Fermentation of Rubus dolichocarpus juice using Lactobacillus gasseri and Lacticaseibacillus casei and protecting phenolic compounds by Stevia extract during cold storage.

This study aimed to investigate the biological activities of Lactobacillus gasseri SM 05 (L. gasseri) and Lacticaseibacillus casei subsp. casei PTCC 1608 (L. casei) in the black raspberry (Rubus dolichocarpus) juice (BRJ) environment, and also the anti-adhesion activity against Salmonella typhimurium (S. typhimurium) in fermented black raspberry juice (FBRJ). Results showed significant anti-adhesion activity in Caco-2 epithelial cells. In the anti-adhesion process, lactic acid bacteria (LAB) improve intestinal health by preventing the adhesion of pathogens. Adding LAB to BRJ produces metabolites with bacteriocin properties. Major findings of this research include improved intestinal health, improved antidiabetic properties, inhibition of degradation of amino acids, and increase in the nutritional value of foods that have been subjected to heat processing by preventing Maillard inhibition, and inhibition of oxidation of foodstuff by increased antioxidant activity of BRJ. Both species of Lactobacillus effectively controlled the growth of S. typhimurium during BRJ fermentation. Moreover, in all tests, as well as Maillard's and α-amylase inhibition, L. gasseri was more effective than L. casei. The phenolic and flavonoid compounds increased significantly after fermentation by both LAB (p < 0.05). Adding Stevia extract to FBRJ and performing the HHP process showed convenient protection of phenolic compounds compared to heat processing.

Oilseed meal proteins: From novel extraction methods to nanocarriers of bioactive compounds.

The global demand for animal proteins is predicted to increase twofold by 2050. This has led to growing environmental and health apprehensions, thereby prompting the appraisal of alternative protein sources. Oilseed meals present a promising alternative due to their abundance in global production and inherent dietary protein content. The alkaline extraction remains the preferred technique for protein extraction from oilseed meals in commercial processes. However, the combination of innovative techniques has proven to be more effective in the recovery and functional modification of oilseed meal proteins (OMPs), resulting in improved protein quality and reduced allergenicity and environmental hazards. This manuscript explores the extraction of valuable proteins from sustainable sources, specifically by-products from the oil processing industry, using emerging technologies. Chemical structure, nutritional value, and functional properties of the main OMPs are evaluated with a particular focus on their potential application as nanocarriers for bioactive compounds.

Biological stabilization of natural pigment-phytochemical from poppy-pollen (Papaver bracteatum) extract: Functional food formulation.

In this study, poppy-pollen extract (as a novel source of pigment and natural phytochemical) was microencapsulated. The spray-drying process maintained the encapsulation efficiency (EE) of phenolic (84-93%), anthocyanins (71-83%), and also antioxidant activity of extract in inhibiting DPPH (68-80%), ABTS (74-95%), OH (63-74%) radicals and reducing power (84-95%). The results of the Photo- and thermal (40-70˚C) stability of the bioactive compounds (TPC and TAC) indicated the thermal degradation and decomposition of particles' surface compounds during storage. The chemical (FTIR) and morphological analyses respectively revealed the insertion of extract compounds in the carrier matrix and the production of healthy particles with wrinkled structures. An increase in the carrier concentration elevated physical-stability, maintained structural properties, reduced hygroscopicity, and formed liquid/solid bridges or deliquescence phenomenon. The evaluation of the color histogram of the fortified gummy-candies indicated the usability of the spray-dried PP extract in producing an attractive red color with high sensory perception.

Metal-binding peptides and their potential to enhance the absorption and bioavailability of minerals.

Minerals including calcium, iron, zinc, magnesium, and copper have several human nutritional functions due to their metabolic activities. Body tissues require sufficient levels of a variety of micronutrients to maintain their health. To achieve these micronutrient needs, dietary consumption must be adequate. Dietary proteins may regulate the biological functions of the body in addition to acting as nutrients. Some peptides encoded in the native protein sequences are primarily responsible for the absorption and bioavailability of minerals in physiological functions. Metal-binding peptides (MBPs) were discovered as potential agents for mineral supplements. Nevertheless, sufficient studies on how MBPs affect the biological functions of minerals are lacking. The hypothesis is that the absorption and bioavailability of minerals are significantly influenced by peptides, and these properties are further enhanced by the configuration and attribute of the metal-peptide complex. In this review, the production of MBPs is discussed using various key parameters such as the protein sources and amino acid residues, enzymatic hydrolysis, purification, sequencing and synthesis and in silico analysis of MBPs. The mechanisms of metal-peptide complexes as functional food ingredients are elucidated, including metal-peptide ratio, precursors and ligands, complexation reaction, absorbability and bioavailability. Finally, the characteristics and application of different metal-peptide complexes are also described.

