Synergistic effects of Ultrasound-Assisted Ultra-High pressure on quality attributes and flavor profile of lactic acid Bacteria-Fermented fresh jujube juice.

Synergistic effects of Ultrasound-Assisted Ultra-High pressure on quality attributes and flavor profile of lactic acid Bacteria-Fermented fresh jujube juice - PMC Skip to main content An official website of the United States government Here's how you know Here's how you know Search Log in Primary site navigation Search Log in Search PMC Full-Text Archive Search in PMC Journal List User Guide As a library, NLM provides access to scientific literature. Inclusion in an NLM database does not imply endorsement of, or agreement with, the contents by NLM or the National Institutes of Health. Learn more: PMC Disclaimer | PMC Copyright Notice Ultrason Sonochem . 2026 May 23;130:107903. doi: 10.1016/j.ultsonch.2026.107903 Synergistic effects of Ultrasound-Assisted Ultra-High pressure on quality attributes and flavor profile of lactic acid Bacteria-Fermented fresh jujube juice Yu Zhang 1, 1 , Qinjie Xue 1, 1 , Yao Yao 1 , Jiachen Xie 1 , Jiaxin Yan 1 , Ruier Yao 1 , Miao Yang 1 , Xiaoyan Sun 1 , Zhihui Yu 1, ⁎ , Ling Ma 1, ⁎ Author information Article notes Copyright and License information 1 College of Food Science and Engineering, Shanxi Agricultural University, Taigu 030801, Shanxi, China ⁎ Corresponding author. zhihuisxau@163.com ml0359@163.com 1 These authors contributed to the work equally and were regarded as co-first authors. Received 2026 Feb 25; Revised 2026 May 7; Accepted 2026 May 22; Collection date 2026 Jul. © 2026 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). PMC Copyright notice PMCID: PMC13235383  PMID: 42208385 Graphical abstract Open in a new tab Keywords: Fermented jujube juice, Ultra-high pressure, Ultrasound, Synergistic mechanism, Antioxidant activity, Volatile flavor compounds Abstract Non-thermal processing technologies have emerged as promising alternatives to preserve bioactive compounds in fruit-based beverages. However, the synergistic effects of ultrasound-assisted ultra-high pressure (US-UHP) combined with lactic acid bacteria fermentation on jujube juice quality remain poorly understood. This study systematically investigated the effects of US-UHP pretreatment on physicochemical properties, bioactive compounds, volatile profiles, and sensory characteristics of fermented jujube juice (FJJ). Results demonstrated that US-UHP pretreated samples exhibited superior quality attributes. At 24‘h fermentation, total phenolic content reached 2.32 mg/mL, ABTS and DPPH radical scavenging activities were 87.30% and 74.86%, respectively, and ascorbic acid content was significantly higher than other groups. After 40 h fermentation, reducing sugar consumption rate reached 54.69%. Notably, US-UHP treatment significantly improved colloidal stability, with particle size maintained at 33.41–53.21 d.nm compared to 1597.67 d.nm in untreated samples, accompanied by lower polydispersity index values. Gas Chromatography-Ion Mobility Spectrometry and Headspace Solid-Phase Microextraction Gas Chromatography-Mass Spectrometry analyses revealed that US-UHP treatment effectively reduced undesirable flavor compounds (butyric acid, hexanal, acetaldehyde) while promoting the accumulation of floral and fruity scent-related ketones. Principal component analysis of electronic nose data confirmed distinct flavor differentiation among treatments, and sensory evaluation demonstrated that US-UHP pretreated samples achieved the highest overall acceptability scores. Correlation analysis indicated significant positive relationships between total phenolic/flavonoid contents and overall acceptance (r > 0.92). These findings demonstrate that US-UHP pretreatment synergistically enhances microbial inactivation, bioactive compound release, and metabolic efficiency, providing theoretical insights and technical guidance for high-quality FJJ development. 1. Introduction Huping jujube ( Ziziphus jujuba Mill. 'Huping zao'), a perennial plant of the Rhamnaceae family, is characterized by its distinctive flask-like morphology and rich functional components, including polysaccharides, phenolic compounds, cyclic adenosine monophosphate (cAMP), triterpenic acids, and flavonoids [1] , [2] . These constituents confer notable biological activities such as antioxidant, anti-inflammatory, hypoglycemic, and neuroprotective effects, underscoring its significant potential within the functional food sector. However, the high moisture content and low acidity of fresh jujube render it susceptible to postharvest deterioration, resulting in substantial losses during storage and transportation [3] . Consequently, its utilization has been largely confined to primary processing methods, which constitutes a critical bottleneck impeding the extension of its industrial value chain. Conventional thermal processing methods effectively inactivate microorganisms and enzymes to extend shelf life; nonetheless, such treatments inevitably compromise both sensory attributes and chemical composition. These adverse effects include vitamin loss, reduction in reducing sugars, flavor deterioration [4] , degradation of biologically active compounds, and undesirable sensory qualities induced by heating [5] . This limitation