| Food sciences |
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| Optimization of chlorophyll extraction from mulberry leaves using response surface methodology |
| CHEN Shao-yuan, LÜ Zhen-er, DONG Feng-li, MAO Bi-zeng |
| College of Agriculture & Biotechnology, Zhejiang University, Hangzhou 310058, China |
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Abstract Chlorophyll plays an important role in our everyday life. It is used not only as an annexing agent in medicinal and calleidic products but also as an unartificial food coloring agent. Chlorophyll and its ramifications have antioxidant and anti-mutation properties. China has abundant mulberry resources. The mulberry leaf is an excellent Chinese herb from ancient times to the present day and contains many kinds of bioactive components, in which the chlorophyll content is an important physiological index. However, the optimization of chlorophyll extraction from mulberry leaf by response surface methodology (RSM) has not been reported so far. Response surface methodology (RSM) is an effectively statistical technique for optimizing complex processes. The greatest advantage of RSM is that we need fewer experimental trials. It is widely used in optimizing variables of the extraction process, such as polysaccharides, alkaloids, phenolic compounds and protein from various materials. Box-Behnken design (BBD) is one of RSM, which only has three-levels (low, medium and high, coded as −1, 0 and +1), and needs fewer experiments. It’s more efficient and easier to arrange and interpret experiments compared with others. This study focused on the optimal conditions of extracting mulberry leaf chlorophyll using RSM. The mixture of acetone and ethanol was used as extraction solvent. The chlorophyll content was calculated by Arnon’s method. Extraction time, temperature, ratio of liquor to material and volume ratio of acetone to ethanol in extraction buffer were selected as the experiment factors according to the results of single factor experiments. The extracting parameters were optimized by RSM. Under the optimal extraction conditions described above, the confirmatory experiment was conducted. The single factor experiment results indicated that extraction time of 5 h (Fig. 2) , extraction temperature of 50 °C (Fig. 3), ratio of liquor to material of 100∶1 (Fig. 4) and volume ratio of acetone to ethanol of 3∶1 (Fig. 1) were adopted for following experiment. Three independent variables were extraction time (C), extraction temperature (B) and ratio of liquor to material (A), according to the influence∶1. Under the optimal extraction condition, the chlorophyll productivity was 5.376 mg/g, which was in consistent with the predicted value of 5.451 mg/g.s of the four factors on the chlorophyll productivity. The statistical analysis indicated that the three variables (Table 4) and the quadratic of extraction time (C) and ratio of liquor to material (A) (Fig. 6) had significant effects on the yields, and there had the significant interaction effects between the variables of extraction temperature (B) and extraction time (C) (Fig. 7). A high determination co-efficient mathematical model was gained that could be used to optimize chlorophyll extraction. Design Expert 7.1.6 software showed the optimal extraction condition was as follows: extraction time of 5.25 h, extraction temperature of 56.5 °C, ratio of liquor to material of 103 In conclusion, the optimization of chlorophyll extraction by RSM is convenient, feasible and highly efficient
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Published: 20 November 2012
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