Picrate paper kits for determination of total cyanogens in cassava roots and all forms of cyanogens in cassava products

The simple semiquantitative picrate method for the determination of total cyanogens in cassava flour has been modified by increasing the concentration of the picrate solution used to make up the picrate papers, such that a linear Beer's Law relation between absorbance and cyanogen content is obtained over the range 0–800 mg HCN equivalents kg⁻¹ cassava. The method has been adapted to determine the total cyanogen content of cassava roots and the results compared using the picrate method and the acid hydrolysis method for six different roots from five cultivars. The agreement between the results is satisfactory. The simple method for determination of total cyanogens in cassava roots in the field is available in kit form. The methodology has been modified to allow determination of the three different forms of cyanogens present in cassava flour, viz HCN/CN⁻, acetone cyanohydrin and linamarin. HCN/CN⁻ is determined by the picrate method in which cassava flour is reacted with 0.1 M sulphuric acid for 3 h at room temperature. HCN/CN⁻ plus acetone cyanohydrin is also determined by the picrate method after treating cassava flour with 4.2 M guanidine hydrochloride at pH 8 for 3 h at room temperature. A comparison has been made of the amounts of the three cyanogens present in six cassava flour samples using the semiquantitative picrate and the acid hydrolysis methods. The agreement between the two methods is satisfactory, which shows that the new methodology works well. The picrate method for determination of the three cyanogens in cassava flour is also available as a kit. © 1999 Society of Chemical Industry     

A survey of total hydrocyanic acid content in ready-to-eat cassava-based chips obtained in the Australian market in 2008

Cassava (Manihot esculenta Crantz) is a widely consumed food in the tropics that naturally contains cyanogenic glycosides (cyanogens, mainly composed of linamarin, acetone cyanohydrin, and hydrocyanic acid). If cassava is not adequately processed to reduce the level of cyanogens prior to consumption, these compounds can lead to the formation of hydrocyanic acid in the gut. Exposure to hydrocyanic acid can cause symptoms ranging from vomiting and abdominal pain to coma and death. In 2008, a survey of ready-to-eat (RTE) cassava-based snack foods was undertaken to determine levels of cyanogens measured as total hydrocyanic acid. This survey was undertaken in response to the New South Wales Food Authority being alerted to the detection of elevated levels of cyanogens in an RTE cassava-based snack food. This survey took 374 samples of RTE cassava chips available in the Australian marketplace. Significant variation in the levels of total hydrocyanic acid were observed in the 317 samples testing positive for cyanogens, with levels ranging from 13 to 165 mg of HCN equivalents per kg (mean value, 64.2 mg of HCN eq/kg for positive samples). The results from this survey serve as a timely warning for manufacturers of RTE cassava chips and other cassava-based snack foods to ensure there is tight control over the levels of cyanogens in the cassava ingredient. Evidence from this survey contributed to an amendment to the Australia New Zealand Food Standards Code, which now prescribes a maximum level for hydrocyanic acid in RTE cassava chips of 10 mg of HCN eq/kg, which aligns with the Codex Alimentarius Commission international standard for edible cassava flour.     

Cyanogen content of cassava roots and flour in Indonesia

A survey has been made of the total cyanogen content of cassava roots and products from the cassava growing provinces of Lampung and East, Central and West Java, in Indonesia. Twenty five samples of cassava products were analysed for cyanogens by the acid hydrolysis method and also by the simple picrate kit method. The mean percentage difference between the results was 17%. Thirty samples of cassava starch and other specialised products had a mean cyanogen content of only 5 ppm, whereas 29 samples of cassava flour, chip and gaplek gave a much higher mean cyanogen content of 54 ppm (SD 51). The WHO safe value for cassava flour is 10 ppm and the Indonesian level is 40 ppm. There are four outliers of cyanogen content 140–200 ppm, which would be dangerous to human health. The cyanogen content of starch/chips/gaplek needs to be reduced by using cultivars of lower cyanogen content and by using improved processing methods. Twenty seven samples of cassava roots gave a mean cyanogen content of 19 ppm (SD 14). ©     

Processing Techniques to Reduce Toxicity and Antinutrients of Cassava for Use as a Staple Food

