Thus, the analytical curve is presented as −log (I/I0) vs. the cop- per concentration.

For this,
a proper R G or B channel was used. Other mathematical manipulation could be performed
such as the R, G, or B values directly, or its combination as R × G × B generating
exponential, polynomial or linear behavior versus concentration. However, −log (I/ I0) is
commonly used once it is linearly correlated with the concentra- tion being very employed
for colorimetry and spectrophotometry [30, 37,38]. By using this calculation, it is possible
to use a reference value (Blank) and to avoid performing the subtraction from the analytical
sig- nal, being similar to the Lambert-Beer law for colorimetry. The signal captured from a
digital camera is due to reflective, however, this signal is correlated with the absorbed
signal, and so, the logarithm signal can be used without loss of performance.

1.1. Optimization of Operating Parameters of Imaging System

To evaluate the effect of light intensity on the analytical response it was employed a
luximeter and the light intensity incident varied at five levels: 20, 35, 90, 240 and 980
LUX. To evaluate the effect of sample volume on the analytical response, it was varied the
volume of the solu- tion of the dye Brilliant Blue 1.0 × 10−5 mol L−1 of 100 to 800 μL with
100 μL of increment on the spot. To verify the RGB channels linear re- sponse due to the
analyte concentration, analytical curves were con- structed employing the dyes Brilliant
Blue, Tartrazine and Direct Red 80, whose concentration ranges were tested
from 1.00
× 10−6 mol L−1 to 1.00 × 10−4 mol L−1, using the light intensity and
the volume determined in earlier studies. The curves were constructed in triplicate, an
image for each curve was captured and the coefficient of determination was evaluated.
The effect of exchanging cameras on the analytical response was assessed by comparison
of two standard curves constructed using Bril- liant Blue dye, and the images were
captured by a camera Sony Cyber- shot DSC-W610 with resolution 14.1 Megapixels (MP)
and also through camera Sony Cyber-shot DSC-S730 with 7.2 MP. Assays were performed
in replicate (n = 7) with only one image being captured for each test. The results were
compared using the paired t-test with 95% confidence.
1.2. Copper(II) Determination in Sugar Cane Spirit

For the copper determination in sugar cane spirit, it was employed the cuprizone, a
bidentate organic reagent, which forms with copper a blue chelate in a slightly alkaline
medium [39], as shown in Fig. 2.
Initially, studies were conducted to select the concentration of cuprizone that would
generate a range of suitable colors for screening analysis type. The concentration of
cuprizone was evaluated in four levels using a fixed concentration of Cu(II) (1.25
mg L−1; 0.0019 × 10−3 mol L−1): 44 (0.16 × 10−3 mol L−1), 88 (0.32
× 10−3 mol L−1), 132 (0.48 × 10−3 mol L−1) and 176 mg L−1 (0.64 × 10−3 mol L−1) all of
them with a fixed volume of 100 μL. Furthermore, to verify the effect of the kinetic of
reaction on the analytical response, the reaction was monitored at 1 min intervals for 10
min employing a solution of Cu(II) 1.25 mg L−1 (0.0019 × 10−3 mol L−1) at a volume of 400
μL. The analytical blank was obtained by replacing the Cu(II) so- lution for distilled water.
After reaction optimization, the volumes were fixed in: 100 μL phosphate buffer pH 9, 400
μL solution of Cu(II)1.25 mg L −1 or sample and 100 μL of cuprizone. The tests were per-
formed in triplicate, and an image was captured for each test.
Effects of possible interferents were evaluated for determining Cu(II) in different proportions
of analyte/interferent. The study of the matrix effect was done by comparing the parameters of
the analytical curve obtained for standard aqueous solutions of Cu(II) with the param- eters of
analytical curve using a sugar cane spirit free of copper(II). The mean responses were compared
using statistical test F (Snedecor) of homogeneity of variances and through the paired t-test with
95% confidence.
Recovery studies were tested in unaged and aged sugar cane spirit samples employing
standard addition method using three concentra- tion levels: 0.75; 2.88 and 5.00 mg L−1.
The accuracy of the method was evaluated by comparing the results obtained by digital
analysis method with the reference method (Flame Atomic Absorption Spectro- photometry)
[40]. The results obtained by both methods were com- pared using the paired t-test at 95%
confidence.
The DIB method was applied for determination of Cu(II) in samples of aged and unaged
sugar cane spirit collected in supermarkets in the cities of Viçosa and Bom Jesus do
Amparo - Minas Gerais/Brazil. No treatment of samples was need before the analysis.