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DETERMINATION OF OPTIMAL PARAMETERS FOR THE PROTEIN INCREASE IN QUINOA (Chenopodium quinoa Willd.) BY THE GERMINATION PROCESS

DETERMINACION DE LOS PARAMETROS OPTIMOS PARA EL INCREMENTO PROTEICO EN LA QUINUA (Chenopodium quinoa Willd.) POR EL PROCESO DE GERMINACIÓN

Kim Eduardo Torres Huanca 1 , Lucía Vanessa Cruz Morales 1 , Paola Michelle Molina Díaz 1 Juan Lopa Bolivar 2 , Erika Pachari Vera 2

Key

words:

SUMMARY Quinoa is a grain with high nutritional value used by the old inhabitants of the Highlands of Titicaca Lake mainly for the protein content. Peru is the first producer with 42000TM, but the consumption is 1.14kg per person/year. It is not only necessary increase the production, also it is important increase the quality of the product. The purpose of the research is to determine the optimum parameters to induce the increase of quinoa proteins through technological processes as the germination, where best protein increase was 38.68% for Pasankalla quinoa in 24h followed by Real quinoa with 26.76% in 48h, both at 30°C. The protein content of these samples are 19.42% and 15.75% respectively. This increase is influenced mainly by variety of the quinoa and the germination time, being the best time 24h for Pasankalla quinoa and 48h for Real quinoa, finally the optimum temperature was at 30°C.

RESUMEN La Quinua es un grano con alto valor nutricional usado por los antiguos habitantes del altiplano del lago Titicaca principalmente por el contenido proteico. Perú es el primer productor con 42000 TM, pero su consumo es de 1.14kg por persona/año. Siendo no solo necesario incrementar la producción, sino también es importante incrementar la calidad del producto. La propuesta de la investigación es determinar los parámetros óptimos para inducir el incremento proteico a través de procesos tecnológicos como la germinación, donde el mejor incremento proteico fue 38.68% para la quinua Pasankalla en 24 horas, seguida por la quinua Real con 26.76% en 48 horas, ambos a 30°C. El contenido proteico de estas muestras son 19.42% y 15.75% respectivamente. Este incremento es influenciado principalmente por la variedad de quinua y el tiempo de germinación, siendo el mejor tiempo 24h para la quinua Pasankalla y 48 horas para la quinua Real, finalmente la temperatura óptima fue de 30°C.

Germination

process,

quinua,

protein

increase,

protein.

Palabras claves:

Proceso

de

germinación,

quinua,

incremento

proteico,

proteínas.

INTRODUCTION Quinoa is a grain with high nutritional value, the inhabitants of the Highlands of Titicaca Lake, in the process of adaptation of their habitat, domesticated it, achieving more than two thousand varieties of quinoa with protein content responding to certain nutritional functions and each variety was produced in specific geographical and climatic conditions. Quinoa is one of the most viable alternative for the food crisis in the world and malnutrition especially of children, for its easy digestibility and essential amino

1. Student from Professional School of Food Industrial Engineering of San Agustin National University Arequipa Peru.

2. Professor from Professional School of Food Industrial Engineering of San Agustin National University Arequipa Peru.

3. Professor from Academic Department of Chemistry of San Agustin National University Arequipa Peru.

acids content, such as lysine which has the property of regenerating neural cells and help the transport of iron in the blood. Peru is the first producer of quinoa in the world. In 2011 the production was 42 thousand MT, of which 19% are destined for export (ADEX 2013) [1] and the rest for the domestic market, the consumption is 1.14 Kg per person in one year. Therefore, it is necessary not only to promote the increase of its production on the basis of the demand, but also opt to increase their protein content through technological processes such as germination arising through this research.

Taking into account the nutrition of the population and the current demand of vegetable protein and germinated seeds, the purpose of the research is to determine the optimum parameters to induce the increase of quinoa proteins through technological processes as the germination.