The enzymatic modification of whey-proteins for spray drying encapsulation of Ginkgo-biloba extract.

Ginkgo biloba extract (GBLE) contains many bioactives including flavonoids and terpene trilactones that play some pharmacological roles. These compounds are sensitive to operating conditions; so, encapsulation is a suitable approach to protect them. In this study, different carriers including maltodextrin (MD), and its combination with gum-Arabic (MD-GA), whey protein concentrate (MD-WPC), and whey-protein hydrolysate (MD-HWPC) were used to encapsulate GBLE. Powder production yield, physicochemical/functional characteristics, physical stability and flowability of particles were affected by the type and composition of carriers. FTIR results indicated the placement of phenolic compounds in the carrier matrix. The SEM images also showed the morphological changes of particles (especially the size, indentation and surface shrinkage) under the influence of various carriers. Microencapsulated powders formulated using MD-HWPC showed the highest values of TPC, DPPH, and ABTS and a lighter color which determined the suitability of this wall material (due to the improvement of surface activity and emulsifying properties of protein as a result of partial enzymatic hydrolysis) to protect the antioxidant properties of GBLE during spray-drying, improving the production yield and preserving physical and functional characteristics of the encapsulated powders.

Nutritional, biological, and structural properties of bioactive peptides from bellflower, Persian-willow, and bitter-orange pollens.

In this study, the composition of amino acids, nutritional characteristics, degree of hydrolysis (DH), antioxidant properties, and antibacterial activity of proteins and hydrolysates of bellflower (Campanula latifolia), Persian willow (Salix aegyptiaca), and bitter orange (Citrus aurantium L.) were investigated under the influence of different proteases (Alcalase: Al, trypsin: Tr, pancreatin: Pa, and pepsin: Pe). Evaluation of the structural features of the proteins showed amide regions (amide A, B, I-III) and secondary structures. Hydrophobic amino acids (∼38%), antioxidants (∼21%), and essential types (∼46%) form a significant part of the structure of flower pollen. The digestibility and nutritional quality (PER) of the hydrolyzed samples (CP: 1.67; CA: 1.89, and PW: 1.93) were more than the original protein. Among proteins and peptides, the highest degree of hydrolysis (34.6%: Al-PWH), inhibition of free radicals DPPH (84.2%: Al-CPH), ABTS (95.2%: Pa-CPH), OH (86.7%: Tr-CAH), NO (57.8%: Al-CPH), reducing power (1.31: Pa-CPH), total antioxidant activity (1.46: Pa-CPH), and chelation of iron ions (80%: Al-CPH and Al-CAH) and copper (50.3%: Pa-CAH) were affected by protein type, enzyme type, and amino acid composition. Also, the highest inhibition of the growth of Escherichia coli (25 mm) and Bacillus cereus (24 mm) were related to CP and PW hydrolysates, respectively. The results of this research showed that hydrolyzed flower pollens can be used as a rich source of essential amino acids as well as natural antioxidants and antibacterial in food and dietary products. PRACTICAL APPLICATION: Enzymatic hydrolysis of Campanula latifolia, Persian willow, and Citrus aurantium pollen proteins was performed. The hydrolyzed ones had high nutritional quality and digestibility (essential amino acids and PER index). Antioxidant properties and chelation of metal ions of peptides were affected by the type of protein and enzyme. The hydrolysates showed inhibitory activity against the growth of Esherichia coli and Bacillus cereus.

Influence of different nano/micro-carriers on the bioavailability of iron: Focus on in vitro-in vivo studies.

