Abstract
Diplocyclos palmatus (L.) C. Jeffrey is an important medicinal plant used in several reproductive medicines. It serves as a wide source of tetracyclic triterpens called cucurbitacins. Response surface methodology (RSM) with Box-Behnken design (BBD) was studied to optimize the production of cucurbitacins. RSM put forth the ideal conditions such as 1:30 SS ratio (g/mL), 80 rpm (mixing extraction speed), 150 µm mean particle size, 30 min extraction time and 50 °C using chloroform in continuous shaking extraction (CSE) and showed the highest cucurbitacin I (CUI) content (2.345 ± 0.1686 mg/g DW). Similarly, the highest yield of cucurbitacin B (CUB) (1.584 ± 0.15 mg/g DW) was recorded at ideal conditions (1:40 g/mL SS ratio and 60 min time and others similar to CUI). Among the tested extraction methods, the highest CUI, CUB, and CUI + B yield (1.437 ± 0.03, 0.782 ± 0.10, 2.17 ± 0.35 mg/g DW, respectively) as well as promising DPPH radical scavenging activity (25.06 ± 0.1 µgAAE/g DW) were recorded from the SBAE (steam bath assisted extraction). In addition, MAE and UAE revealed the highest inhibition of α-amylase (68.68%) and α-glucosidase (56.27%) enzymes, respectively. Fruit extracts showed potent anticancer activity against breast (MCF-7) and colon (HT-29) cancer cell lines (LC50 – 44.27 and 46.88 µg/mL, respectively). Our study proved that SS ratio, particle size and temperature were the most positively influencing variables and served to be the most efficient for the highest recovery of CUI and CUB. Based on the present study, the fruits of D. palmatus were revealed as a potent antioxidant, anti-diabetic and anticancer bio-resource that could be explored further to develop novel drug to manage diabetes, cancer and oxidative stress related disorders.
Highlights
The plants from the family Cucurbitaceae are commonly called as melons, squashes, and gourds which are traditionally used in the human diet
The design matrix of 46 runs was depicted by employing Box-Behnken design (BBD) and their respective responses (CUI, cucurbitacin B (CUB) and cucurbitacin I (CUI) + B, miligram per gram of dry weight (mg/g DW)) are presented in Table A1 (The results are shown in Supplementary Table A1 and Fig. S1)
Higher CUB (1.5840 ± 0.15 mg/g DW) and CUI + B (3.8021 ± 0.31 mg/g DW) were recorded using 1:40 g/mL SS ratio and 60 min extraction time, while keeping remaining parameters as mentioned in CUI. Several extraction methods such as continuous shaking extraction (CSE), steam bath assisted extraction (SBAE), microwave assisted extraction (MAE) and ultrasound assisted extraction (UAE) were studied wherein the highest cucurbitacins content was reported from SBAE
Summary
The plants from the family Cucurbitaceae are commonly called as melons, squashes, and gourds which are traditionally used in the human diet. Most of the drugs used in the cancer therapy have been extracted from the natural resources but at the same time separation and purification of these bioactive compounds is the most crucial process[15]. Various extraction techniques such as continuous shaking extraction (CSE), microwave assisted extraction (MAE), Soxhlet extraction (SE), ultrasound assisted extraction (UAE) and steam bath assisted extraction (SBAE) have been used for the isolation of bioactive compound. Fruit extracts of D. palmatus were studied to investigate in vitro antioxidant, anti-diabetic and anticancer potential as well as identification of bioactive metabolites using LC-MS. The present study forms the first report on the optimized extraction of the cucurbitacins, some important bioactivities and other metabolites using LC-MS
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