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Color analysis of Andean Blackberry (Rubus Glaucus) by the addition of Chlorpyrifos through colorimetry.


Background: The blackberry has a wide acceptance between the people, because his nutritional value, his pleasant aroma and flavor. However, these good properties make of blackberry a target for insects.

Objective: In this paper, it was used the Chlorpyrifos in order to control the pest in crops and reduce the loses of the same. This compound is toxic, so can cause serious damage in the human body when this is ingested, due to the intrinsic toxicity of this class of molecules. It was applied Chlorpyrifos on the variety Andean Blackberry or Rubus Glaucus with a concentration of 0.024. In order to verify the presence of this substance, it was used the colorimetry as measure method. Through this methodology, it was characterized the Andean Blackberry, the commercial version of the pesticide used and the relationship between them in a period of time of 0 to 107 min. It was obtained an average in the base color of 22.58 and 171.3 degrees for this variety of blackberry and the pesticide.

Conclussions: In the addition of the pesticide on the fruit, it was found that principal component of the base color was the fruit, while for the Cab chroma parameter was the pesticide, obtaining a reduction of 50% in this parameter, taking as base the fruit without pesticide. Therefore, it was found a relation inversely proportional for the parameters of the CIE Lab space color and the time of the fruit from the exposition with the pesticide.


Rubus Glaucus, CIE Lab, Pesticide, Hue Angle.


The blackberry fruit is one of the most popular fruits in the market for his pleasant aroma, flavor and nutritional properties, which is convert into a fruit widely harvested [1]. The variety most commonly grown in mountainous areas is the Castilla blackberry (Rubus Glaucus), because it requires between 12 [degrees] and 19 [degrees] C and high humidity for proper development [1], [2]. However, these plants are affected by different crop pests, which attack the fruit and their leaves, as fruit flies, ants, mites, among others [3], [4]. Recently it has achieved a decrease in the amount of lost crops through the use of pesticides [5].

A lot of pesticides used currently can generate adverse effects on humans when are ingested. Due to this, their use is regulated by different government authorities like the Agricultural Colombian Institute (ICA) or the Environmental Protection Agency of United States (EPA) [5]-[7]. The intrinsic toxicity of these compounds are used to control pests and increase the efficiency of crops. In crops of Andean blackberry is commonly used the insecticide chlorpyrifos [8], [9]. Which it is a synthetic compound, reason why is not normal to find residues of them in the environment; This coupled with a poor application at the same crop, has been producing an increase in people poisoned by pesticides mainly in rural areas [5], [10], [11].

The O, O-diethyl O-3,5,6-trichloropyridin-2-yl phosphorothioate, is commonly known as Chlorpyrifos. Its main active site is the organophosphate area. When this compound is ingested attacks the nervous system of the host, because it blocks the acetylcholinesterase (AChE), which is an enzyme used in the proper functioning of the nervous system [12], [13]. In high doses it can produce damage to the muscular system due to the interruption in the communication between the brain and muscles, which can lead to tachycardia, cramps, excessive salivation, among others [11], [14].

Due to the development of resistance by insects to the pesticides used, it is required an increasing the concentration and dose used to kill an insect, reason why has increased the number of intoxicated people with this compound mainly in rural areas [15]. This can be avoided by taking measurements of the concentration of this insecticide in the fruit before being distributed [2]. However, most of pesticides quantification techniques are destructive, whereby the sample is subjected to various chemical processes in order to isolate the compound of interest [16]. One of the techniques used for this process in chromatography, which separates each of the compounds in the sample and thus to compare the desired substance with a pattern in order to determine concentration [17], [18]. This paper presents the methodology to use non - destructive techniques to detect the pesticide in fruit.

To do this, the colorimetry was used in food. This technique is based on the analysis, measuring and quantifying the color of an object through the study of light reflected therefrom using a light pattern [13], [19]. The main components of this technique is the base color and saturation thereof, which is related to the amount of reflected light for each wavelength over a surface. In order to standardize the measures, the color spaces CIE Lab and CIE XYZ were used [20]-[22]. Where for the first the "L" indicates the brightness, "a*" is a scale of values between green and magenta colors, and "b*" is the scale between blue and yellow. While the second color space is used to define the colors perceived by the human eye. In this model, "Y" means brightness; "Z" is approximately equal to the blue stimulus in the human eye (S cones), and "X" is a mixture aimed at the sensitivity curve from red to green (cones L and M).