Anemia resulting from iron (Fe) deficiency is a global public health problem. The deficiency of Fe is usually due to insufficient dietary intake of iron, interaction of Fe with other food components, and thus low bioaccessibility/bioavailability. Fe encapsulation has the potential to tackle some major challenges in iron fortification of foods. Various nano/micro-carriers have been developed for encapsulation of Fe, including emulsions, liposomes, hydrogels, and spray-dried microcapsules. They could reduce the interactions of Fe with food components, increase iron tolerance and intestinal uptake, and decrease adverse effects. This article review covers the factors affecting the bioavailability of Fe along with emerging carriers that can be used as a solution of this issue. The application of Fe-loaded carriers in food supplements and products is also described. The advantages and limitations associated with the delivery efficiency of each carrier for Fe are highlighted.

Bioactivity and applications of saffron floral bio-residues (tepals): a natural by-product for the food, pharmaceutical, and cosmetic industries.

Saffron "Crocus sativus" is a plant of the Iridaceae family. Its therapeutic virtues have been known since antiquity; it is used in traditional medicine and culinary preparations. It is also known for its use in cosmetics because of its beneficial pharmacological activities for human skin. In particular, saffron tepals are the main by-product of saffron processing; they contain several bioactive compounds such as mineral agents, anthocyanins, monoterpenoids, carotenoids, flavonoids, and flavonols (kaempferol). This review aims to describe the different properties of saffron flower tepals, including their botanical characteristics, phytochemical composition, biological activities, and cosmetology and perfumery uses.

Cannabidiol-Loaded Nanocarriers and Their Therapeutic Applications.

Cannabidiol (CBD), one of the most promising constituents isolated from Cannabis sativa, exhibits diverse pharmacological actions. However, the applications of CBD are restricted mainly due to its poor oral bioavailability. Therefore, researchers are focusing on the development of novel strategies for the effective delivery of CBD with improved oral bioavailability. In this context, researchers have designed nanocarriers to overcome limitations associated with CBD. The CBD-loaded nanocarriers assist in improving the therapeutic efficacy, targetability, and controlled biodistribution of CBD with negligible toxicity for treating various disease conditions. In this review, we have summarized and discussed various molecular targets, targeting mechanisms and types of nanocarrier-based delivery systems associated with CBD for the effective management of various disease conditions. This strategic information will help researchers in the establishment of novel nanotechnology interventions for targeting CBD.

Stabilization of anthocyanins by simultaneous encapsulation-copigmentation via protein-polysaccharide polyelectrolyte complexes.

This study prepared a series of polyelectrolyte complexes (PECs) composed of heated whey protein isolate (HWPI) and different polysaccharides for simultaneous encapsulation and copigmentation of anthocyanins (ATC) and their ultimate stabilization. Four polysaccharides including chondroitin sulfate, dextran sulfate, gum arabic, and pectin were chosen due to their abilities to simultaneously complex with HWPI and copigment ATC. At pH 4.0, these PECs were formed with an average particle size of 120-360 nm, the ATC encapsulation efficiency of 62-80%, and the production yield of 47-68%, depending on the type of polysaccharides. The PECs effectively inhibited the degradation of ATC during storage and when exposed to neutral pH, ascorbic acid, and heat. Pectin had the best protection, followed by gum arabic, chondroitin sulfate, and dextran sulfate. The stabilizing effects were associated with the hydrogen bonding, hydrophobic and electrostatic interactions between HWPI and polysaccharides, conferring dense internal network and hydrophobic microenvironment in the complexes.

Natural Bioactive Molecules as Neuromedicines for the Treatment/Prevention of Neurodegenerative Diseases.

The brain is vulnerable to different types of stresses, particularly oxidative stress as a result of oxygen requirements/utilization in the body. Large amounts of unsaturated fatty acids present in the brain increase this vulnerability. Neurodegenerative diseases (NDDs) are brain disorders that are characterized by the gradual loss of specific neurons and are attributed to broad evidence of cell-level oxidative stress. The accurate characterization of neurological disorders relies on several parameters along with genetics and environmental risk factors, making therapies less efficient to fight NDDs. On the way to tackle oxidative damage and discover efficient and safe therapies, bioactives are at the edge of NDD science. Naturally occurring bioactive compounds such as polyphenols, carotenoids, essential fatty acids, phytosterols, essential oils, etc. are particularly of interest owing to their potent antioxidant and anti-inflammatory activities, and they offer lots of brain-health-promoting features. This Review focuses on probing the neuroefficacy and bioefficacy of bioactives and their role in supporting relatively low antioxidative and low regenerative capacities of the brain, neurogenesis, neuroprotection, and ameliorating/treating NDDs.

