 47.92 24.69 379.33 63.81 22.85 7.72

Series 11 Cyclohexylamine 10 - - 8.95 2.83
Series 12 Morpholine 10 166.80 41.90 215.54 112.30 3.33 1.17
Series 13 Morpholine 10 104.10 40.04 269.31**) 90.43 3.73**) 2.49
Series 14 Benzylamine 10 139.16 67.08 169.74 100.51 13.98 9.61
Series 15 Octadecylamine 10 53.72 26.84 145.68 26.06 5.64 3.17
Series 16 Octadecylamine 10 6.00 3.62
Series 17 Decylamine 10 5.11 3.12
11. Blank test without amine

Series 18 Steam alone 10 151.81 49.70 186.25 49.94 (Blank for 10, 12 and 14-17)
Series 19 Steam alone 10 - 186.93 47.69 (Blank for 11 and 13)

*) This value was obtained when the autoclave chamber was not sufficiently rinsed after the alco-
holacetic acid cleansing. The next value was found after further rinsings - the pH was also shown to
be natural and no smell of acetic acid could be detected.

**) The Fe inside the envelopes originating from corrosion products from the test samples was not
determined here. This was negligible in all experiments in which the sample was sealed together with
amine in its envelope but could not be neglected when the sample was sealed without amine.
 28 C-M. FAJERS, L. G . IIOLMLUND A N D E. STENMAN

samples were all cleaned according to Holmlund (1963 c ) . A separate pair

of pincers was used for each test sample throughout the whole of the ex-
periments.
After each test, the autoclave was cleaned and the cleaning procedure
employed was dependent on the type of corrosion medium used. When the
medium consisted of only steam and steam condensate, the sterilization
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chamber was cleaned using de-ionized water. When it contained cyclohexyl-

amine or morpholine, the chamber was cleaned twice with a soap solution
and thereafter washed three times with de-ionized water.
When the medium contained decylamine, the chamber was cleaned three
times with 32 % ethyl alcohol and twice with soap solution and thereafter
washed twice with de-ionized water. When it contained octadecylamine, the
chamber was cleaned three times with a mixture of equal parts of 99.5 yo
alcohol and 20 acetic acid, then twice with soap solution and thereafter
washed four times with de-ionized water (Holmlund, 1963 d, e ; Fujers G
Holmlund, 1967).
For personal use only.

RESULTS

The results presented in Table I1 show that

1) the amines tested effectively inhibited corrosion during the autoclaving
of carbon steels both when added directly to the autoclave feed water and
also when sealed with the sample inside its envelope,
2) octadecylamine was somewhat less effective than cyclohexylamine, mor-
pholine and decylamine, when the inhibitor was added directly to the feed
water and
3) morpholine, decylamine and octadecylamine provided a somewhat better
inhibition than cyclohexylamine and benzylamine when the amine was
enclosed in the container.

DISCUSSION

The corrosion process occurring during autoclave sterilization is electro-

chemical in nature and results mainly in a general surface corrosion. In this
type of corrosion there appear anode and cathode surfaces which are very
small and move continuously over the surface of the metal (Uhlig, 1948;
Lutimer 15 Hildebrund, 1951 ; Ljungberg, 1962). The corroding medium is
steam and condensate, and it has been found that its destructive effect can
be reduced by adding an inhibitor to it. The inhibitor must however be
soluble in water and/or possess such a volatility that it is volatilized and con-
densed with the steam (Dreyfw, 1942).
 CORROSION DURING AUTOCLAVE STERILIZATION 29

