Cassava Processing As A Method of Detoxification: Latin American Starchy Tubers

LATIN AMERICAN STARCHY TUBERS
Chapter 3
Cassava Processing as a Method
of Detoxification
Marney Pascoli Cereda1
3.1. Introduction
Many cultivated plants present cyanide in some or all their parts. According to Cagnon,
Cereda and Pantaroto (2002) it is possible to find cyanoglucoside in wheat, barley,
oat, sorghum, bean sprouts, forage grass, apples, cassava and taro, among others.
Linamarin can be found in many vegetables. It is one of cassava’s cyanogenic com-
pounds, and it is spread all over the plant including the root’s edible parts. Cassava is
probably one of the most known cyanogenic plants due to its large distribution in
tropical countries. In Africa, a special conjuncture of underdevelopment, lack of health
and food resources, low schooling levels and lack of information in general lead to a
situation where cassava and its by-products are the main meal, hence making most
health problems be strongly related to the above mentioned root consumption. On the
other hand, in Latin American countries where cassava is largely consumed by humans
and animals, the situation is quite different. Even in Brazil, where there are concentra-
tion sites of malnutrition, the reported cases of intoxication by consumption of cassava
and its derivates remain rare and their evidence is debatable.
The reasons for the cyanide accumulation in the cassava plant have not been well
determined. Although there are theories asserting that the liberated cyanide would be a
protecting agent against pests and diseases, and a repellent for superior animals, there
is no proof of such a thing. Teles et al. (1993) analyzed 8 varieties with susceptibility
and 8 which are resistant to Xanthomonas campestris pv. maniotis. The contents of
HCN in the roots ranged from 14 mg/gm for Saracura variety to 0.49 mg/g for Roxinha.
Despite the different HCN contents found in those varieties, it was not possible to
establish any kind of relation with the plants’ resistance to this bacterial disease.
1- NGO RAÍZES - Av. Manaca, 524 – 18607-170 Botucatu – SP - Brazil. cereda@raizes-

ong.com.br.
50
Book 3 - Technology, use and potentialities of Latin American starchy tubers
Pereira (1986) analyzed the exudates of stalks of cassava’s leaves, and the result was
a composition formed by fructo-furanoses and reducing sugars, with few amino acids
and no cyanides. The fresh exudate revealed to be harmless to houseflies and mosqui-
toes, which might indicate that this exudate is not a protection agent against the insects
attack upon the cassava.
It is not easy to explain the functions of the cyanogenic compounds in the cassava
plant because many discussions about them are still inappropriately documented in the
specialized literature. The most frequently documented effect of these glucosides is
related to the increase of the plant’s sensibility to fungi attacks when it presents high
contents of cyanogenic compounds. This effect is probably the result of the plant’s
little capacity to produce defensive compounds such as the phytoalexins (Bokanga et
al., 1994).
An often-made mistake concerning the cassava plant is to say that it has cyanide, when
the correct way to put it would be the plant could generate cyanide. The lethal dose of
linamarin ingested by superior animals should also be better determined.
3.2. The glucocyanide in the cassava plant

The cassava concentrates two cyanogenic glucosides in its roots and leaves (figure
3.1), the linamarin and the lotaustralin in a proportion of approximately 93:7. These
two glucosides are capable of generating hydrocyanic acid as long as there is hydroly-
sis, i.e., the cyanide contents and the use of cassava as food are conditioned by the
enzymatic hydrolysis of these two toxic glucosides.
Source: http://www.sdu.dk/Nat/Chem/kemi_net/LostFast/Cassanova1.gif (1993).

Figure 3.1.
Structural formulae of linamarin and lotaustralin.
According to Cagnon, Cereda and Pantaroto (2002), the enzyme that causes catalysis
and determines the biosynthesis speed in cyanogenic glucosides is known and docu-
mented, and it is called linamarase. The high specificity of this enzyme in relation to
the substratum determines which glucoside is formed in each plant separately. There is
51
serious evidence suggesting that the enzymes found in different cyanogenic plants have
the same characteristics.
The insulation of the enzyme’s codifying gene may provide a potential tool for the
modulation of the cyanogenic glucosides in plants. In spite of all advanced steps, the
use of such information is still far from bringing outstanding practical results for the
development of cassava plants without cyanogenic glucosides.
Bokanga (1992) points out two food and nutrition research areas, which could benefit
in Africa from the advances made in biotechnology. The first area refers to the fer-
mented products improvement, of which a large part is cassava prepared, and the
second area concerns cassava’s detoxification. The author justifies their importance
by means of the fact that the presence of cyanogenic glucosides potentially generates
HCN, a toxic substance. The development of varieties with little linamarin contents
would benefit from a better understanding of the molecular mechanisms involved with
the biosynthesis of this compound.
Numerous researches and efforts have been made in order to develop free from cyanide
cassava plants. The cyanogenic glucosides in cassava can be found in any of these
plants’ parts. The results presented by Bokanga et al. (1994) reveal that the biggest
portion of cyanogenic glucosides is produced in the leaves. Therefore, it is impossible
to obtain cassava roots without cyanide just by blocking cyanogenic glucosides trans-
port. Besides the selection of varieties with low cyanide contents, another way to pre-
vent intoxication is the detoxification.
Although they are found in every cassava plant, linamarin content levels vary accord-
ing to each organ separately. Gomez and Valdivieso (1984) studied two cassava-culti-
vated hybrids, the CMC40 (low cyanide) and the CMC84 (high cyanide). Plants with
9 to 12 months of age were analyzed. The highest cyanide concentration was found in
the root, but the separated parenchyma (pulp) showed the lowest values. The leaves
presented intermediary concentration levels when compared with the roots’ tissue. The
CMC40 roots showed large cyanide concentration in the inner layer or cortex, spe-
cially the 9 month plants. The cyanide concentration in the pulp was the most stable
parameter and came out almost unchanged along with the aging of plants. The cyanide
content levels in the CMC84 root’s pulp were approximately 3 times higher than that
in the CMC40 (623 mg/kg on a dry base). The cyanide content changes in the leaves of
CMC40 were more remarkable than those in the leaves of CMC84. Over 90% of the
total cyanide found in the tissues of the analyzed cassava plants came out as linamarin,
also called fixed or bound cyanide.
Even if we could develop or select cassava varieties with very small linamarin con-
tents, we would never be sure that these plants might not generate bigger contents later.
52
Several factors interfere with the linamarin content levels such as the plants’ variety,
the soil nitrogen contents, the weather and the age of the plant.
In order to exemplify the importance of cultivation to the generation of linamarin
contents, Obigbesan (1984) verified the effects of fertilization with N on the linamarin
contents in leaves and roots with accumulated starch. He analyzed cassava plants cv
TMS 30001, 30211, 30395 and 30572, grown in-field experiments in Africa. These
cultivated hybrids were selected for their low contents levels of linamarin. Obigbesan
concluded that the fertilization with N had increased significantly the linamarin con-
tent in the plants. In making gari with the roots of these cultivated hybrids, each bear-
ing different linamarin contents, the final products’ residual contents showed little
variation.
The age of the roots also influences linamarin contents, but this influence is smaller in
the pulp. Cereda et al. (1990a) analyzed the cultivated hybrids called Pioneira (Pio-
neer), recommended for cooking purposes, with 12 and 24 months of age, although
usually the culinary use cassava plants are harvested earlier, at the age of 9 to 10
months. Only the edible parts were analyzed since the inner layer is taken off when
processed. The 12 months old roots’ pulp showed 63.70% of moisture and 61.80 mg/
kg of linamarin. In the 24 months old roots there was 58.52% of moisture, being little
different from the linamarin content of 63.40mg/kg. These results align with the ones
provided by Gomez and Valdivieso (1984).
Obigbesan (1984) comments on the mistaken ideas of the damage caused to human
beings and animals by cassava consumption. The poor knowledge shown by the peo-
ple who use cassava as a means of subsistence is one responsible factor for the exag-
gerated descriptions of intoxication symptoms. Also the doctors who attend patients
lack this kind of knowledge. The conditions that lead to poisoning and death are re-
stricted to some specific regions marked by malnutrition and social conflicts as well.
There is a lot of misunderstanding as for the way analysis’ results are registered. They
can be set out in 5 ways as follows: (a) linamarin (with 87% of potential cyanide), (b)
CNp- : total, bound or potential cyanide, (c) free cyanide CN-, (d) acetone cyanidrine,
and (e) HCN or hydrocyanic acid. The only intoxicating agent is free cyanide CN-, or
its acid, the HCN. The selection of the right methodology is equally important. Bound
linamarin is dosed as total cyanide, and analysis of free cyanide (in other words, the
toxic fraction) is possible only with the hydrolysis of linamarin. There is no reliable
method other than to use the plant’s linamarase enzyme or a commercial preparation.
To make this nomenclature definitely uniform, WOCAS (Working Group on Cassava
Safety) (1992) suggested that CNp, which corresponds to the potential cyanide, repre-
sent linamarin content.
53
3.3. Cyanide liberation in the cassava

