Meat Science 92 (2012) 274–279

Contents lists available at SciVerse ScienceDirect

Meat Science
journal homepage: www.elsevier.com/locate/meatsci

Review

Sodium nitrite: The “cure” for nitric oxide insufficiency

Deepa K. Parthasarathy, Nathan S. Bryan ⁎
Texas Therapeutics Institute, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston 1825 Pressler St. Houston, TX 77030, United States

a r t i c l e i n f o a b s t r a c t

Article history: This process of “curing” food is a long practice that dates back thousands of years long before refrigeration or
Received 18 January 2012 food safety regulations. Today food safety and mass manufacturing are dependent upon safe and effective
Received in revised form 28 February 2012 means to cure and preserve foods including meats. Nitrite remains the most effective curing agent to prevent
Accepted 1 March 2012
food spoilage and bacterial contamination. Despite decades of rigorous research on its safety and efficacy as a
curing agent, it is still regarded by many as a toxic undesirable food additive. However, research within the
Keywords:
Nitrate
biomedical science community has revealed enormous therapeutic benefits of nitrite that is currently
Curing being developed as novel therapies for conditions associated with nitric oxide (NO) insufficiency. Much of
Nutrition the same biochemistry that has been understood for decades in the meat industry has been rediscovered
Epidemiology in human physiology. This review will highlight the fundamental biochemistry of nitrite in human physiology
Diet and highlight the risk benefit evaluation surrounding nitrite in food and meat products. Foods or diets
enriched with nitrite can have profound positive health benefits.
© 2012 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 274
2. The chemistry of meat curing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275
2.1. Nitrite and N-nitrosamines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276
3. Nitric oxide biochemistry and physiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276
4. Nitrite in human physiology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 276
5. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
Disclosure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278

1. Introduction
Salting as means of preserving meat, poultry, fish, seafood, and
vegetables predates written history and was essential in ancient
times for providing nutrient-dense foods during scarcity or popula-
tion migration and before refrigeration was an option. Meat curing
is historically defined as the addition of salt to fresh meat cuts to re-
move moisture and reduce the water activity of the tissues to prevent
spoilage. However salt is poorly defined as there are many salts that
⁎ Corresponding author at: Molecular Medicine, Texas Therapeutics Institute, Brown
Foundation Institute of Molecular Medicine, Department of Integrative Biology and
Pharmacology, The University of Texas Graduate School of Biomedical Sciences at
Houston, The University of Texas – Houston Health Science Center, 1825 Pressler St. 530C,
Houston, TX 77030, United States. Tel.: +1 713 500 2439(office); fax: +1 713 500 2447.
E-mail address: Nathan.Bryan@uth.tmc.edu (N.S. Bryan).
can fulfill this activity although some better than others. In ancient
times, salt was obtained from crystalline deposits left by evaporating
water from brine pools, seawater, or mining directly from the earth.
As a consequence, it often contained natural contaminants such as so-
dium or potassium nitrate or nitrite that contributed directly to the
curing reaction and the preservation process, although unrecognized
at the time. These contaminants, nitrite and nitrate, as it was later
learned were the primary components in curing reactions. The reduc-
tion of nitrate (NO3−) salts to nitrite (NO2−) and then to gaseous NO
and its subsequent reaction with myoglobin, to form the nitrosyl-
myoglobin complex forms the basis for cured meat flavor and color.
It was also later realized that it was bacteria that first converts nitrate
to nitrite, which is the mechanism underlying the preservation of
food by nitrate (Binkerd & Kolari, 1975).
Today most cured meats contain added sodium nitrite or cultured
celery extract where the naturally contained nitrate is reduced to

