Pyrazine flavoring from

In this study the physical parameters involved in interaction of a major class of meat flavorants, methyl pyrazines, with soy proteins were determined at meat roasting temperatures. Beef diffusate, the water soluble, low molecular weight fraction that constitutes about IX of beef, was shown to contain the necessary precursors to obtain a desirable, thermally generated meat aroma (8). Diffusate was heated under controlled conditions and generated volatiles were transferred to a gas chromatograph for separation and quantitation. Methyl pyrazines, either from heated diffusate or from standard solutions, were measured in the presence of purified soy proteins and the thermodynamics of binding were determined. 

Although most of the alkylpyrazines are formed through thermal Interactions of components in food, methoxy-substltuted pyrazlnes are mainly derived from biosynthetic pathways. 2-Isobutyl-3-methoxy-pyrazine Isolated from bell pepper by Buttery at al. (14) is one of the most significant flavor compounds discovered. This characteristic bell pepper aroma compound has an extremely low odor threshold of 0.002 ppb in water (15). 

Minuscule quantities of naturally-occurring pyrazines have been found in some foodstuffs and are largely responsible for their flavor and aroma. For example, 3-isopropyl-2-methoxypyrazine is isolated from green peas and wine and a seasoned wine connoisseur can identify a ppt quantity. In addition, 2-methyl-6-vinylpyrazine exists in coffee. 

In the case of diketopiperazine 13 and related compounds, dehydrogenation of the preceding diketopiperazine occurs in the side chain. A shift of the double bond into the central ring and dehydration may result in the formation of substituted pyrazines. Simple pyrazines are known as signaling compounds from animals. The pyrazines 16 and 17 have also been isolated from marine Streptomycetes [106]. GC/MS investigations of bacterial flavor components [951 indicate that these and others are very wide-spread. 

Musty or potato-like flavor and aroma have been observed as a defect in milk (Hammer and Babel 1957) and Gruyere de Comte cheese (Dumont et al. 1975). This off-flavor results from the production of nitrogenous cyclic compounds by Pseudomonas taetrolens and P. perolens (Morgan 1976). Musty-flavored compounds produced by these organisms include 2,5-dimethylpyrazine and 2-methoxy-3-isopropyl-pyrazine. The Gruyere de Comte with potato off-flavor contained 3-methoxy-2-propyl pyridine, as well as alkyl pyrazine compounds (Dumont et al. 1975). Murray and Whitfield (1975) postulated that alkyl pyrazines are formed in vegetables by condensation of amino acids such as valine, isoleucine, and leucine with a 2-carbon compound. Details of the synthetic mechanism in pseudomonads are unknown. 

The formation of pyrazines in foods has been reviewed extensively by Mega and Sizer (50). Temperature and pH are very important factors in the formation of specific pyrazines. Forty-two pyrazines have been identified in meat from various sources by these authors. MacLeod and Seyyedain-Ardebili (20) listed 49 pyrazines found in beef by various investigators. Ching (19) identified 28 pyrazines in her studies of sugar-amine reactions simulating beef flavor. 

Molasses. A large number of volatile and nonvolatile compounds have been identified in the flavor fractions of various types of molasses (51-621. Compound classes identified include aliphatic and aromatic acids, aldehydes, phenols, lactones, amines, esters, furans, pyrazines, and sulfides. Most of these compounds can arise from carbohydrate degradation through a number of traditional pathways especially because residual nitrogen-containing sources are present. 

Interest in the influence of lipids on pyrazine formation has recently been generated by the identification of long-chain alkyl-substituted heterocyclic compounds in foods and in model systems. Pyrazines in this category include 2-heptylpyrazine isolated from french fried potato flavor (7), and 2-methyl-3(or 6)-pentylpyrazine and 2,5-dimethyl-3-pentylpyrazine, isolated from extruded zein/corn amylopectin/corn oil systems (8, 9). Only the involvement of lipids or lipid-decomposition products in the formation of these compounds could account for the long-chain alkyl substitution on the pyrazine ring. 

