Study of liquid smoke toxicity cocoa shell with different purification methods

Cocoa shell is an underutilized agricultural trash. Cocoa shells can be used to make liquid smoke and cocoa charcoal as basic materials. The charcoal made from cocoa pod husks can be used as a dye adsorbent. The study will look at the effects of different purification and filtering procedures on the toxicity of liquid smoke from cocoa husks in order to determine which purification process is best for making liquid smoke from cocoa husks that is not harmful. The results demonstrated that several procedures for purifying and filtering liquid smoke from cocoa peels were non-toxic. Cocoa shell is an underutilized waste from plantations. Cocoa shells can be used as raw materials in the manufacture of liquid smoke and cocoa charcoal. The burning of cocoa pod husks charcoal can be used as a dye adsorbent. The research intends to observe the different purification and filtering methods on the liquid smoke toxicity from cocoa husks to find out which purification method is more appropriate for liquid smoke from cocoa husks so that it is not toxic. The results showed that the purification and filtering of liquid smoke from cocoa peels using various methods were non-toxic. The best approach with the highest LD50 value is zeolite purification, which has an LD50 value of 3019.95 ppm. The activated charcoal technique, activated charcoal + zeolite method, and distillation method were then used, with LD50 values of 2,951.21 ppm, 2,398.83 ppm, and 1,412.54 ppm, respectively.


Introduction
Cocoa has become one of the plantation commodities that show a very important role in economic activities in Indonesia.Cocoa is also one of Indonesia's export commodities which is quite important as a foreign exchange earner.Indonesia has become the third-largest cocoa exporter and producer in the world after Ghana and Ivory Coast [1].According to research [2], the cocoa husk makes liquid smoke and cocoa charcoal.The pods of cocoa can be utilized as a Rhodamine dye adsorbent [3].The use of liquid smoke as a food preservative has several advantages, including controlling the color and taste of food products.The concentration of usage used is quite small and the application of its use is quite easy and fast, namely by soaking or spraying and mixing directly into food ingredients.
Purification is the process of removing unwanted materials, this process aims to improve the quality of water smoke to a higher selling value.The known purification methods are chemical and physical purification.The purification of the chemical can be performed by adding certain adsorbents or complex compounds [4].
The result of liquid smoke from the pyrolysis process has to be purified, this process determines the type of liquid smoke that is developed into three types of liquid, namely: Grade 1 liquid smoke is liquid smoke resulting from the distillation process and filtering with zeolite which is then followed by fractional distillation followed by filtering with activated charcoal.This liquid smoke has a pale yellow color and is used for ready-to-eat foods such as wet noodles, meatballs, and tofu [5].Grade 2 liquid smoke is the quality of liquid smoke that has passed the distillation stage.This liquid smoke has a brownish-yellow color and is oriented to raw food ingredients such as meat, chicken, or fish as a substitute for formalin [5].Grade 3 liquid smoke is the purification of liquid smoke from tar by using a distillation process.Distillation is a way to separate mixtures based on differences in their boiling points some components can evaporate faster than others.At the time steam is produced from the mixture, the steam contains more components that are more volatile so that the process of separating components from the mixture can occur [5].
The best quality from liquid smoke can be found on the cinnamon material treatment at the temperature of 400 ± 100 o C, namely the performance of the pyrolysis equipment of 16.29 ml/hour [6].According to [2] performing Differential Thermal Analyzes (DTA), the dried cocoa pods were put into a steel pyrolysis reactor with an electric heater, three capacitors, and a distillate reservoir with a combustion temperature of 115-500°C for 5 hours.Liquid smoke can have the role of an antioxidant and antimicrobial.Antioxidants are either electron-donating compounds or reductants.Although the compound has a small molecular weight it can inactivate the development of oxidation reactions, by preventing the formation of radicals.Antioxidants are also compounds that can inhibit oxidation by binding free radicals [7].
Research about liquid smoke purification has been performed by [8], examining the value of antioxidant activity only, while the toxicity value of purified liquid smoke has not been tested.Toxicity is the level at which the substance is damaged when exposed to organisms.Therefore, it is important to measure the toxicity to see the effect on the substructure of the organism.The process of making cocoa shell liquid smoke produces liquid smoke with grade 3 quality.For this reason, purification is necessary to reduce the number of toxic compounds in cocoa shell liquid smoke.The study aimed to determine the different purification and filtering methods for the toxicity properties of cocoa shell liquid smoke and to find out which purification method was more appropriate for cocoa shell liquid smoke using a purification method that was more appropriate for liquid smoke of cocoa shell so that it was toxic less.
Based on the above description, this study was conducted on the purification of smoke liquid using various ways and examined the value of antioxidants LD50 liquid smoke cocoa skin using various purification methods.The goal of this approach is to assess the toxicity of LD50 liquid smoke of cocoa skin through distillation, purification with zeolite, purification with activated charcoal, and a combination of the two so that it can be used as a natural preservative.

