Accepted Manuscript

Rapid Communication$Analytical Methods$Nutritional and Clinical Methods

Influence of serving temperature on flavour perception and release of Bourbon

Caturra Coffee

Ida Steen, Sandra S. Waehrens, Mikael A. Petersen, Morten Münchow, Wender

L.P. Bredie

PII: S0308-8146(16)31508-4
DOI: http://dx.doi.org/10.1016/j.foodchem.2016.09.113
Reference: FOCH 19893

To appear in: Food Chemistry

Received Date: 14 April 2016

Revised Date: 29 August 2016
Accepted Date: 17 September 2016

Please cite this article as: Steen, I., Waehrens, S.S., Petersen, M.A., Münchow, M., L.P. Bredie, W., Influence of
serving temperature on flavour perception and release of Bourbon Caturra Coffee, Food Chemistry (2016), doi:
http://dx.doi.org/10.1016/j.foodchem.2016.09.113

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 Influence of serving temperature on flavour perception

and release of Bourbon Caturra Coffee

Ida Steen1,2,3, Sandra S. Waehrens1, Mikael A. Petersen1, Morten Münchow1,2,3 and Wender L.P.

Bredie1*

Affiliations:
1
Department of Food Science, Faculty of Science, University of Copenhagen, Rolighedsvej 26,

1958 Frederiksberg C, Denmark

2
Speciality Coffee Association of Europe, SCAE, United Kingdom
3
CoffeeMind, coffee-mind.com, Hansstedvej 35, 2500 Valby, Denmark

Authors with contact e-mail addresses:

Ida Steen: ida@coffee-mind.com

Sandra S. Wæhrens: sandrasn@food.ku.dk

Mikael A. Petersen: map@food.ku.dk

Morten Münchow: morten@coffee-mind.com

Wender L.P. Bredie: wb@food.ku.dk; tlf. +45-35 32 33 42

*Author to whom correspondence should be addressed

1
1 Abstract

2 The present study aimed to investigate coffee flavour perception and release as function of serving

3 temperature to support standardisation in the specialty coffee branch. The coffee cultivar Bourbon

4 Caturra was evaluated at six serving temperatures ranging from 31°C to 62 °C. Coffee samples

5 were analysed by dynamic headspace sampling gas chromatography–mass spectrometry and

6 descriptive analyses using sip-and-spit tasting. The release of volatiles followed mostly the van ´t

7 Hoff principle and was exuberated at temperatures above 40ºC. Aliphatic ketones, alkylpyrazines,

8 some furans and pyridines increased most notably at temperatures at ≥50 °C. The changes in

9 volatile release profiles could explain some of the sensory differences observed. The flavour notes

10 of ‘sour’, ‘tobacco’ and ‘sweet’ were mostly associated with the coffees served at 31 °C to 44 °C,

11 whereas coffees served between 50 °C and 62 °C exhibited stronger ‘overall intensity’, ‘roasted’

12 flavour and ‘bitter’ notes.

13

14 Keywords

15 Specialty coffee; flavour release; sensory perception; serving temperature; Maillard

2
16 1 Introduction

17 Coffee has a complex flavour composition with 846 compounds identified in different cultivars,

18 roasts and brews (VCF, 2016). Although the flavour compounds in coffee are manifold they are

19 mainly generated during complex reactions in the roasting process. During coffee brewing, volatile

20 and non-volatile flavour compounds are extracted from the ground coffee with hot water and

21 dispersed over the water, oil and solids phases. The hot water extraction in regular coffee provides

22 0.2 g/L lipids to the brew (USDA, 2015), which can be considered as a significant reservoir for

23 flavour compounds.

24

25 Besides different grinding and brewing techniques, serving temperature is an important factor for

26 the flavour quality of the coffee beverage. Changing the serving temperature will alter the free

27 energy and thus the enthalpy and entropy of the coffee brew. The temperature thus influences the

28 partitioning of flavour compounds between the dispersed phases and the evaporation at the coffee-

29 to-air interface. Since coffee is a highly complex mixture of dispersed components with different

30 physico-chemical properties, changes in flavour release as a result from altering the serving

31 temperature is difficult to predict from first principles in thermodynamics. The release as function

32 of temperature may therefore be better approached by an experimentally measurement of a dynamic

33 release under controlled conditions. Furthermore, flavour release during drinking of coffee may be

34 influenced by interactions with saliva (Denker et al. 2006), added milk (Bucking and Steinhart,

35 2002) and oro-nasal instability of volatiles such as furanthiols (Buettner, 2002; Buettner &

36 Beauchamp, 2010).

37

38 Studies investigating preferences of drinking temperatures of black coffee have revealed a preferred

39 temperature between 52.8 °C to 72.1 °C (Borchgrevink, Susskind, & Tarras, 1999;

3
40 Pipatsattayanuwong, Lee, Lau, & O'Mahony, 2001; Lee & O’Mahony, 2002; Brown & Diller,

41 2008). The Norwegian Coffee Association and National Coffee Association of USA (NCA),

42 recommend a serving temperature ranging between 80 to 85 °C and 82.2 to 85 °C, respectively

43 (Norsk Kaffeinformasjon, 2010; NCA, 2015). These elevated temperatures exceed the thermal

44 orosensory pain threshold (Green, 1985; Margarida, Hardy, & Hammel, 1962) and seem only to be

45 applicable in the cupping method. In this method a turbulent flow of ambient air is rapidly mixed

46 with the intake of a small portion of coffee, creating a rapidly falling temperature and a reduced air-

47 liquid surface pressure, allowing a range of volatiles to be released in the vapour phase. The

48 cupping method is commonly used in the specialty coffee branch to evaluate flavour quality.