MATERIALS AND METHODS

RAW MATERIAL Commercial grains are used, all of these were purchased at fairs and markets in Juliaca city, Puno - Peru, taking into account three varieties shown by Mujica, 2006 [2] and Tapia, 2007 [3]: White Quinoa, Real Quinoa and Pasankalla Quinoa.

PROCEDURE TO PREPARE GERMINATED QUINOA Reception and selection of raw materials: The raw material obtained was selected by size, color and variety. Wash: The grains should be washed to remove saponin in the shell of the quinoa, in addition this operation is performed in order to remove some impurities present in the quinoa. Drying: Previously washed, the grains are dried by the Sun, being spread on a white tablecloth. Weighed: With the help of a scale was determined the quantity of raw material used in the process. Soaked: The quinoa grains should achieve the humidity required for starting the sprouting process. The grains were immersed in a glass container with water at temperatures of 20-30 °C for each variety of quinoa for approximately 6 hours (chaparro et al, 2010) [4], taking care that the water exceed the surface of the grains in 3:1 ratio. Germination: After soaking the samples were distributed in circular trays inside a camera, then it’s necessary to control the temperature for 24 and 48 hours. The change of water takes place every 6 hours. Drying: This was done to remove the water in the germinated grains, store samples and process them for protein determination.

DETERMINATION OF CONTENT AND INCREASE OF PROTEINS The proteins were determined by modified micro-Kjeldahl method proposed by Lopa, 2011[5]. The protein content was calculated as follows:

The calculation of the percentage of protein was determined by the following formula:

1)

14

=

1000

= = ℎ (0.1) = ℎ = (1.1417 1 ó 1.0425 2 3) 14 = ℎ

2)

% = 6,25100%

= = ℎ ℎ 6.25 =

The calculation of the protein increase was carried out with the following formula:

3)

∆% =

100%

= () =

STATISTICAL ANALYSIS The statistical software used for the design was "STATGRAPHIC centurion XV" for Windows. The statistical method applied was the “Completely Randomized Design (CRD), which discussed the statistical difference between established treatments (product of the interaction of the factors), then It was made the Tukey test to determine the best treatments with regard to viability and the size of the radicle, later the viable seeds were selected and used to determine protein content after the germination process. It was used Microsoft Office Excel 2013 for making the comparison between the protein content of each treatment, the increase of proteins too. Finally the optimal parameters and the influence of factors (temperature: 20 and 30 ° C, germination time: 24 and 48 hours, and variety of the quinoa: White Quinoa, Real Quinoa and Pasankalla Quinoa) were analyzed by a block design.

Table 1. Treatments

 

FACTORS

     

TEMPERATURE (° C)

TIME (HOURS)

VARIETY

TREATMENT

CODE

20° C

24

hours

White quinoa

TREATMENT 1

QB2011

20° C

24

hours

Real quinoa

TREATMENT 2

QR2011

20° C

24

hours

Pasankalla quinoa

TREATMENT 3

QP2011

20° C

48

hours

White quinoa

TREATMENT 4

QB2021

20° C

48

hours

Real quinoa

TREATMENT 5

QR2021

20° C

48

hours

Pasankalla quinoa

TREATMENT 6

QP2021

30° C

24

hours

White quinoa

TREATMENT 7

QB3011

30° C

24

hours

Real quinoa

TREATMENT 8

QR3011

30° C

24

hours

Pasankalla quinoa

TREATMENT 9

QP3011

30° C

48

hours

White quinoa

TREATMENT 10

QB3021

30° C

48

hours

Real quinoa

TREATMENT 11

QR3021

30° C

48

hours

Pasankalla quinoa

TREATMENT 12

QP3021

Source: Homemade

RESULTS AND DISCUSSION

DETERMINATION OF VIABLE QUINOA GRAINS

VIABILITY OF THE QUINOA GRAINS To determine viable grains, it is necessary to perform a test allowing to determine if they sprout in its entirety, so it has germinated each variety of quinoa to the temperatures given and at the time established, obtained the following results:

