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[Benzalkonium chloride: unacceptable to sterilize or disinfect medical or dental

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Benzalconium chloride is unacceptable to sterilize or desinfect BRIEF COMMUNICATION

Benzalkonium chloride is unacceptable

to sterilize or disinfect medical
or dental instruments
Enrique Acosta-Gío, CD, PhD,(1) Aurelio Herrero-Farías, CD,(2)
Víctor Hugo Mata-Portuguez, CD.(2)

Please use Spanish citation:

Acosta-Gío E, Herrero-Farías A, Mata-Portuguez VH. El cloruro de benzalconio: inaceptable para “esterilizar o desinfectar”
instrumental médico o dental. Salud Publica Mex 2001;43:570-573.

Abstract
Objective. To compare the sporicidal activity of benzalkonium chloride (BKC) with that of glutaraldehyde. Material
and Methods. A comparative study was conducted at the microbiology laboratory of Facultad de Odontología,
Universidad Nacional Autónoma de México. Bacillus subtilis ATCC 9372 spores were exposed to these germicides
(1 spore x mL) on a 0.22 mm filter. After completing the contact time the spores were washed and the filters were
incubated on nutrient agar for 72 h at 37 ºC. Results. BKC did not eliminate B. subtilis spores at the concentration
used, not even after increasing contact time to 15 h (900-fold the recommended time). Two percent glutaraldehyde
destroyed spores only after 10 h of contact. Urea and sodium chloride showed no sporicidal activity. Conclusions.
The results confirm that BKC lacks sporicidal activity and confirm that this quaternary ammonium compound is
not able to “sterilize” or “disinfect” medical and dental instruments.

Key words: benzalkonium compounds; sterilization; disinfectants; sporicide; germicide; Mexico

Benzalkonium chloride (BKC) is a quaternary acceptable substances.6 However, BKC is widely

ammonium compound (QAC), whose condensed
formula is n-alkyl methyl benzyl ammonium
chloride.1
BKC has a poor history as a disinfectant.
Various gram-negative bacteria can develop in BKC
and such bacteria have been associated with
nosocomial infections, some of which are fatal, from
instruments immersed in BKC.2-5
In 1978, the American Dental Association (ADA)
eliminated QACs, including BKC, from its list of
used in Latin America.7
Table I shows some products with similar
concentrations to BKC, that are indiscriminately
advertised as being antiseptics for external use,
disinfectants for instruments (at cold temperatures),
and effective sterilizers after one minute.8
The World Health Organization (WHO)9 and the
Pan American Health Organization (PAHO)10 use the
terminology “high-level sterilizer, intermediate-
level, and low-level germicide”. The World Health

(1) Laboratorio de Microbiología y Control de Infecciones. División de Estudios de Posgrado e Investigación, Facultad de Odontología, Universidad
Nacional Autónoma de México. UNAM. México, D.F.
(2) PhD candidates, Facultad de Odontología, UNAM. México, D.F.

Received on: November 27, 2000 • Accepted on: May 23, 2001
Address reprint requests to: Dr. Enrique Acosta-Gío, Apdo. 70 – 421, Ciudad Universitaria, 04510 México D.F., México.
E-mail: acostag@servidor.unam.mx

salud pública de méxico / vol.43, no.6, november-december, 2001 1

 BRIEF COMMUNICATION Acosta-Gío E et al

Organization defines sterilization as the destruction
of all microorganisms, including bacterial spores.9
A sterilizing germicide should destroy Bacillus
subtilis and Clostridium sporogenes spores.11
The objective of this study, conducted in
November 2000, was to evaluate the sporicidal
activity of benzalkonium chloride.
Material and Methods
The germicide Timsen (United Chemical Corp. Los
Angeles, CA) was used according to the instructions
on its label (4 g in 1 l of water, with a final
concentration of 0.16% of BKC and 0.24% urea, inert
ingredient). Another germicide, Krit (Farmacéuticos
Altamirano, México) was used at 0.12 % of BKC.
2 % glutaraldehyde (Atosteril, A.T.O. Zizine,
París), a 0.24 % urea solution (Sigma Chemicals. St
Louis, MO) and a saline solution with 140 mM of
sodium chloride (Sigma Chemicals) were used as
control antispetics. All the solutions were prepared
in distilled and deionized water.
Each evaluation of the sporicidal activity was
carried out by quintuplying (5x104 ) B. subtilis
ATCC 9372 spores (SPS, Medical, Rush, NY) on a
0.22 mm polyvinylidene filter (Millipore Corp.
Bedford, MA), according to the method described by
Collins.12 In brief, the spores were suspended in
saline solution and deposited on top of the filter.
Then the saline solution was removed by suction and
the spores were suspended in 50 ml of the
disinfectant solution (1 spore x ml) for evaluation at
1, 20, 360, 600 o 900 minutes at room temperature.
Upon completion of the exposure time, the
disinfectant was removed and the spores washed in
5 ml of 1 % sodium bisulfite solution to inactivate the
glutaraldehyde, followed by a rinse in 300 ml of
saline solution. Finally, the filters were aseptically
placed on an agar nutrient (Difco Labs, Detroit, MI)
and incubated for 72 hours at 37ºC. The results were
described as growth or no growth.
Results
Table II presents the results of the sporicidal
evaluation. BKC did not destroy B. subtilis spores at
the concentration used and the contact time
stipulated on the label, nor when the exposure time
was increased to 15 hours. The 2 % glutaraldehyde
solution destroyed the spores after 10 hours of
contact. The urea and the saline solution did not
demonstrate any sporicidal activity.
Discussion
The results confirmed that BKC lacks sporicidal
activity and sterilizing and disinfecting properties.
Instrument disinfectants should be sporicides.13
Products used in dental health care should
comply with national and international norms.14 The
Food and Drug Administration (FDA) of the United
States regulates the high-level disinfectants/
sterilizers that are used to reprocess instruments.15
The FDA doesn’t approve any BKC as a sterilizer or
disinfectant for instruments.
Low-level germicides, like BKC, can be used to
clean superficial environments,1,6,10 while wearing
protective clothing, since hypersensitivity to BKC
exposure does exist.16-18