49 However, it requires expert training and may be less suitable in formalised descriptive sensory

50 analysis with trained panels. The Specialty Coffee Association of Europe uses a brewing standard,

51 which does not include a guideline for the serving temperature, however there is an interested in

52 well-documented recommendations for the serving temperature of specialty coffees. There is

53 therefore a need for understanding of the relationship between coffee tasting method, serving

54 temperature and flavour perception and release.

55

56 In the present study roasted beans from the Bourbon Caturra variety were chosen as an example of

57 specialty coffee. Bourbon is a variety of the species Coffea Arabica cultivars, which is known for

58 its sweetness making it prized and desirable in the speciality coffee industry. Caturra is a natural

59 mutation of Bourbon, known for its quality when grown at high altitude (Hoffmann, 2014).

60

61 The aim of the study was to investigate the relationship between the serving temperature of coffee

62 and the flavour properties of speciality coffee in a sip-and-spit tasting. The experimental setup

63 combined volatile analysis by headspace sampling and descriptive sensory panel evaluations to

4
64 determine which volatile compounds and flavour properties were susceptible for changes in the

65 serving temperature. This will provide an objective basis for recommendations of serving

66 temperatures and tasting procedures for evaluation of coffees in the speciality coffee branch.

67

68 2 Materials and methods

69 2.1 Coffee beans

70 Roasted beans from the Bourbon Caturra variety were selected as an example of specialty coffee.

71 The beans of Bourbon Caturra were grown at an altitude of 1100-1800 meter, harvested in the

72 period January to Marts 2014, washed and dried after harvest. The variety Caturra is a natural

73 mutation of Bourbon, known for its good quality that increases with high altitude (Hoffmann,

74 2014). The beans were grown at Finca La Perla in Guatemala where the farmers have proven to

75 produce high quality coffee beans as they won the prestigious Presidential award at the 2006 Cup of

76 Excellence competition and have made the auction final four times (2001, 2002, 2006 and 2008)

77 (Mercanta, 2014).

78

79 2.2 Roasting and storage

80 The beans were roasted in a 1 kg Probatino coffee roaster at Kontra Coffee (Hvidovre, Denmark).

81 The roasting was stopped at the #80.7 colour on the Agtron Gourmet Scale. After the roasting

82 process, the beans were placed in a metal container and rotated to cool down. The roasted coffee

83 beans were packed in portions of 1 kg in dark, stand-up, odour-free plastic bags with CO2 degassing

84 valves. The coffee beans were stored at -18 °C maximum 60 days before analyses. One day prior to

85 the analyses, the beans were thawed to 21 °C.

86

5
 87 2.3 Coffee preparation

88 The coffee beans were ground to a slightly coarse particle size in a Mahlkönig (Hamburg,

89 Germany) grinder. Grinding was performed six hours prior to brewing to ensure freshness. The

90 coffee was brewed by adding 50 g of coarse coffee to a French press brewer (Bodum Chambord

91 French Press, coffee maker) and adding 900 ml of hot water (≈ 92 °C). To minimize dregs, coffee

92 filters (INNOFORS Aps) were placed in the press brewers. The mixture was gently stirred ten times

93 with a spoon, and after 4 minutes extraction the plunger was pressed to the bottom. A 115 ml coffee

94 was poured into thermos cups (Gastro Line 18 – 200 ml) and stored in Termaks cabinets at 21 °C

95 until reaching the required temperature. Immediately thereafter, the sample was served to the

96 sensory panel. All temperatures were measured with a wireless k-type thermometer (FLUKE CNX

97 t3000) and FLUKE 62 MAX+ IR thermometer.

98

99 2.4 Coffee serving temperatures

100 In total six different serving temperatures were evaluated; 31 °C, 37 °C, 44 °C, 50 °C, 56 °C and 62

101 °C (±1 °C). These temperatures covered a relevant range suitable for sensory taste evaluations with

102 a trained panel based on pre-trials.

103

104 2.5 Volatile compound analysis

105 Dynamic headspace sampling (DHS) followed by gas chromatography–mass spectrometry (GC–

106 MS) analyses, were performed on the coffee samples held at 31 °C, 37 °C, 44 °C and 50 °C,

107 respectively. Since the DHS could only be carried out up to 50 °C, the headspace volatiles at 56 °C

108 and 62 °C were estimated (see section 2.7.2). All analyses were carried out in duplicate.

109

6
110 2.5.1 Dynamic Headspace Sampling

111 At each sampling temperature a 20 mL coffee with 1 mL of internal standard (5 ppm, 4 Methyl-1-

112 pentanol in water) was placed in a 100 ml gas washing flask and then closed with a purge head. The

113 samples were equilibrated to 31 °C, 37 °C, 44 °C or 50 °C in a circulating water bath and then

114 purged with nitrogen (100 mL min-1) for 20 min under magnetic stirring (200 rpm). The volatile

115 compounds were collected on traps containing 250 mg of Tenax-TA with mesh size 60/80 and a

116 density of 0.37 g mL-1 (Buchem BV, Apeldoorn, The Netherlands). After purging, water was

117 removed from the traps using a flow of dry nitrogen (100 mL min-1 for 10 min).