Based on the above, in this paper is presented the analysis of the Andean blackberry (Rubus Glaucus), as pulp and peel, and its variation by using the insecticide Chlorpyrifos on it, through colorimetry. Obtaining the quantization parameters of the insecticide based on color.



In the experiment Chlorpyrifos commercial was used, recommended for crops of Pineapple, Blackberry, Tomato and Banana. In order to ensure product quality, the values found in the safety data sheet were compared with values taken from the literature. In addition, the search for the main properties of the insecticide was performed.

The experimental work was conducted on the campus of New Granada Military University in Cajica, Colombia. This area of work was chosen, because it was an area of blackberry and strawberry production. Furthermore, it took into account the ambient temperature at which the experiment was performed, that the day was not rainy, and it was not very sunny, in order to ensure that the pesticide application was similar to that made by a farmer.

The molar ant the ppm concentration of commercial solution was determined using the equations 1 and 2, wherein MW was the molecular weight and concentration p was the density. The initial concentration of chlorpyrifos was 480 g / L.



Chlorpyrifos solution added to the blackberries was prepared using the commercial version of the product and distilled water. In order to obtain the concentration of the solution used, it begins with equation 3, where C was the concentration, m was the mass and V was the volume.

C = (m/v) [m.sub.before] = [m.sub.after] = C * V (3)

For a solute in a solution, the mass before and after was the same.Due to, more solute was not added. With the above, equation 4 gives the concentration ratio of the solution made.

[C.sub.1] * [V.sub.1] = [C.sub.2] * [V.sub.2] [([C.sub.1]*[V.sub.1])/[V.sub.2]] = [C.sub.2] (4)

[(1.335 M * 5 mL)/275 mL] = 0.024 M

The same procedure for the concentration in ppm was done. A result of 8065 ppm was obtained. This solution was chosen to work with the blackberries because it was the way in which farmers use the pesticide, when was applied on crops.

Andean Blackberry:

To perform the tests on the fruit, a sampling of 5 different markets in Bogota was made, where it was collected 500 g of freshly harvested fruit by place. For transfer to the campus, the fruit was packed in a Styrofoam box filled with ice and with the fruit properly labeled.

On campus, of the 2.5 kg of fruit, 500 g of Andean Blackberry were chosen (Fig. 1). Taking into acount that the fruit was not damaged and that not had presence of pests. Three similar piles were made, of which the first was used for the analysis of the shell surrounding the mora, the second, was used to examine the pulp when the shell was removed, while for the third analyzed the pulp when this was cut in half.

Was added 0.1 mL of the solution of pesticide on each cut blackberry, in order to analyze the behavior of the fruit when is in presence of pesticide. For which the blackberry was exposed to the environement and the colorimetry measurement was made thorough time.


The colorimetry was made with a non-contact spectrophotometer Flame-S of Ocean Optics (Fig. 2, No. 2), which had a DET2B detector with a detection range from 200 to 1100 nm, with a slit INTSMA-025 and quartz cells of 1 cm. The light source used was a DH-2000-BAL with deuterium and tungsten lamp from Ocean Optics (Fig. 2, No. 1). The reflectance fiber used had a premium grade of 400 pm with solarization-resistant, with a wavelength range of 200 to 1000 nm. For the equipment calibration, a standard diffuse of reflectance from Ocean Optics was used, made of polytetrafluoroethylene (PTFE), with cover of anodized aluminum, hydrophobic, chemically inert and very stable, with a reflectivity higher to 98%.

All measurements of colorimetry were taken in triplicate, in order to ensure the accuracy in each sample.

Analysis results:

One of the most commonly used parameters for measuring color is the hue angle, which indicates the base color of the sample, the wavelength that reflects more light, this parameter is related to the Cab chroma, which indicates the quality of the intensity or the color saturation, for example, a gray color gives a low value of Cab chroma while a bright red will have high values.