0309-1740/$ – see front matter © 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.meatsci.2012.03.001
 D.K. Parthasarathy, N.S. Bryan / Meat Science 92 (2012) 274–279
nitrite by a starter culture of bacteria. The fundamental utility of ni-
trite in the food industry has been in its ability to inhibit the growth
of a number of aerobic and anaerobic microorganisms and especially
suppress the outgrowth of spores from Clostridium botulinum (Hustad
et al., 1973). Nitrite also retards the development of rancidity during
storage, develops the meat flavor and color, stabilizes the oxidative
state of lipids in meat products and it also preserves the spicy,
smoky flavor (Binkerd & Kolari, 1975; Shahidi & Hong, 1991). Nitrite
in combination with other salts and curing factors may also control
the growth of other pathogens such as Bacillus cereus, Staphylococcus
aureus and Clostridium perfringens (Duncan & Foster, 1968; Pierson &
Smoot, 1982; Pradhan et al., 2009). Under conditions of prolonged
temperature fluctuations in a range that promote bacterial growth,
however, it does not prevent pathogen outgrowth and may allow
toxin production or spoilage (Tompkin, Christiansen, & Shaparis,
1978). Other favorable properties of nitrite other than food safety is-
sues include its reduction and reaction with myoglobin to produce
the characteristic reddish-pink cured color. Therefore, nitrite is abso-
lutely essential to food safety, quality and public health. Interestingly,
now there are recognized health benefits for nitrite that go well be-
yond the food industry and into the heart of cardiovascular medicine.
We will present a brief historical account for the use of nitrite in meat
curing and then review the chemistry within the human body and
how this simple anion affects physiology through the preservation
of nitric oxide activity.
2. The chemistry of meat curing
Meat curing is a process by which substances like salt, nitrite and
sometimes nitrate are added to meat products to preserve meat and
to get the characteristic color of the meat. Historically, salt has been
used for meat preservation, which was first practiced in the deserts
of Asia, according to studies by Binkerd and Kolari (1975). In 1863,
Edward Smith described salt as “the oldest and best known of preserv-
ing agents……its chief action appears to be due to its power of attracting
moisture, and thus extracting fluid to harden the tissues” (Bryan &
Loscalzo, 2011). One of the early studies by Polenske (1891) demon-
strated that nitrite was formed by adding nitrate to a pickling
Fig. 1. Changes in color of red meat indicate chemical oxidation changes.
275
solution, due to the action of microorganisms. Haldane (1901) dem-
onstrated that the desirable bright pink color of cured meat is due
to the nitrite reaction with myoglobin to form nitrosylmyoglobin.
We now know that reaction is through NO and not nitrite. The char-
acteristic cured meat color is due to the concentration of heme pig-
ments (myoglobin and hemoglobin) and their chemical states.
Haldane also demonstrated that nitrite combines with hemoglobin
to form nitrosylhemoglobin, which subsequently gets converted to
nitrosylhemochromogen when heated (Haldane, 1901). The naturally
occurring myoglobin reacts with the nitrite or nitrate derived com-
pounds, to form nitrosylmyoglobin which is bright red in color, and
an axial ligand NO is coordinated to the central heme iron (Moller &
Skibsted, 2002). Ascorbate or ascorbic acid is also used as a curing ac-
celerator, which works by reducing Fe 3 + to Fe 2 +. Ascorbate reacts
with nitrous acid (the protonated form of the nitrite ion) to form
NO (Fox & Ackerman, 1968). The NO derived from nitrous acid can
bind to myoglobin (Fe 2 +), forming NO-Mb, which upon heating
yields pink nitroso-hemochrome, with the NO-porphyrin ring intact,
but with heat the globin protein portion of the molecule becomes de-
natured. In the absence of ascorbate, nitrite ion can bind to myoglo-
bin, forming brown NO-MetMb. Under anaerobic conditions, NO-
MetMb can be slowly reduced to red NO-Mb, then converted to
pink NO-hemochrome during cooking (Bryan, 2009). When cured
meat is spoiled or exposed to light and oxygen, the heat stable pink
color changes to brown, due to oxidation. These color changes help
the consumers identify spoiled meat (illustrated in Fig. 1). The pH
of raw meat is 5.4–6.3 and in that pH range, the nitrite and nitrate
are highly soluble in aqueous solutions. In the presence of oxygen,
NO is oxidized back to nitrite (NO2−), or nitrate (NO3−).
Rancidity in nitrite-cured meats is of course inhibited by vacuum
packaging (exclusion of oxygen). Various other mechanisms are re-
sponsible for the antioxidant effect of nitrite in cured meats. NO
may have antioxidant properties, by its ability to act as a free radical
acceptor (Sato & Hegarty, 1973). However, the primary antioxidant
action of NO is its ability to bind and stabilize heme iron of meat pig-
ments, lowering the amount of free iron released during cooking. Free
iron is a potent catalyst of lipid oxidation (Love & Pearson, 1974). In
fact, the major positive attributes of nitrite in meat curing (pink
276 D.K. Parthasarathy, N.S. Bryan / Meat Science 92 (2012) 274–279
color, inhibition of rancidity, inhibition of clostridial growth) are all
explained by the same chemical mechanism; the tight binding of
NO to heme pigments or essential bacterial iron–sulfur proteins, re-
spectively (Reddy, Lancaster, & Cornforth, 1983). As we will highlight
in later sections, nitrite acts in a similar antioxidant fashion in
humans and its chemistry is controlled by oxyhemoproteins.
2.1. Nitrite and N-nitrosamines
N-nitrosamines are potent carcinogens and can be formed from
nitrite in the presence of low molecular weight amines. Hence, the
public health concerns are actually related to the formation of carci-
nogenic N-nitrosamines rather than to the nitrite itself. About 30 dif-
ferent animal species are responsive to the carcinogenic effects of
approximately 300 different N-nitrosamines (Hecht, 1997). Nitrite is
not the only source of N-nitrosamines and the exposure can occur
in occupational settings, through the diet, cosmetics, tobacco prod-
ucts and agricultural chemicals. N-nitrosamine formation in cured
meat products occurs when amines react with nitrogen oxides such
as nitrite in an acid environment (such as the stomach) or when heat-
ed to very high temperatures (which happens during bacon frying).
During an infection or inflammation, products of nitric oxide in-
cluding nitrite are generated that react with the amines by nitrosation
reactions to form N-nitrosamines thus causing human exposure to N-
nitrosamines possibly linking chronic inflammation to certain can-
cers. The nitrosation of amines in the body can also be catalyzed by
