Int J App Pharm, Vol 11, Issue 2, 2019, 152-155Original Article


SUPERCRITICAL CARBON DIOXIDE EXTRACTION OF MANGO GINGER (CURCUMA MANGGA ROXB.): PROCESS OPTIMIZATION USING TAGUCHI METHOD

RINALDI SALEA, ERWAN Y. DARUSSALAM, STEVANUS HIENDRAWAN, BAMBANG VERIANSYAH, RAYMOND R. TJANDRAWINATA*

Dexa Laboratories of Biomolecular Sciences (DLBS), Industri Selatan V Block PP No. 7, Jababeka Industrial Estate II, Cikarang 17550, West Java, Indonesia
Email: raymond@dexa-medica.com

Received: 31 Nov 2018, Revised and Accepted: 30 Jan 2019

ABSTRACT

Objective: Extraction of Curcuma mangga (C. mangga) using supercritical carbon dioxide (SC-CO2) was investigated to provide information about the optimum extraction condition.

Methods: A Taguchi method with L9 orthogonal array design was used to determine the optimum extraction conditions. Effects of extraction pressure, temperature, CO2 flow rate and dynamic extraction time on C. mangga oil yield were investigated at levels ranging between 150-350 bar, 40-60 °C, 10-20 g/min and 120-240 min, respectively.

Results: The highest C. mangga oil yield (5.223%) from SC-CO2 extraction was obtained at a pressure of 350 bar, temperature of 60 °C, CO2 flow rate of 20 g/min and dynamic extraction time of 240 min. The experimental C. mangga oil yield at optimum condition was in a good agreement with the values predicted by computational process using Taguchi method. Based on S/N ratio calculation, the most influencing parameters in maximizing C. mangga oil yield is extraction temperature, followed by extraction pressure, dynamic extraction time and CO2 flow rate.

Conclusion: In this study, Taguchi method was successfully applied to optimize SC-CO2 extraction of C. mangga. Taguchi method was able to simplify the experimental procedure of SC-CO2 extraction.

Keywords: Taguchi, Supercritical carbon dioxide extraction, Curcuma mangga, optimization

© 2019 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijap.2019v11i2.31079

INTRODUCTION

Curcuma genus (Zingiberaceae family) consists of more than 80 species and some are widely used as spices and medicinal preparations [1]. It is native to the warm and humid environments of South and Southeast Asia. Curcuma mangga, which also known as the mango ginger, is one of the significant species in the Zingiberaceae family cultivated for food and medicinal purposes. The rhizomes of C. mangga are similar to ginger but possess a raw mango taste mainly due to the presence of car-3-ene and cis-ocimene [2, 3]. Medicinally, C. mangga rhizomes have been known for its uses to treat stomachache, fever and chest pain. It is also used in postpartum care, specifically to aid the healing of wounds [1, 4]. Literature has shown diverse therapeutic effects of C. mangga extract, such as antibacterial, antifungal, insecticidal, aphrodisiac, antipyretic, anti-inflammatory, anti-mycobacterial, anti-hyper-cholesterolemic and antioxidant properties [1, 3].

Pharmacologically-active compounds in plants are usually present in low concentrations. In recent years, our laboratories have developed and investigated various extraction techniques and biological assays on natural products [5-8]. The most widely-used method is extraction based on organic solvent [9]. Organic solvent extraction methods have been known to possess several drawbacks such as the production of hazardous wastes and few adjustable parameters to control the selectivity of extraction process. Therefore, the development of more reliable, simpler, less chemical-intensive and less expensive technique is necessary for commercial-scale production. In recent years, supercritical carbon dioxide (SC-CO2) extraction has widely been utilized for the extraction of oil and bioactive compounds from plant materials [8,10]. SC-CO2 extraction offers several advantages due to its non-toxic nature and unique physicochemical properties which include low viscosity, fast diffusion and tunable physical properties with temperature and pressure. The low critical temperature and pressure of carbon dioxide (Tc=31 °C and Pc=7.38 MPa) offer the possibility to operate extraction process at low temperature (below 80 °C) and moderate pressure (100–450 bar), which may be an ideal condition for the extraction of thermolabile compound [11, 12]. Despite the high potential of SC-CO2 for the extraction of oil and bioactive compounds, various parameters of SC-CO2 extraction (i.e. temperature, pressure, CO2 flow rate and time of extraction) have become the major obstacle in finding the optimum extraction condition for each plant material [13, 14].

