Ultrasonic-assisted Enzymatic Extraction Research Articles

Green ultrasonic-assisted enzymatic extraction of polysaccharides from Flammulina velutipes residues by response surface methodology

Flammulina velutipes residues, the by-products of Flammulina velutipes industry, cause environmental pollution. The aim of the current work was to evaluate the extraction efficiency of polysaccharides from Flammulina velutipes residues by an ultrasonic assisted snailase enzymatic extraction (UAEE) method, following the principles of green chemistry, mainly solvent and energy saving. The optimized parameters obtained by Response Surface Methodology (RSM) were determined as follows: ultrasonic power at 150 W, extraction time of 90 min, and extraction temperature of 55 °C. The experimental extraction yield (109.54 ± 5.44 mg/g) was closely matched the predicted yield (110.92 mg/g). Polysaccharides from Flammulina velutipes residues (UE-FVRP) had the molecular weight of 170 kDa and a composition comprising 8 monosaccharides, primarily xylose (Xyl, 24.71%), mannose (Man, 17.29%), arabinose (Ara, 11.48%), and galactose (Gal, 15.63%). FT-IR analysis suggested that UE-FVRP likely contained pyranose with α-glycoside bonds. In vitro antioxidant activity assays revealed that UE-FVRP displayed potent DPPH radical scavenging activity and reducing ability. Additionally, UE-FVRP demonstrated the ability to promote the growth of two Lactobacillus strains, indicating its potential as a novel polysaccharide with high in vitro prebiotic properties. Therefore, these findings proposed a novel, green, and efficient extraction method for extracting polysaccharides from Flammulina velutipes residues, highlighting the potential of utilizing Flammulina velutipes residues as a valuable ingredient in both food and feed applications, contributing to the sustainable reuse of agricultural by-products.

Multi-frequency ultrasonic-assisted enzymatic extraction of coconut paring oil from coconut by-products: Impact on the yield, physicochemical properties, and emulsion stability

Extraction of coconut paring oil (CPO) from processing by-products adds value to the product and reduces resource wastage. This study aims to assess the impact of 20 kHz, 20/80 kHz and 20/40/80 kHz of multi-frequency ultrasonic-assisted enzymatic extraction (MFUAEE) on the yield, physicochemical properties, fatty acid composition, total phenolic content, antioxidant activity, and emulsion stability of CPO derived from wet coconut parings (WCP). Results revealed that the CPO extraction yield with MFUAEE was 32.58 % − 43.31 % higher compared to AEE. The tri-frequency 20/40/80 kHz mode of multi-frequency ultrasound pretreatment exhibited the highest CPO extraction yield (70.08 %). The oil extracted through MFUAEE displayed similar fatty acid profiles to AEE, but had lower peroxide value, K232 and K270 values. Particularly, MFUAEE oil contained higher total phenolic content and exhibited potent DPPH free radical scavenging capacity. Results observed by SEM indicated that the pretreatment with multi-frequency ultrasound more efficiently disrupts the cellular structure of the WCP. Additionally, MFUAEE enhanced emulsion stability through the cavitation effect of ultrasound. These findings suggest that MFUAEE is a valuable approach for method for obtaining CPO with elevated extraction yield and superior quality, thereby enhancing the utilization of coconut by-products.

Optimization of Protein Extraction from Halopteris scoparia Macroalgae by Ultrasonic-Assisted Enzymatic Extraction (UAEE): Bioactive, Chemical, and Technological Properties

Optimization of Protein Extraction from <i>Halopteris scoparia</i> Macroalgae by Ultrasonic-Assisted Enzymatic Extraction (UAEE): Bioactive, Chemical, and Technological Properties

Box-Behnken Design for Optimizing Ultrasonic-Assisted Enzymatic Extraction of Soluble Dietary Fiber from Pleurotus citrinopilestus

Pleurotus citrinopilestus contains a variety of physiologically and pharmacologically active compounds. A key active component among these compounds is dietary fiber, a polysaccharide that exhibits several biological properties. The objective of this study was to assess how soluble dietary fiber (SDF) from Pleurotus citrinopilestus responded to ultrasonic-assisted enzymatic (UAE) extraction. The response surface method (RSM) combined with the Box-Behnken design method (BBD) was used to optimize the yield of SDF. The effects of the liquid-solid ratio (35–55 mL/g), α-amylase concentration (0.5–2.5%), complex protease concentration (0.4–2.0%), and ultrasonication time (15–55 min) on the yield of SDF were examined. The RSM results revealed the optimal liquid-solid ratio (45 mL/g), α-amylase concentration (1.5%), complex protease concentration (1.2%), and ultrasonic time (35 min). The SDF yield was 10.25%, which is close to the predicted value (10.08%).

