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0308-8146(95)00104-2 ELSEVIER

Food Chemistry, Vol. 55, No. 3, pp. 203-207, 1996 Copyright 0 1996 Elsevier Science Limited Printed in Great Britain. All rights reserved 0308-8146/96 515.00+.00

Composition of green coffee water-soluble fractions and identification of volatiles formed during roasting
C. A. B. De Maria,” L. C. Trugo,“* F. R. Aquino Neto,b R. F. A. Moreira” & C. S. Alviano’
“Department of Biochemistry, Institute of Chemistry, Universidade Federal do Rio de Janeiro (UFRJ), 21044-500 Rio de Janeiro, Brazil ‘Department of Organic Chemistry, institute of Chemistry, UFRJ, C.T. Bloc0 A, Cidade Universitciria, 21044-500 Rio de Janeiro, Brazil L’Department of General Microbiology, Institute of Microbiology, UFRJ, 21044-500 Rio de Janeiro, Brazil

(Received 31 August 1994; revised version received and accepted 11 April 1995)

Flavour precursors from water-soluble green coffee low molecular weight (B) and high molecular weight (C) fractions were investigated and the volatiles formed during roasting were identified by high resolution gas chromatography/mass spectrometry (HRGC/MS). Roasting promoted extensive degradation of trigonelline, sucrose and amino acids in fraction (B) and of arabinogalactan in fraction (C). The analyses of the roasted isolated fractions showed that furans are not only formed by sucrose degradation but also by arabinogalactan pyrolysis. Also, pyrazines appear to be mainly formed by pyrolysis of hydroxy amino acids from fraction (C). The results also showed that pyridine found in roasted coffee is not exclusively formed by trigonelline degradation but also by protein pyrolysis.

INTRODUCTION Aroma is an important attribute to define consumer acceptance of coffee products. The reactions involved in coffee aroma formation are very complex with Maillard and Strecker reactions, and the degradation of sugar, trigonelline, chlorogenic acids (C and A), proteins and polysaccharides are some examples (Clifford, 1985; Dart & Nursten, 1985). One approach to study the mechanisms of aroma formation is the use of sugar or sugar degradation products together with one or two selected amino acids under coffee roasting conditions (Baltes & Bochmann, 19874e, 1992; Baltes & Knoch, 1993). Although these models are very useful, they do not take into consideration other aroma precursors, particularly macromolecular constituents (e.g. polysaccharides). In a previous work (De Maria et al., 1994) we reported a procedure to study coffee aroma, based on the isolation and roasting of low and high molecular weight water-soluble fractions of green coffee. We used this procedure and high resolution gas chromatography/ mass spectrometry (HRGC/MS) to achieve the identification of volatiles formed in the roasted water-soluble fractions. The distribution of flavour precursors in these fractions before and after roasting were also investigated. *To whom correspondence should be addressed.
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MATERIALS ‘AND METHODS Materials Green arabica coffee was obtained from a local industry. All reagents were of analytical-reagent grade. All analyses were carried out with freeze-dried samples. Methods
Isolation of fractions

Ground defatted green coffee (100 g) was extracted with 4 litres of hot bidistilled water (SO’C) for 15 min. Twenty grammes of freeze-dried water-soluble fraction (A) was then dissolved in ethanol water (80:20 v/v, 50°C) with shaking for 1 h and centrifuged at 100 g for 10 min (De Maria et al., 1994). Both filtrate (B) and residue (C) were freeze-dried for further analyses.
Amino acids, crude protein, sugars, CGA and

trigonelline

Amino

acid

analysis

was

based

on

the

procedure

described by Coelho et al. (1985). Crude protein (N x 6.25) was determined by the Kjeldahl method (Pearson, 1976). The analysis of sugars was carried out using the method described by Albersheim et al. (1967). Total CGA and trigonelline were determined by methods described elsewhere (Trugo et al., 1983, 1991).