A maltol-ammonia browning reaction produced thirteen pyrazines, two pyrroles, two oxazoles, and one pyridine (12). The major products of this system were 2-ethyl-3-hydroxy-6-methylpyridine and 2-ethyl-3,6-dimethylpyrazine. It is difficult to construct possible formation mechanisms for these compounds from maltol and ammonia. All the carbon atoms must come from maltol. It is possible, then, that maltol degrades into smaller carbon units and that these fragments recombine to form larger carbon units, producing these compounds. Recently, the formation of thiophenones and thiophenes from the reaction of 2,5-dimethyl-4-hydroxy-3(2H)-furanone and cysteine or cystine was reported (13. 14). All these reaction mixtures were reported to possess a cooked meat-like flavor. 

Even though imidazoles comprise the second largest fraction of the volatile products obtained from Maillard reaction after pyrazines, they do not contribute any characteristic flavors to cooked foods (23). 

In a recent review on coffee flavor Clarke (3J reported that there are many publications on qualitative data, but information on their actual quantities in coffee is difficult to find. Many of the coffee aroma components are present in high concentrations and these data were used to predict so called consumation ratios for heterocyclic components (7, 9). According to data presented by these authors the consumption of pyrazines from coffee is 380 fold higher compared to pyrazines added as flavoring substances to all other foodstuffs. 

The flavor constituents of plain and roasted cashew nuts have not been previously reported in the literature. In the present study, aroma compounds have been isolated from plain, oven-roasted and oil-roasted cashew nuts by simultaneous distillation extraction and by steam distillation followed by selective extraction, after pH adjustment. Compound identification was carried out by GC and GC-MS analyses. Esters and lactones are present in plain cashews whereas roasted samples also contain pyrazines. 

The pH of the steam distillate was 6.2 in all cases. The total flavor isolate (by SDE) of plain cashews had, on the whole, a strong pungent and green aroma, reminiscent of the cashew nut testa and cashew shell, whereas the flavor isolates from roasted samples had the characteristic mildly nutty aroma also. The flavor fractions obtained by selective extraction method gave some information about the chemical nature of compounds responsible for the characteristic flavor notes. Accordingly, the basic fraction of roasted nuts, (both oven-and oil-roasted), had the typical nutty aroma associated with pyrazine. compounds. The basic fraction of plain cashews did not have any characteristic flavor in particular. 

To sum up, the flavor constituents of plain and roasted cashew nuts are reported here for the first time. The mild flavor of cashew nuts can be attributed to the carbonyls, esters and lactones, especially to 5-heptene-2-one and 1,3-propanediol diacetate. Upon roasting, 2,6-dimethyl pyrazine, 2,6-diethyl pyrazine and the furanone are formed in larger amounts and from flavor profile also these compounds are likely to play a significant role in the characteristic aroma of roasted cashew nuts. 

The flavor industry has introduced, over the years, methods of developing meat flavors by processing appropriate precursors under carefully controlled reaction conditions. As a result, meat flavors having a remarkably genuine meat character in the beef, chicken and pork tonalities are available for the food industry. It has repeatedly been stated that the Maillard reaction is particularly important for the formation of meat flavors. However, of the 600 volatile compounds isolated from natural beef aroma, only 12% of them find their origin in sugar/amino acid interactions and of these 70% are pyrazine derivatives. 

There is increasing evidence that the interaction of lipids with the Maillard reaction is relevant to the generation of flavor in many cooked foods. For instance, the removal of lipids from coconut has been shown to cause flavor changes in the roast material (12). Uncooked coconut contained significant amounts of lactones as the main aroma components on roasting pyrazines, pyrroles and furans were also found in the aroma volatiles which added a strong nut-like aroma to the sweet aroma of the unroasted coconut. When ground coconut was defatted and then roasted, the sweet aroma due to lactones disappeared and the product possessed a burnt, nut-like aroma. A marked increase in the number and amount of Maillard reaction products, in particular pyrazines, was found. 