118

119 2.5.2 Gas Chromatography–Mass Spectrometry

120 The trapped volatiles were desorbed using an automatic thermal desorption unit (TurboMatrix 350,

121 Perkin Elmer, Shelton, Connecticut, USA). Primary desorption was carried out by heating the trap to

122 250 °C with a flow (50 mL min-1) of carrier gas (H2) for 15.00 min. The stripped volatiles were

123 trapped in a Tenax TA cold trap (30 mg held at 5 °C), which was subsequently heated at 300 °C for

124 4 min (secondary desorption, outlet split 1:10). This allowed for a rapid transfer of volatiles to a

125 7890A GC-system interfaced with a 5975C VL MSD with Triple-Axis detector (Agilent

126 Technologies, Palo Alto, California, USA). The transfer line was heated at 225 °C.

127

128 Separation of volatiles was carried out on a DB-Wax capillary column 30 m long x 0.25 mm

129 internal diameter, 0.50µm film thickness. The column pressure was held constant at 2.3 psi

130 resulting in an initial flow rate of 1.4 mL min-1 using hydrogen as carrier gas. The column

131 temperature programme was: 10 min at 30 °C, from 30 °C to 240 °C at 8 °C min-1, and finally 5

132 min at 240 °C. The mass spectrometer was operating in the electron ionisation mode at 70 eV.

133 Mass-to-charge ratios between 15 and 300 were scanned.

7
134

135 Identifications were based on the agreement of spectra with those in the mass spectral database

136 Wiley275.L (HP product no. G1035A) and Linear Retention Indices for authentic compounds run

137 under similar GC conditions. Peak areas were calculated using the MSD Chemstation (Version

138 E.02.02, Agilent Technologies, Palo Alto, California, USA). The peak areas across replicates were

139 checked against the internal standard and expressed as peak areas relative to peak areas obtained

140 from the lowest DHS sampling temperature (31 °C).

141

142 2.6 Descriptive sensory analysis

143 The descriptive sensory analysis took place over a four days period. Two days were used for panel

144 training and two days for the sensory profile assessments. The evaluation of the coffee samples took

145 place in the sensory evaluation booths (ISO 13299, 2003) at the University.

146

147 2.6.1. Sensory panel

148 The sensory panel consisted of eight trained panellists (5 males and 3 females; age range 24-30

149 years) from the external panel at the University of Copenhagen. The panel was recruited according

150 to the selection criteria of ISO 3972 (2011) and had participated in other descriptive tests.

151

152 2.6.2. Coffee evaluation

153 During the training sessions, the sensory panel was instructed in tasting the coffee immediately after

154 serving. The panel developed a sensory vocabulary consisting of ‘overall flavour intensity’, the

155 basic tastes ‘bitter’, ‘sweet’ and ‘sour’, and the flavours ‘tobacco’, ‘roasted’, ‘nutty’ and ‘chocolate’

156 (Table 1). The vocabulary was restricted to the main eight attributes to ensure a short evaluation

157 time in order to keep the temperature changes of the coffee in the mouth to a minimum. The

8
158 panellists took a 10 ml sample in the mouth and swirled the coffee slowly in the mouth before

159 spitting it out. They assessed the respective coffee within 1 min after serving.

160

161 Insert Table 1 approximately here

162

163 The coffee samples were served in four replicates, herein 2 replicates per day. The sample order

164 was randomised within each replicate. However, due to sample preparation logistics, the panellists

165 received the same sample order within each replicate. This choice was a compromise between the

166 errors in batch-batch variation and systematic sample carry-over effects. However, since the

167 between-sample interval was four minutes, the risk of carry-over effects was considered less

168 important. Samples were presented with a 3-digit coded label. The sensory attributes were rated on

169 a 15 cm line-scale with the anchor points ‘absent’ to ‘a lot’. In between each sample the panellists

170 cleansed their palate with plain toast bread, skimmed milk and water at 37 °C. They were given a

171 recuperation time of four minutes.

172

173 2.7 Data analyses

174

175 2.7.1 Sensory data

176 The effect of serving temperature on the flavour intensities was analysed by a mixed model

177 ANOVA in R v2.15.2 (R Development Core Team 2012) using the lmerTest package (Kuznetsova,

178 Brockhoff & Christensen, 2015a). This ANOVA procedure automates optimal model selection of

179 random and fixed effects in linear mixed effects modelling (Kuznetsova, Christensen, Bavay &

180 Brockhoff, 2015b). The R-code used was:

9
181 modelCoffee <lmer(Intensity∼Temperature+Replicate+(1Judge)+(1ServingOrder),data=Coffee),

182 anova(modelCoffee), step(modelCoffee).

183

184 Principal Components Analysis (PCA) was performed on the panel averaged data using the

185 Unscrambler X v10.2 (Camo Software A/S, Oslo, Norway). The data were mean centred without

186 autoscaling using the NIPALS algorithm.

187

188 2.7.2 Volatile analyses

189 The effect of temperature on the size of the peak area for each identified compound was analysed

190 by one-way ANOVA in R v2.15.2 (R Development Core Team 2012). The peak areas relative to

191 T=31 °C were 2 log-transformed and plotted against 1/T according to the van ‘t Hoff approach. A

192 linear regression was performed on the data of each compound using SPSS software (IBM SPSS

193 Statistics, v22, USA). The regression equations were used to predict the peak ratio’s for T=56 °C

194 and T=62 °C, respectively.

195

196 3 Results and discussion

197

198 3.1 Effect of temperature on release of volatiles

199 A total of 76 volatile compounds were identified in the headspace of the Bourbon Caturra coffee

200 brews (Table 2). The majority of the volatiles were furans (26%), pyrazines (14%), ketones (11%),

201 aldehydes (9%), thiophenes (8%), pyrroles (7%) and esters (7%) and had been previously reported

202 in other types of coffee (Sanz, Ansorena, Bello & Cid, 2001; Flament, 2002; Cheong, Tong, Ong,

203 Liu, Curran & Yu 2013; Sunarharum, Williams & Smyth, 2014). Several of these compounds were

204 heterocyclic and typically formed in Maillard reactions at high temperatures and low moisture

10
205 content (e.g. Bredie, Guy, & Mottram, 1998; Bredie, Guy, & Mottram, 2002). An overlay of the

206 chromatograms obtained at 31 °C, 37 °C, 44 °C and 50 °C is presented in Figure 1. A clear

207 difference in the total peak area could be observed with the higher temperature the larger the peak.