Taguchi method is one of the simple and systematic approaches for designing an experiment with less number of experimental runs. It could be employed to optimize the performance characteristics of process parameters, which is proved to be a powerful tool that differs from the traditional full factorial method. This approach could economically satisfy the needs of problem-solving and design optimization. Thus, it is possible to reduce the time and cost for the experimental investigations [15-17]. Taguchi method has been employed in some applications related to SC-CO2 extractions. Ansari et al. (2012) employed Taguchi method to optimize supercritical extraction of oil from spearmint (Menthaspicata L.) leaves [18]. Previously, we compared Taguchi and full factorial method to optimize various parameters on the supercritical extraction of black cumin (Nigella sativa) seeds. It was shown that Taguchi method was able to simplify the experimental procedure and maintain experimental cost at a minimum level without affecting the quality of the result. On the other hand, the oil yield difference between full factorial and Taguchi method is insignificant (±0.1%) [19]. In previous study, we also utilized Taguchi method to optimize SC-CO2 extraction oil from Javanese turmeric (Curcuma xanthorrhiza Roxb.) and emprit ginger (Zingiber officinale var. Amarum) rhizome [8, 11]. The objective of this present study was to investigate the optimum condition for C. mangga oil extraction through SC-CO2 extraction and to investigate the effect of extraction pressure, temperature, CO2 flow rate and dynamic extraction time on C. mangga oil yield using Taguchi method.

MATERIALS AND METHODS

Materials

Dried rhizomes of C. mangga were obtained from CV Cipta Pratama (Magelang, Central Java, Indonesia). The dried rhizomes were comminuted in a milling machine (Quadro Comil, Canada) with 040 G screen and square impeller at 3500 rpm. The moisture content of the sample before extraction was 12.5% wet basis (w. b). Food-grade liquid CO2 (purity of 99.99%) was supplied in a cylinder tube by PT Inter Gas Mandiri (Cikarang, Indonesia).

Supercritical carbon dioxide (SC-CO2) extraction

SC-CO2 extraction was carried out using a customized supercritical fluid extractor with CO2 cycle system (KIST-Korea) described in previous study [19]. Extractor vessel with 1 L capacity was loaded with 50 g of grounded C. mangga. Food grade liquid CO2 was delivered to the extraction vessel using a high-pressure pump (Thar, USA). Extraction pressure was varied from 150 to 350 bar and temperature from 40 to 60 °C. The pressure in extraction vessel was controlled by a back pressure regulator (Tescom, USA). Heat exchangers (Lab. Companion, USA) were provided in the system to maintain the temperature in extractor and separator vessel. The CO2 flow rate was varied from 10 to 20 g/min. Static time was 60 min for all experiments and dynamic time was varied from 120 to 240 min. After experiments had been finished, the extract was separated from SC-CO2 by pressure reduction and collected from the separator.

Taguchi method

Taguchi method utilizes orthogonal array design (OAD) to study a large number of parameters with a small number of experiments. It significantly reduces the number of experimental configurations to be studied without affecting the quality of results and maintains the experimental cost at a minimum level [20, 21]. Taguchi method employs a special set of OAD in each number of experimental conditions to systematically look for favorable operating conditions [22]. A Taguchi method with L9 OAD was used to investigate the optimum condition in SC-CO2 extraction since it is the most suitable design for the investigated conditions, i.e. four factors with three levels (values). The factors and levels are listed in table 1, whereas the structure of Taguchi method with L9 OAD is shown in table 2. S/N ratio calculation is an evaluation of output performance which measures the level of performance and effect of noise parameters on performance. A target value of ‘larger is better’ was used since the purpose of this study was to obtain the highest C. mangga oil yield. In this study, the main effects of the three parameters were studied excluding the interactions between variables.