organic acids). CGA) than fraction (C) which is mainly formed by polysaccharides and proteins. ash. Table 1 shows the distribution of carbohydrates.3 +190 + 8.204 Headspace analysis C. In the course of this work. Arabinogalactan was the major polysaccharide found in fraction (C) and it was affected by roasting with the consequent loss of arabinose (82.6%) than in fraction (C) (13. blot molecular weight water-soluble fraction. trigonelline. The volatile compounds of roasted coffee and of the roasted fractions (B) and (C) are listed in Table 3.1 6.25) Fractions (g%) 80% ethanol-insoluble’ (C) variation (%) non-roast 21.9 1. ‘High molecular weight water-soluble fraction.4 5. Consequently. There was a great loss of trigonelline (70%) and protein (67%).5 g of ground defatted green coffee and fractions (B) and (C) were put in test tubes with screw caps and septa and submitted to vacuum.3 0. In fraction (B). nd. not detected.0 1. On the assumption that the glutamic acid content does not change during roasting. HRGCIMS HRGC was carried out using a Carlo Erba 4300 with FID and Supelcowax fused silica capillary column (30 m x 0. The linear flow rate of hydrogen carrier gas was 40 cm/s.6 29. Serine and threonine were the most affected amino acids in fraction (C) indicating their importance as flavour precursors (Table 2). There was an increase of mannan which may be due to its lesser susceptibility to roasting (Clifford. sucrose was completely degraded on roasting.8%).2 variation (%) -34. RESULTS AND DISCUSSION Fractions were isolated from green defatted coffee based on water solubility and molecular weight. Distribution of carbohydrates. De Maria et al. The oven temperature was held at 40°C for 6 min and programmed to 180°C at 3”C/min. amino acid results may also be expressed after normalizing for glutamic acid. 1994): 0. HRGC/MS was carried out using an HP 5987-A mass spectrometrer with a RTE-6/E data system with a cyclic scan of 1 s and mass range of m/z 50-500.5 -100 -70 -10 -67 Results are averages of duplicate determinations on defatted and dry basis.1 nd nd nd 50 roast nd nd nd nd nd nd 1.2 20. total CGA and protein in the low and high molecular weight fractions (B) and (C) before and after roasting. The distribution of amino acids in fractions (B) and (C) before and after roasting is shown in Table 2.3%). The major compounds in fraction (C) were protein and polysaccharide which was expressed as the monosaccharides produced on hydrolysis (Table 1). Calculation of modified Kovatz indices was based on the method of Van den Do01 & Kratz (1963). a 4 ml headspace sample was injected into the chromatographic column. 1965) the arabinogalactan has a linked galactan main chain with galactose and mainly arabinose-containing side chains. trigonelline. Immediately after roasting. caffeine. trigonelline. Therefore. 1985). these compounds together with humic acids should make up the balance of the fractions in Table 1.5 roast 14. This was expected since fraction (B) contains more labile flavour precursors (sucrose.. total CGA and protein after roasting of water-soluble fraction’ obtained from green coffee Compounds 80% ethanol-soluble* (TV non-roast Galactose Arabinose Mannose Glucose Xylose Sucrose Trigonelline Total CGA Protein (N x 6. the greater susceptibility of arabinose to roasting is due to its arrangement in the arabinogalactan. amino acids. Since roasted fraction (B) showed only trace amounts of free amino acids.25 mm i. More than 90% of the volatile compounds The headspace analysis was based on a previously described technique (De Maria et al. Each sample was roasted at 220°C l 2”C (14 min).6 nd nd nd nd nd 17. A.d.3 -18 -7. Table 1. “Low and high molecular weight water-soluble fractions were obtained by ethanol extraction as described in Methods.3 33.2 1. According to literature data (Wolfrom & Patin. any protein nitrogen detected appears to come from ethanol-soluble peptides.9 2.3 -02. Loss of weight during roasting is higher in fraction (B) (33.0 3.9 nd nd nd 46.2 7.e. Some compounds found in the roasted fractions were not considered because they are not as relevant as aroma precursors (i. about 30 volatiles were identified by HRGC/MS. Specific groups of compounds were then considered in the fractions for the studies of aroma formation under coffee roasting conditions. It is supposed that this polysaccharide is the major carbohydrate aroma precursor in fraction (C).). .2 18. B.

