The instrumental measurement of

colour and lycopene in tomato and

tomato products

Luca Sandei*, A. Maselli°, G. Visentini°, Virgilio Pieracci# and Carlo Leoni*

* SSICA Stazione Sperimentale per l’Industria delle Conserve Alimentari in Parma

Viale F.Tanara,31/a 43100 Parma ITALY
e-mail : luca.sandei@ssica.it; carlo.leoni@ssica.it
Internet : www.ssica.it
° Maselli Misure SpA Via Baganza 4/3 43100 Parma
e-mail: tech@masellimisure.com
# UNIPR Facoltà di Agraria Corso di laurea in Scienze e Tecnologie Alimentari
 KEYWORDS: Carotenoids, Lycopene,
Lycopene instrumental measurements.
The past few years have been
witnessing an increasing
interest in consumers’
health as well as in the nutritional
aspects of tomato and tomato products,
with special regard to the
antioxidant properties of carotenes
(lycopene and other carotenoids specifically
contained in tomato products).
Carotenoids, Lycopene, Lycopene
instrumental measurements

As is well known, the most important dietary source

of natural lycopene for humans are tomato and
tomato products. During the last ten years, several
studies have been carried out at SSICA in Parma in
an attempt to investigate many issues regarding to
the antioxidant “lycopene”, in terms of amounts
present in the raw material, control during the
processing and packaging phases in all the tomato
“filiera” (production chain), with a particular quality
“from field to fork” approach.
 Verba Volant, Scripta Manent……
Lycopene studies at SSICA Parma
 Sandei, L. ; Battilani, A.; Leoni, C.; (2005) Exploitation of by-products (solid
wastes) coming from tomato industry to obtain antioxidants for hi-value
business. Effect of matrix structure in carotenoids (lycopene) extraction from
wastes (peels and seeds) with CO2 SFE. Proceedings of the 9th International
Symposium on the Processing Tomato. Acta Horticolturae
 Sandei, L. ; Cabassi, A.; Leoni, C.; (2003) Effects of Technological Operations
and storage conditions on color and carotenoids content of tomato powder.
Proceedings of the 8th International Symposium on the Processing Tomato. Acta
Horticolturae N. 613 415 – 421.
 Sandei, L.; Siviero, P.; Zanotti, G.; Cabassi, A. (2002). Valutazione del
contenuto di licopene nel pomodoro. L’Informatore Agrario n. 3 2000 59 – 63 .
 Sandei, L.; Siviero, P.; Zanotti, G.; Leoni, C. (2003) Evaluation of the lycopene
content in processing cultivars claiming “High pigment content” Proceedings of
the 8th International Symposium on the Processing Tomato. Acta
Horticolturae N. 613 331.
 Leoni, C.; (2003) Focus on Lycopene. Proceedings of the 8th International
Symposium on the Processing Tomato. Acta Horticolturae N. 613 357 – 363.
 Lycopene structure

Lycopene content is one of the most important

“nutritional” parameter for the evaluation of tomato
and tomato products.

C40H56(,,carotene)

Lycopene content is now becoming a trade

reference parameter which is as important as
Brix degrees ,pH, a/b …. ( Ex. Arizona, India,
Japan and China)
Lycopene characheristics

 Lycopene is the pigment principally responsible for

the charachteristic deep-red color of ripe tomato
fruits and tomato products.
 The color of lycopene is due to its many conjugated
carbon double bonds. Each double bond reduces
the energy required for electron transition to higher
energy states, allowing absorption, by the
molecule, of visible light of progressively lower
frequencies.
 Lycopene absorbs most of the visible spectrum, so
it appears red.
 Aim of the work

 The purpose of this paper is to assess

whether the proposed instrument can
be used to have a quick indication on
lycopene content in fresh tomato fruits,
industrial tomato products and control
during the processing phases.
The technical explanation of
the instrument

Technical explanation of the theory used

to assemble the spectro-colorimeter
Maselli Misure LC01 studied for lycopene
analysis will be presented by one of the
know how researchers:
Andrea Maselli.
Spectrophotometers

Transmittance:
The light goes
through the sample

Reflectance:
45°
The light is
reflected back by
the sample
Sensor
product
interface
 LC01 Maselli - Spectrophotometer
 Measures the diffuse reflectance
 LED machine

LC01 controls 7 visible LED + 1

NIR LED for the Scattering
Compensation

Large area photodiode

light sensor

Light sources
 LC01 Maselli - Spectrophotometer
 Measures the diffuse reflectance
 LED machine

LED
Collimation
- For each LED there is a lenses
Intereference
specific interference filter in Filter
Focusing lens
order to have 5 nm spectral
resolution and a series of
lenses to maximize the light
power.
 LC01 Maselli - Spectrophotometer
 Measures the diffuse reflectance
 LED machine

Each LED is switched

on sequentially

optical
core
 LC01 Maselli - Spectrophotometer
 Measures the diffuse reflectance
 LED machine

-The KM values
(Absorption) are calculated
from the diffuse reflectance
measurements where
S ( scattering ) is a function
of the NIR LED.
 LC01 Maselli - Spectrophotometer
 Measures the diffuse reflectance
 LED machine

The X,Y,Z and L,a,b coordinates are

indirectly calculated starting from the
visible reflectances measured.