Optimization of the ultrasonic-assisted enzymatic extraction of polysaccharides from Dendrobium offcinale Kimura et Migo and bioactivity study

ABSTRACT Ultrasonic-assisted enzymatic extraction (UAEE) technology was employed to extract the polysaccharides from Dendrobium offcinale Kimura et Migo (D. offcinale). The extraction conditions were optimized by central composite design (CCD) found in the response surface methodology (RSM). The polysaccharide yield (34.05 ± 1.03%) was well matched with the predicted value, as obtained under the optimum parameters: enzyme activity of 2040 U/g, a liquid-solid ratio of 71 mL/g, and a time of 84 min. Compared to other extraction methods, UAEE was able to yield D. offcinale polysaccharide (DOP) highly efficiently in a shorter extraction time period and at a lower extraction temperature. The extracted polysaccharide (DOPUAEE) showcased a better nitrite scavenging ability, a high antioxidant activity, and more potency in inhibiting α-glucosidase, expanding its versatility in food applications.

Effects of ultrasonic-enzymatic-assisted ethanol precipitation method on the physicochemical characteristics, antioxidant and hypoglycemic activities of Tremella fuciformis polysaccharides

This investigation involved the extraction of a novel polysaccharide from the spore fermentation broth of Tremella fuciformis using a method that combined ultrasonic and enzymatic assistance with ethanol precipitation. It was then compared with enzymatic and ultrasonic extraction methods. The objective of this research is to offer a reference point for expanding the application of ultrasonic-assisted enzymatic extraction technology in T. fuciformis polysaccharides (TFPs). Based on single-factor experiments, Box-Behnken was used to optimize the extraction conditions of TFPs by ultrasonic-enzymatic-assisted ethanol precipitation extraction. The results revealed an optimal combination of enzymes, with a cellulase-to-papain ratio of 2:1, an enzyme addition of 4000U/100 mL, an enzymolysis temperature of 49 °C, ultrasonicpower at 3 W/mL and an ultrasonictime of 20 min. The extraction rate of TFPs and α- amylase inhibition rates were 23.94 % and 61.44 %, respectively. Comparing the physicochemical properties, structural characterization and in vitro activity of TFPs extracted through different methods, the results showed that ultrasonic treatment significantly influences the apparent morphology of polysaccharide and could enhance its in vitro biological activity. However, different extraction techniques exhibit insubstantial impact on the chemical composition, glycosidic bonds or glycosidic ring configurations within the polysaccharides. Among them, ultrasonic-enzymatic-assisted ethanol precipitation extraction of polysaccharide has the highest extraction rate and the lowest viscosity. It has significant effects on ABTS+ scavenging activity, α- amylase inhibition rate and glucose dialysis retardation index, polysaccharide treated with ultrasonic-enzymatic showed the best performance. These findings suggest that ultrasonic-enzymatic-assisted ethanol precipitation extraction can enhance the activities of TFPs, thereby providing a valuable insight for their future development and application.

Hybrid RSM-GA approach to optimize extraction conditions for blueberry anthocyanins with high antioxidant activity

BACKGROUND: Blueberry contains an abundance of anthocyanins, which are a bioactive component of this fruit. Anthocyanins can be extracted via various methods, and each has pros and cons. OBJECTIVE: This current study reported the optimal conditions for the ultrasonic-assisted enzymatic extraction of blueberry anthocyanins simulated using response surface methodology (RSM) coupled with a genetic algorithm (GA). METHODS: The Box–Behnken design (BBD) was used for the RSM, and the extraction conditions were as follows: temperature, 42°C; ultrasonic power, 310 W; enzyme volume, 0.25%; and extraction time, 42 min. RESULTS: The maximum predicted extraction yield was 6.67 mg/g. The antioxidant activity of anthocyanins extracted via RSM and GA was based on the hydroxyl free radical activity and supersonic anion free radical activity of 230.50±12.76μg/ml and 4.41±0.36μg/ml, respectively. Anthocyanins exracted by the proposed method has stronger free radical removal capacity than that of Vc. CONCLUSIONS: This study shows that the combination of RSM with GA represents an optimized method for extracting blueberry anthocyanins for use in the food industry. This method can maintain high antioxidant potential and can be used as an alternative strategy for high-value products.