Significant cases of poisoning by cyanide are restricted to the regions where cassava
represents a large fraction of the food diet. One needs to know better about cyanide
liberation mechanisms in order to find out the reasons for misunderstanding detoxifi-
cation, acute intoxication and diseases related to the continuous consumption of cas-
sava and its by-products, which also contain residual linamarin.
Linamarin is a water-soluble glucoside that potentially gives out hydrocyanic acid
(HCN). Specialized literature cites some values like 15 to 400 ppm (CN mg/kg of
weight) of cyanide, yet the most frequently found values are between 30 and 150 mg/
kg.
Detection of the bitter taste in raw cassava roots has been used in the fields to differen-
tiate the roots with high linamarin levels (bitter) from those that have low linamarin
levels (soft, sweet), although this method for detecting is not safe. Sundaresan et al.
(1987) studied the cyanide content’s relation with cassava roots’ bitter taste. In the 38
cassava genotypes classified by sensory assessment, linamarin contents ranged from
320 to 1.100 mg CN/kg in the very bitter roots, and from 27,5 to 77,5 mg/kg in the
ones with no bitter taste. Linamarin content levels in the varieties classified as bitter
dropped down to values between 6,9 and 16,7 mg of the substance when the roots were
boiled in water during 30 minutes, or when grated and dried in the sun during 6 hours.
Simultaneously, roots classified as bitter were then considered not bitter, even the very
bitter ones. The authors came to the conclusion that the bitter taste is associated with
linamarin high contents, but only in the raw cassava roots.
Although linamarin has bitter taste, cassava roots pulp shows high contents of sugars
in some periods of the year, these sugars being able to disguise the bitter taste. Moreo-
ver, the sensitivity to bitter taste is genetically controlled, and because people have
different sensitivity degrees, they may or may not consider the same cassava plant
bitter. The cassava external characteristics are not reliable criteria to identify the plants
with high linamarin contents, either. These characteristics vary largely according to
environmental and nutritional conditions, as well as to the type of soil, hence making
the visual recognition impossible.
When tissue is torn apart, linamarin enzymatic hydrolysis occurs by β-glucosidase
(linamarase), which separates from the glucoside present in the intact tissue due to the
fact that it is located at a cell’s distinct place (White et al. 2002). Cleaving produces
glucose and α-hydroxinitrile. The latter turns spontaneously into HCN and correspond-
ing cetones in the catalyses caused by hydroxinitrile-lyase. This process, known as
Cyanogenesis, is described in figure 3.2.
54
Source: Cagnon et al. (2002).

Figure 3.2.
Cyanide liberation by Autochthonous Enzymes Process.
Cyanide toxic effects in superior animals are explained by its affinity with iron, com-
bining itself with the hemoglobin in order to form cyanohemoglobin. In superior plants
and microorganisms, cyanide interferes with the oxidation phosphorylation, combin-
ing itself with cytochrome oxidase, thus inhibiting the electronic transport and conse-
quently the ATP formation (Cereda, 2000).
The limit value that characterizes intoxication by toxic substance is called Lethal Dose
(LD), established during experimental tests with animals. The LD50 represents the
amount of toxic substance that in a single dose can cause death to 50% of individuals
who have taken the substance. In case of cassava containing linamarin, this limit value
is discussible. FAO/WHO (1998) established a limit of 10mg HCN/kg of live weight
(Dantas, 2000), but for inhaled HCN.
However, most part of the cyanide found in the cassava products is eaten, not inhaled,
and will show up as bound or potential cyanide, not as HCN. Since stomach has acid
pH, the bound cyanide hydrolysis speed becomes considerably reduced. Besides that,
few tests have been made so far to measure linamarin content levels in animals. Most
of these tests focus on HCN or on potassium cyanide. In this case, cyanide disposal
differs from the availability of bound organic cyanide in linamarin. In other words,
since the intoxication limits set out by specialized literature are obtained through the
assessment of different substances, no one should rely upon them, not even exaggerate
about their danger levels.
There is a detoxification mechanism that occurs before Lethal Dose limiting level is
reached. It is described in figure 3.3. Through such mechanism, cyanide is transformed
in thiocyanate in the presence of rodanase enzyme and cysteine, the latter being a
sulphur donor amino acid. The thiocyanate obtained, no longer toxic, is eliminated
with urine. For this reason, a way to find out if there was linamarin in the consumed
product and if its metabolism occurred is through thiocyanate detection and its dosage
in urine.
55
Source: Oke (1969).

Figure 3.3.
Linamarin Detoxification Mechanism.
Maduagwu (1989) tested linamarin metabolism in cassava, linen and lima bean, found
in Wistar albine mice, using rats’ liver microsomal preparations. The incubation in
vitro of linamarin, with various concentrations, pH between 6.0 and 6.5, using rats’
liver microsomal preparations, resulted in the substratum’s fast degradation with no
concomitant production of any detectable amount of hydrocyanic acid or thiocyanate,
considered its derivative in detoxification. The same boiling incubation method no
longer degraded linamarin. The mathematical treatment applied to the results could
determine for the in vitro linamarin degradation 3.3 mM as the apparent Km and
degradation maximum speed of 0.017 HCN/minute, expressed in protein milligrams.
The author says that in vivo, during intervals between oral doses ranging from 10 to
350 mg/kg of body weight, linamarin excretion was detected in the rats’ urine in con-
centration levels depending on the administrated dose. On the other hand, the HCN and
thiocyanate excretion in urine was not correlated with the injected dose. After admin-
istrating 10, 50 or 100 mg of linamarin in the animal’s vein, the substance exponential
elimination was verified through assessment of the rat’s blood. The dose half-life was
approximately 90 minutes for all applied dose levels.
Based on the limiting value adopted by FAO/WHO (1998), of 10 mg HCN/kg of live
weight, Lorenzi and Dias (1993) adopted for cassava roots a 100 mg HCN/kg of raw
pulp, considering that after their cooking, this value would drop down to 10 mg. There
is no doubt that this value needs to be discussed.
Lopes (2001) used linamarin obtained from a cassava-cultivated hybrid in toxicologi-
56
cal tests performed on rats. She administrated the substance directly into the animals’
stomach with a probe. This way, the author established for linamarin oral ingestion the
LD50 value of 324.86 mg of linamarin/kg of live weight, or considering the CN-
content in a linamarin molecule, 36 mg HCN/kg of live weight. Other relevant results
were verified such as the fact that linamarin is not cumulative, and that it shows no
residual effects on doses under LD50 limiting value.
Also, cyanogenic glucosides are not the only cassava potentially toxic compounds.
Specialized literature reports the presence of phenolic compounds and scopoletin. These
highly toxic compounds might be underestimated due to the importance given to
linamarin.
Obido and Obasi (1991) described the scopoletin insulation and identification in gari,
a kind of Brazilian flour commonly consumed in Nigeria. The substance content present
in such product did not change during processing like sun drying, refrigerating and
storing. Scopoletin was also identified as the active principle of the Tetrapleura tetraptera
detected in a certain kind of tea used on medicinal purposes in Western Africa. The
substance is a strong hypotension and a non-specified spasmodic agent. Scopoletin
pharmacological effects are probably subjacent factors in tropical neuropathy, which
slowly develops the characteristic endemic symptoms such as optical atrophy, deaf-
ness and neural ataxy in populations subsisting on cassava diet such as gari. Those
toxic characteristics were attributed to the linamarin (cyanide) found in cassava.
As for intoxication caused by linamarin, we can conclude that:
HCN amounts above Lethal Dose limit cause respiratory inhibition and
heart attack. However, linamarin, acetone cyanidrine and HCN are not cumu-
lative.
Animals’ organisms with acid stomachs, including human beings, slowly
eliminate HCN absorbed below Lethal Dose. Cystein amino acid plays an
important role in the elimination mechanism, which explains why rich protein
diets help the organism during detoxification process. Destroyed linamarin is
excreted in urine as thiocyanate. It can also be eliminated intact by urine (Brimer
and Rosling, 1993).
Acute intoxication symptoms appear 4 to 6 hours after ingestion. The
main symptoms are nausea, vomit, headache and finally death. An intrave-
nous injection of thiosulphate can be applied as an antidote. Death will no
longer occur once the symptoms disappear, and recovery can be expected in
the next few hours.
Cases of diseases caused by constant ingestion of linamarin which contents are infe-
rior to Lethal Dose value are still being studied. The quoted diseases in such cases are
57
the optical nerve ataxy, tropical diabetes, konzo paralysis and kidney diseases.
3.4. Acute and Sub-acute intoxication symptoms