208 No obvious differences were found in the qualitative profiles of the volatiles with increasing

209 temperatures.

210

211 Insert Figure 1 approximately here

212

213 The peak area of the vast majority of the volatile compounds increased significantly (p<0.05) with

214 increasing sampling temperature. The total area count for the identified compounds as function of

215 the sampling temperature is shown in Figure 2. The joint release of the identified volatiles increased

216 exponentially with temperature. In general small differences in the joint area count was seen

217 between the 31 °C and 37 °C samples, while at temperatures from 44 °C and above a significant

218 increase the joint area count was observed compared to the 31 °C sample. This indicated that

219 differences in coffee flavour between serving temperatures of 31 °C and 37 °C may be very small.

220 However, at higher temperatures some changes in the flavour profile of the coffee may be expected

221 due to the larger changes in volatile release.

222

223 The increase in peak area with increasing temperature was anticipated since physical-chemical

224 properties such as molecular weight, vapour pressure, boiling point and partition coefficients

225 influence the volatility of aroma compounds (e.g. Van Ruth, O’Connor & Delahunty, 2000). The

226 partial pressure of the compounds is dependent on the temperature, meaning that increasing

227 temperatures lead to greater evaporation, and consequently a higher release of volatile compounds.

228 These findings correspond to a study by Covarrubias-Cervantes, Mokbel, Champion, Jose

11
229 & Voilley (2004) who found that the saturated vapour pressure of aroma compounds increased with

230 increasing temperature. In the present study, volatiles were sampled under DHS conditions as a

231 model for flavour release in the mouth. Under these conditions, mass transfer from oil-to-water and

232 water-to-air may be also be influencing flavour release.

233

234 Insert Figure 2 approximately here

235

236 Insert Table 2 approximately here

237

238 When comparing the relative release with increasing temperature for the individual compounds a

239 large variation was observed. Between 31 °C and 37 °C small changes occurred with the largest

240 increases (1.4 to 1.5 fold) for benzaldehyde, 3-hexanone, phenylethanal, 2-methyl-1-propanal, 3-

241 ethyl-2,5-dimethylpyrazine and 2-methyl-3,5-diethylpyrazine. The latter two compounds have

242 roasted flavour notes, whereas the others elucidate sweet, fruity, floral and almond-like notes (Table

243 2). Between the sampling temperatures of 31 °C and 44 °C most marked increases (2.3 to 2.6 fold)

244 were observed for 3-hexanone, benzaldehyde, 4-methylpyridine and the alkylpyrazines, i.e. 3-ethyl-

245 6-methylpyrazine, 2-ethyl-5-methylpyrazine, vinylpyrazine, 3-ethyl-2,5-dimethylpyrazine and 2-

246 methyl-3,5-diethylpyrazine. These pyrazines possess besides the roasted character also cocoa, nutty

247 and baked potato odour notes.

248

249 When the sampling temperature was raised from 31 °C to 50 °C a total of 21 compounds were

250 increased in the headspace sampling by a factor 2.5 or higher. Among these compounds were 7

251 pyrazines (out of 11) and only 3 furans (out of 20) and one thiophene (out of 6). At the higher

252 serving temperatures (56 °C and 62 °C) the relative increases of the compounds were predicted by

12
253 the van ’t Hoff regression. At these temperatures, alkylpyrazines were predicted to increase up to

254 8.2 fold and ketones up to 6.7 fold. Besides the alkylpyrazines, the compounds predicted to be

255 released most at 62 °C were 4-methylpyridine (6.5 fold), phenylethanal (7.7 fold), 2-methyl-1-

256 propanal (8.1 fold) and benzaldehyde (9.4 fold) (Table 2).

257

258 3.2 Effect of serving temperature on flavour perception

259 The descriptive sensory analysis was performed using the main sensory attributes of the Bourbon

260 Caturra coffee served between 31 °C and 62 °C. The panel mean ratings and significance levels are

261 presented (Table 3). Irrespective of the temperature, the Bourbon Caturra coffee was perceived

262 with ‘bitter’, ‘tobacco’, ‘roasted’ top notes with ‘chocolate’, ‘sour’, ‘nutty’ and ‘sweet’ undertones.

263 The serving temperature range showed clear but relatively small differences in attributes on the

264 sensory scale. Significant (p<0.05) temperature effects were found for ‘overall intensity’, ‘bitter’

265 and ‘sweet’ taste and ‘roasted’ flavour. The relationships between serving temperature and the

266 sensory attributes were evaluated in PCA with all sensory attributes included. A total of 84% of the

267 variance was explained in the first two components (Figure 3).

268

269 Insert Table 3 approximately here

270 Insert Figure 3 approximately here

271

272 PC1 (57%) described the difference between the lower (31 °C to 44 °C) versus the higher (50 °C to

273 62 °C) serving temperatures. PC2 mainly described the difference between the serving temperature

274 of 44 °C versus the 31 °C and 37 °C. The correlation of the samples with attribute loadings showed

275 a clear separation of the samples shifting from more ‘sour’, ‘chocolate’, ‘sweet’ characters (31 °C

276 to 44 °C) towards more clear ‘bitter’, ‘roasted’ flavour notes and increased ‘overall intensity’ (50

13
277 °C to 62 °C). The serving temperature of 44 °C gave the most marked sweetness character in the

278 coffee, whereas the 56 °C coffee serving was perceived as most bitter.