Ultrasonic-assisted enzymatic extraction of sulfated polysaccharide from Skipjack tuna by-products

The effect of ultrasound pretreatment on extraction efficiency of sulfate polysaccharides (SPs) using alcalase from different by-products of Skipjack tuna including head, bone and skin was evaluated. Structural, functional, antioxidant and antibacterial properties of the recovered SPs using the ultrasound-enzyme and enzymatic method were also investigated. Ultrasound pretreatment significantly increased the extraction yield of SPs from all the three by-products compared with the conventional enzymatic method. All extracted SPs showed high antioxidant potential in terms of ABTS, DPPH and ferrous chelating activities where the ultrasound treatment enhanced antioxidant activities of the SPs. The SPs exerted strong inhibiting activity against various Gram-positive and Gram-negative bacteria. The ultrasound treatment remarkably increased antibacterial activity of the SPs against L. monocytogenes but its effect on other bacteria was dependent on the source of the SPs. Altogether, the results suggest that ultrasound pretreatment during enzymatic extraction of SPs from tuna by-products can be a promising approach to improve extraction yield but also bioactivity of the extracted polysaccharides.

Ultrasonic-assisted enzymatic improvement of polyphenol content, antioxidant potential, and in vitro inhibitory effect on digestive enzymes of Miang extracts

The aims of this research were to optimize the ultrasonic-assisted enzymatic extraction of polyphenols under Miang and tannase treatment conditions for the improvement of antioxidant activity of Miang extracts via response surface methodology. Miang extracts treated with and without tannase were investigated for their inhibitory effects on digestive enzymes. The optimal conditions for ultrasonic-assisted enzymatic extraction of the highest total polyphenol (TP) (136.91 mg GAE/g dw) and total flavonoid (TF) (5.38 mg QE/g dw) contents were as follows: 1 U/g cellulase, 1 U/g xylanase, 1 U/g pectinase, temperature (74 °C), and time (45 min). The antioxidant activity of this extract was enhanced by the addition of tannase obtained from Sporidiobolus ruineniae A45.2 undergoing ultrasonic treatment and under optimal conditions (360 mU/g dw, 51 °C for 25 min). The ultrasonic-assisted enzymatic extraction selectively promoted the extraction of gallated catechins from Miang. Tannase treatment improved the ABTS and DPPH radical scavenging activities of untreated Miang extracts by 1.3 times. The treated Miang extracts possessed higher IC50 values for porcine pancreatic α-amylase inhibitory activity than those that were untreated. However, it expressed approximately 3 times lower IC50 values for porcine pancreatic lipase (PPL) inhibitory activity indicating a marked improvement in inhibitory activity. The molecular docking results support the contention that epigallocatechin, epicatechin, and catechin obtained via the biotransformation of the Miang extracts played a crucial role in the inhibitory activity of PPL. Overall, the tannase treated Miang extract could serve as a functional food and beneficial ingredient in medicinal products developed for obesity prevention.

Protective Effect of Flavonoids from Mulberry Leaf on AAPH-Induced Oxidative Damage in Sheep Erythrocytes.

To evaluate the antioxidant activity of flavonoids extracted from Chinese herb mulberry leaves (ML), flavonoids from mulberry leaves (FML) were extracted and purified by using ultrasonic-assisted enzymatic extraction and D101 macroporous resin. Using LC-MS/MS-Liquid Chromatography with tandem mass spectrometry analysis, hesperidin, rutoside, hyperoside, cyanidin-3-o-glucoside, myricitrin, cyanidin, and quercetin were identified, and NMR and UV were consistent with the verification of IR flavonoid characteristics. The antioxidant activity of FML has also been evaluated as well as the protective effect on 2,2 0-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress. The results showed that FML exhibited powerful antioxidant activity. Moreover, FML showed dose-dependent protection against AAPH-induced sheep erythrocytes' oxidative hemolysis. In the enzymatic antioxidant system, pretreatment with high FML maintained the balance of SOD, CAT, and GSH-Px; in the non-enzymatic antioxidant system, the content of MDA can be effectively reduced after FML treatment. This study provides a research basis for the development of natural products from mulberry leaves.

Purification, structural characterization and immunological activity of Sibiraea laexigata (L.) Maxim polysaccharide.