Methodology
This study was carried out at Ekasakti University's Agricultural Product Technology Laboratory in November and December of 2019.This study employs an exploratory strategy to investigate the LD50 content toxicity of liquid smoke using various purifying processes.The pyrolysis procedure can produce liquid smoke.The cocoa skin utilized in this study was taken from Lubuk Minturun Village in Padang City, and it was pyrolyzed till liquid smoke of cocoa skins was formed.The primary substance in this study is liquid smoke grade 3. A. Salina Leach eggs, seawater, methanol, zeolite, and activated charcoal are also required for chemical analysis.This research was conducted at the Agricultural Product Technology Laboratory of Ekasakti University in November -December 2019.This study uses an exploratory design by looking at the LD50 content toxicity of liquid smoke with several purification methods.Liquid smoke can be obtained by the pyrolysis method.The material used in this study was cocoa skin obtained from Lubuk Minturun Village, Padang City, and pyrolysis was carried out until liquid smoke of cocoa skins was obtained.The main material used in this research is liquid smoke grade 3.In addition, the materials needed for chemical analysis are A. Salina Leach eggs, seawater, methanol, zeolite, activated charcoal.A set of pyrolysis equipment, micropipette (Scilogex), A. Salina Leach egg incubator, incandescent lamp (5 watts), dropper (Dropping), 50 ml measuring cup, test tube, petri dish, IOP Publishing doi:10.1088/1755-1315/1306/1/0120033 measuring cup (Pyrex), vortex, magnifying glass, distillation apparatus, electric balance, filter paper (Mettler), UV-VIS spectrophotometer are the tools used to make cocoa shell liquid smoke.

Exploratory research
The purpose of this research is to make the new topic better known to the wider community, to provide a basic overview of the topic of discussion, to generalize ideas and develop a tentative theory, to open the possibility for further research on the topic discussed, and to determine the techniques and directions to be used in subsequent research.The research implementation will be carried out on several purifications such as the treatment below: A= 10% activated charcoal B= Zeolite 10% C= 5% Activated Charcoal + 5% Zeolite D=Distillation temperature140 o C 2.2 Purification of liquid smoke 2.2.1.Distillation [9] This procedure is used to separate components from a mixture because some components evaporate faster than others.Because the resulting steam contains more volatile components, separation of the components from the mixture is possible.Distillation of smoke liquid can be done at temperatures ranging from 100oC to 150oC.Unwanted chemicals, such as tar compounds and aromatic polycyclic hydrocarbons, can also be removed during the liquid smoke distillation process.

Active charcoal [10]
Active charcoal generated from activated cocoa shell charcoal is used to filter the liquid smoke filtrate from active zeolite.The liquid smoke filtrate is passed through an activated carbon column before being collected in a container for testing.

Zeolites [11]
The filtered liquid smoke is then filtered using active zeolite by passing the distillate liquid smoke through the active zeolite column to obtain the purified liquid smoke filtrate using zeolite.The filtrate is kept in a container for testing purposes.[11] Liquid smoke produced from the pyrolysis process is added to active charcoal and zeolite, then allowed to stand overnight.After 24 hours, the liquid smoke is filtered using filter paper, and the antioxidant activity is measured.[12] BLST is the first bioassay for natural material research and one of the procedures that uses poisonous compounds.The advantages of the BLST test are that it does not take long, the method is easy and quick, and it does not cost a lot of money.It also does not require smog procedures, special equipment, or a large number of test material.A bioassay is a test that uses living organisms to measure the efficacy of organic and inorganic materials.The Artemia Salina leach larvae are used as experimental animals in the BLST method, which is a toxicity test because a compound is found in a short period of time (for 24 hours).The BLST method was used to examine the compound's potential to in case shrimp larvae.