ABSTRACT: Cassava is a valuable source of food for developing countries, but it contains highly toxic cyanogen compounds and antinutrients. Cyanogens are found in 3 forms in cassava: cyanogenic glucoside (95% linamarin and 5% lotaustratin), cyanohydrins, and free cyanide. Different processing techniques exist to remove cyanogens and their effectiveness depends on the processing steps and the sequence utilized, and it often is time‐dependent. Pounding or crushing is the most effective for cyanogenic glucoside removal because it ruptures cell compartments, thus allowing direct contact between linamarin and the enzyme linamarase that catalyzes the hydrolytic breakdown. Crushing and sun‐drying cassava roots made into flour removes 96% to 99% of total cyanogens, whereas soaking and sun‐drying into lafun or fufu, or soaking and fermenting and roasting into gari or farina, removes about 98% of cyanogens. For cassava leaves, which have 10 times more cyanogens than roots, pounding and boiling in water is an efficient process to remove about 99% of cyanogens. Other strategies to reduce toxicity include development of low‐cyanogen cassava varieties and cassava transgenic lines with accelerated cyanogenesis during processing. Although phytate and polyphenols have antioxidant properties, they interfere with digestion and uptake of nutrients. Fermentation and oven‐drying are efficient processing methods to remove phytate (85.6%) and polyphenols (52%), respectively, from cassava roots. Sun‐drying the leaves, with or without prior steaming or shredding, removes about 60% phytate. Cassava is a nutritionally strategic famine crop for developing countries and, therefore, reducing its toxicity and improving its nutritional value is crucial.     

Cyanogenicity of cassava varieties and risk of exposure to cyanide from cassava food in Nigerian communities

BACKGROUND: High‐cyanogenic cassava varieties are cultivated in many tropical communities that are free of neurological syndromes attributed to consumption of cassava foods. This study was done in four geographical areas of Nigeria (northern, southwestern, southeastern and an area endemic for ataxic polyneuropathy) to determine if cyanogenicity of cassava is associated with geographical area, altitude or level of cyanogenic compounds in gari, a popular cassava food in West Africa. RESULTS: Mean levels of cassava cyanogens were 153, 127, 68 and 65 mg HCN equivalents (eq.) kg^?1 dry weight (DW) in the endemic, southeastern, southwestern and northern areas respectively (P < 0.0001), while mean levels of gari cyanogens were 9, 4, 7 and 13 mg HCN eq. kg^?1 DW in the respective areas (P < 0.0001). The mean altitude was 35 m in the endemic area, 55 m in the southeastern area, 220 m in the southwestern area and 273 m in the northern area (P < 0.0001). Altitude was associated with cyanogenicity of cassava in univariate and multivariate models (P < 0.0001). One hundred and twenty‐six (93%) farmers and 255 (77%) processors did not perceive cassava or its food products as toxic. CONCLUSION: The findings indicate that cyanogenicity of cassava is determined by environmental factors rather than by conscious selection of varieties by farmers. Farming high‐cyanogenic cassava is not associated with high levels of residual cyanogens in gari. Cassava is not perceived as toxic by farmers and processors. Copyright © 2008 Society of Chemical Industry     

Characterization and comparison of key aroma-active compounds of cocoa liquors from five different areas

The volatile constituents of cocoa liquor and the differences between cocoa liquors from different origins were studied. Direct solvent extraction-solvent assisted flavor evaporation and gas chromatography-olfactrometry-mass spectrometry in conjunction with aroma extract dilution analysis were used to identify the key flavor compounds in five cocoa liquors. There were significant differences of specific compounds between cocoa liquor from different areas of origins. Then, the purge and trap method was used to gathering the aroma-active components of five cocoa liquors from different origins, and one internal standard was used during this process for the further quantitative analysis by gas chromatography-olfactrometry-mass spectrometry. The results indicated that 3-methylbutanal, acetic acid, tetramethylpyrazine, and 3-methylbutanoic acid were the components with high concentrations. The contents of most compounds in the five kinds of cocoa liquor were dramatically different. The content of odorants of cocoa liquors from Papua New Guinea was higher than that of the others and that from Indonesia was the lowest. For sensory evaluation, the overall odor outlines of the five cocoa liquors were very similar, the Papua New Guinea cocoa liquor had higher preference than those of the other four samples. Principal component analysis showed that the characteristics of cocoa liquor from Papua New Guinea, Indonesia, and Ivory Coast were very significant.     

