Extraction Purification Research Articles

Extraction and Partial Purification of Glucoamylase from Rhizopus oryzae by Solid State Fermentation using Agro-Industrial Residues

Background: Filamentous fungi are used to produce many industrial enzymes because these fungi have a great ability to produce such enzymes, especially when using solid substrates that are usually low in cost. The widespread use of this enzyme has made it very important to work on improving its production to achieve the maximum possible benefit.Methods: Five fungal isolates were obtained from soil and decaying fruits and vegetables. All isolates were identified as Rhizopus spp. and cultured on Potato Dextrose Agar (PDA) plates escorted by starch 1%. The appearance of clearance zones around the fungal colonies represents the ability of fungi to produce Glucoamylase. Different solid substrates, nitrogen sources, and temperatures were used to improve enzyme production.Results: Wheat bran gave the highest enzyme production with specific activity 6.25 U/mg, and yeast extract was the potent inducer for enzyme production with specific activity 9.28 U/mg. The optimal temperature for enzyme production was 30°C with specific activity10.27U/mg. Maximum specific activity 18.40U/mg was recorded at 55% saturation with ammonium sulfate precipitation. The result showed increased specific activity 24.84U/mg with dialysis. Partial purification of the enzyme revealed that pH5 was optimum for enzyme-specific activity 9.2U/ml; higher enzyme-specific activity was found with MnSO4 (11.83U/ml) as the best enzyme inducer.Conclusion: This investigation attempted to examine R. oryzae as an effective maker of GA. Innovative work into the nearby creation of GA for industrial use utilizing local resources has demonstrated financial effectiveness. It requires improvement into consistency as needed by worldwide associations dealing with industrial enzymes. Keywords: Glucoamylase; Production; Partial purification; Rhizopus oryzae; Solid state fermentation

Inhibitive Mechanism of Loquat Flower Isolate on Tyrosinase Activity and Melanin Synthesis in Mouse Melanoma B16 Cells.

Melanin naturally exists in organisms and is synthetized by tyrosinase (TYR); however, its over-production may lead to aberrant pigmentation and skin conditions. Loquat (Eriobotrya japonica (Thunb.) Lindl.) flowers contain a variety of bioactive compounds, while studies on their suppressive capabilities against melanin synthesis are limited. Loquat flower isolate product (LFP) was obtained by ethanol extraction and resin purification, and its inhibitory efficiency against TYR activity was investigated by enzyme kinetics and multiple spectroscopy analyses. In addition, the impact of LFP on melanin synthesis-related proteins' expression in mouse melanoma B16 cells was analyzed using Western blotting. HPLC-MS/MS analysis indicated that LFP was composed of 137 compounds, of which 12 compounds, including flavonoids (quercetin, isorhamnoin, p-coumaric acid, etc.) and cinnamic acid and its derivatives, as well as benzene and its derivatives, might have TYR inhibitory activities. LFP inhibited TYR activity in a concentration-dependent manner with its IC50 value being 2.8 mg/mL. The inhibition was an anti-competitive one through altering the enzyme's conformation rather than chelating copper ions at the active center. LFP reduced the expression of TYR, tyrosinase-related protein (TRP) 1, and TRP2 in melanoma B16 cells, hence inhibiting the synthesis of melanin. The research suggested that LFP had the potential to reduce the risks of hyperpigmentation caused by tyrosinase and provided a foundation for the utilization of loquat flower as a natural resource in the development of beauty and aging-related functional products.

Kinetic study of a conventional and ultrasound-assisted extraction of pectin from different plant-based side streams: Impact on pectin extraction yield, purity and molecular pectin structure

The time-dependency of both a conventional and ultrasound-assisted extraction (UAE) was studied for three biomasses, carrot pomace (CP), broccoli stems (BS) and pumpkin pomace (PP). Both extraction processes were performed completely analogous without and with ultrasound waves to unequivocally evaluate the effect of ultrasonication. Besides the pectin extraction yield (considering the yield and pectin purity), the composition and molecular structure of the extracted materials were also studied as a function of time. For all biomasses, UAE resulted in a(n) (slight) increase of the final pectin extraction yield, giving rise to the extraction of an additional 4.2, 5.8 and 10.7% of the amount of pectin initially present for CP, BS and PP, respectively. Prolonged extraction times resulted in pectin structures comparable to conventional extraction (with a higher protein content). Compared to conventional extraction, shorter UAE processes (1, 5 and 2.5 min for CP, BS and PP, respectively) resulted in comparable pectin extraction yields and slightly more branched pectin structures with a higher protein content. Both observations show the potential of UAE for the intensification of the industrial pectin extraction. The three biomasses differed mainly in the amount of non-pectic compounds co-extracted, and the sensitivity by which the UAE process was accelerated.

