Obtention, yields, chemical-microbiological properties and amino acids pro ile in a lour from Sacha Inchi (Plukenetia volubilis l.)

Priscilla. Mora-Aguirre1, Luis. Romero-Hidalgo1, Edgar. Landines-Vera1, Roberto. Ordoñez-Araque*2,3, Mario. Valdez-Díaz1 1Department of Chemical Engineering, Chemical Engineering Faculty, Universidad de Guayaquil (UG), Av. Delta y Av. Kennedy, Guayaquil, Ecuador 2Department of Nutrition and Dietetics, Faculty of Health and Welfare, Universidad Iberoamericana del Ecuador (UNIB.E), Av. 9 de Octubre N25-12 y Colón, Quito, Ecuador 3Gastronomy School, Universidad de las Américas (UDLA), Av. de los Granados y Colimes, Quito-Ecuador


INTRODUCTION
Sacha Inchi (SI) (Plukentia volubilis L.), also known as "Maní del Inca" is an oleaginous plant of the Euphorbiaceae family, native to the rain forest of the Andean region of South America (Maurer, 2012). This plant is also cultivated in other countries, like Colombia, Japan and Indonesia (Rawdkuen et al., 2016), as well as in Ecuador, a small country that is giving the irst steps in the production of SI and the study of the chemical composition is very essential.
This seed is a good source of protein (~27%) and oil (~42%), in fact the quality of this oil is one of the best in this kind of seeds (Hamaker, 1992), has been reported that polyunsaturated fatty acids (PUFAs) has positive bene its regarding the cardiovascular in lammatory diseases and cancer, this has led people to think that these PUFAs are more bene icial than other dietary supplements (Gogus and Smith, 2010), for this reason many small oil seed factories have started the production of this product.
A by-product in the oiling extract process of SI is the pressed cake, the yield of this cake can reach amounts of more than 50% according to the extraction method used (Rawdkuen et al., 2016), for example, in a cold press extraction, the yield of the cake is around 68% (Valdiviezo et al., 2019). In the seed, the percentage of protein is approximately 33%, compared to others oilseeds, is relatively high (Hanssen and Markus, 2011) , and this percentage is better in the pressed cake with an approximate value of 59% (Ruiz, 2013), the aminoacidic composition in the SI protein could reach or exceed the recommended amino acid scoring patterns by FAO ( FAO, 2013), the SI press cake could give us a good contribution, especially in quality protein (Vásquez, 2017).
Since the SI cultivation is being developed in Ecuador, the residue from the seed after the oil extraction could be used for lour production, and allow the development of foods with composite lours, which are characterized by the quality of the amino acids (Duodu and Minnaar, 2011).
The purpose of this research was to obtain lour from the press cake obtained after the oil extraction in SI seeds, watching each parameter to obtain the yields in the process. In addition, a chemical and microbiological analysis were also determined, and a characterization of the protein in the inal product. These results will be useful for SI oil seed producers and manufacturers.

Obtaining Sacha Inchi lour
In this irst step, dry cake samples are received after having extracted the most oil from the seed; as a result, a dry cake with minimal oil remnants is obtained. It must be on sealed plastic containers and avoid the presence of air inside the container.
The cake must not have impurities or contamination. The seeds were harvested in the San Vicente canton -Ecuador Once the raw material is obtained, it is passed to a dryer, in this phase the excess moisture that could have been left in the raw material of the previous extraction of the oil will be removed. This step will help reduce the possibilities of proliferation of pathogens and mycotoxins that could develop in lour. This dryer is tunnel type, and the temperature was controlled at 65 • C for a period of two hours.
After having removed the excess moisture from the raw material, it is taken to a grinding equipment, for this an endless screw mill is used to achieve a good degree of ineness (100 mesh). The time the dried cake degreased spends in the mill is short, about 2 minutes. Finally, to achieve the uniformity of the lour, it was sifted, separated in different lour fractions by difference in size through meshes with controlled porosities from 100 mesh to 600 mesh. For this step an AS 200 analytical sieve vibrator was used for 10 minutes, the desired degree of ineness is between 100 to 150 mesh, in case of obtaining a degree of ineness greater than this the material will be recirculated to the grinding process.
The following equation will be used to calculate the average particle size, Where APS is the average particle size, Ms is the Mesh size and Aw is the percentage withheld of lour.

Moisture
This parameter was determined by the method: AOAC 930.15.

Protein
The method used was AOAC 17th 984.13, to determine the percentage of protein present in the cake and lour of Sacha Inchi.

Lipids
The content of lipid material was determined using the Modi ied Folch method.

Ash
In this parameter the AOAC 942.05method was used, to know the presence of inorganic material.