Table I
SOME PRODUCTS AVAILABLE IN M EXICO WITH
BENZALKONIUM CHLORIDE ( N- ALKIL METHYL BENZYL Table II
AMMONIUM CHLORIDE ) AS THE ACTIVE INGREDIENT
SPORICIDAL ACTIVITY OF BENZALKONIUM CHLORIDE

Timsen Krit Glut CH4N2O NaCl

Name Reg. SSA Use Presentation Dilution Time
Minutes 0.16 % 0.12 % 2% 0.24 % 140 mM
Timsen 0576C99 S+D 4g/1 l 0.16 % 1 min
1 + + + + +
Krit 75303 D+A 12 % 0.12 % 5 min
20 + + + + +
Antibenzyl 62949 D+A 1% 1% ¿?
360 + + + + +
Merthiolate 175M86 A 0.13 % 0.13 % *
600 + + - + +
Benzyl 45108 A 5 g/500 ml 0.02 % **
900 + + - + +
S “sterilizer”, D “desinfectant”, A “antiseptic”
* method of use= topical += growth, ineffective
** meted of use= douche -= no growth, effective

2 salud pública de méxico / vol.43, no.6, november-december, 2001

Benzalconium chloride is unacceptable to sterilize or desinfect
The labels on products available in Mexico
contain contradictions and are lacking, given that
they describe BKC as an antiseptic, a disinfectant and
a sterilizer. In order to disinfect, the dilution of Krit
should be 0.12% BKC and applied for 5 min, while
for Antibenzil it is 1 % BKC, and the contact time is
not established.
Contact time should not be arbitrary. The
PAHO10 and the FDA13 indicate that instrument
immersion can be for up to 10 hours. Atosteril only
showed sporicidal activity after 10 hours, which is
more contact time than what is accepted in France (6
hours) for this product.
BKC was not evaluated against micobacterias or
vegetative bacterias associated with nosocomial
infections such as Serratia marcensens, Staphylococcus
aureus, Enterococcus sp. or Pseudomonas aeruginosa.
The QACs showed little or no activity against
bacterial spores, fungus and the majority of viruses,1
and they didn’t show any tuberculocidal acitivity,19
thus they are inappropriate for the disinfection of
clinical surfaces.
BKC is inadequate, even dangerous, for
reprocessing instruments.20-22 Recently, it was learned
that BKC at 0.1% is not effective as a mouth rinse for
reducing the accumulation of dental bacteria which
causes plaque.23
The evaluation technique described by Collins,12
and used in this study, is a modification of official
methods.11 By using a filtering membrane, false
negative results are avoided, due to the loss of viable
spores. Sodium bisulfite only neutralizes the
glutaraldehyde. It was observed that when the BKC
remains in the membrane, for an insufficient wash,
the spores did not germinate; this bacteriostatic effect
has been known for thirty years24 and should not be
confused with true sporicidal activity.
In this investigation, the germicidal liquids were
evaluated only under optimal conditions. In order to
be approved, the disinfectants should be subjected to
worse case scenarios, such as the presence of organic
or inorganic contamination, and under simulated use
conditions.13
The evident lack of sporicidal activity by BKC
does not require confirmation with more attempts or
with C. sporogenes. The criteria for sporicidal activity
(disinfection by a high-level germicide) only allows
for two failures in 120 attempts. In order to pass the
test for sterilization, no failures are tolerated.13
To avoid the poor use of disinfectants, which put
human lives at risk, the WHO9 and the PAHO10
guidelines for disinfect ion and sterilization should
be in agreement with the concepts and the
salud pública de méxico / vol.43, no.6, november-december, 2001
BRIEF COMMUNICATION
terminology employed by the FDA,13,15 and by
experts in infection control.1,6,25
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