certain bacteria with nitrite to form the N-nitrosamines. The carcino-
genicity of N-nitrosamines is due to a very critical cytochrome P450-
mediated metabolic activation step (Bartsch, Hietanen, & Malaveille,
1989). Although there is evidence to support a plausible biological
mechanism for formation of N-nitrosamines, there are also numerous
effective inhibitors of N-nitrosation reactions in biological systems
(d'Ischia, Napolitano, Manini, & Panzella, 2011). It was discovered
that ascorbic acid (vitamin C) very potently inhibits N-nitrosamine
formation (Mirvish, 1975). Another antioxidant, alpha-tocopherol
(vitamin E), has also been shown to inhibit N-nitrosamine formation
(Mirvish, 1996). Ascorbic acid, erythorbic acid and alpha-tocopherol
inhibit N-nitrosamine formation due to their oxidation–reduction
properties. For example, when ascorbic acid is oxidized to dehydroas-
corbic acid, nitrous anhydride, a potent nitrosating agent formed from
sodium nitrite, is reduced to NO, which is not a nitrosating agent.
Stoichiometrically, one molecule of ascorbic acid can reduce two mol-
ecules of acidified nitrite to NO (Archer, Tannenbaum, Fan, &
Weisman, 1975; Licht, Tannenbaum, & Deen, 1988). However, in the
presence of dissolved oxygen, NO can be oxidized back to nitrite/ni-
trous acid. This recycling means that more than half the molar equiv-
alent of ascorbic acid compared to nitrite is required to prevent
formation of N-nitroso compounds. In other words, for every mole
of nitrite, one mole of ascorbate is needed to yield one mole of nitric
oxide, plus another 0.5 mole ascorbate to prevent the back reaction.
The ratio of ascorbic acid to nitrite is recognized to be a major deter-
minant of the generation of N-nitroso compounds within the acidic
lumen of the stomach (Archer et al., 1975). Contemporary meat-
curing methods use ascorbic acid or erythorbate to prevent N-
nitrosation reactions and to facilitate the curing process.
3. Nitric oxide biochemistry and physiology
Before we review the underlying biochemistry of nitrite in human
physiology, it is first necessary to describe the fundamental roles and
production pathways for NO and its implications in health and dis-
ease to better appreciate the new-found role of nitrite. The mamma-
lian biosynthesis of NO discovered in the 1980's for its roles in the
immune (Hibbs, Taintor, & Vavrin, 1987; Stuehr & Marletta, 1985),
cardiovascular (Arnold, Mittal, Katsuki, & Murad, 1977; Furchgott &
Zawadzki, 1980; Ignarro, Buga, Wood, Byrns, & Chaudhuri, 1987)
and nervous (Garthwaite, Charles, & Chess-Williams, 1988) systems
established a startling new paradigm in the history of cellular signal-
ing mechanisms. Prior to this discovery, NO was widely recognized as
a toxic molecule; NO was considered a common air pollutant, a con-
stituent of cigarette smoke, and a toxic gas, which appears in the ex-
haust of motor cars and jet airplanes, causes acid rain, and destroys
the ozone layer. Thus it was essentially inconceivable that cells
would intentionally produce a toxic gas as part of normal physiology.
This same theme is now evolving around nitrite; a once considered
harmful toxic food additive to now a beneficial molecule with medic-
inal properties. NO is now recognized as one of the most important
signaling molecules in the body, and is involved in virtually every
organ system where it is responsible for modulating an astonishing
variety of effects. The primary targets for NO are thiols (Stamler et
al., 1992) or iron/copper-containing proteins (Arnold et al., 1977).
NO can bind to soluble gualylyl cyclase (sCG) and cause an increase
in second messenger cGMP (Arnold et al., 1977) and mediate a num-
ber of physiological functions. This pathway was considered the basis
of NO based signaling until it was recognized that NO elicited a num-
ber of physiological and biological effects that were not dependent
upon cGMP production. It is now recognized that NO can react direct-
ly with thiyl radicals to form nitrosothiols or other reaction products
of NO e.g. nitrite, N2O3 or N2O4 that can post-translationally modify
thiols to affect protein structure and function (Foster, McMahon, &
Stamler, 2003). NO has been shown to be involved in and affect prac-
tically every organ system in the body (Moncada, Palmer, & Higgs,
1991). One can then imagine the host of diseases or conditions and
multi-systemic symptoms that may be caused or affected by the
body's dysregulation of NO production/signaling. Maintaining NO ho-
meostasis is critical for optimal health and disease prevention. Nitrite
may be fundamental in maintaining NO homeostasis through reduc-
tion to NO and through maintenance of nitrosothiols.
The consequences of NO insufficiency are broad and profound. The
continuous generation of NO is essential for the integrity of the car-
diovascular system, and decreased production and/or bioavailability
of NO is central to the development of many disorders (Moncada et
al., 1991). Aging is considered the single largest risk factor related
to cardiovascular related diseases and deaths. Cardio-protection de-
creases with increasing age and is attributed to a decline in NO. The
lack of NO production can lead to hypertension, atherosclerosis, pe-
ripheral artery disease, heart failure, and thrombosis leading to
heart attack and stroke, the leading cause of death for all Americans.
Remarkably, all of these conditions have been shown to be positively
affected by dietary nitrite interventions (Bryan & Loscalzo, 2011;
Lundberg, Weitzberg, & Gladwin, 2008; Lundberg et al., 2009)
4. Nitrite in human physiology
Although the L-arginine–NO pathway was the first to be discov-
ered, it does not necessarily mean that it is the primary pathway for
the endogenous production of NO. In fact nitrite may be central in
the maintenance of NO homeostasis. The activation and metabolism
of nitrite in human physiology require hemoproteins (Gladwin,
Crawford, & Patel, 2004) as in meat curing. Research performed
over the past decade realized that nitrite is physiologically recycled
in blood and tissues to form NO and other bioactive nitrogen oxides
(Benjamin et al., 1994; Bryan et al., 2005; Lundberg, Weitzberg,
Lundberg, & Alving, 1994; Zweier, Wang, Samouilov, & Kuppusamy,
1995). Nitrite is an oxidative breakdown product of NO that has
been shown to serve as an acute marker of NO flux/formation
(Kleinbongard et al., 2003). Nitrite is in steady state equilibrium
with S-nitrosothiols (Angelo, Singel, & Stamler, 2006; Bryan et al.,
2005) and has been shown to activate sGC and increase cGMP levels
in tissues (Bryan et al., 2005), activities very similar to NO. In the
early 1980's it was shown that, in addition to dietary exposure, nitrite
is also generated endogenously (Green, Tannenbaum, & Goldman,
 D.K. Parthasarathy, N.S. Bryan / Meat Science 92 (2012) 274–279 277