Relationship between thermomechanical properties and baking expansion of sour cassava starch (Polvilho azedo)

The factors involved in the baking expansion of native and sour cassava starch doughs were compared with those of native corn starch. Unlike corn starch dough, native and sour cassava starch doughs showed expansion properties during baking. The storage modulus E ′ decreased for cassava starch doughs before baking expansion, but remained unchanged for corn starch dough. Expansion during the baking of sour cassava starch was attributed to water vaporisation and the fluidity of starch paste. The fact that temperature and weight loss variations at adequate water contents were significantly greater for cassava than corn starch dough is indicative of the important role played by starch melting in expansion. Expansion ability could be correlated with changes in dough–crumb thermomechanical properties when close to the starch melting temperature. © 2001 Society of Chemical Industry     

Improved enzymic assay for cyanogens in fresh and processed cassava

The assay for cassava cyanogens developed at the Natural Resources Institute has been modified to overcome some of the problems encountered when the assay is applied to cassava products. Inclusion of 25% ethanol in the extraction medium increased the volume of recovered extract from heat-processed cassava products, eliminated the need for centrifugation and did not interfere with any aspect of the assay. Greater cyanohydrin recovery was noted and the calculation for cyanogen contents was changed to take into account the total extract volume. The separate assay of the three cyanogens (glucosides, cyanohydrins and free cyanide) was achieved by buffering aliquots of the extract followed by appropriate treatment. The importance of assaying for free cyanide (HCN) at pH 4 was demonstrated. Above this pH, cyanohydrin degradation also produces free cyanide, giving rise to misleading values. The efficiency of the extraction medium in recovering added linamarin and cyanohydrin from cassava foods was determined. Recoveries of cyanohydrin were improved using the ethanol/acid medium. The stability of the cyanogens in the ethanol/acid extraction medium was tested at ambient and refrigeration temperatures. Over a two-month period, refrigerated extracts showed acceptable variation as compared with normal variation within the assay (5%) for total and non-glycosidic cyanogens but the levels of free cyanide showed heavy losses (15–56% lost). Since the relative toxicities of the three cyanogens have yet to be ascertained, the relative amount of each cyanogen may be important when assessing the safety of cassava products.     

Viscoelastic properties of reconstituted cassava dough

The influence of water (60–70%) and salt (1–2%) content on the viscoelastic properties of cassava dough, reconstituted from cooked flour, was studied using a controlled strain rheometer. Reconstituted cassava dough behaved as a solid-like material with the storage modulus (G') predominant over the loss modulus (G"). As the water content was increased, G' decreased and G" increased; but tan δ was practically independent of the water content. This behaviour suggested that water had plasticising effects, but probably did not change dough structure. The effect of salt content on the dynamic rheological properties of cassava dough was not significant, except for G" values at water contents close to 60%.     

Mild methods of processing cassava leaves to remove cyanogens and conserve key nutrients

Current methods for processing cassava leaves to remove cyanogens involve pounding followed by boiling in water or boiling intact leaves for 30 min or longer. Boiling in water rapidly removes cyanogens but also breaks down vitamins, proteins and S-containing amino acids, which are necessary to detoxify ingested cyanide. Two methods have been developed to remove cyanogens whilst conserving these key nutrients present in cassava leaves. The first method involves pounding leaves in a pestle and mortar for a minimum of 10 min until the leaves are well macerated, followed by washing the pounded leaves twice in twice their weight of water at ambient temperature, which reduces the total cyanide remaining to 8%. Two further washes reduce the total cyanide to 3%. The second method is to immerse cassava leaves in ten times their weight of water at 50 ± 3 °C for 2 h followed by one change of water and further immersion for 2 h at 50 °C which reduces the total cyanide remaining to 7%.     

Processing factors affecting the level of residual cyanohydrins in gari

Intake of cyanogens in gari, a food processed from cassava roots, is implicated in the causation of tropical ataxic neuropathy (TAN). This neurological syndrome is endemic in some communities in south‐western Nigeria. Studies have shown that methods of processing cassava roots determine the quantity of cyanogens in gari. This study was conducted to investigate the effects of the method of dewatering and the duration of fermentation on cyanogens in gari. Cassava roots (400 kg) were peeled, washed, grated and divided into 14 woven polyethylene sacks. The mash in seven of the sacks was dewatered continuously during fermentation, while the mash in the remaining seven sacks was fermented without dewatering, but dewatered at the end of fermentation. Cassava mash from each treatment was roasted into gari at 24 h intervals up to 168 h. Mean cyanohydrin content in gari roasted from cassava mash dewatered continuously during fermentation was 10.8 mg HCN eq kg^?1 dw (CI 9.7–11.9), while mean cyanohydrin content in gari roasted from cassava mash dewatered after fermentation was 6.3 mg HCN eq kg^?1 dw (CI 5.3–7.4). Mean linamarin content was 4.0 mg HCN eq kg^?1 dw (CI 3.1–4.9) and mean HCN content was 1.6 mg kg^?1 dw (CI 1.3–1.9) in gari roasted from cassava mash dewatered continuously, while mean linamarin content was 3.2 mg HCN eq kg^?1 dw (CI 2.3–4.0) and mean HCN content was 1.2 mg kg^?1 dw (CI 0.9–1.5) in gari roasted from cassava mash dewatered after fermentation. The method of dewatering cassava mash and the duration of fermentation were significantly associated with the level of cyanohydrin in gari (p < 0.001). This study shows that dewatering of cassava mash continuously during fermentation contributes to the dietary cyanide load in TAN‐affected communities. © 2002 Society of Chemical Industry     