Sibiraea laexigata (L.) Maxim (SLM) has been used as an herbal tea for treating stomach discomfort and indigestion for a long time in china. Polysaccharides have been identified as one of the major bioactive compounds in the SLM. In the present paper, ultrasonic-assisted enzymatic extraction (UAEE) method was employed in polysaccharides extraction derived from SLM using polyethylene glycol (PEG) as extraction solvent, two SLM polysaccharides (SLMPs) fractions (SLMPs-1-1 and SLMPs-2-1) were purified by DEAE Cellulose-52 and Sephadex G-100 chromatography in sequence. Then, the preliminarily structure of the two factions were characterized by chemical composition analysis, molecular weight measurement, UVS, HPLC-PMP, FT-IR, nuclear magnetic resonance (NMR) spectra analysis and SEM. The results showed that SLMPs-1-1 and SLMPs-2-1 with different molecular weights of 1.03 and 1.02 kDa, mainly composed of glucose (46.76 and 46.79%), respectively. The results of structural characterization from FT-IR, 1H NMR, and SEM revealed that SLMPs-1-1 and SLMPs-2-1 contained the typical pyranoid polysaccharide with α-glycosidic bond and β-glycosidic bond. Furthermore, it was found that SLMPs-1-1 could increase the levels of tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2), and alleviated the immune organs tissue damage of cyclophosphamide (Cy)-treated mice. RT-qPCR and Western-Blot analysis showed that SLMPs-1-1 could significantly up-regulated the levels of NF-κB, TLR4, which revealed that SLMPs-1-1 could participate in immunosuppressive protection of Cy-treated mice. These findings suggested that the potential of SLMPs-1-1 as an alternative immunostimulator could be used in food and pharmaceutical industries.

Structure characterization and antioxidant activity analysis of polysaccharides from Lanzhou Lily.

Lanzhou Lily (Lilium davidii var. unicolor) is a traditional medicinal plant and popular edible vegetable bulb in China. In this study, the polysaccharides of Lanzhou Lily (LLPs) were extracted by polyethylene glycol-based ultrasonic-assisted enzymatic extraction method (PEG-UAEE). The optimum process conditions were obtained by single-factor experiments and response surface methodology (RSM). Then, the preliminarily structure of LLPs was characterized by HPLC, FT-IR, and SEM, and its antioxidant activities were evaluated. The results showed that LLPs yield reached 14.75% under the optimized conditions: E/S ratio 1,400 U/g; pH 5.0, ultrasonic time 30 min; and ultrasonic temperature 50 °C. The LLPs has pyranoid ring, uronic acid, and the characteristic absorption peaks of -OH, C = O, and C-H. The results of scanning electron microscope indicated that the LLPs had irregular distribution, dispersed structure, and many holes. The HPLC analysis showed that the LLPs were heteropolysaccharide containing galactose (6.36%), glucose (76.03%), rhamnose (2.02%), and arabinose (7.09%). Moreover, the LLPs showed obvious antioxidant effect in vitro.

An integrated study using ultrasonic-assisted enzymatic extraction of hydrolysates from rice based distillery byproduct and its characterization

The present study aim is to establish a greener approach by integrating an ultrasonic process that is amalgamated with enzymes for distillers dried grains with solubles (DDGS), a distillery by-product, into an improved value-added product. A good synergistic effect is recorded by applying ultrasonication in conjunction with enzymatic extraction of protein from rice-based DDGS. The ultrasonic-assisted extraction (UAE) method was examined for optimum conditions using Box-Behnken design (BBD) based response surface methodology (RSM). The optimized parameters viz. amplitude, solid loading and extraction time, were obtained as 90.01%, 10% and 18.06 min, respectively which yielded 65.76% of DDGS protein. The coefficient of determination (R2) for extracted protein (%) and degree of hydrolysis (DH) were 0.981 and 0.976, respectively validating a good fit of the model with the experimental data. The protein hydrolysates obtained after ultrasonic-assisted enzymatic hydrolysis were found to exhibit significantly (P < 0.05) improved functional properties in comparison to untreated DDGS. Additionally, the effects of applied treatment was evaluated by a reduction in the particle size, structural changes (XRD and FE-SEM) and their functional properties (solubility, water holding capacity, oil holding capacity, emulsification, and foaming properties). The obtained results reveal that ultrasonic-assisted enzymatic hydrolysis was capable of enhancing the functional properties of extracted protein hydrolysate. Therefore, ultrasonic-assisted enzymatic hydrolysis can be applied as a promising technology for improving protein yield and functionalities of rice DDGS protein, which could facilitate the potential application of distillery by-products in the food industry.