Volatile organic amines have been found to be effective as inhibitors in

power plants and industrial steam heating systems. Holmlund (1963) and
Fajers & Holmlund (1966 ; 1967) have tested compounds systematically for
instrument sterilization and found them to be very efficacious. The mecha-
nism responsible for the inhibition obtained with different types of volatile
amines has not yet been established. The amines have previously been divided
into two groups, viz. neutralizing and filming, depending on the manner in
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which they are supposed to exert their inhibiting effect. Cyclohexylamine,

morpholine and benzylamine are usually considered to act mainly by neutral-
izing the acid radicals in the corroding medium. They are all strongly basic
and can be classified chemically as cyclic hydrocarbons (Carswell & Morrill,
1937; Dreyfus, 1942; Berk & Nigon, 1948; Muguire, 1954). Octadecyl-
amine is the best known of the filming amines which are usually straight
chain primary amines containing 10 to 18 carbon atoms and an amine group.
It is non-basic and is considered to inhibit corrosion by forming a mono-
molecular chemisorbed layer on the metal surface which functions as a bar-
rier between the metal and the corroding medium (Kuhler & Brown, 1954;
For personal use only.

Kajanne, 1957). Decylamine also possesses a straight chain with 10 carbon

atoms and an amine group but is, in contrast to octadecylamine, strongly
basic.
The inhibiting action of amines on electrochemical corrosion is dependent
on, among other things, the nature of the corrosive medium and the length
of the corrosion reaction. Under the special conditions which exist during
autoclaving, viz. a short attack which because of the high temperature is
particularly intense, it seems that the neutralization of acid radicals in the
medium is of prime importance for the inhibition of the corrosion (Fajers,
Holmlund & Stenman, 1967).
The autoclave chamber was cleaned thoroughly between experiments
using a special previously tested washing procedure for the removal of any
possible traces of amine. Octadecylamine was however found to be harder
to remove than the other inhibitors employed. The cleansing of the auto-
clave chamber was therefore checked after each of these experiments by
performing several consecutive tests using steam alone (Series 5).
All of the amines examined were found to be effective as inhibitors in-
dependent of how they were added to the corrosive medium (Table 11). In
certain experiments (Type A and B), the amine was mixed with the auto-
clave feed water and volatilized with the steam and was proved to inhibit
corrosion both when the sample was freely supported in the autoclave chamber
and also when it was enclosed in an envelope. In other experiments (Type C),
the amine was added by sealing it together with the sample in the envelope.
 30 C-M. FAJERS, L. G . HOLMLUND AND E. STENMAN

This procedure eliminates primarily the corroding effect of the steam and
condensate inside the envelope. It is becoming increasingly common now-
adays to autoclave instruments packed in bacteria-tight envelopes so that
they can be easily stored subsequently under sterile conditions (Wallen-
berg & Jarnhall, 1956; Savas, 1958; Brigden, 1960; Prince & Mogg, 1960;
Fajers & Holmlund, 1966).
The results for, for example, Series 10 and 15 (Table 11) show that a con-
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siderable reduction in the corrosion also occurs for the freely exposed sam-
ples. This indicates that a certain amount of the amine diffuses out through
the envelopes. This is particularly conspicuous in Series 10 in which the
amine was introduced into the envelope by impregnating its paper section
with cyclohexylamine. This amine is very volatile and thus quickly diffuses
out through the envelope and hence the amine concentration inside the
envelope becomes too low for complete corrosion inhibition. However, by
impregnating the paper section with a less volatile amine (Series 13), it was
found that the inhibition was nearly complete even if a certain amount of
the amine diffused out and inhibited the corrosion of the freely exposed
For personal use only.

samples.
This method in which one introduces into the envelope a quantity of
amine, chosen whh regard to the amount of corrosive medium in it, means
both that the total quantity of amine required is greatly reduced and also
that the highest amine concentration is obtained very close to the corrosion-
sensitive surface (See also Fajers & Holmlund, 1967). One should aim at
using the smallest possible amount of amine for complete inhibition since
most amines in high concentrations are toxic. Traces of amines and/or their
salts may remain on the instrument surface after autoclaving and, when the
instrument is used, these substances may be transferred to the body or into
the tissues and there produce toxic reactions. However, the surface products
obtained from carbon steel samples after corrosion inhibition using cyclo-
hexylarnine, decylamine and morpholine did not produce any cytotoxic
effects (Holmlund & Tarnvik, 1963; Holmlund & Tarnvik, 1967). No toxic
or undesirable dermal effect could moreover be found with octadecylamine
(Maguire, 1954).