6 1.9 5.2 -1. the furans present in roasted coffee appear to be formed mainly by sucrose degradation but also by arabinogalactan pyrolysis.9 5.1 4.1 non-roast 8. 1973). Dart & Nursten. Distribution of amino acids before and after roasting of water-soluble fractions” obtained from green coffee Compounds 80% ethanol-solubleh (B) non-roastd Lysine Histidine Arginine Aspartic acid Threonine Serine Glutamic acid Proline Glycine Alanine Valine Isolerncine Leucine Tyrosine Phenylalanine I.e.8 + 8.3 I.1 -11. in g% of each amino acid x 100 g of encountered in roasted coffee were also found in the roasted fractions (B) and (C) which shows a good recovery of original coffee volatiles after fractionation. The absence of pyrazines in fraction (B) may be owing to the low content of amino acids. rafter normalizing for glutamic acid. pyridine may also be formed by hydroxyamino acid pyrolysis (Baltes & Bochmann.0 -44. acylpyrroles and furanones) were not found in this fraction. sugar degradation is more pronounced via Maillard reactions rather than pyrolysis. 1985.5 -12.0 -1. Seven identified furans are listed in Table 3.1 4. 1987a. as dRoasted fraction (B) showed only trace amounts of amino acids.fractions Table 2.2 + 13. These results indicate that pyridine of roasted coffee is formed not only by trigonelline degradation.0 -92. Kato et al.2 7. pyrazines of roasted coffee are mainly formed by hydroxy amino acid pyrolysis. Consequently.1 8. however. However.4 3. 11 alkylpyrazines were detected. but also by protein pyrolysis. This may explain why furans were found in lesser quantity in roasted fraction (C) than in fraction (B) (Table 3). Furthermore. 1985).5 28. trigonelline was not found in roasted fraction (C) (Table 2) but pyridine was identified after roasting (Table 3). as already mentioned by Baltes & Mevissen (1988).8 2. It has been well documented that the presence of furans in roasted coffee is due to sucrose degradation (Clifford. Cyclotene (2-hydroxy-3-methyl-2-cyclopentene1-one) was readily detected in fraction (B) (Table 3) but only a .d.3 -40. 1987a) which are present in this fraction.6 -38.4 2.0 -16. only hydroxy amino acids produce pyrazines by pyrolysis (Wang & Odell. 1985.8 4.3 4.4 7. In roasted fraction (C). The higher amount of volatiles found in the roasted fractions as compared to the whole coffee is due to the concentration of aroma precursors after fractionation.0 -33.5 2.9 -89 -67 + 38. 1967).9 5.2 2. which were not present in sufficient amounts to form detectable pyrazines. Results are averages of duplicate determinations on defatted and dry basis. 1970.0 3.5 -12. Therefore.3 4. However. It is well documented that pyrolysis of hydroxy amino acids produce alkylpyrazines (Baltes & Bochmann. sucrose is the principal carbohydrate present in fraction (B) which contains furans. none of them were found in the roasted fraction (B) (Table 3).5 5.8 3. It is also suggested that aroma compounds from fraction (C) are mainly formed via pyrolysis due to the predominance of high molecular weight material. In this study there is strong evidence that threonine and also serine are the most important alkylpyrazine precursors since they were the most intensely degraded amino acids in fraction (C).5 5.5 3.5 9. In fact.Composition of green coflee water-soluble. In fact. typical Maillard reaction compounds (i.6 -17. in comparison to sucrose. data of amino acids are expressed total protein. On the other hand.6 3.5 14.6 20. Based on these observations it is reasonable to conclude that high molecular weight water-soluble compounds of fraction (C) are the most relevant precursors of alkylpyrazines in roasted coffee. Wang & Odell.9 -1.4 3.5 roast 8. Pyrazines are another group of powerful aroma compounds present in roasted coffee (Dart & Nursten.5 17.3 Fractions (g%) -___ 80% ethanol-insoluble’ (C) variation -2. ‘In relation to non-roast. high loss of trigonelline during roasting (Table 2) was related to the high amounts of pyridine found in the roasted fraction (B) (Table 3).6 4. Goldman et al.0 0. Furans of the fraction (C) should be formed via arabinogalactan pyrolysis.9 3. 1974).3 -20 -22 -2.8 14.9 3.0 -33.4 (%)d -29. 1973). Pyridines are the most important group of volatiles obtained by heating of trigonelline (Viani & Horman..3 I.5 +21.4 -22.8 205 o~h~c in Table I. Consequently.1 4.1 6.8 5.. On the other hand.4 -30.1 3.5 21.9 2.0 -36. furans were also identified in fraction (C) which did not contain sucrose (Table 1).7 -28.