Color must be calculated without

Scattering compensation
 Lycopene : transmittance and
diffuse reflectance

Absorbance from transmittance KM (absorbance) from diffuse reflectance

1. Lycopene 505 nm characteristic

absorption peak is at around 1. Lycopene characteristic
505 nm absorption peak absorption is at
565 nm
565 nm
2. Lycopene is measured
2. Lycopene has to be without exctraction, sample
exctracted with a solvent to preparation is not request
be measured

SADLER LC01
 Transmittance - Sadler Reflectance – LC01 - Maselli
Tomato sample

Solvent Lycopene extraction

Tomato as it is into the becker

Surnatant phase in the cuvette

Reflectance Measurament

Transmittance measurament

Lycopene Concentration
 Samples used for the trial

The trials were performed on 2 different groups, the

former consisting of samples with highly homogeneous
Group
profile Homogeneous
1 :(from 2: Non-
Groupthe
puree to paste products), latter of
samples homogeneous samples
samples with heterogeneous shape (pulp – diced pdcts
and raw matter). This was done in order to cover almost
 Tomato puree  Diced tomatoes (dice or
all possible variability of the physico-chemical
cubes)
(5 x 3= 15 samples)
characteristics of tomato products (5 x 15 = 75 samples)
 Tomato concentrates (range- (5 x 15 = 75 samples)
12 – 38 °Bx) (5 x 15 = 75  Raw tomatoes (homogenized
samples) under vacuum)
 “Crushed” tomatoes -passata (5 x 3= 15 samples)
(1,2 and 0,8 mm refining
sieve)
(5 x 10 = 50 samples)
Correlations
LHPLC LLC01 L__HUNT A__HUNT B__HUNT a/b Hunt L_LC01 A_LC01 B_LC01 a/b LC01
Lycopene HPLC Pearson Correlation 1,0000 0,9926 -0,9334 0,2859 -0,7490 0,8127 -0,8544 0,6300 -0,2719 0,8626
Sig. (2-tailed) . 0,0000 0,0000 0,0001 0,0000 0,0000 0,0000 0,0000 0,0001 0,0000
N 202 192 177 177 177 177 192 192 192 192
Lycopene LC01 Pearson Correlation 0,9926 1,0000 -0,9217 0,2444 -0,7696 0,8121 -0,8642 0,6168 -0,4008 0,8619
Sig. (2-tailed) 0,0000 . 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000
N 192 320 305 305 305 305 320 320 320 320
L__HUNTER Pearson Correlation -0,9334 -0,9217 1,0000 -0,3752 0,7727 -0,8752 0,9304 -0,6516 0,6445 -0,8987
Sig. (2-tailed) 0,0000 0,0000 . 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000
N 177 305 305 305 305 305 305 305 305 305
a__HUNTER Pearson Correlation 0,2859 0,2444 -0,3752 1,0000 0,0129 0,5461 -0,1997 0,7561 0,2334 0,5349
Sig. (2-tailed) 0,0001 0,0000 0,0000 . 0,8231 0,0000 0,0004 0,0000 0,0000 0,0000
N 177 305 305 305 305 305 305 305 305 305
b__HUNTER Pearson Correlation -0,7490 -0,7696 0,7727 0,0129 1,0000 -0,8275 0,7674 -0,3860 0,7633 -0,7186
Sig. (2-tailed) 0,0000 0,0000 0,0000 0,8231 . 0,0000 0,0000 0,0000 0,0000 0,0000
N 177 305 305 305 305 305 305 305 305 305
a/b HUNTER Pearson Correlation 0,8127 0,8121 -0,8752 0,5461 -0,8275 1,0000 -0,7732 0,7451 -0,5283 0,9123
Sig. (2-tailed) 0,0000 0,0000 0,0000 0,0000 0,0000 . 0,0000 0,0000 0,0000 0,0000
N 177 305 305 305 305 305 305 305 305 305
L_Maselli LC01 Pearson Correlation -0,8544 -0,8642 0,9304 -0,1997 0,7674 -0,7732 1,0000 -0,5079 0,5914 -0,8459
Sig. (2-tailed) 0,0000 0,0000 0,0000 0,0004 0,0000 0,0000 . 0,0000 0,0000 0,0000
N 192 320 305 305 305 305 320 320 320 320
a_Maselli LC01 Pearson Correlation 0,6300 0,6168 -0,6516 0,7561 -0,3860 0,7451 -0,5079 1,0000 0,2648 0,8411
Sig. (2-tailed) 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 . 0,0000 0,0000
N 192 320 305 305 305 305 320 320 320 320
b_Maselli LC01 Pearson Correlation -0,2719 -0,4008 0,6445 0,2334 0,7633 -0,5283 0,5914 0,2648 1,0000 -0,2949
Sig. (2-tailed) 0,0001 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 . 0,0000
N 192 320 305 305 305 305 320 320 320 320
a/b Maselli LC01 Pearson Correlation 0,8626 0,8619 -0,8987 0,5349 -0,7186 0,9123 -0,8459 0,8411 -0,2949 1,0000
Sig. (2-tailed) 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 0,0000 .
N 192 320 305 305 305 305 320 320 320 320
** Correlation is significant at the 0.01 level (2-tailed).
Statistical analysis (Bivariate Correlation) of data was
performed using the program SPSS Release 10.0 for
Windows (1999,SPSS Inc.).
 Results of Statistical Analysis
Total Samples - Lycopene HPLC data VS Lycopene with LC01 data