Application of essential oils as natural biopesticides; recent advances.

There is an urgent need for the development of sustainable and eco-friendly pesticide formulations since common synthetic pesticides result in many adverse effects on human health and the environment. Essential oils (EOs) are a mixture of volatile oils produced as a secondary metabolite in medicinal plants, and show activities against pests, insects, and pathogenic fungi. Their chemical composition is affected by several factors such as plant species or cultivar, geographical origin, environmental conditions, agricultural practices, and extraction method. The growing number of studies related to the herbicidal, insecticidal, acaricidal, nematicidal, and antimicrobial effects of EOs demonstrate their effectiveness and suitability as sustainable and environment-friendly biopesticides. EOs can biodegrade into nontoxic compounds; at the same time, their harmful and detrimental effects on non-target organisms are low. However, few biopesticide formulations based on EOs have been turned into commercial practice upto day. Several challenges including the reduced stability and efficiency of EOs under environmental conditions need to be addressed before EOs are widely applied as commercial biopesticides. This work is an overview of the current research on the application of EOs as biopesticides. Findings of recent studies focusing on the challenges related to the use of EOs as biopesticides are also discussed.

Microencapsulation of Yerba mate extract: The efficacy of polysaccharide/protein hydrocolloids on physical, microstructural, functional, and antioxidant properties.

Effects of hydrolyzed whey protein concentrate (WPC) and its combination with polysaccharides as wall material in spray-drying microencapsulation of Yerba mate extract (YME) have not been investigated yet. Therefore, it is hypothesized that the surface-active properties of WPC or WPC-hydrolysate may improve different properties of spray-dried microcapsules (such as physicochemical, structural, functional and morphological properties) compared to neat MD and GA. Thus, the objective of current study was to produce microcapsules loaded with YME by different carrier combinations. Effect of maltodextrin (MD), maltodextrin-gum Arabic (MD-GA), maltodextrin-whey protein concentrate (MD-WPC), and maltodextrin-hydrolyzed WPC (MD-HWPC) as encapsulating hydrocolloids was studied on physicochemical, functional, structural, antioxidant and morphological characteristics of the spray-dried YME. The type of carrier significantly affected spray dying yield. Enzymatic hydrolysis by improving the surface activity of WPC increased its efficiency as a carrier and produced particles with high production yield (about 68 %) and excellent physical, functional, hygroscopicity and flowability indices. Chemical structure characterization by FTIR indicated the placement of phenolic compounds of the extract in the carrier matrix. FE-SEM study showed that the microcapsules produced with polysaccharide-based carriers were completely wrinkled, whereas, the surface morphology of particles was improved when protein-based carriers were applied. Among the produced samples, the highest amount of TPC (3.26 mg GAE/mL), inhibition of DPPH (76.4 %), ABTS (88.1 %) and hydroxyl (78.1 %) free radicals were related to microencapsulated extract with MD-HWPC. The results of this research can be used to stabilize plant extracts and produce powders with appropriate physicochemical properties and biological activity.

Nano-delivery systems for encapsulation of phenolic compounds from pomegranate peel.

Pomegranate fruit is getting more attention due to its positive health effects, and pomegranate peel (PP) is its main byproduct. PP has the potential to be converted from environmentally polluting waste to wealth due to its rich phenolic compounds such as ellagitannins, proanthocyanidins, and flavonoids with antioxidant, antimicrobial, and health effects. These phenolics are susceptible to environmental conditions such as heat, light, and pH as well as in vivo conditions of gastrointestinal secretions. Some phenolics of PP, e.g., ellagitannins could interfere with food ingredients and thus reduce their beneficial effects. Also, ellagitannins could form complexes with salivary glycoproteins, then a feeling of astringency taste. In this article, nano-delivery systems such as nanoparticles, nanoemulsions, and vesicular nanocarriers, designed and fabricated for PP bioactive compounds in recent years have been reviewed. Among them, lipid-based nano carriers i.e., solid lipid nanoparticles, nanostructured lipid carriers, and vesicular nanocarriers have low toxicity, large-scale production feasibility, easy synthesis, and high biocompatibility. So, it seems that the extraction and purification of bioactives from pomegranate wastes and nanoencapsulating them with cost effective and generally recognized as safe (GRAS) materials can be a bright prospect in enhancing the quality, safety, shelf life and health benefits of pomegranate products.