. 4.206 C. Res. 5. Model reactions on roast aroma formation. large amounts still remain in roasted fraction (C) (Table 3). (1970). De Maria et al. Table 3..A. 1990) and this appears to be the route of cyclotene formation in roasted coffee.<’ Ethylpyrazine”” 2. 340-S. Bakes. & Bochmann. ?dentification via a published I. 1963). P. Curbohydr. D.8 8 178 +++ (+) (+) 278 3 8 8 I. Agric. 7. 35. 6. English. Kato et al. Food Chem. J. UFRJ.P Phenol”’ Results are averages of duplicate 530 541 571 585 599 633 639 678 883 809 897 729 732 737 737 766 774 783 786 804 820 845 860 883 886 934 1014 1049 II41 determinations. (1987~).5. A method for the analysis of sugars in plant cell-wall polysaccharides by gas liquid chromatography. Baltes and Bochmann (I 98743. I: Reaction of serine and threonine with sucrose under the conditions of coffee roasting and identification of new coffee aroma compounds.3-Dimethyl-pyrazined. UFRJ. .’ 2-Methyl-3-[2H]-furanone’ 2.340-6. (1969). This may in fact be expected because fraction (C) contains high molecular weight aroma precursors which are more difficult to interreact. + + low concentration. modified spectrum: retention 80% ethanol-insoluble (C) I (Y (+) ++ +++ ++++ ++++ +++ ++ ++ + ++ ++ ++ +++ ++ + +++ 2 I 3. 1987~).. (1967).6 5 7 (+) 278 279 I I ++ 2. A (1967). + very low concentration.5 4. 8.“.5. Da Silva (Institute of Microbiology.. . & Barr.6 2 4. Brazil) for technical assistance in the mass spectrometry analysis.4 4. P.5.6 4. +++ +++ ++ +++ ++ +++t + + ++ (+I (+) (+I + ++’ +++’ ++++ +++ ++++ (+I +++ ++ ++++ ++++ ++ +++ +++ ++ +++ + +++ +++ ++ ++++ ++ ++ ++ index (Van den Dool & Kratz.536 4. Nevins. We are grateful to Lucia M.536 2 4.I. + + + high concentration. Baltes and Bochmann (1987e). The results found in this work showed that fractions (B) and (C) are relevant and representative of coffee aroma precursors. ( +) Trace compound. Goldman et al. IK.e 2-Furfurald. P. Baltes and Bochmann pound available. Baltes and Bochmann (19876).’ Furfuryl-methanoate’ Pyrrole’ 2-Methyl-pyrrole’ 5-Methyl-2-furfural’ 2-FormylI -methylpyrrole’ 2-Acetyl-I-methylpyrrole’ 2-Furfuryl-alcohol’ Cyclotene’ 2-Methoxy-phenold. B. The approach used to collect the headspace fraction formed after roasting proved to be rapid and appropriate for the analysis of the aroma compounds produced. Although pyrrole is extremely reactive.6-Diethyl-pyrazine’ 2. W. We gratefully acknowledge Luis R. Stenhagen et al. (1967).*.5 476 5 4. Sucrose degradation is the main source of carbocyclic compounds (Nishimura & Mihara. It has been reported that alkylpyrroles can be formed via hydroxyamino acid pyrolysis (Baltes & Bochmann. 3. In the present work.536 4. six pyrroles were identified.c 2-Ethyl-6-methylpyrazine”” 2-Ethyl-S-methylpyrazine”’ Trimethyl-pyrazined.3-Diethyl-pyrazine”’ Acetic acidd.5-Dimethyl-pyrazined’ 2. + + + + extremely high concentration. 5. 2. Stall er al. * Estimate of abundance in the co-eluted compounds.6-Dimethyl-pyrazined. trace amount was found in fraction (C). Thus. dreference comRef.’ as in Table (1987~). Brazil) for helping in the amino acids analysis. Damasceno and Isabela Wanderley (Institute of Chemistry. Some compounds identified after roasting of coffee and of the water-soluble fractions” Fractions TK Roasted coffee 80% ethanol-soluble (B) I-Methyl-pyrroled~’ S-Methyl-Z-vinylfuran’ I-Ethyl-pyrrole’ Pyridine”’ Pyrazined’ 2-Methyl-tetrahydro-3-furanone’ Methylpyrazine”’ 2. D. ACKNOWLEDGEMENTS We acknowledge the financial support from CNPq and CAPES (Brazil). REFERENCES Albersheim. G. the formation of alkylpyrroles in fraction (C) appears to follow this route.