Table 2. Evidence of Multiple ranges for viability by treatments (Method: 95.0 percent Tukey HSD)

TREATMENTS

Cases

Average

Homogeneous

(%)

groups

QB3011

2

3.0

X

QB2011

2

4.5

X

QB3021

2

5.0

X

QB2021

2

6.5

X

QR2011

2

90.0

X

QR3011

2

93.0

XX

QP2011

2

96.5

XX

QP3011

2

99.0

X

QR3021

2

99.0

X

QR2021

2

99.5

X

QP3021

2

99.5

X

QP2021

2

100.0

X

Source. Homemade

The picture shows White Quinoa seeds germinated below 10%, this would indicate that for every 100 quinoa grains, less than 10 germinated. Real quinoa and Pasankalla quinoa have germinated almost in its entirety, taking the best result in Pasankalla quinoa germinated at 20°C. The quantity of germinated seeds become homogeneous after two days because the seeds have passed by an imbibition process, where the quinoa grains absorbed water. To supplement the information contained in table, below it is presented an analysis of the radicle size of the germinated grains, showing different varieties of quinoa, after subjecting them to temperatures and times given and comparing this information with the bibliography.

RADICLE SIZE

35 30 25 20 15 10 5 0 WHITE QUINOA WHITE QUINOA PASANKALLA PASANKALLA REAL
35
30
25
20
15
10
5
0
WHITE QUINOA
WHITE QUINOA
PASANKALLA
PASANKALLA
REAL QUINOA
REAL QUINOA
20°C
30°C
QUINOA 20°C
QUINOA 30°C
20°C
30°C
-
-
-
MUJICA 24 H
9.05
7.75
-
-
MUJICA 72 H
21.45
20.7
-
-
-
RESEARCH 24 H
0
0
11.80
12.60
10.60
9.20
RESEARCH 48 H
0
0.4
21.80
23.00
26.80
29.40
SIZE OF RADICLE
(MILIMETERS)

Figure 1. Radicle Size for each treatment and comparison with bibliography (Mujica et al, 2006)

Pasankalla Quinoa subjected at 30°C for 48 hours presented greater average radicle (29.4 mm), while White Quinoa subjected to each established treatments does not increase radicle, which contrasts with the information provided by Mujica et al, 2006 [2]; where indicates that White quinoa germinated at 20°C for 24 hours have a radicle size of 9.05 mm, which is greater than the data obtained in the research (0 - 0.5 mm), possibly due to the grain is not viable. Mujica et al, 2006 [2] also show that Pasankalla Quinoa have 7.75 mm of radicle size at 20°C sprouted for 24 hours, data lower than those obtained in the research (11.88mm at 20° C and 12.6mm at 30°C).

The data obtained in the research with 48 hours of germination in Pasankalla Quinoa (26.8mm at 20° C and 30° c 29.4mm) are higher than those shown by Mujica et al, 2006 [2] with 72 hours of germination (20.7mm at 20° C). Also there is a correlation between the days of germination and radicle size: if the germination is longer, the radicle is larger, this is given in the case of Real Quinoa and Pasankalla Quinoa. Furthermore the radicle grows more with temperature at 30°C, because the heat is a form of energy, when the water in contact with the seed increases the temperature, part of the supplied energy is invested in increasing the diffusion of water, therefore, increases the rate of water absorption, within certain limits. It has been found experimentally that a 10°C increase in temperature doubles the absorption rate at the beginning of the process of imbibition (UNALM LIMA, 2012) [6].

The viable seeds for the given treatments are Pasankalla Quinoa and Real Quinoa.

DETERMINATION OF THE PROTEIN INCREASE IN QUINOA AFTER THE GERMINATION

After the analysis of the viable grains and the radicle size, Pasankalla Quinoa and Real Quinoa were selected for the determination of the proteins and the protein increase, this results are shown below in figure 2a and 2b, the treatments were temperatures at 20 - 30°C, and 24 48 hours of time.