1981). Shortly thereafter, the entire L-arginine–nitric oxide synthase
(NOS)-system was discovered and was found to be the major endog-
enous source of nitrite, since NO is rapidly oxidized to nitrite (Hibbs
et al., 1987). Once nitrite is produced and circulated, it is taken up
by peripheral tissues and can be stored in cells. The one-electron re-
duction of nitrite can occur by ferrous heme proteins (or any redox
active metal) through the following reaction:
ðIIÞ þ ðIIIÞ − jury from heart attack (Bryan et al., 2007). Published studies also
NO2 þ Fe þ H ↔NO þ Fe þ OH :
This is the same biologically active NO as that produced by NOS,
with nitrite rather than L-arginine as the precursor and is a relatively
inefficient process (Feelisch et al., 2008). It is now clear that nitrite is
recycled in vivo and again forms bioactive nitrogen oxides, including
NO (Lundberg & Weitzberg, 2005; Lundberg, Weitzberg, Cole, &
Benjamin, 2004; Lundberg et al., 2008; Zweier, Samouilov, &
Kuppusamy, 1999). Thus, instead of simply wasting the products of
NO oxidation, mammals store and actively recycle it. Human nitrite
reduction to NO was first described in the stomach, where salivary ni-
trite forms NO non-enzymatically via acid-catalyzed reduction
(Benjamin et al., 1994; Lundberg et al., 1994). Soon after this observa-
tion, Zweier described NOS-independent nitrite reduction in the is-
chemic and acidic heart (Zweier et al., 1995). In the last 10 years it
has become evident that blood and tissue nitrite is reduced under
physiological conditions to form NO and modulate blood flow
(Cosby et al., 2003; Gladwin et al., 2000). Subsequent studies show
that a variety of enzymes and proteins can catalyze the one-electron
reduction of nitrite to NO in blood and tissues (Feelisch et al., 2008).
This cycle is illustrated in Fig. 2.
Much of the recent focus on nitrite physiology is due to its ability
to be reduced to NO during ischemic or hypoxic events (Bryan, 2006;
Bryan et al., 2004; Zweier et al., 1995). Nitrite reduction in mammali-
an tissues has been linked to the mitochondrial electron transport
system (Kozlov, Staniek, & Nohl, 1999; Walters, Casselden, & Taylor,
1967, protonation (Zweier et al., 1995), deoxyhemoglobin (Cosby et
al., 2003), and xanthine oxidase (Li, Samouilov, Liu, & Zweier, 2004;
Webb et al., 2004). Nitrite can also transiently form nitrosothiols
(RSNOs) under both normoxic and hypoxic conditions (Bryan et al.,
2004) and a recent study by Bryan et al. demonstrates that steady
state concentrations of tissue nitrite and nitroso are affected by
changes in dietary nitrite intake (Bryan et al., 2005). Furthermore
enriching dietary intake of nitrite translates into significantly less in-
demonstrated that nitrite therapy given intravenously prior to reper-
fusion protects against hepatic and myocardial ischemia/reperfusion
(I/R) injury (Duranski et al., 2005). Additionally, experiments in pri-
mates revealed a beneficial effect of long-term application of nitrite
on cerebral vasospasm (Pluta, Dejam, Grimes, Gladwin, & Oldfield,
2005). Moreover, inhalation of nebulized nitrite selectively dilates
the pulmonary circulation under hypoxic conditions in vivo in
sheep (Hunter et al., 2004). Topical application of nitrite improves
skin infections and ulcerations (Hardwick, Tucker, Wilks, Johnston,
& Benjamin, 2001). In meat products, the oxidation of lipids is pre-
vented by nitrite, primarily by NO binding to heme proteins, lowering
the amount of free iron available to catalyze initiation and propaga-
tion steps of lipid oxidation. Studies by Carr and Frei (2001) have
shown that myeloperoxidase mediated low density lipid modification
is inhibited by nitrite. Furthermore, in the stomach, nitrite-derived
NO seems to play an important role in host defense (Duncan et al.,
1995) and in regulation of gastric mucosal integrity (Bjorne et al.,
2004). Nitrite has also been shown to completely prevent microvas-
cular inflammation and endothelial dysfunction due to a high fat,
high cholesterol diet (Stokes et al., 2009) and reduce C-reactive pro-
tein. Chronic sodium nitrite therapy augments ischemia-induced an-
giogenesis and arteriogenesis demonstrating that sodium nitrite
therapy is a recently discovered therapeutic treatment for peripheral
artery disease and critical limb ischemia (Kumar et al., 2008). Most
recently, a dietary supplement formulated with dietary nitrite and