42. Unters Forsch. & Bochmann. Winter.. C. 141-5. Macrae. C. B. III: Mass spectrometric identification of pyrroles from the reaction of serine and threonine with sucrose under the conditions of coffee roasting. C.. Kurata. 305-74. Chem. R. Model reactions on roast aroma formation. Plant & Soil. De Maria. Pearson. Helv. H. Stall. Winter. De Maria. 628-93. 366-71. 184.. Model reactions on roast aroma formation. & Knoch. 50. Thermal behavior of trigonelline. Food Sci. & Dick. Lebensm.478-84. Churchill Livingstone. J. Model reactions on roast aroma formation. Croom Helm.. I. A. M. Moreira. IV: Mass spectrometric identification of pyrazines from the reaction of serine and threonine with sucrose under the conditions of coffee roasting. C. Z.. Linhares.. IV. S. I. C. Elsevier Applied Science. XII: Reaction of glucose with aspartic acid or asparagine at three different temperatures.. J. Lebensm. & Mevissen. L. D. J. J. Unters Forsch. I 1-l 2. 3rd edn. Trugo. C. C. M. 30.. Macrae. M. 39. Wiley. In Cofie: Chemistry (Vol. 185.. 194. M. H. Chem... Gautschi. and carboxyclic compounds from the reaction of serine and threonine with sucrose under the conditions of coffee roasting. Viani.4060-83. R. Van den Dool. N. Baltes. pp.463-71. p. Formation of pyrazines from thermal treatment of some amino-hydroxy compounds. C. Food. Baltes. P. Ishitsuka. C. & Mihara. Z. L. 184.. Lebensm. (1973). . B. Flament. 179-88. (1969). 485-93. & McLafferty. I. (1991). E. Food Chem. Clifford & K.. W. (1970). (1985). 34. Abrahamsson. Ltd. Willhalm. (1987~). Chemical and physical aspects of green coffee and coffee products. S.. Chim. (19876). 34.. C. & Martin. L. & Horman. & Odell.. E. Coelho. Trugo.. Z. I. oxazoles.. V. W. 21. M. S. Lebensm. Agric. & Willhalm.. Simultaneous determination of total chlorogenic acid and caffeine in coffee by high-performance gel filtration chromatography. Investigation of 2hydroxy-2-cyclopenten-l-ones in roasted coffee. 11: Mass spectrometric identification of furans and furanones from the reaction of serine and threonine under the conditions of coffee roasting. 81. Agric. (1983). & Bochmann. S. B. W. New York. Food Chem. F. R. Willson. F.. D. Sur I’arome de cafe. 30&6. V: Mass spectrometric identification of pyridines. A. M. Biol. Flament. Z. 343-9. 264. L. London. Chim. P.. & Fujimaki. G. Model reactions of roast aroma formation. (1967).. (1967). Lebensm. J.. J. 87.-7. Lebensm. J. Nishimura. J. L. G. Helv. An arabinogalactan. Dart. 33746. (1965). L. Clarke & R. II: The formation of some uncommon N-heterocyclic compounds and furans after roasting of tryptophan with reducing sugars and sugar degradation products.868-70. pp. (1990). 0. Trugo.Composition qfgreen coffee water-solubkfractions 207 Baltes. Baltes. (1963). 121617. W. Kato. Z. T. Acta. A. & Bochmann. M.. Model reactions on roast aroma formation. Composition of green coffee fractions and their contribution to the volatile profile formed during roasting.. Carbohydrates of the coffee bean. (1987e). R. G. & Wernerck. London. Food Agric. Unters Forsch. W. (1985). G. (1993).. Atlas of’ Mass Spectral Data (Vol. 50. Chromatogr. Model reactions of roast aroma formation. The Chemical Analysis of Foods. Baltes. & Patin. & Nursten.. & Bochmann. Unters Forsch. W. L. R. Wolfrom. (1988). Food Chem. Clifford. N. Volatile components. (19874. Bohnenstengel. 48. VI: Volatile reaction products from the reaction of phenylalanine with glucose during cooking and roasting. I. Baltes. Model reactions on roast aroma formation. 209-14. P. (1976). C. Wang. M. (1994). Acta. pp. M. Seibl. Gautschi. Pyrolysis of B-hydroxy-amino acids. A generalization of the retention index system including linear temperature programmed gas-liquid partition chromatography. 694 705. Agric. Stenhagen. Unters Forsch. F. Z. eds R. R. R. E. (1985). especially L-serine. (I 2) 182632. 49. E. In Cqfir: Botany. Food Chem. I). W.5-9. London.. 187.. & Stall. J. & Krdtz. Biochemistry and Production of Beans and Beverage. G. J. 11. & Werneck. B. F. R. Goldman. (1992). (1974). Sur I‘arome de cafe. Unters Forsch. 22365.. eds M. 184... Organic Chem. 38. Amino acid distribution in some fungal melanins and of soil humic acids from Brazil. Sci. 1038-41. 7th edn. Determination of purine alkaloids and trigonelline in instant coffee and other beverages using high-performance liquid chromatography. D. & Baltes. I). 11: Pyrazines et pyridines. S.