Fiber
Using the AOAC 978.10 method the iber content was determined.

Microbiological and toxicological analysis in Sacha Inchi lour
For these analyzes the following parameters were determined:

Total aerobes
The BAM-FDA CAP # 3 2001 method was used.

Total coliforms
The BAM-FDA CAP # 4 2002 method was used.

Escherichia coli
The BAM-FDA method CAP # 4 2002 was used.

Amino Acid Pro ile in Sacha Inchi Flour
For this analysis the AOAC 994.12 method was used. The ACQUITY UPLC system (Waters, Milford, MA, USA) consisting of thermostat, autosampler, highpressure binary pump and photodiode array detector PDA (an optical detector in the range ultravioletvisible light that operates between 190 nm and 700 nm) was used for the analysis of 17 AAs. Chromatographic separation was obtained with the AccQ-Tag Ultra C-18 column (2.1 mm 9100 mm; 1.7 lm).

Statistics analysis
For the analysis three repetitions of each one were made, and they obtained the means with their respective deviations standard. In addition, a statistical analysis was made using Tukey's ANOVA test using the Real Statistics program to determine the possible signi icant differences at p <0; 05 level.

Yields in the obtention of Sacha Inchi lour
For the drying of Sacha Inchi cake, the results were successful, as shown in Table 1, a sample of 466,525 g were dried, which reduced its moisture by 6.81%, according with (Fortin, 2009) this parameter should not exceed from 13% Figure 1: Drying speed Figure 1 shows the moisture loss of the wet cake per unit of time and surface area (Ocon and Tojo, 1998).
Using the Equation (1) the average particle size of the Sacha Inchi lour is calculated. As show in Table 2 a greater retention in the 1mm size mesh is evidenced, the average particle size is 1837 µm.
The yield of lour obtained at the end of the process is 91,6% ±1,45 the remaining percentage is due to the loss of moisture during the drying process and the particles retained in each equipment (Giraldo-Gómez et al., 2019).

Physicochemical analysis in Sacha Inchi cake and lour
As show in Table 3 the moisture decreases by 3,7% by the effect of drying. An increase of 3% is observed for the protein resulting in a lour with a high protein content, (FAO, 1980) , indicating that the lower limit for a lour to be considered proteinic is 35%. Comparing these results with the obtained by (Rawdkuen et al., 2016), which hydrolyzed the cake protein giving a result of 45.9% indicates that this parameter is of high value.
The difference of lipids between both compounds does not represent a signi icant value. The remaining oil (10,5%) has a high content of unsaturated fatty acids (Maurer, 2012), which   have anti-in lammatory properties and offer protection against heart disease and high blood pressure (Williams, 2011).
The ash have a minor percentage for the Sacha Inchi lour (3,1%) and is within the limit recommended by (FAO, 1980).
The presence of iber in Sacha Inchi lour (19.89%) makes it a good food supplement, because it generates an increase in peristaltic movements of the intestine, facilitates transit and intestinal distention (Badui, 2006).

Microbiological and toxicological results in Sacha Inchi lour
The results of the microbiological tests as shown in Table 4, determine that according to the limits established by (CODEX-FDA), the samples satisfy the parameters necessary for their use as an ingredient in the production of various food products.

Amino acid composition and comparison with other studies
The amino acid content in the lour from the Sacha Inchi cake is higher in these four essential amino acids Threonine, Valine, Methionine and Isoleucine compared to the samples from the seed and protein extract shown in Table 5 , (Rawdkuen et al., 2016(Rawdkuen et al., , 2018, however the content of other essential amino acids such as Leucine, Lysine, Tryptophan and Histidine was much higher in the samples of protein extract and Sacha Inchi seeds. The comparison of the essential amino acids present in different samples of Sacha Inchi not only indicate their differences, it also allows to demonstrate the high content of essential amino acids they have and determine that they can be used as ingredients of food products such as granola bars, cookies, noodles and drinks as described by (Jagersberger, 2013) (Geneva, 2002). Finally, regarding the content of non-essential amino acids, Sacha Inchi cake lour has a total of 601.32 g.kg −1 , compared to the total content of Sacha Inchi protein isolate 153 g.kg −1 and the seed of Sacha Inchi 370.7 g.kg −1 ; These results demonstrate that the Sacha Inchi cake lour has a greater amount of non-essential amino acids than the other two types of products of Sacha Inchi.

CONCLUSIONS
The process of obtaining Sacha Inchi lour gives a high yield (91.6%) which provides a high content of proteins, iber and lipids with a high percentage of unsaturated fatty acids. The proportion of essential amino acids such as Leucine, Lysine, Tryptophan and Histidine is higher compared to other studies. This product meets the daily requirements of most essential amino acids.