Food processing causes linamarin contents to decrease drastically in relation to those
of product in natura. Regardless of this fact, Oke (1969) reminds that when residues
are in large amount, even with doses that are below lethal limit, they can compromise
some based cassava foods. According to the author, the regular consumption of these
products can result in chronicle diseases. This theory had been well accepted until it
became again the subject of new discussions in the last few years, and still needs
confirmation.
Significant cases of intoxication by cyanide are restricted to regions where meals con-
sist almost exclusively of cassava. They are also related to malnutrition problems.
Comparing with Africa, South America, where cassava is largely consumed, has few
confirmed intoxication cases by cyanide as well as less malnutrition problems.
Mathangi et al. (2000) reminded that cassava is the most consumed product in some
developing countries. The introduction of cassava as food was connected with several
diseases including pancreatic diabetes (tropical pancreatic calcification). There are
some studies made with rats about the prolonged ingestion of cassava. The authors
obtained results that proved cassava food was not the cause of diabetes in rats, even
after one year of continuous consumption. Temporary changes happened in the serum
insulin contents and lipase levels, but the meaning of such changes could not be ex-
plained. No histopathological evidences of acute or chronicle pancreatic problems were
verified, but there were toxic hepatic changes in the liver. As a conclusion, the continu-
ous cassava ingestion for up to one year did not lead to chronicle diabetes or pancre-
atic diseases in experimental tests made with rats.
There are only a few confirmed intoxication cases. In general, they relate to children
suffering from acute malnutrition. Akintonwa and Tunwashe (1992) describe one fatal
intoxication case in which 3 patients (2 women, 17 and 18 years of age, and one 8-year
old boy) were admitted in a hospital in Lagos, Nigeria, after eating food made with
gari. The annual average consumption of gari per person in Nigeria is 67 kg. The
patients had vomited and reported abdominal pain right after the meal. They were all
unconscious at moment of examination. Kidney decease was then diagnosed and all
three died 24 hours after being admitted in hospital. Their blood and urine average
cyanide contents were 1.12 and 0.54 mg/liter, respectively. The authors emphasize the
need for acceptable maximum limits of cyanide in gari and other cassava products.
The 24 hours period between the patients’ admission in hospital and death, leaves
58
some doubt about the latter’s real cause, provided that death caused by cyanide usu-
ally happens from 2 to 4 hours after the substance ingestion.
The reports of diseases related to prolonged consumption of cassava-derived products
with high linamarin residual contents are more frequent, but equally confusing. Cliff et
al. (1997) described one outbreak of konzo, kind of symmetric spasmodic paralysis,
associated with the cassava consumption and exposure to cyanide. The number of
patients treated in rehabilitation clinics was 384. The prevailing epidemic rate in the
most affected area was 30 patients per 100 inhabitants. The majority was women and
children. Due to the war, the communities were forced to consume more cassava prod-
ucts, but had simplified their processing so to prepare the meals in less time than
usually. After the end of war, those communities still depended on the cassava that
contained high levels of linamarin, and continued to prepare it inadequately. The out-
break lasted 2 years (during war’s last year and first year of peace), reaching higher
levels in the cassava harvest periods. Although most of the cases were verified in rural
areas, there were some patients from the cities and the coast. Young students’ urine
showed high thiocyanate and linamarin contents, and low inorganic sulfate concentra-
tions. According to the author, there were enough conditions for the disease to appear:
intensive growing of cassava with high linamarin content levels; its insufficient process-
ing, an easy way to ingest large amounts of cyanide; and a low consumption of protein
foods.
Akanji and Famuyiwa (1993) studied the diabetes development associated with the
consumption of cassava and residual cyanide. They made tests of glucose resistance
via intraperitoneal every 4 weeks, in groups of non-milking rats, before and after they
had been fed corn and cassava, with or without sufficient protein and a supplementary
sub-lethal dose of cyanide. After 4 weeks had passed, the monitored animals’ weight
(taking protein supplemented corn starch), as well as other protein supplemented diets,
increased about 50 grams. The weight increase of the rats with protein-deficient diets
was much smaller (22 grams or 50%), a standard kept by this group until they were 12
weeks old. Plasma thiocyanate contents were identical in all rats at the moment they
stopped being milked, and stayed like that after 8 weeks of controlling diet. For cas-
sava diet and diet supplemented with cyanide, plasma thiocyanate contents increased
from 200 to 300% after 4 weeks from the beginning of the tests. The values returned to
normal in all groups after 4 additional weeks during which the animals were fed with
the controlling diet. The rats’ tolerance to glucose was influenced in variable propor-
tion after 4 weeks of being fed different diets. As a conclusion, the authors state that
feeding growing rats with cassava and other non-protein diets caused effects that were
independent, and can produce addictive effects on glucose tolerance. The protein defi-
59
ciency cassava diet supplemented with cyanide didn’t show any effects on the glucose
endurance level.
The article’s weakest point is the comparison between two kinds of diet, the one with
cassava, poor in protein, and the one with corn, which is richer in protein. In this case,
if protein is so important, cassava without any kind of supplementation will always be
worse, independently of the cyanide content, because it is influenced by the low protein
level.
Once linamarin distribution in the plant, and the danger it represents when ingested are
determined, the next important step will be to establish the effects of reduction proc-
esses on glucosides level and the amounts of linamarin retained in the product to be
consumed.
3.5. Cyanide Detoxification in Cassava