Synthesis and characterization of selenium nanoparticles stabilized with cocamidopropyl betaine.

In this work, selenium nanoparticles (Se NPs) stabilized with cocamidopropyl betaine were synthesized for the first time. It was observed that Se NPs synthesized in excess of selenic acid had a negative charge with ζ-potential of -21.86 mV, and in excess of cocamidopropyl betaine-a positive charge with ξ = + 22.71 mV. The resulting Se NPs with positive and negative charges had a spherical shape with an average size of about 20-30 nm and 40-50 nm, respectively. According to the data of TEM, HAADF-TEM using EDS, IR spectroscopy and quantum chemical modeling, positively charged selenium nanoparticles have a cocamidopropylbetaine shell while the potential- forming layer of negatively charged selenium nanoparticles is formed by SeO32- ions. The influence of various ions on the sol stability of Se NPs showed that SO42- and PO43- ions had an effect on the positive Se NPs, and Ba2+ and Fe3+ ions had an effect on negative Se NPs, which corresponded with the Schulze-Hardy rule. The mechanism of coagulating action of various ions on positive and negative Se NPs was also presented. Also, influence of the active acidity of the medium on the stability of Se NPs solutions was investigated. Positive and negative sols of Se NPs had high levels of stability in the considered range of active acidity of the medium in the range of 1.21-11.98. Stability of synthesized Se NPs stability has been confirmed in real system (liquid soap). An experiment with the addition of Se NPs stabilized with cocamidopropyl betaine to liquid soap showed that the particles of dispersed phases retain their initial distributions, which revealed the stability of synthesized Se NPs.

Encapsulation of Vitamin B12 by Complex Coacervation of Whey Protein Concentrate-Pectin; Optimization and Characterization.

Vitamin B12 (VB12) is one of the essential vitamins for the body, which is sensitive to light, heat, oxidizing agents, and acidic and alkaline substances. Therefore, the encapsulation of VB12 can be one of the ways to protect it against processing and environmental conditions in food. In this work, the influence of pectin concentration (0.5−1% w/v), whey protein concentrate (WPC) level (4−8% w/v) and pH (3−9) on some properties of VB12-loaded pectin−WPC complex carriers was investigated by response surface methodology (RSM). The findings showed that under optimum conditions (1:6.47, pectin:WPC and pH = 6.6), the encapsulation efficiency (EE), stability, viscosity, particle size and solubility of complex carriers were 80.71%, 85.38%, 39.58 mPa·s, 7.07 µm and 65.86%, respectively. Additionally, the formation of complex coacervate was confirmed by Fourier-transform infrared (FTIR) spectroscopy and atomic force microscopy (AFM). In addition, it was revealed that the most important factor in VB12 encapsulation was pH; at a pH < isoelectric point of WPC (pH = 3), in comparison with higher pH values (6 and 9), a stronger complex was formed between pectin and WPC, which led to an increase in EE, lightness parameter, particle size and water activity, as well as a decrease in the zeta-potential and porosity of complex carriers.

Bioactive-loaded nanodelivery systems for the feed and drugs of livestock; purposes, techniques and applications.

Advances in animal husbandry and better performance of livestock results in growing demands for feed and its nutrients, bioactive compounds (bioactives), such as vitamins, minerals, proteins, and phenolics, along with drugs/vaccines. To protect the feed bioactives in unintended circumstances, they can be encapsulated to achieve desired efficacy in animal feeding and nanoencapsulation gives more potential for better protection, absorption and targeted delivery of bioactives. This study reviews structures, properties, and methods of nanoencapsulation for animal feedings and relevant drugs. Essential oil (EOs) and plant extracts are mostly encapsulated bioactives and phytochemicals for poultry diets and chitosan is found as most effective nanocarrier to load EOs and plant extracts. Nanoparticles (NPs) and nanocapsules are frequently studied nanocarriers, which are mostly processed by using the ionotropic/ionic gelation. Nanofibers, nanohydrogels and nanoemulsions are not found yet for their application in feed bioactives. These nanocarriers can have an improved protection, stability, and controlled release of feed bioactives which benefits to additional nutrition for the growth of livestock regardless of the low stability and water solubility of bioactives. For ruminants' feeds, nano-minerals, vitamins, phytochemicals, essential fatty acids, and drugs are encapsulated by NPs to facilitate the delivery to target organs through direct penetration, to improve their bioavailability, to generate more efficient absorption in cells and tissues, and protect them from rapid degradation. Furthermore, safety and regulatory issues, as well as advantages and disadvantages of nanoencapsulation application in animal feeds are also discussed. The review shows an accurate design of NPs can largely mask safety issues with straightforward approaches and awareness of safety concerns is fundamental for better designing of nanoencapsulation systems and commercialization. This review gives an insight of understanding and potential of nanoencapsulation in ruminants and poultry feedings to obtain a better bioavailability of the nutrients and bioactives with improved safety and awareness for better designing of nanoencapsulating systems.