279

280 Although flavour release was clearly shown to increase with higher serving temperature,

281 temperature itself may have influenced the sensory perception of the coffee due to peripheral and

282 perceptual mechanisms at a higher level, e.g. sensory masking effects by inputs from

283 termoreceptors. For instance, it is well known that basic tastants possess lower taste threshold

284 values close to 30 °C (McBurney, Collings & Glanz, 1973) than at higher temperatures. However,

285 at supra-threshold concentrations, temperature effects on basic taste perception become less

286 apparent. For caffeine, a bitter tastant in coffee, temperature had an influence at taste recognition

287 level, but this effect was completely diminished at the terminal taste threshold (Paulus & Reisch,

288 1980). The effect of temperature on taste perception in the Bourbon Caturra coffee cannot be

289 excluded but may be relatively small since taste intensities were perceived at moderate to strong

290 levels.

291

292 Relationship between volatiles release and coffee flavour perception

293 Studies on coffees have shown that furans and pyrazines quantitatively represent the largest share of

294 the volatiles. However, sulfur-containing compounds together with pyrazines are considered to be

295 most important contributors to the roasted coffee flavour (Nijssen, Visscher, Maarse, Willemsense,

296 & Boelens, 1996). Furans have relatively high sensory thresholds compared to other groups of

297 coffee volatiles (Burdock, 2010) but have been found in concentrations where they likely contribute

298 to burnt sugar, nutty, roasted and coffee-like aroma notes (Akiyama et al., 2007; Bicchi et al., 2011;

299 Burdock, 2010; Cheong et al 2013; JECFA, 2005). In the present study on the release of coffee

300 volatiles from Bourbon Caturra coffee, furans were quantitatively less influenced by increasing

14
301 temperatures than the release of alkylpyrazines, ketones, and other Maillard-derived compounds

302 like some pyrroles and pyridines. However, qualitatively, some furans like 2-

303 [(methylthio)methyl]furan (coffee-like), 2,3-dihydro-6-methieno[2,3c]furan (kahweofuran roasted,

304 coffee-like) and 2-methylfuran (spice smokey) may be important for the overall coffee flavour.

305 They have been reported as important flavour compounds in Arabica coffee (Ribeiro, Augusto,

306 Salva, Thomaziello & Ferreira, 2009; Akiyama et al., 2007).

307

308 Pyrazines arise from Maillard reactions and Strecker degradation of amino acids in the coffee

309 roasting process. Several alkylpyrazines possess relatively low sensory thresholds and are

310 considered of key importance to coffee flavour, e.g. contributing to roasted, nutty and cocoa aromas

311 (Acree & Arn, 2004; JECFA, 2005). Since their release in Bourbon Caturra was significantly

312 increased at the serving temperatures above 44 °C, they were most likely involved in the increased

313 ‘roasted’ notes and ‘overall flavour intensity’ observed in these samples. Furthermore,

314 alkylpyrazines are known to possess bitter taste properties and the greater release to the headspace

315 may also have imparted the bitter taste of the coffee, which was most noticeable at the higher

316 serving temperatures. This may be due to a greater activity and solubility in the aqueous phase

317 relatively to the lipid phase of these and other bitter compounds in the coffee.

318

319 The sulfur-containing compounds were mainly Maillard-derived heterocyclic compounds including

320 thiazoles, thiopenes, and the earlier mentioned furans. Even present in relatively low concentrations

321 these compounds, due to their sensory potency, contribute significantly to coffee flavour (Maga,

322 1975; Ribeiro et al., 2009). The thiophenes and thiazoles showed a moderate release with increasing

323 temperature but to a lower extend than pyrazines and furans.

324

15
325 Phenylethanal (floral, honey), 3-hexanone (sweet, fruity) and 2-methyl-1-propanol (winey) were

326 among the few compounds that increased already notably at the lower serving temperature (44 °C).

327 In the sensory profile of the coffee at this serving temperature, sweetness was perceived at the

328 highest intensity level. This illustrated that lower temperatures (44 °C or lower) may have potential

329 to assess subtle flavour notes more clearly from the dominating roasted flavour.

330

331 Other studies have indicated an effect of product temperature on flavour perception. Engelen, de

332 Wijk, Prinz, Janssen, Weenen and Bosman (2003) and Ventanas, Mustonen, Puolanne and Tuorila

333 (2010) reported that subjects perceived higher flavour intensities from food served at higher

334 temperature. In these studies it was not clear whether the temperature effect was due to increased

335 release of flavour compounds. However, the present work suggests a contribution of the release of

336 volatiles at elevated temperatures to the changes in flavour notes of coffee.

337

338

339 Although the present study only investigated the Bourbon Caturra speciality coffee, the results

340 indicate that coffees from similar roasting conditions will taste more similar at high temperatures. In

341 order to perceive the more subtle differences and delicate aromas, such as fruity and flowery notes,

342 coffee should be evaluated at lower serving temperatures. Furthermore, the study revealed that

343 serving temperature and assessment methodology needs proper standardisation to make fair

344 comparisons and labelling of product quality.