The Ocean View software with which the samples were measured, gives us the parameters of color space CIE Lab and CIE XYZ, where the first indicates relationships of color, while the second indicates relations taking as starting point the colors that can be detected by the human eye. However, the last color space was not used, instead the normalized parameters through Equation 5 were obtained, and a combination of these parameters with the originals were used.

x = [X/(X + Y + Z)] y = [Y/(X + Y + Z)] z = [Z/(X + Y + Z)] (5)

The CIE Lab and CIE xyz parameters were visualized in the software Matlab R2016a[R], where the average and the standard deviation of each sample was performed. In order to obtain the dispersion of the data, and thus the change of each sample in the analysis.

Additionally, the spectra of wavelength vs. intensity of each sample were taken. In order to obtain the characteristic peaks of each sample, besides looking for the pattern of light reflection of the fruit.

In order to verify the data dispersion in each one of the spectra taken by triplicate, an analyzes of the standard deviation was performed. Wherein each point of each sample take for triplicate was compared, and then, the average of all standard deviations were obtained.


Initially, to each blackberry was added the insecticide Chlorpyrifos such as described in the section of methodologies and materials. In the experiment were taken four samples through the time. For each one of the measurements made, an alphanumeric code was designated as indicated in

Table 1. Due to the Hue angle and Cab chroma parameters were intrinsically correlated in the base color of each sample, an analysis on the whole was performed. In the

Table 1 was observed the standard deviation for the Cab chroma represented with horizontal lines.

For both parameters (Cab chroma and Hue angle) a high standard deviation of the data for blackberry pulp was found, when it was slice. This was due to the change of color in the inner part of the fruit, as evidenced in

Fig. 1. Where, the outside has a dark red color, which indicates high values of color saturation or Cab chroma, while the heart of the fruit was reddish white with low saturation values.

The other properties not shown a high deviation in the samples. The insecticide Chlorpyrifos pure (commercial concentration) obtained the lowest value in the standard deviation, ensuring an adequate characterization of the analyte. The Andean blackberry used in experiments was characterized. It was found the average of the base color on the color wheel was 22.58 [+ or -] 10.40, these values were among the orange and red colors.

The samples contaminated with Chlorpyrifos do not present a clear pattern as a function of time, however, by adding Chlorpyrifos on the blackberry (Mt0, MT1, MT2, Mt3) the color saturation decreases about 50% of the value of blackberry unpolluted (Pulcm). Hue angle value follows the expected behavior, in which the pesticide did not affect the color base of the sample, due to the minimal amount of Chlorpyrifos applied.

The initial analysis was complemented with the 3D color space CIE Lab in the Figure 3. In the figure there the points of characterization for the blackberry, the insecticide Chlorpyrifos in his commercial form and the variation over time for the fruits contaminated with the pesticide.


Fig. 3, the colors observed with the colorimeter for each point were shown (True color). In the characterization of the blackberry were obtained values in line with the expectations based on the color seen by the human eye. The color of slice blackberry had more light than the peeled blackberry for the reasons already exposed. For the fruit polluted with the pesticide, exist a clear increase in brightness due to the addition thereof.

In order to obtain the color variation through the time when the pesticide was on the fruit were determined the relationships of the CIE Lab color space, which were shown in

Fig. 4.

The parameters involved had the same descendant behavior versus time. As time increases, the L *, a * and b * tend to take the same values that had the unpolluted fruit. One reason for this was the decrease in the concentration of pesticide in the fruit, due to the possible metabolism of this compound with a component of the organic matrix, to volatilization of the compound in the environment, the difference concentration between the surface of the fruit and the air surrounding it.

In addition to the measurements of the space color CIE Lab, the values for the CIE XYZ color space were made. These values were normalized as shown in the section of methods and materials, obtaining the values reported in

Table 2.

In this space of standardized color, it can observe the changes that occur in the different samples. In the changes was a clear decrease in the "x" parameter between the fruit without pesticides and contaminated fruit, so it reduce the change in the color red was an important factor, in order to determining the presence of the insecticide Chlorpyrifos on the Andean blackberry. Instead, the "z" parameter representing the blue color seen by the human eye, has a significant increase, due to the color of the insecticide. On the contrary, the parameter "y" was not significant.

In order to complement the analysis, the reflection spectra was taken for each sample analyzed, these were shown in Fig. 5. The first part of the figure shows the characterization of Andean blackberry and the commercial insecticide used..