Concentrations of thermally generated meat flavor components are diminished by protein adsorption when soy extenders are added to fresh meat products before heating. The amounts of individual alkyl pyrazines, thermally generated by heating beef diffusate, decreased linearly as the amount of whole soy, soy 7S or soy 11S proteins were increased in a model system. Similar recoveries were obtained when pyrazines were mixed with soy either as chemical standards or from diffusate. Stoichiometry and energetics of interaction were determined for methyl pyrazine congeners with soy proteins at 120° and 145°C. Results of this study suggest that flavorants can be added in readily determined amounts to compensate for losses due to adsorption in meat-soy products. 

There was no loss in flavorant at a 100 mg level when either whole soy or soy 7S protein was used, but there was a 14-24 percentage loss when 11S protein was used. Only soy 11S protein affected the substituted pyrazine content of the mixture at all addition levels. The higher the substituted pyrazine congener, the smaller was the percentage loss with any of the protein types. At the 500 mg level, only about 50% of the amount of any of the methyl pyrazines in the control was recovered from either soy 7S or 11S protein while 70% was recovered from whole soy protein. These results greatly extend the initial work reported by Palkert and Fagerson (11J who determined that about 75% of dimethyl thiazole, a sulfur-nitrogen heterocycle, was recovered from dry, textured soy protein. 

The problem of developing desirable meat flavor in the presence of vegetable protein has been clearly demonstrated in the literature. Physical measurements after heating a meat model system with soy proteins have shown a dramatic reduction in the concentration of alkyl pyrazine compounds due to interaction with the soy proteins. These interactions have been defined in terms of stoichiometry and binding energies from measurements on pure standards of the methyl pyrazines. 

Nearly 1000 compounds have so far been identified in the volatile constituents of meat from beef, chicken, mutton and pork (6). The largest number of volatiles has been determined in beef and these were representative of most classes of organic compounds. Hydrocarbons, alcohols, aldehydes, ketones, carboxylic acids, esters, lactones, ethers, sulfur and halogenated compounds as well as different classes of heterocyclic substances (Figure 1) namely furans, pyrldlnes, pyrazines, pyrroles, oxazol(in)es, thiazol(in)es, thiophenes were present in cooked meat flavor volatiles as shown in Table I. Many of these compounds are unimportant to the flavor of meat and some may have been artifacts (16). 

Three extremely odorous pyrazines, 3-isopropyl-2-methoxypyrazine, 2-s-butyl-3-methoxypyrazine, and 2-isobutyl-3-methoxypyrazine have been shown to be present in green peas, and are likely to be of major significance in the flavor of peas (59). The volatile oil of green bell peppers has been found to contain 2-isobutyl-3-methoxypyrazine as a major component (60,61). The alkylpyrazines in potato chips (62,63) and roasted peanuts (63) have been examined. 2-Isopropyl-3-methoxypyrazine has been characterized in the vacuum steam volatile oil of potatoes (64), 2-ethyl-3-methoxypyrazine in cooked potato (65), and 3-ethyl-2,5-dimethylpyrazine and 2-ethyl-3,5-dimethylpyTazine as the components important to the aroma of baked potato (66). A variety of alkylpyrazines have been identified in roasted sesame seeds (67) and 21 pyrazines have been identified in the aroma components isolated from roasted green tea (68). 

There are many reports of Pseudomonas cultures producing musty, earthy, and potato-like odors ( l-6). The work of Morgan et al. (T) established 2-methoxy-3-isopropyl pyrazine to be partially responsible for these odors. Subsequently, 2-methyoxy-3-isopropyl pyrazine was found in bell peppers (8), a similar compound 2-methoxy-3-secbutyl pyrazine was identified in galbanum oil (9), and several 2-methoxy-3-alkyl pyrazines were identified in various raw botanicals (10). The odor threshold exhibited by 2-methoxy-3-isobutyl pyrazine (1 part in 10 indicates flavor significance for these compounds even at the exceptionally low concentrations in which they occur in foods and other natural products Producing these compounds from microbial fermentations could be an economical source of flavor for the food industry. 

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