Optimization of phenolic substance extraction of some plants by Taguchi method; examination of their antioxidant and antibacterial capacities

Each step of the process interacts with the others in order to produce a specific result. An activity is defined as the activities of a number of factors, including machines, tools, processes, and people. By contrast, input variables are parameters that have an impact on test results, which can be improved in order to provide measurable or unmeasurable results as a result of changes made. Whenever there is a need to improve or increase the quality performance of a product or process, the term 'experimental design' refers to the process of observing the differences between the selected quality characteristics, as a result of adjusting the various factors and levels that influence the product/process, and interpreting those differences with a variety of techniques. To reach the target value as quickly as possible, experimental design minimizes the number of variables while simultaneously allowing factors that affect the design to be evaluated independently. It is common for this method to be used when trying to analyze the effects of phenolic compounds found naturally in plants, which are secondary metabolites which have biological effects. Understanding how these compounds interact with the external environment is crucial to understanding how they function in plant biology. Reduce the number of variables in the design so that the key factors influencing the performance of the phenolic compounds can be identified Therefore, in addition to solving the above problems in the design, this can also bring about more opportunities for increasing the target value in the shortest possible time. Phenolic compounds of different nature and quantity are characteristic for plants. They take part in plants as a defense mechanism. A number of heart-related conditions can be reduced by using these compounds, such as coronary heart disease, cataracts, diabetes and many others. Thus, it is used in many areas of medicine and pharmacology, particularly as a treatment for various diseases. Extraction techniques and conditions are important in obtaining phenolic compounds. To reduce the amount of phenolic substance lost during extraction, factors such as solvent preferences, control over extraction time and temperature should be optimally regulated as well. The Hibiscus (Hibiscus sabdariffa L) plant is used in many industrial food products (jam, confectionery, ice cream, chocolate, sweeteners, etc.) due to its rich content and intense dish leaf color. A number of antioxidant, antibacterial, antipyretic and blood pressure lowering effects have been found in studies conducted with the hibiscus plant. Safflower petals contain gallic acid, chlorogenic acid, syringic acid, quercetin, and epicatechin. It is believed that both phenolic compounds and flavonoids glycosyl quinochalcones are colors. This flower's color pigments are largely derived from safflower. Safflower flowers Since this substance determines the majority of its therapeutic effects, it is included in pharmacological studies. Flowers of safflower contain rich amounts of phenols that are antioxidants, and in addition, because they contain a number of other substances, they are cardiovascular protective, liver cleaners, anticancers, as well as protecting the nervous system and the lungs. It has found wide use in the treatment of phenolic found in safflower color pigments in the structure; irrigation, fertilization, sowing frequency, sowing time, harvest It is affected by many parameters such as time, type, environmental stress and It has been determined by studies that it can vary. Harvest from plants secondary metabolites of flowers obtained after temperature, UV rays, pH, various It changes depending on different factors such as gases, metal ions and chemicals. Reported. Althaea officinalis, known as marshmallow flower, has been reported in pharmacological researches to be used in the procedure of some diseases like inflammation, stomach ulcers, cough and cold. Calendula (Calendula officinalis L.) is a herbaceous plant belonging to the Asteraceae family that can grow as a single or biennial plant. Various fields, such as medicine and pharmacy, as well as the food and cosmetic industries, utilize the calendula's rich chemical content. Calendula consists of a variety of chemical components that are biologically active. These include terpenoids, flavonoids, coumarins, quinine, essential oils, carotenoids, and amino acids. In addition, calendula, vanillic acid, chlorogenic acid, caffeic acid, oleanolic acid, β-amirin acetate, β-amirin, rutin, narcisssin, isoquercitrin, isorhamnetin 3-glucoside, quercetin, protocatechin acid, syringic acid, p-coumaric acid; In addition, high levels of antioxidants such as calendoflacid, quercetin-3-Orutinoside, isoramnetin, calendoflavosit, isorhamnetin-3-O neohesperidoside, isoramnetin 3-O-2G-ramnosyl routineosite, quercetin-3-O glucoside, isoramnetin-3-Orutinoside, and calendoflavobioside detect the presence of many components with properties Due to its rich pharmacological properties, it occupies an important position in the group of alternative medicinal aromatic plants. A number of ailments, such as eczema, ulcers, and irritated skin have been treated with it since time immemorial. To obtain phenolic substances, these plants, which are used as tea and for medicinal purposes, will be used. Factors such as temperature, pressure, and humidity affect the extraction process. In an extraction process, the quality and yield can be affected by factors such as the solid-liquid ratio, the solid-solvent contact surface, the temperature, the extraction processing time, or the solvent type. An experimental design based on Taguchi was used in this study to optimize parameters related to extraction processes. For Taguchi modeling, 4 different factors were selected from the parameters affecting the extraction process: extraction method, solvent, temperature and duration. 