Practical application of nanoencapsulated nutraceuticals in real food products; a systematic review.

In recent decades, due to the increase in awareness, most consumers prefer foods that not only satisfy their primal urge of hunger but also include health-promoting effects on the body. Therefore, the food industry has an increasing tendency to apply the nutrients (like vitamins, essential fatty acids and minerals) and replace synthetic additives with natural bioactives (like phenolics and essential oils) to produce functional products. However, low dispersibility and shelf-stability as well as presenting unpleasant taste and odor are the most critical barriers for direct incorporation of these useful compounds into foods. In this context, nanoencapsulation has been proposed as a relatively new solution to overcome the mentioned limitations. However, fewer studies have focused on incorporating the bioactive-loaded nanocarriers into the food matrices. This study intends to help the development of functional food production by doing an exhaustive review on the incorporation of nanoencapsulated ingredients into the real food system and resulted interaction of nanocarriers and food products. According to the literature, incorporation of the nanoencapsulated bioactive ingredients into foods can be effectively used to enhance their stability during the processing and storage stage and their bioavailability as well as to delay lipid oxidation and microbial growth in food, without negatively affecting physicochemical, organoleptic and qualitative properties. However, some published results to date declared that food matrix might adversely affect the bioavailability and antimicrobial activity of nanoencapsulated ingredients. It seems that further studies are required to contribute to the choice of appropriate healthy ingredients and wall materials for incorporating into a given food structure.

Anti-Depressant Properties of Crocin Molecules in Saffron.

Saffron is a valued herb, obtained from the stigmas of the C. sativus Linn (Iridaceae), with therapeutic effects. It has been described in pharmacopoeias to be variously acting, including as an anti-depressant, anti-carcinogen, and stimulant agent. The therapeutic effects of saffron are harbored in its bioactive molecules, notably crocins, the subject of this paper. Crocins have been demonstrated to act as a monoamine oxidase type A and B inhibitor. Furthermore, saffron petal extracts have experimentally been shown to impact contractile response in electrical field stimulation. Other research suggests that saffron also inhibits the reuptake of monoamines, exhibits N-methyl-d-aspartate antagonism, and improves brain-derived neurotrophic factor signaling. A host of experimental studies found saffron/crocin to be similarly effective as fluoxetine and imipramine in the treatment of depression disorders. Saffron and crocins propose a natural solution to combat depressive disorders. However, some hurdles, such as stability and delivery, need to be overcome.

Ultrasound-assisted extraction of saffron bioactive compounds; separation of crocins, picrocrocin, and safranal optimized by artificial bee colony.

In this work, a four-factor five-level full factorial central composite design (CCD) was used to optimize the ultrasound-assisted extraction (UAE) of saffron major components, namely picrocrocin, safranal and crocin. The process parameters included ethanol concentration (0-100%), extraction time (2-10 min), duty cycle (0.2-1.0) and ultrasonic amplitude (20-100%). The extracted compounds were measured both by spectrophotometry and chromatography techniques. The results revealed that the middle concentrations of ethanol and relatively long process durations along with high duty cycles and ultrasonic amplitudes had the most profound impact on the yields of the extracted bioactives. UAE was optimized using response surface methodology (RSM) and artificial bee colony (ABC); a comparison between these methods indicated their optimization power was approximately the same. According to the RSM analysis, an ethanol concentration of 58.58%, extraction time of 6.85 min, duty cycle of 0.82 and amplitude of 91.11% were the optimum extraction conditions, while the optimal conditions resulting from ABC were 53.07%, 7.32 min, 0.93 and 100% for the UAE variables respectively. Finally, HPLC analysis was carried out on the UAE optimum extract resulting from RSM. Four crocetin esters were detected in the optimal extract.