345

346 4 Conclusion

347 The serving temperature of coffee influenced the release of volatile compounds and

348 correspondingly the intensity of flavour characters. The dynamic volatile release as function of

16
349 serving temperature followed in most instances a van ´t Hoff-type pattern, allowing the prediction

350 of release for different volatile compounds. The release of volatiles from the Bourbon Caturra

351 coffee was exuberated above 40 °C. Maillard-derived heterocyclic compounds, especially

352 alkylpyrazines released most marked at 50 °C or higher temperature. The shift towards greater

353 levels of Maillard-derived volatiles coincided with more intense ‘roasted’, ‘bitter’ and ‘overall

354 flavour intensity’ properties of the coffee. Serving the coffee below 44 °C facilitated the

355 translucence of non-‘roasted’ coffee flavours. When evaluating specialty coffees in a sip-and-spit

356 tasting setting with a trained sensory panel, serving at lower temperatures should be considered to

357 reduce the overruling impact of roasted flavours on more subtle flavour character notes.

358

359 Acknowledgment

360 The staff of the sensory laboratory at the University of Copenhagen is thanked for assistance in the

361 sensory analyses.

362

363 Statement on conflict of interest

364 This study was carried out as part of the Food Innovation and Health study programme. The authors

365 declare not conflict of interests with any parties involved in the study.

366

17
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465

466

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467 Figure captions (brief title + description)

468 Figure 1. Ion Chromatograms. Joined total ion chromatograms of headspace analysis of

469 Bourbon Caturra coffee at sampling temperatures of 31°C, 37°C, 44°C and 50°C.

470

471 Figure 2. Ratio of increase. The relationship between serving temperature and the ratio increase

472 of the total volatiles in the headspace sampled from Bourbon Caturra coffee. Data points at 56ºC

473 and 62ºC were estimated based on van ´t Hoff regression. The relationship indicates the

474 importance of serving temperatures above approximately 40ºC for the accelerated release of

475 volatile compounds.

476

477 Figure 3. PCA bi-plot. PCA bi-plot of the average descriptive sensory panel data showing the

478 relationships between sensory variables and serving temperatures of the Bourbon Caturra Coffee.

23
Figure 1. Joined total ion chromatograms of headspace analysis of Bourbon Caturra coffee at
sampling temperatures of 31°C, 37°C, 44°C and 50°C.

24
Figure 2. The relationship between serving temperature and the ratio increase of the total volatiles
in the headspace sampled from Bourbon Caturra coffee. Data points at 56ºC and 62ºC were
estimated based on van ´t Hoff regression. The relationship indicates the importance of serving
temperatures above approximately 40ºC for the accelerated release of volatile compounds.

25
Figure 3. PCA bi-plot of the average descriptive sensory panel data showing the relationships
between sensory variables and serving temperatures of the Bourbon Caturra Coffee.

26
Table 1. Sensory vocabulary and definitions used in the coffee flavour evaluation.

Group Descriptor Definition Reference material

Overall impression Flavour intensity Overall flavour intensity when -

tasting the sample
Basic taste Sour Sour taste associated with citric Citric acid solution: 0.6 g
acid solution citric acid/L

Bitter Bitter taste associated with bitter Coffee with 15 minutes

coffee extraction time

Sweet Sweet taste associated with sucrose Sucrose solution: 7.2 g

solution sucrose/L
Retronasal aroma Roasted Aroma associated with Roasted toast bread
roasted/burned toast

Nutty Aroma associated with roasted Roasted hazelnuts

nuts

Tobacco Aroma associated with pipe Red Orlik Tobacco

tobacco

Dark chocolate Aroma associated with dark Dark chocolate (100%)

chocolate (not the bitter taste)

27
Table(s)

OTcd Appr. Peak ratio relative to 31 ºC

LRIa IDb
Compound Flavour descriptorsc (µg/L) R2 e conc. at p-value
31°C
37 ºC 44 ºC 50 ºC 56 ºC 62 ºC
(µg/L)f

Aliphatic alcohols
2-Methyl-1-propanol 1104 MS + LRI Winey*, *** 360 – 3300 0.993 4 1.4 2.4 3.8 5.5 8.1 0.005
1-Hexanol 1369 MS + LRI Sweet. green*,***,fruity*** 200 – 2500 0.713 5 0.8 1.2 1.8 -g - 0.017

Aldehydes
Propanal 792 MS + lri Sharp, pungent*,*** 9.5 - 35 0.849 28 1 1.4 2.1 - - 0.523
2-Methylbutanal 909 MS + LRI Green, fruity**,*** roasted cocoa**** 1 0.575 1530 0.9 1.1 1.3 - - 0.015
3-Methylbutanal 916 MS + LRI Fruity, fatty/creamy, almond*.**. *** 0.2 - 7 0.773 10 0.9 1.6 1.8 - - 0.012
2-Butenal (E or Z) 1034 MS + lri - - 0.919 9 1.1 1.4 2.1 2.5 3.2 0.017
Hexanal 1084 MS + LRI Grassy*,***, fatty, green, fruity*** 4.1 - 22.8 0.714 51 0.8 1.4 1.6 - - 0.004
Benzaldehyde 1533 MS + LRI Sweet, strong almond*,** 100 – 4600 0.875 36 1.5 1.7 4.4 6.1 9.4 <0.001
Phenylethanal 1653 MS + lri Hawthorne**, floral, honey*,**,*** 4 0.991 17 1.4 2.3 3.7 5.3 7.7 0.001

Ketones
2-Butanone 903 MS + lri Chemical, slightly fruity*** - 0.617 872 0.8 1.4 1.6 - - 0.011
2-Pentanone 975 MS + lri Ethereal, fruity*, sweet, fruity*** 70 0.665 122 0.8 1.3 1.5 - - 0.001
2,3-Butanedione 980 MS + LRI Buttery*,*** 0.3 – 15 0.376 1190 0.9 0.8 1.8 - - 0.338
3-Hexanone 1051 MS + lri Sweet, fruity, waxy, rum notes*** 41 – 81 0.983 24 1.5 2.6 3.1 4.8 6.7 0.004
2,3-Pentandione 1068 MS + LRI Buttery*,** - 0.86 1420 0.9 1.7 2.2 3 4.1 0.001
(E)-3-Penten-2-one 1129 MS + lri - - 0.883 26 1 1.8 2.2 3 4 0.003
Cyclopentanone 1182 MS + lri Like peppermint*,*** - 0.909 12 1 1.7 2.4 3.2 4.2 0.007
2-Methyl-2-cyclopenten-1-one 1378 MS + lri - - 0.975 7 1.2 1.8 2.4 3.2 4.1 0.001