Comparison of different extraction methods on the physicochemical, structural properties, and in vitro hypoglycemic activity of bamboo shoot dietary fibers

The effect of alkali extraction (AE), enzymatic extraction (EE), ultrasonic-assisted enzymatic extraction (UAEE), and shear homogeneous-assisted enzymatic extraction (SHAEE) on the physicochemical, structural properties, and in vitro hypoglycemic activity of bamboo shoot dietary fibers (BSDF) were investigated and compared. BSDF obtained by AE had the lowest protein content and crystallinity index. The lowest oil holding capacity (OHC) and highest protein content were observed in EE. BSDF generated highest OHC and glucose adsorption capacity by UAEE. SHAEE obtained the highest SDF content (17.89%), water-holding capacity (8.81 g/g), and α-amylase activity inhibition ratio (19.89%) and the smallest particle size (351.33 μm). BSDF extracted by SHAEE and UAEE presented a porous and loose structure. Furthermore, the in vitro hypoglycemic activity of the four BSDF samples generally followed the order of SHAEE > UAEE > EE > AE. Results show that SHAEE is an innovative and promising method to obtain BSDF with its excellent physicochemical and functional properties.

Effect of Extraction Procedures with Ultrasound and Cellulolytic Enzymes on the Structural and Functional Properties of Citrus grandis Osbeck Seed Mucilage.

The structural and functional properties of Citrus grandis Osbeck (CGO) seed mucilage by different extraction practices, including conventional citrate buffer, ultrasonic-assisted (UAE), enzymatic-assisted extraction (EAE) with cellulase or Celluclast® 1.5 L and various ultrasonic-assisted enzymatic extraction (UAEE) procedures were investigated. It was found that CGO seed from agricultural and processing byproducts is an excellent new source of high methoxyl pectin with quite high intrinsic viscosity (about 108.64 dL/g) and molecular weight (about 1.9 × 106) as compared with other pectin sources. UAEE with Celluclast® 1.5 L enhanced the extraction yield most pronouncedly (about 2.3 times). Moreover, the monosaccharide composition of CGO seed mucilage is least affected by EAE with Celluclast® 1.5 L. In contrast, EAE with cellulase dramatically reduces the galacturonic acid (GalA) content to less than 60 molar%, and increases the glucose (Glc) content pronouncedly (to about 40 molar%), which may be considered as an adverse effect in terms of pectin purity. Though extraction procedures involved with ultrasound and cellulolytic enzymes generally show a decrease in GalA contents, weight average molar mass and intrinsic viscosity, EAE with Celluclast® 1.5 L is least affected, followed by UAE and UAEE with Celluclast® 1.5 L. These features can be leveraged in favor of diversified applications.

Optimization of ultrasonic-assisted enzymatic extraction of kiwi starch and evaluation of its structural, physicochemical, and functional characteristics

A new ultrasound-assisted enzymatic extraction (UAEE) method of starch from kiwifruit was established and optimized using response surface methodology (RSM). Under optimal conditions (the pectinase-to-cellulase-to-papain ratio = 1:2:1 g/kg, solid/liquid ratio = 1:6.68, extraction pH = 5.23, ultrasound power = 300 W, and extraction temperature = 52 °C), the kiwi starch (KS) yield was about 4.25%, and the starch content of KS was 873.23 mg/g. Compared to other extraction methods, UAEE can obtain KS with high yield and purity with a shorter extraction time and less solvent and enzyme. The extracted KS has a low gelatinization enthalpy (8.02 J/g) and a high peak viscosity (7933 cP), with obvious particle properties and low adhesion. In addition, KS is rich in polyphenols, has strong antioxidant activity, and has higher contents of amylose starch (30.74%) and resistant starch (60.18%). This study established a novel and highly efficient method for KS extraction and suggest several possible applications for KS in the food industry.

Optimization of Ultrasonic-Assisted Enzymatic Extraction of Tannic Acid from Chromolaena Odorota sp.