Diet

Bacterial nitrate
reductases NO3- Oxidation

NO2- Oxyhemoglobin NO2-

Neuroglobin
Deoxyhemoglobin/myoglobin NO Ceruloplasmin
Xanthine oxidoreductase (XOR) oxygen
Respiratory chain enzymes
Plant based nitrite reductase
Aldehyde oxidase
Carbonic anhydrase
Protons NOS
Vitamin C, polyphenols

Fig. 2. A schematic presentation of a mammalian nitric oxide (NO) cycle. NO is generated by nitric oxide synthases (NOS) in most cells of the body and participates in regulation of
numerous physiologic functions. The bioactivity of NO is partly regulated by its rapid oxidation to nitrite (NO2−) or, in the presence of oxyhemoglobin, to nitrate (NO3−). Nitrate is
the predominant NO oxidation product in the circulation. In our bodies, nitrate can undergo reduction to nitrite, and this process is strongly dependent on commensal bacteria. In
blood and tissues, nitrite can be further reduced to NO and other bioactive nitrogen oxides. There are several enzymatic and nonenzymatic routes that can catalyze this reduction,
most of which are greatly enhanced under hypoxic conditions. This mammalian nitrogen cycle can be fueled by the diets rich in nitrite and nitrate.
278 D.K. Parthasarathy, N.S. Bryan / Meat Science 92 (2012) 274–279
natural product chemistry to reduce nitrite to NO has been shown to
modify cardiovascular risk in patients over the age of 40, reduce blood
pressure and reduce markers of inflammation (Zand, Lanza, Garg, &
Bryan, 2011). All of these studies together along with the observation
that nitrite can act as a marker of NOS activity (Kleinbongard et al.,
2003) opened a new avenue for the diagnostic and therapeutic appli-
cation of nitrite, especially in cardiovascular diseases, using nitrite as
marker as well as an active agent. In fact a report by Kleinbongard et
al. (2006) demonstrates that plasma nitrite levels progressively de-
crease with increasing cardiovascular risk. Since a substantial portion
of steady state nitrite concentrations in blood and tissue are derived
from dietary sources (Bryan et al., 2005), modulation of nitrite and/
or nitrate intake may provide a first line of defense for conditions as-
sociated with NO insufficiency (Bryan, 2006).
5. Conclusion
The emerging health benefits of nitrite represent a profound
change in paradigm from the past 50 years. Until now, scientists
have operated under the paradigm of the L-arginine–NO pathway by
NOS enzymes as the only pathway to produce NO. There are a number
of recycling pathways to regenerate NO from dietary nitrite. The
emergence of a redundant pathway for maintenance of NO homeo-
stasis by dietary nitrite provides a new mode of intervention and a
new paradigm for restoring NO homeostasis. The provision of nitrite
as sources of NO may then be viewed as a system of redundancy. Ni-
trite therapy or supplementation may restore NO homeostasis from
endothelial dysfunction and provide benefit in a number of diseases
characterized by NO insufficiency (Bryan, 2009; Bryan & Loscalzo,
2011). If so, this will provide the basis for new preventive or thera-
peutic strategies and new dietary guidelines for optimal health.
There are currently a number of clinical trials using sodium nitrite
as a therapeutic agent (www.clinicaltrials.gov). From a public health
perspective, we may be able to make better recommendations on
diet and dramatically affect the incidence and severity of cardiovascu-
lar disease and the subsequent clinical events. Replenishing nitrite
through dietary means may then act as a protective measure to com-
pensate for insufficient NOS activity under conditions of hypoxia or in
a number of conditions characterized by NO insufficiency. In fact, use
of a rationally designed nitrite and nitrate enriched dietary supple-
ment has been shown in a clinical trial to restore NO homeostasis
and modify cardiovascular risk factors such as hyperlipidemia (Zand
et al., 2011). There are a host of diseases which are associated with
decreased NO availability as measured by nitrite. Becoming more ev-
ident is the enormous benefit of exogenous dietary nitrite in a num-
ber of disease models in animals and even in humans. The active
agent of some medicinal foods may very well be nitrite. The historical
classification of nitrite as a “cure” may now have new meaning.
Disclosure
N.S. Bryan and UTHSC-H have financial interests in Neogenis, Inc.,
a company that develops, produces and sells nitric oxide related
products intended to improve health, develop diagnostics for nitric
oxide related metabolites, and perform commercial measurement of
nitric oxide metabolites in biological samples.