Rising African cassava production,diseases due to high cyanide intake and control measures

Cassava is the staple food of tropical Africa and its production, averaged over 24 countries, has increased more than threefold from 1980 to 2005, and the population has more than doubled over that time compared with a 1.5 times increase worldwide. Agriculturally, cassava performs very well but the roots and leaves contain cyanogenic glucosides that are dangerous to human health. These cyanogens sometimes produce acute intoxication leading to death, they exacerbate goitre and cretinism in iodine‐deficient regions, cause konzo and are implicated in the occurrence of tropical ataxic neuropathy and stunting of children. Konzo is an irreversible paralysis of the legs with many thousands of cases, mainly amongst children, in Mozambique, Tanzania, Democratic Republic of Congo, Cameroon, Central African Republic and probably other tropical African countries. Attempts to alleviate cassava cyanide toxicity have included the development of an information network and distribution in developing countries of picrate kits, which measure total cyanide in cassava and urinary thiocyanate. A simple wetting method that reduces total cyanide in cassava flour three‐ to sixfold has been successfully field tested and is being introduced in Mozambique. Transgenic technology shows promise in increasing the rate of loss of cyanide from roots during processing. World health and agricultural bodies should pay more attention to emerging health problems associated with toxicity of cyanogens in cassava. Copyright © 2008 Society of Chemical Industry     

Souring and breakdown of cyanogenic glucosides during the processing of cassava into akyeke

The population and composition of the lactic acid bacteria microbiota as well as the content of cyanogenic glucosides occurring at various stages of fermentation and subsequent processing of cassava roots into akyeke, a steamed sour cassava meal, were investigated. The number of lactic acid bacteria and percentage titratable acidity increased during 5 days of fermentation, but decreases were observed in the subsequent operations of 'washing' the dough with water followed by partial drying and steaming. In field and laboratory samples, Lactobacillus plantarum accounted for 59.3% and 52.3%, Lactobacillus brevis 23.3% and 22.8% and Leuconostoc mesenteroides subsp. cremoris 14.5% and 15.8%, respectively, of all lactic acid bacteria isolated at various stages of fermentation and processing. A reduction of about 98% occurred in the total cyanogens (CN) content of cassava roots during processing, from 69.3 to 1.4 and 110.3 to 2.8 mg CN equivalent/kg dry weight for laboratory and field samples of akyeke, respectively.     

Fermentation of cassava into foofoo: Effect of time and temperature on processing and storage quality

The preparation and distribution of foofoo comprises several stages, namely soaking, dewatering and storage prior to sale. The effects of four different time/temperature soaking processes on the quality of foofoo (pH, microbial load, residual cyanogens) were determined. The processes were chosen as they gave roughly equivalent degrees of retting (the softening of cassava during soaking) by which the progress of the initial fermentation is normally judged. Instrumental measurement of cassava hardness confirmed that the processes were equivalent within experimental limits. The four soaking processes had no significant effect on the water content of foofoo, nor did this change during storage. Processing did affect the pH, total titratable acidity and microbial profiles. Analysis of the aqueous extracts from the processes showed that the glucose present in the fresh cassava was fermented into a mixture of acetic, butyric, isobutyric and lactic acids with some ethanol. The lowering of the pH reduced the number of Enterobacteriaceae to the limit of detection and also affected the types of microbe present. The different processes also affected the removal of cyanogens from the product and the processes at 20°C and 35°C led to unacceptable levels of cyanogens (mainly cyanohydrin) in foofoo.     

Preparation of linamarase solution from cassava latex for use in the cassava cyanide kit

A simple method is described for the preparation of linamarase from cassava latex (sap). Latex, obtained from the end of the petiole (stalk) of cassava leaves, is mixed with water and the solution filtered to give a crude solution of enzyme. This solution may be stored indefinitely in the deep freeze. It may be used without any purification in kits for determination of cyanogens in cassava tubers and cassava products (flour, gari, etc). Immobilisation of linamarase in filter paper discs in the presence of a stabiliser (gelatin/PVP-10) reduces its activity to 25% and in the absence of stabiliser to only 4%.     