Toxicity test
Calculating the toxicity value is used to see the level of safety, one method that can be used is BSLT toxicity testing (Brine Shrimp Lethality Test).Probit analysis was found from the results of testing the toxicity of cocoa shell liquid smoke on Artemia Shalina Leach test animals (Mayer et., al,1982. ).Testing the toxicity activity of the four treatments can be presented in Figure 1. Figure one explains the toxicity properties of the filtered treatment B (zeolite), it is not toxic because zeolite can absorb CO2 gas and harmful metal materials so the liquid smoke produced is not toxic [13].
While in treatment D it also does not contain toxic substances because distillation is one of the distillation methods that can remove odors, color changes, and other impurities so that they are pure and non-toxic [14].
The BSLT is a method for testing harmful materials that was developed as the first bioassay for natural material research.Artemia Salina Leach larvae are used as experimental animals in this procedure.The BSLT technique of toxicity testing is an acute toxicity test in which the harmful effect of a substance is measured in a short period of time, which is 24 hours after the test dosage is administered.The approach is designed to evaluate the LD50 value of the plant's active components' activity against Artemia Salina Leach larvae.If the LD value of an extract is 1000 g/ml, it is hazardous according to the BSLT technique [15].Figure 2 depicts the value of the toxicity activity of activated charcoal purification.The curve above explains the sum of the y values or the linear equation Y= 0.616x + 2.864 at the value of R2= 0.9096.The level of accuracy shows that the toxicity activity does not contain toxic because the activated charcoal filtering material used in the test does not affect the growth of artemia Salina leach although activated charcoal has an active absorption.
Its toxic effect on Artemia Salina Leach using different ppm concentrations This method is a simple, fast, easy, cheap, and simple biological test method, which usually uses shrimp larvae (Artemia Salina Leach).The toxicity of a compound can be determined by calculating the number of deaths.shrimp larvae with the Lethal Concentration parameter of 50 mortality percentage of Artemia Salina leach on activated charcoal treatment [16].The value of the zeolite purification toxicity activity can be presented in Figure 3.The curve above explains the sum of the values of y or the linear equation Y= 0.984x + 1.568 at the value of R2= 0.9096.The level of accuracy shows that it does not contain toxic because zeolites can absorb CO2 gas and dangerous metal materials, so the liquid smoke that is emitted produced is nontoxic [13].The mortality test was carried out with a lethal concentration of 50% (LD50).LD50 is a value that indicates the concentration of toxic substances that can cause the death of the organism until the curve above shows the percentage of death of Artemia Salina leach in zeolite treatment.The value of the toxicity activity of activated charcoal + zeolite purification can be presented in Figure 4.The curve above explains the total value of y or the linear equation Y= 1.392x +0.3 at the value of R2= 0.9629 to the level of accuracy, indicating that the toxicity activity does not contain toxic, because + zeolite which is both negatively charged, has no effect on the growth of Artemia Salina leach.
For the safety of the liquid smoke, it is necessary to research the toxicity test of methanol extract of liquid smoke on Artemia Salina Leach shrimp larvae using the Brine Shrimp Lethality Test (BSLT) method.As in the toxicity activity test in the treatment of mixed filtering of activated charcoal and zeolite.The value of the distillation purification toxicity activity can be presented in Figure 5.The curve above explains the number of values or linear equations Y = 1.448x+0.442at the value of R2 = 0.9605 to the level of accuracy, indicating that the toxicity activity does not contain toxins, because the distillation method is one of the distillation methods that can remove odors, color changes, and substances other impurities so that they are pure and non-toxic [14].
Separation of particles by heating did not affect the growth of the test animals used to identify the percentage of death in the activity test of distillate liquid smoke.LD50 toxicity calculation data can be presented in Table 1.
Table 1.LD50 toxicity calculation data.From the table above, it can be seen that the toxicity value of cocoa shell liquid smoke obtained from the results of different treatments in each animal test sample of Artemia Salina leach that did not contain toxic compounds although the number of ppm was different between A, B, C and D treatments but both had an LD50 value > 1000ppm [12].Based on the toxicity test above, it can be seen that the average percentage of deaths in the cocoa shell liquid smoke toxicity test with different purifications did not cause death to the test animals.
Toxicity is a condition that indicates a toxic or toxic effect on a substance.A toxicity test looks at the number of deaths from artemia Salina leach once in 24 hours.Brine Shrimp Lethality Test (BSLT) is a screening method to determine the toxicity of a compound using Artemia Salina experimental

Figure 2 .
Figure 2. Toxicity activity of liquid smoke filtered with activated charcoal.

Figure 4 .
Figure 4.The toxicity activity of activated charcoal + zeolite filtering.
Non-ToxicThe data was obtained from the average results of each test.