References
Angelo, M., Singel, D. J., & Stamler, J. S. (2006). An S-nitrosothiol (SNO) synthase func-
tion of hemoglobin that utilizes nitrite as a substrate. Proceedings of the National
Academy of Sciences of the United States of America, 103(22), 8366–8371.
Archer, M. C., Tannenbaum, S. R., Fan, T. Y., & Weisman, M. (1975). Reaction of nitrite
with ascorbate and its relation to nitrosamine formation. Journal of the National
Cancer Institute, 54(5), 1203–1205.
Arnold, W. P., Mittal, C. K., Katsuki, S., & Murad, F. (1977). Nitric oxide activates guany-
late cyclase and increases guanosine 3′:5′-cyclic monophosphate levels in various
tissue preparations. Proceedings of the National Academy of Sciences of the United
States of America, 74(8), 3203–3207.
Bartsch, H., Hietanen, E., & Malaveille, C. (1989). Carcinogenic nitrosamines: Free rad-
ical aspects of their action. Free Radical Biology & Medicine, 7(6), 637–644.
Benjamin, N., O'Driscoll, F., Dougall, H., Duncan, C., Smith, L., Golden, M., et al. (1994).
Stomach NO synthesis. Nature, 368(6471), 502.
Binkerd, E. F., & Kolari, O. E. (1975). The history and use of nitrate and nitrite in the cur-
ing of meat. Food and Cosmetics Toxicology, 13, 655–661.
Bjorne, H. H., Petersson, J., Phillipson, M., Weitzberg, E., Holm, L., & Lundberg, J. O.
(2004). Nitrite in saliva increases gastric mucosal blood flow and mucus thickness.
The Journal of Clinical Investigation, 113(1), 106–114.
Bryan, N. S. (2006). Nitrite in nitric oxide biology: Cause or consequence? A systems-
based review. Free Radical Biology & Medicine, 41(5), 691–701.
Bryan, N. S. (2009). Food, nutrition and the nitric oxide pathway: Biochemistry and bioac-
tivity (pp. 238). Lancaster, PA: DesTech Publishing.
Bryan, N. S., Calvert, J. W., Elrod, J. W., Gundewar, S., Ji, S. Y., & Lefer, D. J. (2007). Dietary
nitrite supplementation protects against myocardial ischemia–reperfusion injury.
Proceedings of the National Academy of Sciences of the United States of America,
104(48), 19144–19149.
Bryan, N. S., Fernandez, B. O., Bauer, S. M., Garcia-Saura, M. F., Milsom, A. B., Rassaf, T.,
et al. (2005). Nitrite is a signaling molecule and regulator of gene expression in
mammalian tissues. Nature Chemical Biology, 1(5), 290–297.
Bryan, N. S., & Loscalzo, J. (2011). Nitrite and nitrate in human health and disease. In A.
Bendich (Ed.), Nutrition and health. New York: Humana Press.
Bryan, N. S., Rassaf, T., Maloney, R. E., Rodriguez, C. M., Saijo, F., Rodriguez, J. R., et al.
(2004). Cellular targets and mechanisms of nitros(yl)ation: An insight into their
nature and kinetics in vivo. Proceedings of the National Academy of Sciences of the
United States of America, 101(12), 4308–4313.
Carr, A. C., & Frei, B. (2001). The nitric oxide congener nitrite inhibits myeloperoxida-
se/H2O2/Cl − -mediated modification of low density lipoprotein. Journal of Biolog-
ical Chemistry, 276(3), 1822–1828.
Cosby, K., Partovi, K. S., Crawford, J. H., Patel, R. P., Reiter, C. D., Martyr, S., et al. (2003).
Nitrite reduction to nitric oxide by deoxyhemoglobin vasodilates the human circu-
lation. Nature Medicine, 9(12), 1498–1505.
d'Ischia, M., Napolitano, A., Manini, P., & Panzella, L. (2011). Secondary targets of
nitrite-derived reactive nitrogen species: Nitrosation/nitration pathways, antioxi-
dant defense mechanisms and toxicological implications. Chemical Research in Tox-
icology, 24(12), 2071–2092.
Duncan, C., Dougall, H., Johnston, P., Green, S., Brogan, R., Leifert, C., et al. (1995). Chem-
ical generation of nitric oxide in the mouth from the enterosalivary circulation of
dietary nitrate. Nature Medicine, 1(6), 546–551.
Duncan, C. L., & Foster, E. M. (1968). Effect of sodium nitrite, sodium chloride, and so-
dium nitrate on germination and outgrowth of anaerobic spores. Applied Microbiol-
ogy, 16(2), 406–411.
Duranski, M. R., Greer, J. J., Dejam, A., Jaganmohan, S., Hogg, N., Langston, W., et al.
(2005). Cytoprotective effects of nitrite during in vivo ischemia–reperfusion of
the heart and liver. The Journal of Clinical Investigation, 115(5), 1232–1240.
Feelisch, M., Fernandez, B. O., Bryan, N. S., Garcia-Saura, M. F., Bauer, S., Whitlock, D. R.,
et al. (2008). Tissue processing of nitrite in hypoxia: An intricate interplay of nitric
oxide-generating and -scavenging systems. Journal of Biological Chemistry, 283(49),
33927–33934.
Foster, M. W., McMahon, T. J., & Stamler, J. S. (2003). S-nitrosylation in health and dis-
ease. Trends in Molecular Medicine, 9(4), 160–168.
Fox, J. B. J., & Ackerman, S. A. (1968). Formation of nitric oxide myoglobin: Mechanisms