Being water-soluble, most part of glucosides capable of generating HCN such as
linamarin and lotaustralin is removed in the process. However, depending on the process-
ing method, residues of these toxic compounds may still remain (Oke, 1969). The
effective reduction of cyanogenic compounds level requires two consecutive phases, as
Cagnon, Cereda and Pantaroto (2002) emphasized:
1) Grating or shredding roots, which allows the cells’ break up to free linamarase
(β-glucosidase EC 3.2.1.21), enzyme that is capable of hydrolyzing linamarin
into glucose and cyanohydrin;
2) Heating the grated roots mass to remove free cyanide residues (acetone-
cyanohydrin and HCN).
Between these two phases, there is a pressing phase commonly adopted, where water-
soluble glucocyanides are dragged along with the water from roots’ constitution. This
process can cause serious environmental damage due to the cyanide that is washed off
and becomes stationary in residual waters. The residual cyanide detoxification was set
out by Pantaroto and Cereda (2001), in volume 4 of this series.
Detoxification practical methods were adopted in food processing in most part of the
tropical regions. Data from specialized literature, however, show that linamarase con-
tent levels are not high enough to degrade linamarin completely. Hence, the traditional
techniques are effective against free cyanide, but not always so against cyanide bound
glucoside.
To ensure the processing of cassava food or derived products with low or no cyanide
contents, a good method is to search for varieties with high levels of linamarase en-
60
zymes. Unfortunately, this is not a common practice among researchers and breeders.
Varieties with high contents of linamarase are more feasible than varieties with no
linamarin or lotaustralin.
Cassava plantation is well developed between 30 degrees latitude north and south from
the Equator line. The Asian regions located in this strip use cassava mainly as raw
material for starch extraction or to feed animals. South America and Africa are the
continents where cassava is used for cooking purposes, mostly the roots but also the
leaves in smaller proportion. Brazil is an outstanding Latin American country for its
large quantities of prepared foods. In Africa, fermented and dehydrated foods are the
most common and in Brazil, foods cooked in water are common.
The most important factors involved with the detoxification of based cassava foods
are the ones that interfere with the biochemical hydrolysis process of glucosides capa-
ble of generating cyanide. These factors are the pH, water availability or water activ-
ity, and the temperature. Another important factor is the physical state of the food
prepared with cassava. The smaller the particles, the most effective the detoxification.
Grated and ground foods result in safer products.
During the second hydrolysis phase, hydroxynitrile-lyase enzyme will be activated
when the pH is between 3.5 and 6.0, and the temperature is below 65ºC. If the pH is
above 4.0 and the temperature above 30ºC the reaction will be spontaneous. The pH
value in healthy cassava tissues is between 5.5 and 6.0, thus allowing the enzyme to
easily activate.
pH: pH interferes with linamarase activation. If pH value is not in accordance with

the right parameter, detoxification will occur in a slower way. If its values fall out of
the ideal strip, i.e. below 3.5, detoxification process will be blocked and linamarin will
become residual, even if all the other conditions are favorable. The fact that linamarin
can remain residual does not mean that the food is toxic, but the potential cyanide
content will be high.
Foods with highly potential cyanide contents may stay intact when inside monogastric
stomachs, which have low pH that is inadequate for hydrolysis; or their hydrolysis can
be very slow, helping the freed cyanide to be converted into thiocyanate before being
eliminated by urine. In reverse, residual linamarin is a serious problem for polygastric
animals, which have neutral or basic pH in their stomach. The presence of protozoan
in the rumen with high cellulose hydrolysis capacity is another aggravating factor. In
such cases, β−type link of linamarin and lotaustralin is easily broken up, and the free
cyanide may reach the lethal dose limit.
In Africa, fermentation is widely used to prepare cassava foods. The flour named gari
61
is a good example. The paste made of ground roots is pressed between two heavy rocks
during over 24 hours. It is a slow process that can allow the roots’ sugars (about 4%)
to reduce pH, therefore providing cyanide fixation. In Brazil, people prepare the same
kind of flour, but the pressing method occurs faster even in places where it is hand
made. Since the pressing system water drags out linamarin and lotaustralin, com-
pounds which are water soluble, residual cyanide content is inferior to 5%. More than
hydrolysis, it is the pressing which can explain the elimination of most of linamarin
and lautostralin. The fermentation is of little influence, the total process being more
important.
Brazilian fermented food is prepared with starch previously extracted and no longer
containing glucoside, or with whole roots that in the final processing step are washed
and dried in hot oven. Tucupi is the food that may show the highest residual linamarin
content levels. It is a partially fermented sauce, made with the water obtained by press-
ing of grated yellow pulp cassava (roots with high linamarin contents). In this case, the
combination of high linamarin content levels with low pH may result in cyanide fixa-
tion. Food prepared with leaves does not show any risks because despite being the
richest in linamarin, the leaves also have the highest linamarase contents.
Water availability or activity: without water, the biochemical reactions are paralyzed
or too slow, resulting in high residual cyanide levels. This can be even worse when pH
is below 3.5, which leads to linamarin fixation, like in gari. Low water activity can
also occur during cassava grinding process, when drying is too fast. The sun drying
method can lead to the cyanide elimination because it is slower than the artificial
heating means, giving the enzyme more time to react. Following this principle, drying
in the shade can be even better. There is no information in specialized literature refer-
ring to the water activity limits in which linamarase enzyme is activated. One can
conclude that the free water used in food processing is favorable for detoxification.
Temperature: considering the first and second hydrolysis phases, the temperature
should neither overpass 65°C, nor be under 30°C. In order to accelerate detoxification,
the temperature has to change slowly from the tropical countries’ room temperatures
(25 to 30°C) up to the limit of 65°C. Higher temperatures can help acetone-cyanohy-
drin volatilization, although they will have no effects on linamarin and lotaustralin.
Since hydrocyanic acid is volatile at 27°C and up, there is no need to apply higher
temperatures. The water boiling temperature cannot facilitate detoxification, but oth-
erwise causes cyanide fixation.
62
3.5.1. Detoxification by means of Cooking
An example of the right choice among detoxification methods is the way cassava roots
are processed in Brazil for cooking purposes (Table 3.1). First the housewife removes
the cortex from the roots that were previously cut in transversal peaces. By doing this,
she gets rid of 81.0% of the potential cyanide. From the other 19% remaining in the
parenchyma (pulp or central cylinder), the most relevant reduction occurs in cooking
and frying, a process that reduces 56% of the pulp’s potential cyanide contents; in
other words, around 92% of the total cyanide present in the raw root.
Table 3.1.
Yields of cassava used for cooking purposes in Brazil after processing, and its
cyanide contents.
-*- : no data.
In rural areas, the most common processing system adopted by women is to put large
quantity of peeled cassava roots in pans full of cold water, covering them completely.
Cooking is done slowly, with gradual increase of temperature taking place inside cas-
sava pieces, and so making detoxification much more effective. This effectiveness
enhances when the cassava is then fried.
Vitrac et al. (2000) describe an experiment made with 13 cassava varieties from the
CIAT’s germplasm data base, in Colombia, in order to characterize three different
levels of compounds found in the pulp: moisture, cyanide, starch and amylose. Ten and
twelve-month old roots were picked, peeled and cut in 1.5 mm thick cylindrical pieces.
The frying time ranged from 70 to 90 seconds, and the oil temperature between 140
and 160°C. Relation time/temperature was adjusted to provide final moisture of 0.004kg/
kg wet base, corresponding to around 0.3 of water activity. The authors assert that
frying only partially eliminated the cyanide, whose 40% were kept. For this reason,
cultivated hybrids with high cyanide content levels may result in bitter meals.
63
In this aspect, all data align with those provided by Cereda et al. (2000), who also
found out a low removal of cyanide from the hybrid Pioneira when it was fried without
any previous cooking. However, in this case the pieces were cut thicker (1 cm sec-
tions), French fries’ shape. Cyanide retention percentage was much higher, 60%. This
retention level occurs because, immersion in high temperature – a frying process char-
acteristic - inactivates the enzyme that is responsible for linamarin degradation.
Combined treatments can be more effective. Cereda et al. (1999a) assessed the effects
of combined processing methods on 12 and 24 months old Pioneiras used for cooking
purposes. To fit this process, the roots were peeled and the pulp’s central cylinder was
sectioned in 1 cm square sticks by 5 cm long. The sticks were blanched during 3
minutes in boiling water and then cooked for 10 minutes in boiling water. Frying was
made with soy oil at 190°C, temperature monitored by thermostat during variable
periods of time, until the cassava sticks were slightly brown outside. The total cyanide
(linamarin) and moisture were analyzed before and after both treatments (Table 3.2).
Table 3.2.
The Influence of treatments on total cyanide contents found in 12 and 24
months old cv Pioneiras’s pulp sticks
Source: Cereda et al. (1990a).
Table 3.2 shows that cooking followed by frying was a most efficient treatment, al-
though cv Pioneira’s total linamarin content could not be removed. The same table
reveals that moisture decreased along the treatments specially frying, which is dehy-
drating (Cereda et al. 1990a). For this reason, frying associates two negative aspects
64
related to the cassava detoxification: the high temperature with the low water activity.
Table 3.3.
Influence of slow freezing at -20ºC during 60 days, on total residual cyanide
content found in 12 and 24 months old cv Pioneiras’ blanched, cooked and fried
pulp sticks.
Source: Cereda et al. (1990a).
Cereda et al. (1990a) analyzed pulp sticks that had been frozen at -20°C during 60
days (Table 3.3). The simply frozen sticks (witness) showed linamarin content reduc-
tion of 39.25%. These results can be explained by the fact that slow freezing leads to
the formation of large ice crystals, which make their way through the cells facilitating
the contact between enzyme and substratum. When witness (no treatment) was consid-
ered 100% for comparison purposes, the authors verified that cooking + frying combi-
nation had been the cause of the biggest reductions, whereas only frying was responsi-
ble for the highest retention rates (Cereda et al. 1990a).
3.5.2. Fermentation as a Detoxification Method