S/N ratio was calculated using the following Eq. (1):

Where yi represents the experimentally observed value of the ith experiment, and n is the number of trial at the same level. Statistical analysis was performed using MINITAB v.15 (Minitab Inc., USA) statistical software package.

Table 1: Factors and levels used in experimental design

Level Factors
Pressure (bar) Temperature ( °C) CO2 flow rate (g/min) Dynamic time (min)
A B C D
1 150 40 10 120
2 250 50 15 180
3 350 60 20 240

Table 2: Standard for L9 orthogonal arrays

Run Independent variables
A B C D
1 1 1 1 1
2 1 2 2 2
3 1 3 3 3
4 2 1 2 3
5 2 2 3 1
6 2 3 1 2
7 3 1 3 2
8 3 2 1 3
9 3 3 2 1

RESULTS AND DISCUSSION

Effect of SC-CO2 extraction conditions on C. mangga oil yield

Effects of four operating conditions of SC-CO2 extraction, namely pressure, temperature, CO2 flow rate and dynamic extraction time on the extraction of C. mangga oil were investigated using Taguchi method. The experimental responses in terms of oil yield are summarized in table 3. Results showed that the obtained C. mangga oil yield was varied from 0.524-4.889%.

A total of 9 experiments were performed in order to obtain the results of C. mangga oil yield from SC-CO2 extraction. Since a higher oil yield was the desirable purpose, the quality characteristic was set as 'larger is better'. The optimization of C. mangga oil yield from SC-CO2 extraction was considered from S/N ratio in each level of parameters. Fig. 1 depicted the S/N ratio of each individual factor. The higher number of S/N ratio represents a higher oil yield.

Table 3: Results of C. mangga oil using SC-CO2 extraction

Run Pressure (bar) Temperature ( °C) CO2 flow rate (g/min) Dynamic time (min) Yield (%)
1 150 40 10 120 0.524
2 150 50 15 180 2.766
3 150 60 20 240 4.222
4 250 40 10 120 4.280
5 250 50 15 180 3.274
6 250 60 20 240 4.889
7 350 40 10 120 4.128
8 350 50 15 180 4.419
9 350 60 20 240 4.292

Fig. 1: Effects of pressure (a), temperature (b), CO2 flow rate (c), and dynamic time (d) on C. mangga oil yield

Fig. 1 shows the main effect plot of C. mangga oil yield to different extraction factors and levels. Main effect plot was performed based on the average response for each combination of control factor levels. It can be seen in fig. 1a that increased pressure from 150 to 250 bar resulted in a significant increment of C. mangga oil yield, while further increasing pressure from 250 to 350 bar resulted in a smaller oil yield increment. Generally, using a higher pressure at isothermal conditions resulted in an increased solvent density and subsequently solvent power and solubility of the compounds. An increase in CO2 density accelerated the mass transfer between analytes and solvent during the extraction process and thus it improved the extraction yield. As the density increased, the distance between molecules was decreased and the interaction between compounds and CO2 was increased, which led to a greater solubility of compounds in CO2 [14].

The selection of operating conditions depends on the specific compound or compound family to be extracted. Molecular weight and polarity of the compound have to be taken into account case by case. The extraction temperature for thermolabile compound has to be fixed between 35 and 60 °C, e.g. in the vicinity of the critical point and as low as possible to avoid degradation [8]. At a constant pressure, an increase in temperature reduced the density of SC-CO2, hence reducing the solvent power of SC-CO2, but it increased the vapor pressure of the compounds and thus it became easier to extract [14]. These two competing factors changed differently and it depends on extraction conditions. Above the crossover pressure, the effect of temperature on density became less than that of solute vapor pressure. Therefore, the yield tends to increase with temperature. Below the crossover pressure, the effect of temperature on density became more than that of solute vapor pressure [23]. From fig. 1b, it can be seen that as temperature increased from 40 to 60 °C, the oil yield was also increased. This can be explained that the vapor pressure of the oil was increased and its solubility and extraction was improved at higher temperature [24].

