The world is facing a problem regarding the use of petroleum fuels that has led to a search for a suitable alternative fuel source. Researchers have come up with the idea of producing biofuel to overcome this problem. In this study, microalgae were explored as a high potential feedstock to produce biofuel. In order to produce a large quantity of biofuel with low cost at a short time, the manipulation of nutrients is a factor in microalgae cultivation. In this study, Iron (II) Chloride (FeCl2) was added to the nutrients to initiate a stressful condition during growth which contributes to the produce of lipid. Isolated microalgae species were identified as Scenedesmus sp. During mass cultivation, the microalgae cultures were scaled up to 2 L of culture. Three flasks of microalgae culture were labelled with S1, S2, and S3. Flask S1 acts as a control without the addition of FeCl2, while another two flasks acted as experimental flasks. Flask S2 was supplemented with 0.5 mg FeCl2 while Flask S3 was supplemented with 1.0 mg of FeCl2. With the addition of Iron (II) Chloride, microalgae entered a stationary phase at day 9 and day 10 as compared to the control flask which enters the stationary phase at day 7. This also affects the dry weight. Flask 3 produces 0.8658 g of microalgae powder compared to Flask 1 and 2 which produced 0.4649 g and 0.5357 g respectively. Lipid analysis was done by using GCMS and GCFID. Flask 3 produced various types of fatty acids which can be used for biodiesel production compared to other cultivates. In Flask 1, docosanoic acid which is a saturated fatty acid was detected. While in Flask 2 (S2), with the addition of 0.5 mg of FeCl2, docosapentaenoic acid was produced. In the last flask which involved the addition of 1.0 mg of FeCl2, more fatty acid was detected. In GC-FID data, 6 types of fatty acids were detected. Linolein acid, linolenic acid, stearidonic acid, docosapentaenoic acid, docosahexaenoic acid and docosanoic acid were produced at different retention times. Most of the fatty acids produced are polyunsaturated fatty acid (PUFA). In transesterification, the fatty acid reacts with methanol and acid catalyst. The reaction produces fatty acid methyl ester. In Flask 1, the control flask, without the addition of FeCl2, no fatty acid methyl esters (FAME) was produced. However, in Flask 2 and 3 which were added 0.5 mg FeCl2 and 1.0 mg FeCl2, n-hexadecanoic acid methyl ester which is also known as palmitic acid was produced. Palmitic fatty acid can be used for biodiesel production.
Clinacanthus nutans, an herbal shrub belonging to the Acanthaceae family, is traditionally used as a functional food to treat various ailments in Malaysia and Indonesia. Although the polar fraction of this plant shows non-toxic effect, the toxicity of the non-polar extract is not reported so far. The present study aimed to assess the toxic effect and determine the lethal concentration of this non-polar fraction using zebrafish embryos. The n-hexane fraction was partitioned from the crude extract of C. nutans obtained using 80% methanolic solution. After spawning of the adult male and female zebrafish, the eggs were collected, transferred into a 96-well plate and incubated with the n-hexane fraction at concentrations of 15.63 μg/ml, 31.25 μg/ml, 62.5 μg/ml, 125 μg/ml, 250 μg/ml and 500 μg/ml in 2% DMSO. The survival and sublethal endpoint were assessed, the mortality and hatchability rates were calculated based on microscopic observation, while the heartbeat rate was measured using DanioScope software. The median lethal concentration (LC50) of the C. nutans n-hexane fraction, which was determined using probit analysis, was calculated to be 75.49 μg/mL, which is harmful. Moreover, gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of palmitic acid, phytol, hexadecanoic acid, 1-monopalmitin, stigmast-5-ene, pentadecanoic acid, heptadecanoic acid, 1-linolenoylglycerol and stigmasterol in the n-hexane fraction.
Activation of inflammatory pathways via reactive oxygen species (ROS) by free fatty acids (FFA) in obesity gives rise to insulin resistance and endothelial dysfunction. Withaferin A (WA), possesses both antioxidant and anti-inflammatory properties and therefore would be a good strategy to suppress palmitic acid (PA)-induced oxidative stress and inflammation and hence, insulin resistance and dysfunction in the endothelium. Effect of WA on PA-induced insulin resistance in human umbilical vein endothelial cells (HUVECs) was determined by evaluating insulin signaling mechanisms whilst effect of this drug on PA-induced endothelial dysfunction was determined in acetylcholine-mediated relaxation in isolated rat aortic preparations. WA significantly inhibited ROS production and inflammation induced by PA. Furthermore, WA significantly decreased TNF-α and IL-6 production in endothelial cells by specifically suppressing IKKβ/NF-κβ phosphorylation. WA inhibited inflammation-stimulated IRS-1 serine phosphorylation and improved the impaired insulin PI3-K signaling, and restored the decreased nitric oxide (NO) production triggered by PA. WA also decreased endothelin-1 and plasminogen activator inhibitor type-1 levels, and restored the impaired endothelium-mediated vasodilation in isolated aortic preparations. These findings suggest that WA inhibited both ROS production and inflammation to restore impaired insulin resistance in cultured endothelial cells and improve endothelial dysfunction in rat aortic rings.