Pulsed Electric Fields assisted extraction of proteins and phycocyanin from Arthrospira platensis biomass: A kinetic study

The objective of this work was the investigation of the kinetics of protein and C-phycocyanin extraction from Arthrospira platensis (Spirulina) biomass assisted by Pulsed Electric Fields (PEF). Fresh, untreated and PEF-treated (7.8 kV/cm, 0–137.6 kJ/kg) biomass was suspended in water and incubated at 30–50 °C for up to 24 h. Protein and C-phycocyanin recovery kinetics exhibited a sigmoidal behavior. The increase in treatment specific energy and incubation temperature up to 40 °C led to significant extraction acceleration. A treatment of 137.6 kJ/kg prior to incubation at 40 °C led to the minimization of protein characteristic incubation time (2.3 h vs. 14.4 h and 10.4 h of untreated sample at 30 and 40 °C, respectively). At 50 °C a significant reduction of the extraction yield was observed, due to the inactivation of the proteolytic enzymes. Higher treatment specific energy led to increased C-phycocyanin purity of the extracts at 40 °C. The maximization of the C-phycocyanin extract purity factor (1.14 vs 0.66 for untreated samples) was achieved with a PEF treatment at 137.6 kJ/kg and incubation at 40 °C for 3 h.

Study of the microRNA fraction in food waste and other biomasses. Assessment of its antioxidant and anti-inflammatory activity

MicroRNAs (miRNAs) are short non-coding RNAs with important biological functions. Their presence in food and their potential role as new bioactive molecules is generating great expectations. For the first time, the miRNA fraction within food waste and algae has been studied, highlighting their bioactivity and its potential upcycling. MiRNAs from barley malt rootles, brewer’s spent grains, Spirulina, Chlorella, orange peels, pomegranate and apricot seeds, and watermelon waste were extracted using two kits, one with chloroform and a more sustainable alternative. Higher extraction yields were obtained with the chloroform-containing kit. Purity of extracts was confirmed by UV spectroscopy and reversed-phase high performance liquid chromatography (RP-HPLC) with UV detection. Further analysis by RP-HPLC and high-resolution mass spectrometry (MS) demonstrated the potential and feasibility of this technique for the analysis of miRNA although there are still some limitations to overcome before its routine application. Apricot seed miRNAs demonstrated remarkable anti-inflammatory and antioxidant capacity.

Expression of Concern: Simultaneous extraction and preliminary purification of polyphenols from grape pomace using an aqueous two-phase system exposed to ultrasound irradiation: process characterization and simulation.

[This corrects the article DOI: 10.3389/fnut.2022.993475.].

Improved Extraction of Crocin and Comprehensive Evaluation of its Physicochemical, Biological, and Functional Properties

Crocin, as a valuable metabolite of saffron, encompasses numerous nutritional, pharmacological, and medicinal properties suitable for different applications. This study aimed to effectively extract crocin compounds with high yield and quality from saffron stigma and comprehensively investigate their various physicochemical and functional properties. To this end, the modified solvent extraction method was used for crocin extraction and its different functional and physicochemical properties were investigated using various characterization techniques. The functional and biological properties of the resultant crocins in terms of antioxidant activity, antibacterial potency, anti-tyrosinase property, and cytotoxicity were investigated. According to the results, a high extraction yield and purity for extracted samples (19.6% and 13.4% production yield and 78% and 98.4% purity in the first and second crystallization steps, respectively) were obtained using the green, environmentally friendly, and sustainable crystallization method. The coloring index, bitterness, and aromatic power of the extracted crocin were comparable with the standard crocin. Morphological, structural, and thermal properties of the extracted crocin from different analytical methods confirmed its preserved composition and structure through the solvent extraction process. Moreover, a desired antioxidant activity by the DPPH, superoxide, hydroxyl radical scavenging, and reducing power assays were observed for resultant crocin products. In the antibacterial and cytotoxicity assays, a 5 mg/mL minimum inhibitory concentration (against Gram-positive and Gram-negative bacteria) and more than 95% cell viability (toward the NIH 3T3 cell line) were recorded, respectively. A potent anti-tyrosinase activity (IC50 = 73.93 μg/mL) with a significant reduction in the melanin production of the B16F10 cell line also was noticed for the obtained crocin products.