Previous study only implemented the time consuming and low amount of yield technique for extraction from Chromolaena odorata which is conventional method. Nonconventional extraction method with short extraction time and high amount of yield was applied in this study by applying ultrasound-assisted enzymatic extraction (UAEE). UAEE was used to extract tannic acid from Chromolaena odorata. The extraction parameters involved were enzyme concentration, sonication time and duty cycle at constant temperature of 50°C, solid to liquid ratio of 1:10 and sonication power at 60% amplitude. The optimum extraction process was found at cellulase enzyme concentration of 4%, sonication time of 60 minutes and duty cycle of 50% with the obtained concentration of tannic acid at 1.6152 mg/mL. The study showed that the UAEE could be employed to enhance yield of tannic acid, reduce the extraction time and ensuring green extraction method were applied in the study.

Ultrasonic-Assisted Enzymolysis Extraction and Protective Effect on Injured Cardiomyocytes in Mice of Flavonoids from Prunus mume Blossom.

Prunus mume blossom is an edible flower that has been used in traditional Chinese medicine for thousands of years. Flavonoids are one of the most active substances in Prunus mume blossoms. The optimal ultrasonic-assisted enzymatic extraction of flavonoids from Prunus mume blossom (FPMB), the components of FPMB, and its protective effect on injured cardiomyocytes were investigated in this study. According to our results, the optimal extraction process for FPMB is as follows: cellulase at 2.0%, ultrasonic power at 300 W, ultrasonic enzymolysis for 30 min, and an enzymolysis temperature of 40 °C. FPMB significantly promoted the survival rate of cardiomyocytes and reduced the concentration of reactive oxygen species (ROS). FPMB also improved the activities of proteases caspase-3, caspase-8, and caspase-9 in cardiomyocytes. The cardiomyocyte apoptosis rate in mice was significantly reduced by exposure to FPMB. These results suggest that the extraction rate of FPMB may be improved by an ultrasonic-assisted enzymatic method. FPMB has a protective effect on the injured cardiomyocytes.

Characterization, antioxidant and immunomodulatory effects of selenized polysaccharides from dandelion roots

The polysaccharide (DRP) was gained from dandelion roots by ultrasonic-assisted enzymatic extraction (UAEE) followed by two-step column purification. Then selenylation of DRP has been accomplished by HNO3-Na2SeO3 method. sDRP-1 and sDRP-2 with the selenium content of 170 ± 1.13 and 710 ± 4.00 μg/g were prepared for further structural characterization and bioactivity determination. DRP, sDRP-1, and sDRP-2 were composed of the same monosaccharides in different molar ratios, and the molecular weights of DRP, sDRP-1 and sDRP-2 were 8700, 7900, and 5600 Da, respectively. Fourier transform infrared (FT-IR) spectra confirmed that DRP, sDRP-1, and sDRP-2 possessed similar functional groups. The results of Congo red test, X-ray diffraction (XRD) and scanning electron microscopy (SEM) showed that DRP, sDRP-1, and sDRP-2 had no three helix structure, did not form single crystal, and all belonged to amorphous morphology. sDRP-1 and sDRP-2 possessed greater antioxidant activities in vitro than the native polysaccharide DRP. At the same time, the selenized polysaccharides showed better immunomodulatory ability and could be used as new-type immunoenhancer. The present conclusions provided theoretical basis for the new application of dandelion polysaccharides and the development of dandelion resources.

Development and validation of a novel approach based on ultrasonic-assisted enzymatic extraction and RP-UHPLC for simultaneous determination of astaxanthin isomers and alpha-tocopherol in Haematococcuspluvialis derived supplements

A rapid, effective andgreenmethod was developed for simultaneous determination of total (free and esterified) astaxanthin (AX) isomers (all-E, 9Z and 13Z) and alpha-tocopherol(AT) in Haematococcuspluvialis derived supplements. The new method employed a highly efficient ultrasonic-assisted enzymatic extraction (UAEE) techniqueto perform deesterificationwith Cholesterol esterase from Pseudomonas fluorescenspermitting the concurrent detection. The subsequent RP-UHPLC method for separating and measuring was performed on a simple C18 column within 10.5 min by using methanol and ammonium acetate as mobile phase with a gradient elution. The proposed method was validated according to international guidelines and itproved to be highly accurate and robust. The optimized UHPLC method allowed easy transfer to HPLC, and allowed rapid analysis of active ingredients profiling in H. derived supplements.To our knowledge, this is the first quantification approach describing the rapid simultaneous analysis of the functional lipophilic substances including AX isomers in H. derived supplements using UAEE technique combined with RP-UHPLC.Moreover, this holistic approachcan be used to identify whether AX products are of natural origin or chemical synthesis, and may find more applications in new forms of H. derived productswith complexbiological matrix for more research on the bioavailability of AXisomersfrom natural source.