SUMMARY

The volatile organic amines, cyclohexylamine, morpholine, benzylamine,

decylamine and octadecylamine, have been tested as corrosion inhibitors
in the autoclave sterilization of instrument steels. The corrosion experiments
were performed in a high pressure autoclave on carbon steel samples that
 CORROSION DURING AUTOCLAVE STERILIZATION 31

corresponded as far as composition and pretreatment were concerned to

dental and surgical cutting instruments.
The amines were introduced into the corrosive medium, i.e. steam and
condensate, (a) by adding them immediately to the autoclave feed water,
the samples being either freely exposed or enclosed in sealed nylon foil
envelopes in the autoclave chamber, or (b) by placing them inside the en-
velopes which were then sealed. Irrespective of which method was used, the
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amine concentration employed was usually close to the lower boundary for
maximum corrosion inhibition.
The investigation showed that
1) these amines effectively inhibited the corrosion of carbon steels both
when the sample was freely exposed in the autoclave and the inhibitor added
to the feed water and also when the sample and inhibitor are sealed together
in an envelope and then autoclaved,
2) morpholine, cyclohexylamine and decylamine were somewhat more ef-
fective than octadecylamine when the amine was added to the autoclave feed
water,
For personal use only.

3) morpholine, decylamine and octadecylamine were somewhat more ef-

fective than cyclohexylamine and benzylamine when the amine was placed
directly in the envelope.

R~SUM$
CORROSION AU COUHS DE LA ST~RILISATIONd L’AUTOCLAVE
11. ESSAIS CONCERNANT L’EMPLOI DE DIFFCRENTS TYPES D’AMINES
ORGANIQUES VOLATILES COMME INHIBITEURS D E CORROSION

Les amines organiques volatiles suivantes : cyclohexylamine, morpholine,

benzylamine, dCcylamine et octadCcylamine, ont CtC soumises & des essais
concernant leur emploi comme inhibiteurs de corrosion au cours de la stC-
rilisation B l’autoclave des instruments en acier. Les expCriences de corrosion
ont CtC effectuhes dans un autoclave B haute pression sur des Cprouvettes
d’acier au carbone correspondant du point de vue de la composition ei du
traitement prkalable aux instruments coupants dentaires et chirurgicaux.
Les amines ont Ct6 introduites dans le milieu corrosif, c’est-&-direvapeur
et produit de condensation, (a) en ajoutant les amines directement A l’eau
alimentant l’autoclave, les Cprouvettes &ant placCes dans la chambre de
l’autoclave soit librement exposCes soit enfermCes dans des enveloppes de
film de nylon fermkes hermktiquement, ou (b) en mettant les amines di-
rectement dans les enveloppes avant la fermeture hermbtique de celles-ci.
 32 C-U. FAJERS, L. G. HOLMLUND AND E. STENMAN

Quelle que fut la mCthode utilisCe, les amines ont CtC employees en con-
centration voisine de la limite inferieure donnant un inhibition maximum
de la corrosion.
Cette Ctiide a mis en Cvidence que:
1) les amines inhibent efficacement la corrosion des aciers au carbone, tant
lorsque 1’Cprouvette est librement exposCe dans l’autoclave et l’inhibiteur
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ajoutC dans l’eau alimentant l’autoclave que lorsque 1’6prouvette et l’in-

hibiteur sont enferm6s ensembles dans une enveloppe fermCe hermCtique-
ment avant la stkrilisation,
2) la morpholine, la cyclohexylamine et la dCcylamine sont un peu plus
efficaces que 1’octadCcylamine lorsque l’amine est ajoutCe 21 l’eau alimentant
l’autoclave,
3) la morpholine, la dCcylamine et 1’octadCcylamine sont un peu plus ef-
ficaces que la cyclohexylamine et que la benzylamine lorsque l’amine est
placCe directement dans l’enveloppe.
For personal use only.