Functional Tea Extract Inhibits Cell Growth, Induces Apoptosis, and Causes G0/G1 Arrest in Human Hepatocellular Carcinoma Cell Line Possibly through Reduction in Telomerase Activity.

The functional tea CFT-1 has been introduced into China as a nutraceutical beverage according to the "Healthy China" national project. The effects on human hepatocellular carcinoma (HCC) cells remain unclear and were investigated with the functional tea extract (purity > 98%). The morphological changes in the cells were observed with microscopes. Cell proliferation, migration, cycle distribution, and apoptotic effects were assessed by MTT, Transwell assays, and flow cytometry, respectively, while telomerase inhibition was evaluated with telomerase PCR ELISA assay kits. The CFT-1 treatment resulted in cell shrinkage, nuclear pyknosis, and chromatin condensation. CFT-1 suppressed the growth of Hep3B cells with IC50 of 143 μg/mL by inducing apoptosis and G0/G1 arrest in Hep3B cells. As for the molecular mechanism, CFT-1 treatment can effectively reduce the telomerase activity. The functional tea extract inhibits cell growth in human HCC by inducing apoptosis and G0/G1 arrest, possibly through a reduction in telomerase activity. These results indicate that CFT-1 extract exhibited in vitro anticancer activities and provided insights into the future development and utilization of CFT-1 as functional foods to inhibit the proliferation of HCC cells.

The yield and purity of DNA extracts from seeds of eight six accessions of Treculia species using Zymo Research mini-prep DNA extraction kit

The yield and purity of DNA extracts from seeds of eight six accessions of Treculia species using Zymo Research mini-prep DNA extraction kit

Mechanism study of synergistic aluminum extraction and purification by ammonium for high-value conversion of secondary aluminum dross

Secondary aluminum dross (SAD) constitutes a significant volume of hazardous solid waste with huge productions. Balancing extraction efficiency and purity within the current resourcing recovery process proves challenging and impedes high-value conversion. Consequently, the NH4HSO4-H2SO4 system was employed in this study to convert SAD into high-purity alumina nanoparticles, and the effect of ammonium was investigated. The results indicated that the leaching rate reached 85.98 % under the optimum conditions (170 °C, [Htot]=12 mol/L, 16/1,6 h); the interaction of NH4+ and H+ with the oxygen atoms on the surface of Al2O3 was demonstrated to enhance the reactivity of the aluminum atoms by DFT calculations. The incorporation of NH4+ also facilitated the separation of aluminum by leveraging the temperature-dependent solubility characteristic of NH4Al(SO4)2. Molecular dynamics simulation revealed that NH4+ inhibits the diffusion rate of Al3+, thereby regulating the doping impurities such as Mg and Ca, and improving the purity of Al2O3 (95.29–99.64 %). Additionally, Nanoscale regulation was executed through the conversion of the NH4AlO(OH)HCO3 precursor and control. The sintering and agglomeration problems brought about by pyrolysis in atmospheres such as H2O and SO3 were avoided. This research provides reference for the high-value utilization of aluminum-containing solid waste.

Characterization, in vitro bioaccessibility and antidiabetic activities of maltodextrin and lecithin nanomaterials loaded with ellagic acid

Pomegranate peel is one of the wastes generated in large quantities in the food industry and ellagic acid is a natural bioactive phenolic compound in pomegranate peel. In this study, high-purity and low-cost ellagic acid was produced by acid hydrolyze process from pomegranate peel and characterization, in vitro bioaccessibility, antioxidant, and antidiabetic activity of the ellagic acid nanoencapsulated with maltodextrin and soy lecithin were studied. The extract yield and purity of ellagic acid was 6.47 and 88.74%, respectively. Results showed that lecithin coated samples were more stable with − 43.16 mV zeta potential and had higher encapsulation efficiency (99.29%) compared to maltodextrin. However, coated samples with maltodextrin had higher production efficiency (91.40%) and loading capacity (54.70%). The thermal resistance of ellagic acid after the nanoencapsulation process was significantly raised. The ellagic acid nanoencapsulated with maltodextrin had higher bioaccessibility (80.22%) and solubility in biological buffers (64.00-98.20%). The α-amylase and α-glucosidase inhibitiory activities of the ellagic acid decreased after the encapsulation process. However, the α-amylase inhibitory activities of the nanoencapsulated materials were higher than acarbose as a positive control. As a result, thermal resistance, solubility in a biological medium, and bioaccessibility of ellagic acid after nanoencapsulation with maltodextrin can be increased. Thus, the use of ellagic acid nanoencapsulated with maltodextrin in the food, cosmetic, and pharmacological industries can be diversified.