Optimization of the basic formulation of a traditional baked cassava cake using response surface methodology

The optimum formulation for production of a Malaysian traditional baked cassava cake was determined using response surface methodology (RSM). Effects of amount of ingredients such as sugar (10-30%) and coconut milk (15-35%) on the textural characteristics (hardness and chewiness) and sensory qualities (colour, firmness, cassava flavour and overall acceptability) of cakes were investigated. Significant regression models which explained the effects of different percentages of sugar and coconut milk on all response variables were determined. The coefficients of determination, R² of all the response variables were higher than 0.8. Based on the response surface and superimposed plots, the basic formulation for production of Malaysian traditional baked cassava cake with desired sensory quality was obtained by incorporating with 25% of sugar and 20% of coconut milk.     

Fungal colonization of sago starch in Papua New Guinea

Sago starch is an important source of dietary carbohydrates in lowland Papua New Guinea. Over the past 30 years there have been sporadic reports of severe illness following consumption of sago starch. A common assumption is that fungal metabolites might be associated with the illness, leading to the need for a more thorough investigation of the mycoflora of sago starch. Sago starch was collected from areas of high sago consumption in Papua New Guinea for fungal analysis (69 samples). Storage methods and duration were recorded at the time of collection and pH on arrival at the laboratory. Yeasts were isolated from all samples except two, ranging from 1.2 x 10(3) to 8.3 x 10(7) cfu/g. Moulds were isolated from 65 of the 69 samples, ranging from 1.0 x 10(2) to 3.0 x 10(6) cfu/g. Of 44 samples tested for ergosterol content, 42 samples showed the presence of fungal biomass. Statistical analyses indicated that sago starch stored for greater than five weeks yielded significantly higher ergosterol content and higher numbers of moulds than sago stored for less than five weeks. The method of storage was also shown to influence mould numbers with storage in natural woven fibre containers returning significantly greater numbers than present in other storage methods tested. Potentially mycotoxigenic genera of moulds including Aspergillus and Penicillium were commonly isolated from sago starch, and as such storage factors that influence the growth of these and other filamentous fungi might contribute to the safety of traditional sago starch in PNG.     

Degradation of cyanogenic glycosides by Lactobacillus plantarum strains from spontaneous cassava fermentation and other microorganisms

Strains of Lactobacillus plantarum, Leuconostoc mesenteroides, Candida tropicalis and Penicillium sclerotiorum were screened for 19 enzymatic activities using the commercial kit API zym (Bio Mérieux). This activity was compared to the ability of degrading the toxic cyanogenic glycosides amygdalin, linamarin, and linseed cyanogens (a mixture of linustatin and neolinustatin). Good correlation between the beta-glucosidase activity found in the API zym screening and the ability to degrade the cyanogenic glycosides was found for the first three species mentioned. P. sclerotiorum strains exhibited very high activity in the API zym test (substrate: 6-Br-2-naphthyl-beta-D-glucopyranoside), but proved unable to degrade any of the cyanogenic substrates. Among the seven strains of L. plantarum tested, a great variation was seen in the beta-glucosidase activity as well as in the ability to degrade the cyanogens. This was also the case for the strains of C. tropicalis. However, all the glucosidase positive strains of these species were also able to degrade all of the cyanogens tested and at approximately the same rate. A co-culture of the most active strain of L. plantarum and C. tropicalis seemed to degrade linamarin faster than the mono cultures. L. plantarum LPI (originally isolated from fermented cassava) was investigated in further detail. The hydrolytic activity of this strain was intracellular or cell bound, and beta-bis-glycosides such as amygdalin were hydrolysed by a two-stage sequential mechanism as follows: (1) amygdalin to prunasin and (2) prunasin to cyanohydrin. Finally, inoculation of extracted linseed meal (containing linustatin and neolinustatin) with L. plantarum LPI resulted in hydrolysis of the glycosides.     

Rheological and textural properties of lafun,a stiff dough,from improved cassava varieties

We studied the textural and rheological (viscoelastic) properties of fresh lafun dough, a fermented cassava product, and their changes during storage at 45 °C for 5 and 24 h, in order to determine after-cooking storability. Lafun flours were produced from three types of cassava varieties: seven improved white-fleshed varieties, seven improved provitamin A carotenoids (pVAC) varieties and two local white-fleshed varieties; and processed into lafun doughs. Pasting properties of the flours were assessed. Flours from local varieties had pasting profiles with highest viscosities, while pVAC flours had the lowest. The three types of cassava varieties varied significantly in most of their pasting properties. Four promising improved varieties were identified, based on high peak viscosity (55.8–61.5 P) and stiffer texture than local varieties during storage. Undesirable varieties were also found, which softened during storage instead of hardening. Optimum texture of lafun dough was obtained after 5 h of storage.