Acids
3-Methylbutanoic acid 1020 MS + lri Fruit-like, apple, cherry*** - 0.914 9 1 1.2 1.6 1.7 2 0.009

Esters
Ethyl acetate 892 MS + LRI Acetic, ethereal*, fruity, sweet*** 5 – 5000 0.538 597 0.8 1.2 1.5 - - 0.084
Acetic acid methyl ester 825 MS + lri Fruity*,***, sour**, green, ethereal*** 1500 – 47000 0.732 24 0.9 1.2 1.7 - - 0.004
Furancarboxylic acid methyl ester 1330 MS Berry, fruity, winey*, tobacco*** - 0.904 6 1 1.6 2.1 2.7 3.4 <0.001
Furfuryl acetate 1545 MS + lri Ethereal-floral fruity*, fruity*** - 0.92 272 1.3 1.6 2.8 3.5 4.8 <0.001
Ethyl-2(E)-4(Z)-decadienoate 1857 MS - - 0.96 17 1.2 1.8 3 3.9 5.4 0.002

Phenols
Phenol 1423 MS Medicinal, sweet, tar*, phenol** 5.5 0.935 2 1.1 1.7 2.3 3 4 0.001
2-Methoxyphenol 1837 MS + lri Smoke-like, somewhat medicinal* 3 - 31 0.839 6 0.9 1.4 2.6 3.4 4.7 0.006
2-Methylphenol 1906 MS Musty, phenolic aftertaste*** 45 0.704 8 1.2 1.1 1.7 - - 0.07

Furans
2-Methylfuran 868 MS + lri Spicy smoky * 3.5 0.752 537 1 1.1 1.6 - - 0.083
2,5-Dimethylfuran 948 MS + lri Spicy smoky* - 0.716 45 1 1.1 1.6 - - 0.035
2-Vinylfuran 1078 MS + lri Phenolic, coffee-grounds**** - 0.82 49 1.1 1.1 1.5 - - 0.053
2-Vinyl-5-methylfuran 1161 MS + lri - - 0.824 43 1.1 1.1 1.5 - - 0.037
2-(2-Propenyl)furan (E or Z) 1212 MS - - 0.842 11 1.1 1.2 1.6 1.8 2.1 0.011
2-(Methoxymethyl)furan 1249 MS + lri Airy, roasted coffee* - 0.868 116 1 1.3 1.6 1.9 2.2 0.001
2-Methyltetrahydrofuran-3-one 1275 MS + lri Nutty, creamy, almond*** - 0.834 292 0.9 1.6 1.9 2.5 3.2 0.001
2-Furfural 1435 MS + lri Bread, almond, sweet*.**.*** 280 – 8000 0.968 2 1.1 1.6 2.1 2.6 3.2 0.005
2,5-Dimethyl-3-ethylfuran 1484 MS - - 0.934 13 1.1 1.9 2.2 3.1 4 <0.001
2-[(Methylthio)methyl]furan 1495 MS + lri Coffee-like* - 0.848 55 0.9 1.5 1.7 2.2 2.7 0.001
1-(2-Furanyl)-ethanone 1514 MS + lri Coffee-like* - 0.982 383 1.2 1.7 2.2 2.8 3.5 0.001
1-(2-Furyl)-2-propanone 1529 MS + lri Radish*** - 0.961 44 1.1 1.8 2.5 3.3 4.4 <0.001
5-Methyl-2-furfural 1583 MS + lri Almond, caramel, burnt sugar**. *** 6 0.994 400 1.1 1.3 1.5 1.6 1.8 0.001
2,2'-Bifuran 1604 MS + lri - - 0.925 30 1.1 1.3 1.7 1.9 2.2 0.011
2,2'-Methylenebisfuran 1618 MS + lri - - 0.957 44 1.1 1.4 1.8 2.1 2.5 0.021
2-Acetyl-5-methylfuran 1628 MS + lri Strong, nutty*,***, caramel-like**** - 0.98 8 1.3 1.8 2.8 3.6 4.8 0.002
2-(2-Furanylmethyl)-5-methylfuran 1689 MS + lri - - 0.957 13 1.1 1.5 2.1 2.5 3.1 0.017
2,3-Dihydro-6-methylthieno[2,3c]furan 1754 MS Roasted, smoky**** - 0.94 50 1.1 1.6 2 2.5 3.1 0.001
3-Phenylfuran 1834 MS + lri - - 0.947 7 1.1 1.5 2.3 2.8 3.5 0.001
1-(5-Methyl-2-furanyl)-1-propen-3-al 1846 MS - - 0.852 2 1.1 1.5 3.1 4.1 5.8 0.002