Supercritical CO2 Extraction of Terpenoids from Indocalamus latifolius Leaves: Optimization, Purification, and Antioxidant Activity.

This study aimed to investigate the efficacy of supercritical CO2 (SC-CO2) extraction in enhancing the extraction rate, purity, and antioxidant activity of Indocalamus latifolius (Keng) McClure (Poaceae) leaf terpenoids (ILLTs). Crude extracts obtained from leaves were subjected to qualitative and quantitative analyses, revealing neophytadiene, phytol, β-sitosterol, β-amyrone, squalene, and friedelin as the primary terpenoid constituents, identified through gas chromatography-mass spectrometry (GC-MS). Compared with steam distillation extraction (SD), simultaneous distillation extraction (SDE), ultra-high pressure-assisted n-hexane extraction (UHPE-Hex), ultra-high pressure-assisted ethanol extraction (UHPE-EtOH), ultrasound-assisted n-hexane extraction (UE-Hex), and ultrasound-assisted ethanol extraction (UE-EtOH), SC-CO2 exhibited a superior ILLT extraction rate, purity, and antioxidant activity. Scanning electron microscopy (SEM) observations of the residues further revealed more severe damage to both the residues and their cell walls after SC-CO2 extraction. Under optimal parameters (4.5 h, 26 MPa, 39 °C, and 20% ethyl alcohol), the ILLT extraction rate with SC-CO2 reached 1.44 ± 0.12 mg/g, which was significantly higher than the rates obtained by the other six methods. The subsequent separation and purification using WelFlash C18-l, BUCHI-C18, and Sephadex LH-20 led to an increase in the purity of the six terpenoid components from 12.91% to 93.34%. Furthermore, the ILLTs demonstrated cytotoxicity against HepG2 cells with an IC50 value of 148.93 ± 9.93 μg/mL. Additionally, with increasing concentrations, the ILLTs exhibited an enhanced cellular antioxidant status, as evidenced by reductions in both reactive oxygen species (ROS) and malondialdehyde (MDA) levels.

Determination of calcium β-hydroxy-β-methyl butyrate content in milk and dairy products by solid-phase extraction and high performance liquid chromatography

In this paper, an analytical method based on solid-phase extraction (SPE) purification and high-performance liquid chromatography method (HPLC) was developed for the first time to determine the content of calcium β-hydroxy-β-methyl butyrate (CaHMB) in milk and dairy products. The quantitative detection was performed by HPLC with diode array detector (DAD) under external standard method. The limit of quantification (LOQ), calculated as 10 times the standard deviation, was 0.10 g/100 g. The average recoveries were in the range of 92.40–103.00 %, with relative standard deviations (RSDs) of 1.25–3.77 %, measured at three concentration levels. The real sample analysis results showed that CaHMB was detected in infant formula, formulated milk powder, and milkshake samples, the contents of which were in the range of 0.60–8.66 g/100 g, respectively. The developed method features good purification effect, good separation specificity, good repeatability, accuracy and reliability, and can meet the content determination of CaHMB in milk and dairy products.

Sustainable proteins from wine industrial by-product: Ultrasound-assisted extraction, fractionation, and characterization

Plant proteins are increasingly being used in the food industry due to their sustainability. They can be isolated from food industry waste and converted into value-added ingredients, promoting a more circular economy. In this study, ultrasound-assisted alkaline extraction (UAAE) was optimized to maximize the extraction yield and purity of protein ingredients from grapeseeds. Grapeseed protein was extracted using UAAE under different pH (9–11), temperature (20–50 °C), sonication time (15–45 min), and solid/solvent ratio (10–20 mL/g) conditions. The structural and functional attributes of grapeseed protein and its major fractions (albumins and glutelins) were investigated and compared. The albumin fractions had higher solubilities, emulsifying properties, and in vitro digestibilities but lower fluid binding capacities and thermal stability than the UAAE and glutelin fraction. These findings have the potential to boost our understanding of the structural and functional characteristics of grapeseed proteins, thereby increasing their potential applications in the food and other industries.