The Andean blackberry had no peaks or valleys representative, while the commercial pesticide had a unique fingerprint that differentiates it from other compounds. By adding the pesticide to fruit was observed an increase in the signal intensity from 350 nm to 1000 nm, indicating the presence of a foreign agent on the surface of the fruit. This substance can be confirmed by observing the valleys at 760 nm and 820 nm, also there was reflection peak with less intensity at 430 nm. These valleys and peaks were characteristic of the insecticide chlorpyrifos. Through the time, exist a clear reduction of the intensity signals, indicating that the presence of the compound decreases with time.


Chlorpyrifos insecticide was widely used in various fruit crops around the savannah of Bogota, mainly due to its effectiveness to kill different pests. This was achieved by the organophosphorus function of the compound, which causes a blocking to the Acetilcholinesterase enzyme. Therefore, an analysis of characteristic properties of the insecticide was developed, taking into account his commercial version.

The Andean blackberry was widely grown around the savannah of Bogota; reason why recently have increased cases of poisoning with this pesticide. The blackberry used in the experiment was characterized by finding the base color of the different parts of the fruit. In addition, the change was determined in the color by adding the pesticide on the surface of the same, obtaining a wide variation in the red and the blue, this last by the pesticide, while for brightness there no significant change.

The relationship between CIE Lab color space and the time after application of the pesticide was found. Obtaining a decrease in the intensity of the spectra. The lines with major and minor absorption were determined in the color spectrum, finding a reflection peak at 430 nm and two valleys in 760 nm and 820 nm.


The authors would like to offer their special gratitude to the Research Vice-chancellorship of Nueva Granada Military University for financing the research project IMP_ING 1777 titled: "Analisis de residuos de plaguicidas en frutas tropicales en Colombia para la prediccion de posibles efectos en la salud humana", 2016.


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(1) Jorge Eliecer Buitrago Salazar, (1) Olga Lucia Ramos Sandoval, (1) Dario Amaya Hurtado

(1) Engineering in mechatronics, Faculty of Engineering, Nueva Granada Military University, Bogota D. C, Colombia.

Received 22 August 2016; Accepted 1 November 2016

Address For Correspondence:

Dario Amaya, Engineering in mechatronics, Faculty of Engineering, Nueva Granada Military University, Bogota D. C., Colombia.

Copyright [c] 2016 by authors and American-Eurasian Network for Scientific Information.

This work is licensed under the Creative Commons Attribution International License (CC BY).

Table 1: Cab Chroma and Hue angle values for the samples.

Blackberry's Sample    Acronym  Hue angle

Shell                  Casm      19,572 [+ or -] 6,531
Slice Pulp             Pulcm     18,835 [+ or -] 14,731
Peeled Pulp            Pulpm     29,346 [+ or -] 4,953
Pesticide t = 0 min    Mt0       16,251 [+ or -] 3,651
Pesticide t = 18 min   Mt1       15,895 [+ or -] 1,857
Pesticide t = 51 min   Mt2        9,145 [+ or -] 6,225
Pesticide t = 107 min  Mt3       15,231 [+ or -] 4,344
Pure Chlorpyrifos      Pure     171,327 [+ or -] 1,474

Table 2: Values for xyz representation.

Values  x             y             z

Casm    0,618+-0,066  0,330+-0,031  0,052+-0,036
Pulcm   0,560+-0,075  0,358+-0,029  0,082+-0,049
Pulpm   0,632+-0,021  0,328+-0,008  0,040+-0,015
Mt0     0,487+-0,007  0,388+-0,004  0,125+-0,004
Mt1     0,490+-0,012  0,386+-0,006  0,123+-0,006
Mt2     0,479+-0,007  0,389+-0,005  0,132+-0,004
Mt3     0,501+-0,021  0,380+-0,010  0,118+-0,011
Pure    0,418+-0,001  0,435+-0,001  0,147+-0,001
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Author:Salazar, Jorge Eliecer Buitrago; Sandoval, Olga Lucia Ramos; Hurtado, Dario Amaya
Publication:American-Eurasian Journal of Sustainable Agriculture
Article Type:Report
Geographic Code:3COLO
Date:Sep 1, 2016
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