Pyrroles
1-Methyl-1H-pyrrole 1143 MS + lri Smoky-tarry; diluted: sweet, woody**** - 0.559 291 0.8 1.2 1.4 - - 0.011
1-Ethyl-1H-pyrrole 1184 MS + lri - - 0.451 22 0.9 1 1.2 - - 0.132
1H-Pyrrole 1521 MS + LRI Nutty, sweet, warm, ethereal*,*** 20000 – 49600 0.852 149 0.9 1.5 1.8 2.4 3 0.008
1-Ethyl-2-formylpyrrole 1623 MS + lri - - 0.973 15 1.2 1.9 2.5 3.3 4.3 <0.001
2-Formyl-1-methylpyrrole 1634 MS + lri - - 0.979 100 1.2 1.7 2.2 2.8 3.5 0.001
1-(2-Furfuryl)pyrrole 1821 MS + lri Vegetable, earthy-green*,*** 100 0.962 146 1.1 1.8 2.4 3.2 4.2 <0.001
1-(5-Methylfurfuryl)pyrrole 1849 MS - - 0.917 4 1.1 1.7 3 3.9 5.5 0.001

Pyridines
Pyridine 1186 MS + lri Penetrating odour, sharp taste *** 0.079 - 790 0.947 937 1.1 1.5 1.7 2.1 2.4 0.004
4-Methylpyridine 1227 MS + lri - - 0.959 2 1.2 2.3 3.1 4.6 6.5 <0.001

Pyrazines
Pyrazine 1213 MS + lri Sweet, corn-like. nutty*, spinach*** 300 0.959 103 1.1 1.6 2 2.5 3.1 0.005
Methylpyrazine 1278 MS + lri Nutty, cocoa **, roasted, earthy*** 60 – 100000 0.983 566 1.2 1.6 1.9 2.3 2.8 0.001
2,5-Dimethylpyrazine 1337 MS + lri Cocoa, nutty**,***, roasted** 80 – 1800 0.99 157 1.2 1.8 2.5 3.3 4.2 <0.001
2,6-Dimethylpyrazine 1343 MS + lri Cocoa, roasted nutty, roast beef** 200 – 1500 0.992 155 1.3 1.9 2.5 3.3 4.3 <0.001
Ethylpyrazine 1348 MS + lri Musty, nutty, peanut butter*.*** 4000 – 22000 0.985 145 1.3 1.6 2 4.5 7.2 <0.001
2,3-Dimethylpyrazine 1360 MS + lri Nutty, cocoa-like*,*** 2.5 0.974 25 1.2 1.7 2.3 3 3.8 0.001
2-Ethyl-6-methylpyrazine 1396 MS + lri Roasted baked potato*, roast** - 0.99 105 1.3 2.2 3.1 4.4 6.1 <0.001
2-Ethyl-5-methylpyrazine 1401 MS + lri Nutty, roasted, grassy*, roast** 16 - 100 0.989 71 1.3 2.1 2.9 4.1 5.6 <0.001
Vinylpyrazine 1447 MS + lri - - 0.984 13 1.3 2.6 3.7 5.6 8.2 <0.001
3-Ethyl-2,5-dimethylpyrazine 1458 MS + lri Potato, roasted**, nutty, roasted**** 5 0.996 37 1.4 2.4 3.5 5 7.1 <0.001
2-Methyl-3,5-diethylpyrazine 1508 MS Roasted nut, baked potato*, grassy**** - 0.993 8 1.4 2.6 4 6 8.9 <0.001

Thiophenes
Thiophene 1016 MS + lri Garlic** - 0.537 29 0.9 1 1.4 - - 0.034
3-Methylthiophene 1119 MS + lri Fatty, winey****, roasted, meaty***** - 0.78 4 1 1.1 1.4 - - 0.093
2-Methyltetrahydrothiophen-3-one 1540 MS + lri - - 0.928 16 1.1 1.8 2.2 3 3.9 <0.001
2-Thiophenecarboxaldehyde 1707 MS + lri Nutty, spicy meat, coconut-like**** 200 0.975 18 1.1 1.7 2.2 2.7 3.5 0.002
3-Acetylthiophene 1792 MS + lri - - 0.98 5 1.2 1.8 2.5 3.1 4.1 0.001
3-Acetyl-2,5-dimethylthiophene 1816 MS - - 0.944 8 1.1 1.6 2.4 2.9 3.8 0.001

Thiazoles
Thiazole 1259 MS + lri Green, sweet, nutty, tomato *,*** 3.1 0.875 12 1 1.6 2 2.7 3.4 0.007
4-Methylthiazole 1295 MS + lri Nutty, green*, roasted meat**,*** - 0.899 22 1 1.7 2.1 2.9 3.7 0.003
4,5-Dimethylthiazole 1385 MS + lri Nutty, meaty*., roasted, smoky** 470 0.881 6 1.2 1.5 1.5 1.9 2.1 0.053
Table 3. Mean intensity ratings (cm on 15 cm scale) of the sensory descriptors with the significance
level for the factor serving temperatures.

Mean scaling Lowest - highest

Sensory attribute value (±SE) sample mean p-value

Overall intensity 9.3 (0.19) 8.3 – 10.5 0.001

Sour 8.0 (0.20) 7.0 – 9.0 0.068
Bitter 9.7 (0.19) 8.8 – 10.6 0.011
Sweet 3.6 (0.15) 2.8 – 4.6 0.027
Tobacco 9.6 (0.19) 8.8 – 10.4 0.107
Roasted 9.1 (0.19) 7.8 – 9.7 0.001
Nutty 4.2 (0.13) 3.8 – 4.7 0.174
Chocolate 7.4 (0.21) 6.5 – 8.0 0.197

30
Research highlights

- Release of volatiles from Bourbon Caturra coffee is exuberated above 40 °C

- Release of Maillard-derived volatiles is more marked above 50 °C
- Shift in Maillard volatiles concur with roasted, bitter and overall flavour intensity
- Tasting below 44°C increases the translucence of non ‘roasted’ coffee flavours
- Serving temperature should be considered in specialty coffee quality selection

31