3 levels of these factors have been determined. For the extraction method, ultrasonic, classical and mixing methods were chosen. For the ultrasonic extraction, a heated ultrasound bath at a frequency of 45 kHz was used. As a classical extraction method, the process was carried out in the water bath at the specified temperature and time. In the mixed extraction method, a magnetic rod rotating at a speed of 300 rpm was used. Solvents that are considered grass should be selected because of the use of phenolic substances in areas of human health concern, such as medicine and pharmacy. For this reason, 3 solvents were used: ethanol, methanol and distilled water. Since it is known that temperature (25°C, 40°C, 55°C) and duration (30 min., 60 min., 90 min.) phenolic substances undergo thermal degradation due to increased temperature and extraction time, high temperatures and long periods of time were not preferred. There are 4 factors chosen and 3 levels of factors. The degree of freedom for the determined factors and levels is 8. The determined factors and the degree of freedom were modeled by selecting the L9 orthogonol array in the experimental design using the Minitab19 package program. As a result of the modeling, 9 different experimental sets and 3 repetitions were extracted in the program. An analysis of total phenolic matter (TFM) from extraction processes was conducted according to Folin-Ciocalteau method, and the results were reported in gallic acid. The highest value for hibiscus was 24,8±1,01 mg GAE/g, for safflower 3,08±0,21 mg GAE/g, for marshmallow flower and aynuprifa, respectively, 3,50±0,05 mgGAE/g and 39,47±2,06 mgGAE/g. All TFM quantity values for 9 experimental sets are processed in the experimental design as a response to the program as a system output. According to these responses, Taguchi analysis was performed in the program and signal-to-noise ratios for all factors and levels were calculated. Impact graphs of these ratios and the average were obtained as program output. As a result of these calculations, the optimum parameters of hibiscus, marshmallow flower, calendula plant were found as mixed extraction, distilled water 55°C and 60 min. The temperature for this plant was determined to be 40°C at sunflower, unlike the other plants.The percentages at which factors and parameters affect the amount of phenolic substance was investigated so that more knowledge about the modeling could be gained and comments would be easier to make. For this reason, analysis of variance was used. When the effect surfaces for all plants are examined, the effect of the solvent factor is higher than other factors in percentage. For the hibiscus, the safflower, and the marshmallow flower, together with the calendula plant, the percentage contributions of the solvent factor were as follows: 59,09%, 81,82%, 60,70%, and 58,50%, respectively. Analysis of variance also tests whether the results are significant at the specified confidence intervals. Research was conducted on hibiscus, calendula, safflower, and marshmallow flower extraction processes based on various factors. The study found that while the changes in all factors of hibiscus and calendula plant had a mojor efficacy on the extraction process, the changes in temperature and duration factors for safflower and marshmallow flower were found to be insignificant (p<0.05). The extraction process was conducted parallel with these plants in order to maintain the accuracy of the modeling, and the results were compared with those expected by the program. To maintane the correctness of the modeling, the extraction process was conducted in three parallels with these plants, and the results were compared with the expected values in the program. The accuracy of the model was evaluated by analyzing the difference between observed and expected values. Calendula and safflower results were meaningless at 95% confidence intervals. For hibiscus and marshmallow flowers, the results are significant. Following the extraction process performed with optimal parameters, the activity capacity and antibacterial capacity of the antioxidant were determined as a result of the extraction process. Antioxidant capacity determination was performed according to DPPH radical capture method. The % inhibition values of hibiscus, safflower, marshmallow flower and calendula plants were calculated as 60,03%, 43,85%, 53,98% and 35,02%, respectively. In order to verify the results, they were compared to the BHT (butylated hydroxytoluene) antioxidant standard as a benchmark. This study was conducted using the well diffusion method in order to study the antibacterial properties of plants.The gram-negative bacteria Escherichia coli (gram positive bacteria Bacillus cereus (SBT-8) and Staphylococcus aureus (29213) were used for this procedure. Calendula plant provides the highest inhibition on the Bacillus cereus (SBT-8) strain (Hibiscus;10 mm, safflower; 9 mm Marshmallow; no effect). For the bacterium Staphylococcus aureus (29213), the inhibition diameters of hibiscus, safflower and calendula plants were as follows; It measured 12 mm, 9 mm, 10 mm. In the examination of the results, marshmallow flower was not found to exhibit an inhibitory effect on any strain of bacteria. In addition, no plant effect has been found on E. coli bacteria.