A)

25.00

20.00 15.00 10.00 5.00 0.00 PASANKALLA QUINOA PASANKALLA QUINOA REAL QUINOA 20°C REAL QUINOA 30°C
20.00
15.00
10.00
5.00
0.00
PASANKALLA QUINOA
PASANKALLA QUINOA
REAL QUINOA 20°C
REAL QUINOA 30°C
20°C
30°C
0 H
14.00
14.00
12.50
12.50
H
24
15.67
19.42
12.81
12.90
H
48
12.90
12.69
13.40
15.75
PROTEIN CONTENT (%)

B)

50.00 38.68 40.00 26.76 30.00 20.00 11.90 8.00 10.00 3.31 4.00 0.00 0.00 0.00 0.00
50.00
38.68
40.00
26.76
30.00
20.00
11.90
8.00
10.00
3.31
4.00
0.00
0.00
0.00
0.00
0.00
-10.00
-7.85
-9.33
-20.00
PASANKALLA QUINOA
PASANKALLA QUINOA
REAL QUINOA 20°C
REAL QUINOA 30°C
20°C
30°C
Día 0
0.00
0.00
0.00
0.00
Día 1
11.90
38.68
3.31
4.00
Día 2
-7.85
-9.33
8.00
26.76
PROTEIN INCREASE (%)

Figure 2. Protein content in Pasankalla quinoa and Real quinoa germinated at 20 30°C per 24 48 hours (A). Increase of proteins (%) in Pasankalla quinoa and Real quinoa germinated at 20 30°C per 24 48 hours (B).

Pasankalla quinoa has the high protein content (19.42%) when it was subjected to the germination process at 30°C per 24 hours followed for Real quinoa (15.75%) germinated at 30°C per 48 hours (Fig. 2a). Both cases are the result of the protein increase 38.68% and 26% respectively (Fig. 2b), these increases are the highest, but do not concord with the information exposed by Chaparro et al (2010) [4], who indicates that the protein content in germinated quinoa does not increase, the protein found in seeds with two and three days of germination is statistically equal to non-germinated quinoa (day 0). Parillo and Lupo (2009) [7], presented the results of a proximal analysis in White quinoa where in “day 0”, this had 14.71% of proteins, in “day 3” the proteins are 14.98% this response to 3.2% of increase. Researchers as Colmenares de Ruiz and Bressani (1990) [8] and Chirinos and Pachari (2007) [9] indicate an increase in the protein content after germination in kiwicha (grain similar to quinoa) and other seed as corn. Figure 2b also shows that the proteins of Pasankalla quinoa decrease below the percentage of “day 0” on the second day, this reduction is expressed in 7.85%, Chaparro et al (2010) [4] also indicate that there was a decrease of protein in White quinoa at the same time, expressed in 0.61% below the percentage of “day

0”.

Pasankalla quinoa shows increase in the first day, and a decrease in the second day. Real quinoa shows a continuous increase for the two days, this characteristic should be for the variety of the Quinoa.

DETERMINATION OF THE OPTIMAL PARAMETERS AND MORE INFLUENTIAL FACTORS IN THE GERMINATION PROCESS OF QUINOA

A)

16 12 8 VARIETY OF QUINOA PASANKALLA QUINOA 4 REAL QUINOA 0 PROTEIN INCREASE (%)
16
12
8
VARIETY OF QUINOA
PASANKALLA QUINOA
4
REAL QUINOA
0
PROTEIN INCREASE (%)
30 20 VARIETY OF QUINOA PASANKALLA QUINOA 10 REAL QUINOA 0 -10 PROTEIN INCREASE (%)
30
20
VARIETY OF QUINOA
PASANKALLA QUINOA
10
REAL QUINOA
0
-10
PROTEIN INCREASE (%)

20

24 48 30 TIME OF GERMINATION TEMPERATURE (°C) B) 23 19 15 11 7 TIME
24
48
30
TIME OF GERMINATION
TEMPERATURE (°C)
B)
23
19
15
11
7
TIME OF GERMINATION
3
24
hours
48
hours
-1
20
30
C)
TEMPERATURE (°C)
PROTEIN INCREASE (%)

Figure 3. Interaction between protein increase (%) and temperature (°C) over the variety of quinoa (A). Interaction between protein increase (%) and time of germination (hours) over the variety of quinoa (B). Interaction between protein increase (%) and temperature (°C) over the time of germination (hours) (C).