Fermentation is a simple process used in food production. For cassava, fermentation
process consists of putting the whole or sectioned roots, peeled or not, in water with no
inoculating substances or supplementary nutrients. One variant of the process is fer-
mentation of the grated pulp, like in the gari production.
Onabowale (1992) mentions that cassava hydrocyanic acid can be detoxified through
fermentation process, acid hydrolysis or both. Acid hydrolysis proved to be more ef-
fective, removing 98% of the total cyanide from cassava roots, while combined fer-
mentation with acid hydrolysis as well as fermentation alone reduced the cyanide con-
tents in 95 and 87.84% respectively.
According to Vasconcelos et al. (1990), gari, largely consumed by several countries in
the African Westside, can be considered as a fermented cassava product. The authors
65
investigated cassava detoxification during gari processing. By keeping cyanide re-

moval as the scope of the study, they verified that the most important stages in the
process are the product’s initial grating and final roasting. Linamarin hydrolysis de-
pends on the existence of endogenous linamarase as the lactic bacterias detected dur-
ing fermentation were not directly involved with linamarin hydrolysis.
Being a very popular meal, gari is one of the most studied products made in Africa.
Aletor (1993) used 48 albino rats in order to investigate the nutritional implications of
residual cyanide levels in gari. As assessment criteria, he used performance, nitrogen
balance, organs weight, thiocyanate content levels in urine and blood serum, besides
hematological variables. The rats showed no external signs of intoxication, but the
ones fed gari meals containing cyanide had lower growing rates than those under con-
trol (P<0.05). Cassava fermentation improved food assimilation as verified by the
resulting higher diet efficiency rates. Proteinaceous efficiency rates and biological val-
ues increased along with fermentation time. Thiocyanate levels in blood serum and
urine were higher (P<0.001) in gari fed rats than in controlled rats fed based cornstarch
meals. Regression analysis showed a positive correlation (P<0.01, r = 0.97) between
urine thiocyanate and gari’s residual cyanide contents. The studied hematological vari-
ables revealed no consistent tendency relating to the dietetic treatments.
Detoxification by means of fermentation deserves some comments. Originally, the In-
dians in Old Americas used roots fermentation to soften cassava due to the rudimen-
tary grating systems available in those times (Figure 3.4). When fermented, the roots
not only become softer and easy to mince with a pestle, but they also lose soluble
linamarin in the rivers. However, the primary aim of fermentation was not to eliminate
cyanide, but to facilitate food processing. Cassava Brazilian flour processing adapted
very well to Africa’s customs and culture when it was taken to that Continent and
fermentation of the grated pulp then became quality sign criteria.
Figure 3.4.
Primitive grater used by the Amazon Brazilian Indians
to make cassava Brazilian flour.
66
Another cassava detoxifying way found in the specialized literature is the adding of
exogenous linamarase during fermentation. A series of studies published in the last few
years showed that it is possible to reduce the toxic levels (Maher and Hughes, 1971).
Other authors suggest the inoculation of cassava with a microorganism that produces
linamarase (Coock, 1982). Among the studied microorganisms, Brevibacterium line-
age showed good results in degradation in vitro of cyanogenic glucosides (Giraud et al.
1993; Mkpong et al. 1990). Some authors showed that fungi (Ikediobi and Oniyke,
1982) and bacteria that produce lactic acid are capable of generating linamarin hy-
drolysis (Padmaja and Balagopal, 1985). For instance, cells cultures of Lactobacillus
plantarum in cellobiose MRS (a nutrient mean) may degrade cassava extracted linamarin
in less than two hours (Padmaja and Balagopal, 1985).
Padmaja and Balagopal (1985) state that fungus R. oryzae, associated with damages
caused in cassava after its harvest, is able to metabolize cyanide. Cassava cyanogenic
glucoside degradation by R. oryzae was studied through cultivating the organism in
potato dextrose syrup with or without linamarin and potassium cyanide. The influence
of the growing organism adaptation in high and low cyanide concentration, and the
transfer of extra cellular rodanase to the culture containing cyanide were also as-
sessed. The study revealed R. oryzae’s potentiality use in glucoside detoxification of
cassava as human and animal food, as well as in industrial residues treatment.
In spite of their scientific fundament, the fermentation processes, using inoculums or
enzymes, are difficult to be adopted both for commercial and domestic purposes be-
cause of their complexity, qualified labor requirements and high costs.
3.5.3. Grating and pressing, followed by heating, as a Detoxifica-

tion method
Amerindians developed one of the most effective detoxification methods. They domes-
ticated cassava and created empirical as well as pre-scientific systems that still today
are among the safest. Such method consists of grating, pressing, crumbling and drying
(roasting). Considering the cyanide elimination aspect, this process is effective be-
cause the grating step puts enzyme in contact with the substratum under the best reac-
tion conditions, pH between 5.5 and 6.0 (root’s natural pH) and room temperature.
Once the ideal conditions for hydrolysis are provided, pressing step carries linamarin
that may have remained intact, and the acetone cyanidrine, both water-soluble. At last,
the free cyanide, as HCN, is eliminated by means of heating the wet product in an open
oven. Analysis proved that just 1mg/kg of linamarin remains in Paulista type Brazilian
flours (a kind of cassava Brazilian flour produce in Southeast of Brazil).
67
Linamarin residues sometime found in the African flours and fermented products may
result from adaptations made in the processing, or from lack of efficient equipments.
Dufour (1989) describes the methods adopted by Tukanos, Indians who live in the
northwest side of the Brazilian Amazon. These methods are used to process roots and
leaves from cassava varieties with high linamarin content levels. The obtained prod-
ucts are a traditional diet. The reproduction of such methods in laboratory, using 2
high cyanide contents cultivated hybrids (bitter cassavas), showed that the methods
used to make cassava bread reduce quickly and efficiently cyanide contents without
affecting nutritional value.
Aletor (1993) describes a variant of gari’s making process in which the flour is fried
after it has been pressed. He does not explain the process in details, but one can as-
sume that frying was done in the oven with oil or fat. Aletor examined 108 samples of
manufactured gari collected from the main producers in Nigerian communities. These
samples were analyzed for linamarin (total or potential cyanide) and free or bound
hydrocyanic acid (acetone-cyanohydrin). Distribution of Linamarin levels frequency
revealed that approximately 80% of the samples contained less than 20 mg/kg, 13%
contained 20 to 30 mg/kg and the remaining 7%, 30 to 40 mg/kg. Around 90% of all
samples had 10 mg/kg of bound cyanide.
There were differences in and among the communities as for the residual cyanide
levels found in the gari samples. According to Aletor, those differences occurred due to
several factors, some of them related to agriculture (one or more types of cultivated
hybrids used in each or more communities; different cultivation treatments; environ-
mental factors), or technology, like lack of standardization for the processing tech-
niques before and during gari production. The process was studied in laboratory, using
one cassava variety with high linamarin content level. Results showed that grinding,
more than fermentation is decisive in the reduction of residual cyanide bore by the final
product, although these two steps can have complementary effects. Garification (fry-
ing) made right after grating and pressing resulted in high linamarin content levels,
higher than those found in commercialized samples.
Mlingi et al. (1993) described the results of analyses made in one North Tanzania
village affected by konzo, in parallel with its neighbor, another village where the dis-
ease had not been registered. The authors studied the exposure to based cassava food
with high cyanide residual contents, and its connections with the disease. Food short-
age caused a reduction in the traditional process traditionally adopted by both villages.
The intense commerce of these cassava products had also contributed to a process
simplification in the affected village. One could verify that when the cassava roots
were processed in the traditional way, including grating and fermentation of the matter,
68
there was a drastic decrease in terms of cyanide contents. Nevertheless, considerable