ZL‘SAMMENFASSUNG
KORROSION BE1 AUTOKLAVSTERILISIERUNG
11. FLUCHTIGE ORGANISCHE AMINE ALS KORROSIONSHEMMER

Die fliichtigen organischen Amine : Cyklohexylamin, Morpholin, Benzylamin,

Dekylamin und Oktadekylamin wurden als Korrosionshemmer bei Auto-
klavsterilisierung von Instrumentmaterial ausgebrobt. Die Korrosionsteste
wurden in einem Hochdruckautoklaven an Probestucken aus Kohlestahl
ausgefiihrt, der betreffend der Bearbeitung und Zusammensetzung des Ma-
terials zahnantlichen und chirurgischen schneidenden Instrumenten ent-
sprach.
Das korrodierende Milieu, d.h. Dampf und Kondensat, wurde mit Aminen
vermengt, teils dadurch, dass sie dem *feed water$ des Autoklaven hinzuge-
fiigt wurden, wobei die Probestiicke frei im Autoklaven lagen oder verpackt
in Nylonmaterial, teils dadurch, dass der Korrosionshemmer inwendig an
den Verpackungen angebracht wurde, in denen die Probestiicke wahrend
des Testes eingeschlossen waren. Die Aminmengen, die angewandt wurden,
lagen, ungeachtet des Mischungsverfahrens, meistens nahe der unteren
Grenze fur maximale Korrosionsverhinderung.
Die Untersuchung zeigte:
dass alle ausgeprobten Amine effektiv die Korrosien von Kohlestahl hemm-
ten, gleichviel ob das Material frei im Autoklaven lag und der Korrosions-
hemmer dem *feed water, des Autoklaven hinzugefiigt wurde oder ob
 CORROSION DURING AUTOCI.AVE STERILIZATION 33

das Probemoterial in Verpackungen eingeschlossen wurde, in welche

der Korrosionshemmer angebracht war,
dass Morpholin, Cyklohexylamin und Dekylamin etwas effektiver als Okta-
dekylamin als Korrosionshernrner waren, wenn die Amine dem ufeed
water)) des Autoklaven beigefugt wurden,
dass Morpholin, Dekylamin und Oktadekylamin eine etwas bessere Kor-
rosionshemmung ergaben als Cyklohexylamin und Benzylamin, wenn
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die Amine in der Verpackung angebracht wurden, in denen das Probe-

stuck eingeschlossen war.

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in steam heating systems. Bureau of Mines, Technical Paper 714.
Brigden R. J., 1960: Nylon packet system for sterilization. Nursing Mirror 8: 1.
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Carswell T. S. & H . L. Morrill, 1937: Cyclohexylamine and dicyclohexylamine. Properties

and Uses. Industrial and engineering chemistry, 29: 1247.
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-s- 1966: On steam corrosion and steam corrosion inhibition. V. Amount of cyclohexyl-
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steel are exposed. To be published.
Fajers C-M., L. G. Holmlund & A . Turnuik: Cytotoxic investigation of morpholine and
benzylamine. (Unpublished.)
Holmlund L. G., 1963 a: Colorimetric determination of iron with orthophenanthroline at
high electrolyte concentration. Acta Odont. scand. 21: 309.
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 34 C-M. FAJERS, L. G . H O L M L U N D AND E. STENMAN

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amine. Two corrosion inhibitors. Odont. Revy 14: 335.
Kajanne P., 1957: Experiences witl: filming amines in Finland. Corrosion Prevention and
Control 4: 49.
Kahler H . L. & J . K . Brown, 1954: Experiences with filming amines in control of condensate
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Addresses:
C-M. Fajers, L. G. Holmlund,
Department of Pathology I Department of Dental Technology,
University of Umed University of Umed

E. Stenman,
Department of Oral Diagnosis,
University of Ume6