of the reaction with various reductants. Journal of Food Science, 33, 364–370.
Furchgott, R. F., & Zawadzki, J. V. (1980). The obligatory role of endothelial cells in the
relaxation of arterial smooth muscle by acetycholine. Nature, 288(5789), 373–376.
Garthwaite, J., Charles, S. L., & Chess-Williams, R. (1988). Endothelium-derived relaxing
factor release on activation of NMDA receptors suggests role as intercellular mes-
senger in the brain. Nature, 336(6197), 385–388.
Gladwin, M. T., Crawford, J. H., & Patel, R. P. (2004). The biochemistry of nitric oxide,
nitrite, and hemoglobin: Role in blood flow regulation. Free Radical Biology & Med-
icine, 36(6), 707–717.
Gladwin, M. T., Shelhamer, J. H., Schechter, A. N., Pease-Fye, M. E., Waclawiw, M. A.,
Panza, J. A., et al. (2000). Role of circulating nitrite and S-nitrosohemoglobin in
the regulation of regional blood flow in humans. Proceedings of the National Acad-
emy of Sciences of the United States of America, 97(21), 11482–11487.
Green, L. C., Tannenbaum, S. R., & Goldman, P. (1981). Nitrate synthesis in the germfree
and conventional rat. Science, 212(4490), 56–58.
Haldane, J. (1901). The Red colour of salted meat. The Journal of Hygiene, 1(1), 115–122.
Hardwick, J. B., Tucker, A. T., Wilks, M., Johnston, A., & Benjamin, N. (2001). A novel
method for the delivery of nitric oxide therapy to the skin of human subjects
using a semi-permeable membrane. Clinical Science (London, England), 100(4),
395–400.
Hecht, S. S. (1997). Approaches to cancer prevention based on an understanding of N-
nitrosamine carcinogenesis. Proceedings of the Society for Experimental Biology and
Medicine, 216(2), 181–191.
Hibbs, J. B., Jr., Taintor, R. R., & Vavrin, Z. (1987). Macrophage cytotoxicity: Role for L-ar-
ginine deiminase and imino nitrogen oxidation to nitrite. Science, 235(4787),
473–476.
Hunter, C. J., Dejam, A., Blood, A. B., Shields, H., Kim-Shapiro, D. B., Machado, R., et al.
(2004). Inhaled nebulized nitrite is a hypoxia-sensitive NO-dependent selective
pulmonary vasodilator. Nature Medicine, 10, 1122–1127.
Hustad, G. O., Cerveny, J. G., Trenk, H., Deibel, R. H., Kautter, D. A., Fazio, T., et al. (1973).
Effect of sodium nitrite and sodium nitrate on botulinal toxin production and ni-
trosamine formation in wieners. Applied Microbiology, 26(1), 22–26.
 D.K. Parthasarathy, N.S. Bryan / Meat Science 92 (2012) 274–279
Ignarro, L. J., Buga, G. M., Wood, K. S., Byrns, R. E., & Chaudhuri, G. (1987). Endothelium-
derived relaxing factor produced and released from artery and vein is nitric oxide.
Proceedings of the National Academy of Sciences of the United States of America, 84,
9265–9269.
Kleinbongard, P., Dejam, A., Lauer, T., Jax, T., Kerber, S., Gharini, P., et al. (2006). Plasma
nitrite concentrations reflect the degree of endothelial dysfunction in humans. Free
Radical Biology & Medicine, 40(2), 295–302.
Kleinbongard, P., Dejam, A., Lauer, T., Rassaf, T., Schindler, A., Picker, O., et al. (2003).
Plasma nitrite reflects constitutive nitric oxide synthase activity in mammals.
Free Radical Biology & Medicine, 35(7), 790–796.
Kozlov, A. V., Staniek, K., & Nohl, H. (1999). Nitrite reductase activity is a novel function
of mammalian mitochondria. FEBS Letters, 454(1–2), 127–130.
Kumar, D., Branch, B. G., Pattillo, C. B., Hood, J., Thoma, S., Simpson, S., et al. (2008).
Chronic sodium nitrite therapy augments ischemia-induced angiogenesis and
arteriogenesis. Proceedings of the National Academy of Sciences of the United States
of America, 105(21), 7540–7545.
Li, H., Samouilov, A., Liu, X., & Zweier, J. L. (2004). Characterization of the effects of ox-
ygen on xanthine oxidase-mediated nitric oxide formation. Journal of Biological
Chemistry, 279(17), 16939–16946.
Licht, W. R., Tannenbaum, S. R., & Deen, W. M. (1988). Use of ascorbic acid to inhibit
nitrosation: Kinetic and mass transfer considerations for an in vitro system. Carci-
nogenesis, 9(3), 365–372.
Love, J. D., & Pearson, A. M. (1974). Metmyoglobin and nonheme iron as prooxidants in
cooked meat. Journal of Agricultural and Food Chemistry, 22(6), 1032–1034.
Lundberg, J. O., Gladwin, M. T., Ahluwalia, A., Benjamin, N., Bryan, N. S., Butler, A., et al.
(2009). Nitrate and nitrite in biology, nutrition and therapeutics. Nature Chemical
Biology, 5(12), 865–869.
Lundberg, J. O., & Weitzberg, E. (2005). NO generation from nitrite and its role in vas-
cular control. Arteriosclerosis, Thrombosis, and Vascular Biology, 25(5), 915–922.
Lundberg, J. O., Weitzberg, E., Cole, J. A., & Benjamin, N. (2004). Nitrate, bacteria and