linamarin contents remained in the flour even after a 4-night fermentation period.
Linamarin value was higher in the flour obtained by reduced process, i.e. 57.6 mg/kg
of dry weight, compared with 36.7 presented by the products traditionally fermented
during 4 nights. The high linamarin content levels could be confirmed by the high
thiocyanate content levels detected in urine from the affected village, 490.48 µmol/liter
against the 50.39 µmol/liter collected from its healthy neighbor. The exposure to cya-
nide was enhanced due to a very poor diet in terms of protein. This resulted in less
sulphurous amino acids to convert cyanide into thiocyanate, and was confirmed by the
low sulphate content levels shown in the collected urine. The urine inorganic sulphate
contents were 3,802.369 for the affected village, and 7,038.855 µmol/liter for the one
not affected by konzo.
O’Brien et al. (1992) described the influence of two cassava detoxification methods
for cyanide found in meals traditionally prepared in Cameroon, Africa. They assessed
farine de manioc, processed like traditional dry flour, and baton de manioc, obtained
after cassava roots cut in cubes are fermented in water during 2 days. The matter is
then pounded resulting in a paste that is wrapped in leaves and taken to boil in water
for 20 to 30 minutes. In each processing stage, cassava’s three glucoside toxic com-
pounds were dosed: linamarin (total or potential cyanide), cyanohydrin (acetone-cy-
anohydrin) and HCN or free cyanide. In-water traditional fermentation, followed by
pressing and drying in the sun or boiling methods reduced linamarin’s 91 to 515 mg/kg
content values found in the fresh roots to more acceptable values of 0.0 to 11.3 mg/kg
of prepared and ready for consumption meals. Fermentation temperature and the size
of root pieces had an influence on the linamarin hydrolysis rates, which caused a
temporary increase of cyanohydrin contents as an intermediary product of the reac-
tion. Despite the low pH found in the fluid, it was possible to detect cyanohydrin
hydrolysis for free cyanide (hydrocyanic acid) all throughout the different phases of
fermentation, allowing the following removal of volatile cyanide by combination of
pressing, sun drying and cooking.
Analysis of 30 cassava Brazilian flour samples produced in different Brazilian states
by means of several processes revealed that Brazilian flours have separate character-
istics. The studies were made according to techniques dictated by Essers (1995), i.e.
through enzyme controlling. Analyses included the Brazilian Southern states, with fast
processing systems, including mechanical peeling and the removal only of the brown
outside skin of the roots. Processed Brazilian flours were also taken in the North and
Northeast regions, characterized by rudimentary methods. Of all Brazilian flours
analyzed at UNESP (University of São Paulo State – Brazil), 11 had come from the
69
North and Northeast, and 18 from the South and Southeast. Statistic assessment showed
that despite North and Northeast’s Brazilian flour having on average higher linamarin
content levels, i.e. 1.52, these are not very different from South and Southeast’s 0.94.
The results indicate that detoxification does not depend on the technological level ap-
plied to the process, an observation reinforced by the fact that technology used in the
South and Southeast is much more advanced and makes use of most of the roots, in
comparison with the North and Northeast states, where losses are higher. Eight sam-
ples of fermented Brazilian flours (carimã and d’água), besides twenty non-fermented
(dry, paulista, and roasted) were included in the study. The fermented Brazilian flours
presented an average linamarin content of 1.71 mg/kg against the non-fermented Bra-
zilian flours’ 0.99. The high content value of fermented Brazilian flours is based mainly
in a flour that contained 6.6 mg/kg. Nevertheless, the resulting differences were not
significant.
Sources: CERAT/UNESP, unpublished data.

Figure 3.5.
Linamarin content distribution in 29 samples of cassava Brazilian flours from 7
Brazilian states.
The farinha d’ água (fermented Brazilian flour from Amazonian region) samples from
Amazon and Pará are from handmade process, manual peeling of the out skin and the
inner layer, maceration in water, pressing and oven roasting (Chapter 21 of this vol-
ume). This technology is very similar to the ones applied in Africa in gari making. The
results shown in Figure 3.5 reveal very low linamarin contents. Even considering the
limits established by FAO, all these Brazilian flours would be safe for human con-
sumption. Only one type had linamarin content levels higher than 2 mg/kg. The aver-
age results were 1.14 mg/kg, with median (class’s most frequent average) of 0.87 mg/
kg. Such low values confirm the detoxification efficacy obtained through the grating,
70
pressing and drying sequence. Detoxification does not depend on the technological
level applied or whether cassava is or is not fermented. Even with pH being pulled
down by fermentation toward levels that are not ideal for linamarase activation, cas-
sava roots maceration in water may have promoted the dissolution of linamarin. Even
though both regions have adopted similar ways to process cassava, the reasons why
African flours gari type can contain over 20 times more linamarin than the Brazilian
ones have still to be explained.
3.5.4. Dehydration as a Detoxifying Method

One of the techniques used to detoxify cassava products is dehydration. It can be done
by natural or artificial sun drying. Since both ways rapidly decrease the water activity
and use high temperatures, factors that together can damage linamarase enzyme ac-
tion, artificial drying should be used carefully. Specialists wishing to improve Amerin-
dian’s methods with more efficient equipments and faster processes performed several
disastrous experiments.
Ospina and Wheatley (1992) described the process that uses cutting, naturally sun
drying, scraping and pelletization of cassava roots. Drying done with the help of more
sophisticated technologies resulted in better quality chips flours, but with negative
effects on linamarin detoxification. These results can be explained by the use of intense
heat and fast drying, which disables linamarase enzyme and fixes cyanide.
The dried cassava leaves can be given to animals as food. Gomez and Valdivieso
(1985) analyzed the aerial part of 4 cultivated cassava hybrids that were respectively
6, 8, 10 and 12 months old. The sun drying treatment on cemented ground was com-
pared with oven drying at 60°C. Besides cyanide the study analyzed tannin in the
dehydrated leaves. The dehydrated leaves’ composition, including calcium, phospho-
rous and amino acids, was similar to the sun dried lucerne flour. The cassava plants
produced on average 25.30% of dried matter; the dried leaves’ gross protein was 13.20%
and the gross fiber was 16.20%. Proteins and fibers were the compounds that varied
mostly according to the age of the plant. The sun and oven drying removed respec-
tively 82.94% and 69.76% of free cyanide and 24.36% of total cyanide (linamarin)
from the fresh leaves. The samples dried in the sun showed less tannin than the sam-
ples dried in the oven. The obtained results show how important it is to select the best
drying methods for cyanide contents’ reduction. As sun-drying process uses lower
temperatures, it is slower, allowing linamarase enzyme to act. Studies show that cas-
sava leaves picked at night, macerated and dried in the shade can be almost completely
detoxified.
71
Dehydrated leaves’ linamarin contents can vary a lot due to the already mentioned
causes. Figure 3.6 shows a histogram of results from laboratories that analyzed the
total cyanide in dehydrated cassava leaves of different varieties, harvest periods, ages
and processing systems. The highest frequency was for the quantity of 25 mg/kg,
although some samples showed high residual amounts of linamarin, enough to cause
problems in animals, especially polygrastric ones. The dehydrated leaves with higher
linamarin content levels had probably been dried fast.
Source: CERAT/UNESP unpublished data.