Fig. 1c shows the effect of CO2 flow rate on oil yield. C. mangga oil yield was increased at CO2 flow rate of 10-15 g/min, while further increasing CO2 flow rate from 15 to 20 g/min resulted in a smaller oil yield increment. Mass transfer was increased with an increased CO2 flow rate since there was a higher amount of SC-CO2 passed through the extractor and it decreased the film thickness around the particles throughout the bed, which lowered external mass transfer resistance. However, increased CO2 flow rate resulted in a reduced residence time of CO2. In this case, the extraction process deviated from the equilibrium and SC-CO2 left the extractor unsaturated [25-27].

For all experiments, the static time was fixed at 60 min. During the static time, the system in extractor is allowed to reach an equilibrium that involved an interaction between the supercritical fluid and plant matrix. For a system in which the extraction rate is determined by the transfer of a solute from particle surface into the fluid, it can be assumed that the equilibrium between supercritical fluid and the matrix is established in a suitable period of static extraction time and will be released during dynamic extraction time. Fig. 1d shows that C. mangga oil yield was increased along with increased extraction time by up to 180 min and it did not vary with further increase in extraction time. Similar phenomena were also observed in our previous study on C. xanthorrhiza Roxb extraction [8].

Optimum condition

In this study, since the target value was ‘larger is better’, the optimum condition for each factor was defined as the level that gives the highest point of means of each plot for C. mangga oil yield. Based on the results of Taguchi method, the highest C. mangga oil yield from SC-CO2 extraction was obtained at a pressure of 350 bar, temperature of 60 °C, CO2 flow rate of 20 g/min and dynamic extraction time of 240 min. S/N ratio calculation results are shown in table 4. Greater difference value (Δ) of the average S/N ratio indicated that the control factors have a greater substantial effect on S/N ratio. Based on S/N ratio calculation, the most influencing parameters in this process is extraction temperature, followed by extraction pressure, dynamic extraction time and CO2 flow rate.

Table 4: S/N value calculated for each parameter and level

Level Pressure Temperature CO2 flow rate Dynamic time
1 3.712 3.796 3.819 3.865
2 16.761 15.146 15.748 15.902
3 17.389 17.718 16.360 17.450
Δ (max–min) 13.677 13.922 12.541 13.585
Rank 2 1 4 3

Experimental validation

Experimental validation is the final step in the modeling process to investigate the accuracy and robustness of the established model. The final analysis involved a comparison between the predicted values of the established model and experimentally validated values. The predicted value at optimized condition is 5.997%, while the actual value at this condition is 5.223%. Percentage error found in this study was 12.9%. It was found that the average percentage error was below 15%, confirming and concluding that the methodology used in establishing the model in this scientific research was systematic. C. mangga oil yield obtained in this study was higher than previous studies using SC-CO2 extraction. In the previous study, Krishna et al. used SC-CO2 extraction to extract C. mangga oil. They varied the extraction pressure from 100 to 350 bar, extraction temperature from 40 to 60 °C and extraction time from 5 to 15 h. Results of previous study showed that the best C. mangga extraction yield was 2.83% at 350 bar and 60 °C for 15 h [28].

CONCLUSION

Optimization of SC-CO2 extraction on the yield of C. mangga oil was successfully carried out using Taguchi method. The optimum condition to obtain the highest C. mangga oil yield (5.223%) was achieved at a pressure of 350 bar, temperature of 60 °C, CO2 flow rate of 20 g/min and dynamic extraction time of 240 min. The experimental oil yield at optimum condition was in accordance with the predicted computational results (5.997%). S/N ratio calculation identified extraction temperature as the most influencing parameters in maximizing C. mangga oil yield followed by extraction pressure, dynamic extraction time and CO2 flow rate.

ACKNOWLEDGEMENT

The authors acknowledge Dexa Laboratories of Biomolecular Sciences (DLBS)–PT. Dexa Medica for financial support. The authors would like to thank Isabela Anjani for critical review on this manuscript.

AUTHORS CONTRIBUTIONS

All the author have contributed equally

CONFLICT OF INTERESTS

The authors declared no conflicts of interest with respect to the authorship and/or publication

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