The Australian Wine Research Institute

Quantitative analysis of phenolic compounds in red wine fermentation using near infrared spectroscopy
Daniel Cozzolino, Mariola Kwiatkowski, Mango Parker, Bob Dambergs, Wies Cynkar, Mark Gishen and Markus Herderich The Australian Wine Research Institute, PO Box 197, Glen Osmond (Adelaide) SA 5064, Australia Cooperative Research Centre for Viticulture, PO Box 154, Glen Osmond (Adelaide) SA 5064, Australia Daniel.Cozzolino@awri.com.au

Introduction Figure 1. Score plot for the first three principal components for replicate fermentations indicating the time course. During wine fermentation many grape constituents such as tannins and anthocyanins are extracted and modified by (bio) chemical reactions. The analysis of compounds affected by fermentation can in some cases require several steps of sample preparation that can result in delays in obtaining the result. The objective of this work was to determine the feasibility of near infrared (NIR) spectroscopy to rapidly monitor phenolic compounds during the red wine fermentation process.

Materials and methods

Samples taken from several different experimental wine fermentation trials conducted by the AWRI in the years 2001, 2002 and 2003, were scanned (4002500 nm) in transmission mode in a 1 mm path length cuvette, using a monochromator spectrophotometer (FOSS NIRSystems6500). Calibration equations were developed for the concentration of malvidin-3-glucoside (M3G), tannins (T) and pigmented polymers (PP) using an HPLC method as the reference. Spectral data collection and manipulation were achieved using VISION software. Partial least square (PLS) regression models between spectral and HPLC data were developed using The Unscrambler software.

Figure 2. Eigenvectors for the first three principal components.

Results and conclusion

Figure 1 shows the changes that occur during fermentation in the scores for the first three principal
0.2
Anthocyanins

OH

PC1 PC2 PC3 PC4

components. This same type of pattern was observed for all fermentations monitored during the study. Figure 2 shows the eigenvectors for the the first three principal components. The highest eigenvectors were observed around 520 nm (most likely caused by anthocyanins), around 2000 nm (OH tones most likely due to water and alcohol) and around 2200 nm (CH overtones, possibly due to phenolics). High coefficients of determination in calibration (R2) and low standard error in cross validation (SECV) were obtained for M3G, T

Eigenvectors

0.1

0.0

and PP (Figure 3). If the residual predictive deviation (RPD), defined as the ratio between the standard deviation of the population (SD) and the standard error of cross validation (SECV), is greater than three, a calibration model is considered as acceptable for analytical purposes.
CH 500 1000 1500 2000 2500

-0.1

The results indicate that NIR spectroscopy shows promise as a means of rapidly monitoring the concentration of phenolic compounds during red wine fermentation.

Wavelength (nm)

400 350

Malvidin-3-glucoside predicted by NIR

300 250 200 150 100 50 0

800

Pigmented polymers predicted by NIR

Tannin predicted by NIR

n = 492 2 R = 0.91 -1 SECV = 28 mgL RPD = 3.5

1000

n = 270 2 R = 0.83 -1 SECV = 131.1 mgL RPD = 1.8

80 70 60 50 40 30 20 10 0 0 10 20 30 40 50 60 70 80

n = 290 2 R = 0.87 -1 SECV = 5.9 mgL RPD = 3.1

600

400

200

0
0 50 100 150 200 250 300 350 400

200

400

600

800

1000

Malvidin-3-glucoside determined by HPLC

Tannin determined by HPLC

Pigmented polymers determined by HPLC

Figure 3. Correlation between NIR-predicted and reference method determinations for the concentrations of M3G, T and PP in red wine ferments. Note: RPD = SD/SECV

COOPERATIVE RESEARCH CENTRE

for

VITICULTURE