Picture 3.6.
Variation of linamarin content in cassava dehydrated leaves.
The use of inadequate processing is mentioned in specialized literature as another

factor for the appearance of sub chronic intoxication symptoms associated with the
ingestion of food containing high residual linamarin levels. This happen when, in spite
of the existence of appropriate methods to detoxify food, for some reason they are not
applied.
Essers and Nout (1989) discuss the importance of cutting and drying for the detoxifi-
cation of cassava roots. The authors tell about fresh cassava roots cut in pieces with
similar cyanide contents. One piece from each root was immediately frozen and analyzed
to serve as reference. The remaining pieces were dried and stored for 8 months, in
similar conditions to the domestic ones found in Mozambique’s rural areas. Fungi in
varied levels appeared in these pieces that had been sun dried. The pieces that con-
tained fungi were analyzed for moisture, total and free cyanide, pH, luminosity (color)
and presence of aflatoxins. There was counting and identification of the fungi. The
traditional processing system led to a considerable loss of total cyanide, from 92.3 to
99.5%. The free cyanide content varied between 19 and 89% in comparison to the total
cyanide. No relation between the contents of the fresh roots and of the dried and stored
72
ones was found in terms of total cyanide. The darkest flours revealed lower cyanide
contents either total or free, as well as less acid pH. There was no detection of afla-
toxin in the dried product. The authors concluded that for cassava flour made this way,
the consumer’s safety cannot be guaranteed just by the assessment of color and fungi
growing extension.
Almazan (1992) analyzed gari storage at 25°C, inside plastic bags during 2 to 40
weeks. He verified a decrease in cyanide concentration and also in the product’s sticki-
ness.
Monroy et al. (1991) studied the drying process applied to cassava chips and its influ-
ence on linamarin detoxification. The authors assessed the chips thin layer being dehy-
drated in greenhouse, and determined the process timing as well as humidity, which
together can be more adequate to cyanide elimination. They studied three ranges of
temperature, 35, 60 and 85°C. Over 70% of total and bound cyanide were eliminated
at drying temperature of 60°C, air speed of 1.5 m/s, and 0.003 kg humidity /kg of dry
air. The best parameter for linamarin elimination was found at 60°C. These results
show the effect of surrounding tissues’ thermal protection on linamarase activity, and
prove that hydrolysis is the limiting reaction in linamarin elimination.
3.6. Cyanide and natural diets

The use of cassava to feed animals is very important in all continents that can cultivate
it. Despite the fact that both underground (roots) and aerial parts of this plant can be
used, its applications are still limited and little significant. Most of people’s indiffer-
ence comes from fear and lack of knowledge about the detoxification methods. The use
of cassava as animals’ feed is possible and requires attention. Accurate information
about the plant’s composition and the animals to be fed is of great importance. The
adaptation to a new diet is absolutely necessary not only concerning cassava products
and by-products, but for all new foods.
Basic knowledge of the above mentioned findings about cyanide cannot be despised. If
adequately adapted, monograstric animals become less susceptible to accidental in-
toxication than polygrastric animals. This is explained by the acid pH existing in the
stomach of the first, while the latter has neutral pH stomach. In acid mean, linamarase
enzyme stays inactive and there is slow liberation of cyanide, giving time for its elimi-
nation before lethal doses are reached. Combination of neutral pH and bacteria that
induce linamarin hydrolysis is lethal, because cyanide is quickly freed and may reach
the lethal dose limit before being eliminated. Specialized literature suggests some ways
to make animal dieting safer.
73
Panigrahi et al. (1992) describe one cultivated cassava hybrid with high linamarin
content levels, whose roots were cut but not peeled, and taken to slow drying at a
temperature of 25ºC and then to fast drying in the oven. One group of pieces took over
24 hours to dehydrate by fast drying process, whereas the other group was dehydrated
in 72 hours time by slow drying. The dehydrated products were incorporated in semi
synthetic diets to feed one-day old chicks, in 250 to 500 g/kg portions, as paste or
pellets, in separate experiments. Both drying systems produced food containing simi-
lar concentrations of polyphenols, but different concentrations of linamarin. Total cya-
nide varied from 38 mg/kg for the slow drying method to 482 mg/kg for the fast drying
method. The growth rate of chicks fed slowly dried products was higher than of those
fed fast dried products. Comparing with a controlled diet, the gain of weight by chicks
fed diet containing 500 g/kg (258mg/kg of total cyanide) was 77% smaller, although
the performance rate of the birds that had consumed 142mg/kg of total cyanide was
equally low.
Still according to Panigrahi et al. (1992), the relation water-food amounts consump-
tion was higher in groups fed fast dried diets than in those fed slowly dried diets, a fact
reflected in the high content levels of the water excreted. Pancreas weight was smaller
in birds fed fast dried diets than in the groups fed slowly dried diets, but only in
experiments with paste food. The diets did not have any effects on the mortality rate of
the chicks. Although pelletization process generates high temperatures, no significant
effect was verified on cyanide reduction or the birds’ growth. In brief, slower drying
system produces food with lower cyanide content levels that consequently are less
toxic for chicks. Based cassava diets bearing total cyanide contents below 40 mg/kg
can be given to chicks in 500 gm/kg of live weight doses, without incurring any risks.
Onabowale (1992) describes the growth of laying chicken fed diets where 40 to 50%
of corn was replaced by an equivalent product obtained from detoxified cassava. These
chickens’ eggs production was more efficient than of those which had been fed corn
(71.43 against 66.67%). No adverse effect related to the diets was verified as for the
birds’ growth or performance. Growth rate increased and diet’s conversion efficiency
rate remained the same, when compared to corn.
Gomez et al. (1983) described the drying method in the production of cassava chips
used as food for swine. According to these authors, roots of the cultivated cassava
hybrids CMC40 and Llanera, with low linamarin contents, and Mcol 1684, with high
linamarin contents, were scraped and sun dried on cemented or earth ground. The dried
chips’ cyanide content was lower than 100 mg/kg. Based sorghum diets supplemented
or not with 30% of cassava food were given to 48 pigs weighing 17 kg, during 60 days,
until they reached the 96 to 98 kg weight mark. The same type of diet was given to 14
74
female pigs, while a separate diet containing 40% of cassava chips fed 14 days old
piglets. An industrialized diet supplemented or not with 10 to 20% of dried cassava
chips, was given to 735 Arbor Acres chickens. The performance of pigs and chickens
fed with partially replaced cassava diets was similar to the one of those fed with con-
trolled diets, regardless of the high or low cyanide contents found in the cultivated
hybrids.
Gomez and Valdivieso (1988) tell about the effects of silage made with cassava chips
from whole roots of a cultivated hybrid with medium cyanide contents. The 12 months
old plants were kept for 2 weeks inside 2 silos. The initial linamarin contents found in
the fresh chips were 503 and 556 mg/kg, registered in dry matter, for silos 1 and 2
respectively. Of all linamarin content (total cyanide), 40 and 43% were free cyanide.
At the end of the first week stay in silos, most cyanide was in free form, which is more
volatile. When the 2-weeks period stay in silos ended, the cassava chips’ total cyanide
concentrations were 181 and 141 mg/kg of dry matter, for silos 1 and 2 respectively,
corresponding to 36 and 25% of the initial concentrations of total cyanide. Of these
quantities, 95% were in free cyanide form. The authors emphasize that silo storage is
an efficient means of reducing the chips’ cyanide content. The final dry matter amount
resulting from silo storage was approximately 9% higher than that of fresh chips.
After 26 weeks, pH values were 3.82 and 4.13 for silos 1 and 2 respectively, and the
average dry matter of the samples was 40.6 to 41.5%.
3.7. Potential usage of Linamarin