Synergized enzyme-ultrasound-assisted aqueous two-phase extraction and antioxidant activity validation of polysaccharides from tobacco waste

A powerful and innovative technique was developed to efficiently extract and refine polysaccharides from tobacco waste. This was achieved through a combination of enzyme and ultrasound-assisted extraction assisted aqueous two-phase extraction (EUAATPE). The formation of an aqueous two-phase system (ATPS) using various salts and ethanol was investigated. ATPS of 18.0 % (NH4)2SO4 (w/w) and 25.0 % ethanol (w/w) was chosen due to its favorable selection extraction ability and high recovery of 93.68 ± 0.40 %. By utilizing both single-factor experiments and response surface method (RSM), the key factors involved in the EUAATPE process were optimized. The optimal conditions were as follows: ultrasonic power 180 W, ultrasonic time 9 min, cellulase concentration 2.0 %, pectinase concentration 1.0 %, extraction temperature 50 °C, extraction time 1.5 h, and liquid-to-solid ratio 30:1 (g/g). Under the optimal conditions, the extraction yield of polysaccharides was 6.72 ± 0.11 (mg/g), and the purity of the polysaccharides from the bottom phase was 78.73 ± 3.17 %. EUAATPE resulted in a significant enhancement in the extraction yield and purity of polysaccharides from tobacco waste compared to traditional methods such as hot water extraction (HWE), ultrasound-assisted extraction (UAE) and enzyme-assisted extraction (EAE). Polysaccharides of tobacco waste obtained from various methods had analogous monosaccharide compositions, different molar ratio compositions and surface structure characteristics. Moreover, all polysaccharides exhibited a certain extent antioxidant activities and polysaccharides from the bottom phase (PBP) extracted by EUAATPE showed superior antioxidant activity. Hence, EUAATPE, being an innovative and eco-friendly extraction technique, presents a productive substitute for extracting and refining polysaccharides from discarded tobacco resources.

Evaluation of antioxidant potential of crude extract of metabolites from endophytic fungi isolated from <i>Annona senegalensis</i> Pers

Degenerative diseases resulting from free radicals can be prevented through the use of natural antioxidants. This study aimed to assess the free radical scavenging potential of the underexplored endophytic fungal metabolites from Annona senegalensis Pers (wild custard apple). Freshly collected root, midrib, and leaf tissues were planted on malt extract agar following surface sterilization with 70% ethanol for 3 minutes, washed twice with distilled water, immersed in a sodium hypochlorite solution (4%) for 5 minutes, and subsequently rinsed with sterile water. Seven endophytic fungi were isolated, and the antioxidant activity of their metabolites was evaluated using the 2,2- diphenyl-1-picrylhydrazyl free radical scavenging method, with ascorbic acid serving as a reference antioxidant. The crude extract from the seven endophytic fungal isolates demonstrated concentration- dependent antioxidant activities. LB2 exhibited a strong antioxidant activity with an IC50 value of 38.15 μg/ml. Extracts from RT1 and LB1 showed antioxidant activities, with the highest inhibition of 60% and 65%, respectively, observed at a concentration of 1000 μg/ml. This was compared favorably to ascorbic acid, which exhibited 94% inhibition. This indicates that the crude extract from endophytic fungi isolated from A. senegalensis possesses free radical scavenging properties. Further purification and elucidation of the crude extract will reveal the bioactive compounds responsible for the antioxidant activities.

Research Progress on Taxus Extraction and Formulation Preparation Technologies.

Taxus, as a globally prevalent evergreen tree, contains a wealth of bioactive components that play a crucial role in the pharmaceutical field. Taxus extracts, defined as a collection of one or more bioactive compounds extracted from the genus Taxus spp., have become a significant focus of modern cancer treatment research. This review article aims to delve into the scientific background of Taxus extracts and their considerable value in pharmaceutical research. It meticulously sifts through and compares various advanced extraction techniques such as supercritical extraction, ultrasound extraction, microwave-assisted extraction, solid-phase extraction, high-pressure pulsed electric field extraction, and enzymatic extraction, assessing each technology's advantages and limitations across dimensions such as extraction efficiency, extraction purity, economic cost, operational time, and environmental impact, with comprehensive analysis results presented in table form. In the area of drug formulation design, this paper systematically discusses the development strategies for solid, liquid, and semi-solid dosage forms based on the unique physicochemical properties of Taxus extracts, their intended medical uses, and specific release characteristics, delving deeply into the selection of excipients and the critical technical issues in the drug preparation process. Moreover, the article looks forward to the potential directions of Taxus extracts in future research and medical applications, emphasizing the urgency and importance of continuously optimizing extraction methods and formulation design to enhance treatment efficacy, reduce production costs, and decrease environmental burdens. It provides a comprehensive set of preparation techniques and formulation optimization schemes for researchers in cancer treatment and other medical fields, promoting the application and development of Taxus extracts in pharmaceutical sciences.