R2= 0,985

Model Summary
Model R R Square Adjusted R Std. Error of the Estimate
Square
1 0,9926 0,9853 0,9852 24,3997
a Predictors: (Constant), LLC01
b Dependent Variable: LHPLC

Coefficients
Unstandardized Standardized t Sig. Correlations
Coefficients Coefficients
Model B Std. Error Beta Zero-order
1 (Constant) -11,042 2,6058 - 3,52E-05
4,23736
LLC01 0,971 0,0086 1 113 0 1
a Dependent Variable: LHPLC
Results of Statistical Analysis
GROUP 1 (Homogeneous Samples) Lycopene HPLC VS Lycopene with LC01

900,0
y = 1,0452x + 0,1655
800,0 2
R = 0,9918
700,0
600,0
LC 01

500,0
400,0
300,0
200,0
100,0
0,0
0,0 100,0 200,0 300,0 400,0 500,0 600,0 700,0 800,0
HPLC

Model
Summary
Model R R Square Adjusted R Std. Error of the Estimate
Square
1 0,9927 0,9854 0,9852 27,0702
a Predictors: (Constant), LLC01
b Dependent Variable: LHPLC

Coefficients
Unstandardized Coefficients Standardized t Sig.
Coefficients
Model B Std. Error Beta
1 (Constant) 4,092 5,2408 0,78086 0,43698
LLC01 0,945 0,0123 1 77 0
a Dependent Variable: LHPLC
Results of Statistical Analysis
GROUP 2 (Non Homogeneous Samples) Lycopene HPLC VS Lycopene with LC01
140,0
y = 0,672x + 46,573
120,0 2
R = 0,7354

100,0

LC 01 80,0

60,0

40,0

20,0

0,0
0,0 20,0 40,0 60,0 80,0 100,0 120,0 140,0
HPLC

Model Summary
Adjusted R
Model R R Square Square Std. Error of the Estimate
1 0,7223 0,5217 0,5120 20,4294
a Predictors: (Constant), LLC01
b Dependent Variable: LHPLC

Coefficients
Standardized
Unstandardized Coefficients Coefficients t Sig.
Model B Std. Error Beta
1 (Constant) -6,655 12,7909 -0,5203 0,605196
LLC01 0,934 0,1277 1 7 9,72E-10
a Dependent Variable: LHPLC
 Conclusions

The aim of this study was to assess the performance

in terms of feasibility of a new instrument (Maselli
LC01) for quick and reliable measurement of
lycopene in tomato products.

On the total of samples considered (Group 1 and

Group 2 together) a good correlation (R2= 0,98) was
obtained between lycopene amount measured with
Maselli LC01 spectro-colorimeter and that measured
using the HPLC method.
 Conclusions
Notwithstanding the good correlation obtained on total
samples, in the case of non-homogeneous low-
lycopene products, the correlation coefficient shows a
lower precision degree (R2= 0,73) especially in the case
of the lowest lycopene amount.

The colour parameters measured with the HunterLab

Labscan XE (Spectro-Colorimeter) used as reference
instrument at SSICA, shows a positive, although not
exhaustive, correlation between colour parameters L (R2=
0,93), a/b ratio (R2= 0,81) with lycopene concentration
(Bivariate correlation).
Enjoy your red processed tomatoes……
Thank you for your attention.