As we can see, the available specialized literature reveals the disadvantages of residual
linamarin consumption by the food. Aerobic animals, either superior or not, die of
asphyxia and respiratory system paralysis when lethal dose limit is reached. On the
other hand, diseases caused by sub lethal doses have not been well explained yet.
However, it is known that malnutrition and problems related to social instability might
have equal or superior relevance in the scale of factors that are responsible for the
appearance of these diseases.
One scientific issue that has been little explored in specialized literature or by the
media concerns the power of low linamarin residual contents (β-glucoside) to kill car-
cinogenic cells, leaving the healthy ones alive, as a target effect. Brazil is known for
the habit of consuming cassava products that have minimum doses of linamarin. This
practice has not been scientifically assessed yet. According to results of analyzes per-
formed in CETESP, by means of AMES control, P.A. Sigma linamarin showed no
genotoxic activity in the presence of E. coli PQ37 strain, absent metabolic activity, in
75
0.005, 0.05, 0.5 and 1.0 mg/assay trial. Toxicological controls made at UNESP’s
Pharmacological Department of Biosciences Institute, with injection of P.A. linamarin
in mice via peritoneum, indicated DL50 of 750 mg/kg.
In biochemical terms, this selected death is possible. After examining such hypothesis,
Cereda and Mattos (1996) developed one methodology of carcinogenic cells’ death
assessment. Hypothetically, nitrilosides metabolism kills specifically the cells attacked
by cancer. Rats with carcinoma of Walker revealed the highly therapeutic effects brought
by HCN after they had inhaled this substance. Moreover, long-term experiments dem-
onstrated that nitrilosides produce carcinostatic effects. Experiments made with hu-
man beings that suffered from carcinomatous limphadernitis and lung metastases showed
the anti-carcinoma effect of amidalin, injected daily intravenous via, when powered by
complementation with β-glucosidase. The peoples not suffering from cancer like Indi-
ans from Hunza de Karakorum tribe and the Arctic’s Eskimos have a glucoside rich
diet everyday, like cherry seeds and apricot.
According to Cereda and Mattos (1996), the biochemical base of such theory is that
the detoxifying activity in cells occurs thanks to rodanase enzyme. The healthy cells of
human beings have rodanase, and can defend themselves against cyanide; differently
from the carcinogenic cells, that have no rodanase, hence being cyanide’s preferred
target.
Senior (2002) also tells about cyanide’s power to destroy carcinogenic cells. One of
the glucosides that showed good results after being analyzed was amidalin, as Syrigos;
Rowlinson-Busza and Epenetos (1998) mentioned. Not only linamarin produces this
effect; other similar nitriles have an equal ability, as pointed out by Hjarna et al. (1999)
and Hansen et al. (2000). The international specialized literature includes the researches
made by an English scientist, Monica Hughes, from School of Biochemistry and Ge-
netics of New Castle University Medical School, who made evidence of this selective
effect. In order to continue her research, Hughes cloned microorganisms that were
capable of producing β-glucosides. The results are being expected eagerly so that
people can view linamarin in a different perspective.
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84

Cassava Processing As A Method of Detoxification: Latin American Starchy Tubers

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LATIN AMERICAN STARCHY TUBERS

1- NGO RAÍZES - Av. Manaca, 524 – 18607-170 Botucatu – SP - Brazil. cereda@raizes-

3.2. The glucocyanide in the cassava plant

Source: http://www.sdu.dk/Nat/Chem/kemi_net/LostFast/Cassanova1.gif (1993).

3.3. Cyanide liberation in the cassava

Source: Cagnon et al. (2002).

Source: Oke (1969).

3.4. Acute and Sub-acute intoxication symptoms

3.5. Cyanide Detoxification in Cassava

pH: pH interferes with linamarase activation. If pH value is not in accordance with

3.5.1. Detoxification by means of Cooking

Source: Cereda et al. (1990a).

Source: Cereda et al. (1990a).

3.5.2. Fermentation as a Detoxification Method

investigated cassava detoxification during gari processing. By keeping cyanide re-

3.5.3. Grating and pressing, followed by heating, as a Detoxifica-

there was a drastic decrease in terms of cyanide contents. Nevertheless, considerable

Sources: CERAT/UNESP, unpublished data.

3.5.4. Dehydration as a Detoxifying Method

Source: CERAT/UNESP unpublished data.

The use of inadequate processing is mentioned in specialized literature as another

3.6. Cyanide and natural diets

3.7. Potential usage of Linamarin

ALETOR,V.A. Cyanide in gari. 2. Assessment of some aspects of the nutrition, bio-

BOKANGA, M. Constraints in food and nutrition research. In: THOTTAPPILLY, G.;

CAGNON, J.R.; CEREDA, M.P.; PANTAROTTO, S. Glicosídeos cianogênicos da

CEREDA, M.P. Caracterização dos subprodutos da industrialização da mandioca..

CEREDA, M.P. Processing of cassava roots in Brazil. Safety implications. In : M.

CEREDA ,M.P.; SARMENTO,S.B.S.;WOSIACKI,G.;ABBUD,N.S.;ROÇA,R. de O.

CEREDA,M.P.; SARMENTO,S.B.S; WOSIACHI,G.; ABBUD,N.S.; TAKEDA,

CURY,R. Distribuição da diversidade genética e correlações de características em

DUFOUR, D.L. Effectiveness of cassava detoxification techniques used by indigenous

ESSERS, A.J.A. Removal of cyanogens from cassava: studies on domestic sun-drying

GIRAUD, E.; GOSSELIN, L.; RAIMBAULT, M. Degradation of cassava linamarina

GIRAUD, E. Production of a Lactobacillus plantarum starter with linamarase and

GOMEZ, G.; VALDIVIESO, M. Cassava foliage: chemical composition, cyanide con-

GOMEZ, G.; VALDIVIESO, M. Changes in cyanide content of cassava tissues as

GOMEZ,G.; VALDIVIESO,M.; SANTOS,J.; HOYOS,C. Evaluation of cassava root

HANSEN, C.M.; HANSEN, D.; HOLM, P.K.; LARSSON, R.; BINDERUP,L.

HJARNAA, P.J.; JONSSON, E.; LATINI, S.; DHAR, S. ; LARSSON, R. ; BRAMM,

KOJIMA,M.; IWATSUKI,N.; DATA,E.S.; VILLEGAS,C.D.V.; URITANI,I. Changes

LEGRAS, J.L.; JORY, H.; GALZY, P. Detoxification of cassava pulp using

LUTALADIO, N.B.; WAHUA,T.A.T. Variability in leaf HCN, protein, and

MADUAGWU, E.N. Metabolism of linamarina in rats. Food and Chemical Toxicol-

MAHER E.P. Isolation of Linamarin-Lotaustralin from Trifolium repens. Phytochem-

MATHANGI, D.C.; DEEPA, R.; MOHAN, V.; GOVINDARAJAN, M.;

McMAHON, J.M.; WHITE,W.L.B.; SAYRE, R.T. Cyanogenesis in cassava (Manihot

MLINGI,N.V.; ASSEY,V.D.; SWAI,A.B.M.; MCLARTY, D.G.; KARLEN, H.;

MONROY RIVERA,J.A.; LEBERT,A.; MARTY,C.; MUCHNIK,J.; BIMBENET,

MONROY RIVERA,J.A.; LEBERT,A.; MARTY,C.; MUCHNIK,J.; BIMBENET,J.J.

OBIDO,A.O.;OBASI,S.C. Coumarin compounds in cassava diets: 2. Health implica-

OBIGBESAN, G.O. Cyanide content of cassava cultivars in relation to nitrogen ferti-

ONABOWALE, S.O. Constraints and projections for processing and utilization of

cassava. IN: HAHN, S. K.; REYNOLDS,L.; EGBUNIKE, G. N. Cassava as live-

PADMAJA, G; BALAGOPAL, C. Cyanide degradation by Rhizopus oryzae. Cana-

PANTAROTTO, S.; CEREDA, M.P. Linamarina e sua decomposição no ambiente.

PEREIRA, J.F.; Splittstoesser,W.E. Exudate from cassava leaves.

PETROCCIOLI, M.; BRIMER, L.; CICALINI, A.R.; FEDERICI, F. The linamarase

SAYRE, R.T. Cassava research in the Sayre. Disponível em:<http:www.biosci.ohio-

SAYRE. R.T. Purification, characterization and localization of linamarase in cassava.

SELMAR. D. Cyanogenesis, cyanogenic glicosídeos, linamarina, linustatin, transport.

SPENCER, K.C.; SEIGLER, D.S; NAHRSTEDT, A. Phytochemistry, v. 25, p.645,