Extraction of Natural Food Color from Tomato, Carrot and Turmeric

The demand for natural and healthier food products has increased dramatically in recent years due to growing consumer awareness of the impact food has on health as well as evidence of adverse effects from various ingredients, including some additives. This study evaluates the extraction method, solvent effects, precipitation, identification, purification, and lycopene, carotene, and curcumin content from turmeric, carrot, and tomato. Along-side, tomato, carrot, and turmeric samples were subjected to two different extraction and purification processes: solvent extraction for lycopene and carotene, and alkalization for curcumin. The samples were extracted and purified at room temperature (30ºC) and chilled (4ºC) for a period of three weeks. Tomato extracts had lycopene contents ranging from 0.0153 to 0.0362 mg/100g. obtained carotene content, which was then extracted using a solvent in the range of 61.43 to 81.72 mg/100g. The alkalization process produced a curcumin concentration of 91.17 to 110.41 mg/100g. Comparing the ethyl acetate extraction technique to the anti-solvent method, a greater amount of lycopene (red) and carotene (orange) precipitation was obtained. The maximum amount of curcumin precipitation obtained in lower pH solution. Lower pH is important to maintain the stability of curcumin precipitation. The experience of this research work suggested that lycopene from tomatoes, carotene from carrots, and curcumin from turmeric might be an excellent source to meet the increasing need for natural colorants.

A rapid solid-phase extraction purification combined with UPLC[sbnd]MS/MS used for simultaneous determination of eight tetracyclines and three metabolite in chicken and pork

A method for rapid detection and identification of 8 tetracyclines (TCs) and 3 metabolite residues in chicken and pork was established and verified by combining rapid solid-phase extraction (SPE) technology with ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLCMS/MS). After a sample was extracted, it was purified with a PRiME HLB cartridge, and the cartridge did not need to be activated, balanced, or eluted. The filtrate was dried under nitrogen, redissolved, and subsequently analyzed via UPLCMS/MS. The limits of detection for chicken and pork were 0.05–0.48 μg/kg and 0.05–0.52 μg/kg, respectively. The limits of quantification were 0.16–1.61 μg/kg and 0.16–1.75 μg/kg, respectively. The recoveries were 85.15%–107.50% and 84.21%–106.37%, respectively. The established method was suitable for qualitative and quantitative analyses of TCs and their metabolite residues in chicken and pork and has been used for the detection of TC in chicken and pork samples from local markets.

Kinetic parameter for scale-up and γ-oryzanol content of rice bran oil as antioxidant: Comparison of maceration, ultrasonication, pneumatic press extraction

Rice bran oil is one of oryzanol source oils. Oryzanol is an antioxidant compound that is related to the absorption of cholesterol, and is used in hyperlipidemia treatment and menopause problems. RBO extraction, purification and its γ-oryzanol content have been carefully reviewed. The quality and concentration of γ-oryzanol depend on the extraction process and purification. In selecting the extraction method to obtain the highest oryzanol content, in addition to comparing the concentration of oryzanol obtained and it can also be done by comparing the extraction kinetics parameters. Modeling according to physical or empirical kinetics can contribute in increasing the result of extraction. This study aims to determine the highest oryzanol content in rice bran oil, comparing several extraction methods and studies of rice bran oil extraction kinetic is necessary for scale up purposes. In this study is conducted Rice Bran Oil Extraction with n-Hexane solvent using several different methods, such as maceration, ultrasonication, and pneumatic press extractions. Independent variable that is used is the extraction time and yield as dependent variable. The study shows that the best extraction method to get the highest yield is 10.34 % by ultrasonicator and oryzanol content is 5.09 mg/g by a pneumatic press machine. According to kinetic parameter k2 is 0.001546, Cs is 0.0589, and h is 0.4707, R2 = 0.9715 obtained from extraction using ultrasonicator.