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Characterization of the Fermentation Process and Aroma Profile of Carob Brandy

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The possibility of producing carob brandy was investigated, focusing on the fermentation of carob and the preliminary characterization of the volatile components in the obtained distillates. Fermentations were carried out in carob mash with or without added nutrients by five Saccharomyces cerevisiae strains at three different temperatures. The obtained wine was subjected to fractional distillation in a copper still to produce carob spirit. Analysis of sugars and fermentation products was performed by high performance liquid chromatography. Gas chromatography and gas chromatography coupled with MS detection was used to analyze the volatile components of carob wine and brandy. Carob flour and the strains used can be efficiently used for the fermentation process to produce carob wine with ethanol content ranging from 46.4 to 50.5 gL−1 and corresponding yield coefficients ranging from 0.45 to 0.49 gg−1. More than ninety compounds detected in carob spirit; ethyl 2-methyl butanoate, ethyl 2-methyl propanoate, ethyl cinnamate, ethyl hexanoate, beta-ionone, ethyl butanoate and ethyl octanoate, largely contribute to the bouquet of the spirit. Thus, a novel volatile spirit may be an additional product in the carob processing chain and represent a new potential, especially for small carob growers.


  • Fermentation
  • Yeast
  • Carob
  • Carob brandy
  • Distillate
  • Fruit spirit

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  • DOI: 10.1007/978-3-031-04797-8_7
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Authors would like to thank to Valentina Papić Bogadi for helpful comments on the manuscript and for correcting the English style.


The research is funded by the Croatian Science Foundation, grant number: IP-11-2013_3304-TEUCLIC and IP-2018–01-9717- SPB-LCF, and paper was produced as part of the “Atrium of Knowledge” project co-financed by the European Union from the European Regional Development Fund and the Operational Programme Competitiveness and Cohesion 2014–2020.

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Correspondence to Jasna Mrvčić .

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Appendix 1. Conditions of Head Space Gas Chromatography with Flame-ionization Detection (HS and GC-FID)

HS conditions GC conditions
O/N/T 80/100/110 ℃
Thermostating time: 20 min
Pressurization time: 0.2 min
Injection time: 0.05 min
Withdrawal time: 0.1 min
Carrier: He
Pressure: 25 psgi
Injector temperature: 110 ℃
Detector temperature: .0 ℃
Oven program:
35 ℃, 5 min
10 ℃/min 60 ℃
60 ℃, 2 min
10 ℃/min, 180 ℃
180 ℃, 7 min

Appendix 2. Validation Parameters for Analysis of VOC’s by HS-GS-FID Method

Compound Linearity range
R2 over calibration curve range LOD
mean ± sd A (%) CV (%)
Propan-1-ol 3–100 0.9976 2.564 8.548 2.960 ± 0.001 −1.33 0.04
98.260 ± 1.77 −1.74 1.77
Butan-2-ol 1–100 0.9968 0.508 1.692 1.070 ± 0,001 +7.00 0.09
96.446 ± 0.627 −3.55 0.65
Ethyl acetate 1–100 0.9953 0.171 0.572 1.007 ± 0.013 +0.70 1.29
97.387 ± 1.477 −2.61 1.52
2-methylpropan-1-ol 1–100 0.9976 0.474 1.581 1.009 ± 0.044 +0.90 4.36
96.606 ± 0.511 −3.39 0.53
3-methylbutan-1-ol 0,5–100 0.9986 0.472 1.574 0.534 ± 0.037 −10.40 6.80
97.103 ± 1.587 −2.897 1.63
2-methylbutan-1-ol 1–100 0.9978 0.387 1.290 1.085 ± 0.040 +8.5 3.69
95.047 ± 1.515 −4.95 1.59
Ethyl-butanoate 0,5–80 0.9972 0.063 0.208 0.389 ± 0.085 −22.20 21.85
77.497 ± 0.202 −3.13 0.269
Iso-amyl-acetate 0,05–80 0.9960 0.005 0.014 0.051 ± 0.009 +2.00 17.64
79.980 ± 0.584 −0.03 0.73
Ethyl-hexanoate 1–80 0.9919 0.032 0.106 1.164 ± 0.063 +16.40 5.41
77.011 ± 0.415 −3.74 0.54
Ethyl-octanoate 0,5–100 0.9904 0.060 0.200 0.492 ± 0.073 −1.60 14.84
96.436 ± 0.568 −0.36 0.59

Data on GC Method Validation

Method linearity was determined by evaluating the regression curve and it is indicated by the square correlation coefficient (R2). Linearity was achieved with a minimal R2 of 0.990.

Detection limits were determined by replicate HS-GC-FID analysis with the lowest concentration for all tested compounds (Table 1). The limit of detection (LOD) and quantification (LOQ) were calculated using the following equations:

LOD = 3 * S/N ratio * lowest concentration of linear sample

LOQ = 10 * S/N ratio * lowest concentration of linear sample

S/N ratio was determined using TotalChrom GC software (Perkin-Elmer).

Precision was expressed as the coefficient of variation (%CV) of HS-GC-FID method and it was determined in five replicates in concentrations pointed out in Table S4.

Accuracy (A) was calculated as the percentage relative error of the method:

A = (mean calculated concentration – nominal concentration)/nominal concentration * 100%.

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Mrvčić, J. et al. (2022). Characterization of the Fermentation Process and Aroma Profile of Carob Brandy. In: , et al. 10th Central European Congress on Food. CE-Food 2020. Springer, Cham.

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