Real quinoa presents more increase of proteins than Pasankalla quinoa in different temperatures, in this case, temperature does not influence over the protein increase (Fig. 1a), but Real quinoa increase the protein content from 24 hour to 48 hours, and this is different for Pasankalla quinoa, which decrease the protein content at the same time, this difference should be given for the variety mainly (Fig. 3b). When

the germination is per 24 hours, the protein increase is better than 48 hours, this result is influenced by the variety of the quinoa, mainly for Pasankalla quinoa, which present the highest protein increase in 24 hours (this increase is more than Real Quinoa) (Fig. 3c) This observations indicate that the factor with influence over the germination process in quinoa grains are mainly the variety, next the time of germination, being 24 hours the best time for Pasankalla Quinoa and 48 hours for Real Quinoa, finally the temperature being the optimum at 30°C.

CONCLUSIONS

Quinoa grains presented protein increase after the use of the treatments, the best protein increase was 38.68% for Pasankalla quinoa in 24 hours at 30°C followed by Real quinoa with 26.76% in 48 hours at 30°C, also the protein content of these treatments are 19.42% and 15.75% respectively. The increase is influenced by variety of the quinoa and the time of the germination, being the best time 24 hour for Pasankalla quinoa and 48 hours for Real quinoa, finally the temperature being the optimum at

30°C.

BIBLIOGRAPHY

1. ADEX Asociación de Exportadores Perú. Conferencia de planificación y solución de la problemática de siembra, producción e industrialización de la quinua en el Año Internacional de la Quinua. January 2013. Arequipa Perú.

2. Mujica Ángel, Ortiz Rene, Bonifacio Alejandro, Saravia Raúl, Corredor Guillermo, Romero Arturo, Jacobsen Sven Erik. Agroindustria de la Quinua (Chenopodium quinoa Willd.) en los países andinos. Edit.: Universidad Nacional del Altiplano. 2006. Puno Perú.

3. Tapia Mario E.; Fries Ana María. Guía de Campo de los Cultivos Andinos. 2007. Lima Perú.

4. Chaparro Rojas, D.C.; Pismag Portilla, R.Y.; Elizalde Correa, A.; Vivas Quila, N.J. y Erazo Caicedo, C.A. Effect of the germination on the protein content and digestibility in amaranth, quinua, soy bean and guandul seeds. 2010. Colombia.

5. Lopa Bolivar, Juan. Guía de laboratorio de Química Analítica de los Alimentos. Edit.:

Universidad Nacional de San Agustín. 2011. Arequipa Perú.

6. UNALM Universidad Nacional Agraria la Molina. Efectos de la temperatura sobre la germinación. 2012. Lima Perú.

7. Chirino Ch. Isabel Lupo Ch. Rolando. Determinación de Parámetros óptimos: Relación quinua malteada, cascara de platano y agua que influyen de la preparación de una bebida fermentada a partir de quinua (Chenopodium quinoa Willd.) y plátano (Musa cavendishi). Edit.: Universidad Nacional de San Agustín. 2009. Arequipa Perú.

8. Colmena de Ruiz A.S.; Bressani R. Effect of Germination on the Chemical Composition and Nutritive Value of Amaranth Grain. Edit: American Association of Cereal Chemists. 1990. USA.

9. Chirinos Silva, Adjany Yuleimi. Pachari Vera, Erika. Obtención de parámetros óptimos en la elaboración de una bebida alcohólica a partir de Kiwicha (Amaranthus caudatus linnaeus) y Maiz (Zea mays L.) germinados. Edit.: Universidad Nacional de San Agustín. 2007. Arequipa Perú.