human health. Nature Reviews Microbiology, 2(7), 593–602.
Lundberg, J. O., Weitzberg, E., & Gladwin, M. T. (2008). The nitrate–nitrite–nitric oxide
pathway in physiology and therapeutics. Nature Reviews. Drug Discovery, 7(8),
156–167.
Lundberg, J. O., Weitzberg, E., Lundberg, J. M., & Alving, K. (1994). Intragastric nitric
oxide production in humans: Measurements in expelled air. Gut, 35(11),
1543–1546.
Mirvish, S. S. (1975). Blocking the formation of N-nitroso compounds with ascorbic
acid in vitro and in vivo. Annals of the New York Academy of Sciences, 258, 175–180.
Mirvish, S. S. (1996). Inhibition by vitamins C and E of in vivo nitrosation and vitamin C
occurrence in the stomach. European Journal of Cancer Prevention, 5(Suppl. 1),
131–136.
Moller, J. K., & Skibsted, L. H. (2002). Nitric oxide and myoglobins. Chemical Reviews,
102(4), 1167–1178.
Moncada, S., Palmer, R. M. J., & Higgs, A. (1991). Nitric oxide: Physiology, pathophysi-
ology and pharmacology. Pharmacological Reviews, 43(2), 109–142.
Pierson, M. D., & Smoot, L. A. (1982). Nitrite, nitrite alternatives, and the control of Clos-
tridium botulinum in cured meats. Critical Reviews in Food Science and Nutrition,
17(2), 141–187.
279
Pluta, R. M., Dejam, A., Grimes, G., Gladwin, M. T., & Oldfield, E. H. (2005). Nitrite infu-
sions to prevent delayed cerebral vasospasm in a primate model of subarachnoid
hemorrhage. JAMA : The Journal of the American Medical Association, 293(12),
1477–1484.
Polenske, E. (1891). Uber den Verlust, welchen Rindfleisch and Nahrwert durch das
Pokeln erleidet sowie uber die Veranderungen salpeterhaltiger Pokellaken. Arbei-
ten aus dem kaiserlichen Gesendheitsamt, 7, 471–474.
Pradhan, A. K., Ivanek, R., Grohn, Y. T., Geornaras, I., Sofos, J. N., & Wiedmann, M.
(2009). Quantitative risk assessment for listeria monocytogenes in selected cate-
gories of deli meats: Impact of lactate and diacetate on listeriosis cases and deaths.
Journal of Food Protection, 72(5), 978–989.
Reddy, D., Lancaster, J. R., Jr., & Cornforth, D. P. (1983). Nitrite inhibition of Clostridium
botulinum: Electron spin resonance detection of iron–nitric oxide complexes. Sci-
ence, 221(4612), 769–770.
Sato, E., & Hegarty (1973). Warmed over flavor in cooked meats. Journal of Food Science,
36, 1098–1102.
Shahidi, F., & Hong, C. (1991). Evaluation of malonaldehyde as a marker of oxidative
rancidity in meat products. Journal of Food Biochemistry, 15, 97–105.
Stamler, J. S., Simon, D. I., Osborne, J. A., Mullins, M. E., Jaraki, O., Michel, T., et al. (1992).
S-nitrosylation of proteins with nitric oxide: Synthesis and characterization of bio-
logically active compounds. Proceedings of the National Academy of Sciences of the
United States of America, 89, 444–448.
Stokes, K. Y., Dugas, T. R., Tang, Y., Garg, H., Guidry, E., & Bryan, N. S. (2009). Dietary ni-
trite prevents hypercholesterolemic microvascular inflammation and reverses en-
dothelial dysfunction. American Journal of Physiology - Heart and Circulatory
Physiology, 296(5), H1281–H1288.
Stuehr, D. J., & Marletta, M. A. (1985). Mammalian nitrate biosynthesis: Mouse macro-
phages produce nitrite and nitrate in response to escherichia coli lipopolysaccha-
ride. Proceedings of the National Academy of Sciences of the United States of
America, 82(22), 7738–7742.
Tompkin, R. B., Christiansen, L. N., & Shaparis, A. B. (1978). Effect of prior refrigeration
on botulinal outgrowth in perishable canned cured meat when temperature
abused. Applied and Environmental Microbiology, 35(5), 863–866.
Walters, C. L., Casselden, R. J., & Taylor, A. M. (1967). Nitrite metabolism by skeletal
muscle mitochondria in relation to haem pigments. Biochimica et Biophysica Acta,
143(2), 310–318.
Webb, A., Bond, R., McLean, P., Uppal, R., Benjamin, N., & Ahluwalia, A. (2004). Reduc-
tion of nitrite to nitric oxide during ischemia protects against myocardial ische-
mia–reperfusion damage. Proceedings of the National Academy of Sciences of the
United States of America, 101(37), 13683–13688.
Zand, J., Lanza, F., Garg, H. K., & Bryan, N. S. (2011). All-natural nitrite and nitrate con-
taining dietary supplement promotes nitric oxide production and reduces triglyc-
erides in humans. Nutrition Research, 31(4), 262–269.
Zweier, J. L., Samouilov, A., & Kuppusamy, P. (1999). Non-enzymatic nitric oxide
synthesis in biological systems. Biochimica et Biophysica Acta, 1411(2–3),
250–262.
Zweier, J. L., Wang, P., Samouilov, A., & Kuppusamy, P. (1995). Enzyme-independent
formation of nitric oxide in biological tissues. Nature Medicine, 1, 804–809.