constrains their commercial viability and health-promoting potential. Consequently, there is growing interest in alternative processing approaches capable of preserving or even enhancing the nutritional and sensory properties of these products while ensuring safety and stability [6] . The use of emerging non-thermal technologies, such as ultra-high pressure (UHP), enables microbial inactivation while preserving nutritional integrity and organoleptic characteristics [7] . UHP utilizes hydrostatic pressure of 100–600 MPa to disrupt microbial cell membranes, achieving efficient sterilization while promoting the release of phenolic and flavonoid compounds, thereby significantly enhancing the antioxidant activity of juice [8] . Ultrasound (US) technology, relying on the cavitation effect (the localized high temperature and pressure generated by the rapid formation and collapse of microbubbles), achieves cell wall disruption, enhances mass transfer, and enables efficient extraction of bioactive components [9] . US intervention applied to lactic acid bacteria during the logarithmic growth phase can increase the total ester content in fermented jujube juice (FJJ) by fourfold, significantly optimizing the aroma profile [3] . When US technology is combined with fermentation, it can markedly improve the release efficiency of phenolic and flavonoid compounds by promoting the decomposition of plant cell walls and enhancing enzyme activity [10] . Furthermore, the synergistic application of UHP and US in treating jujube juice significantly increases the total phenolic content and antioxidant activity, while effectively inhibiting the Maillard reaction and preserving the original color of the juice [8] . The synergistic integration of these technologies offers promising avenues for maximizing extraction efficiency while maintaining functional integrity and sensory characteristics, thereby addressing both market expectations and manufacturing requirements. Fermenting fruit juices with lactic acid bacteria (LAB) holds significant potential and promising market prospects in the food industry. LAB fermentation can alter the composition and characteristics of bioactive compounds, improve the sensory quality of fermented juices, extend shelf life, and enhance the bioavailability of vitamins and minerals. This process has been proven to enrich the phenolic profile and bioactive components of fruit juices [11] . Lactiplantibacillus plantarum can produce pectinases and cellulases through its metabolism, degrading the cell walls of Huping jujubes and releasing bound phenolic compounds. Concurrently, it converts sugars into organic acids such as lactic acid and acetic acid, reducing the sweetness of the juice and imparting a refreshing taste [12] . Studies have shown that fermenting jujube juice with L. plantarum LP-CWC-Zao can increase the total phenolic content, enhance the DPPH radical scavenging rate by 28.56%, and synthesize esters like ethyl benzoate, enriching the fruity aroma profile [13] . However, single-strain fermentation of Huping jujube juice has certain limitations: its high sugar and high viscosity characteristics can easily lead to a decline in LAB viability during the later stages, resulting in incomplete sugar consumption. Furthermore, the disproportionate balance of alcohols and aldehydes in traditionally fermented juices can easily mask the inherent light, sweet flavor of Huping jujubes, leading to a sour and astringent aftertaste [14] . Co-fermentation with LAB can effectively promote the release of polyphenols, drive the structural transformation of flavonoids, and induce a reconstruction of the phenolic profile. Previous studies have separately investigated single non-thermal technologies on jujube juice quality, such as UHP-assisted phenolic release [15] or US intervention on L. plantarum during fermentation [16] . Mixed-strain LAB fermentation has also been shown to reshape the phenolic profile of fruit juices [17] , enhance their antioxidant capacity and overall quality, improve their nutritional components and volatile flavor compounds, and further boost their functional properties [18] . However, to the best of our knowledge, the synergistic effects of US-UHP treatment combined with mixed-strain LAB fermentation on fresh jujube juice have not been systematically investigated, and the dynamic changes in flavor, colloidal stability, and bioactive compounds under this combined treatment remain unknown. This novel approach was systematically compared with the traditional pasteurization process. Multidimensional analytical techniques, including Gas Chromatography-Ion Mobility Spectrometry (GC-IMS), Headspace Solid-Phase Microextraction Gas Chromatography-Mass Spectrometry (HS-SPME-GC–MS), and electronic nose, were employed to systematically investigate the effects of different processing methods on the physicochemical properties, functional components, and volatile flavor compound profiles of fermented fresh jujube juice. This study aimed to establish a theoretical foundation and technical framework for the high-value utilization of fresh jujube resources, while simultaneously providing scientific insights for quality assurance and industrial scale-up of fermented specialty fruit and vegetable beverages. 2. Materials and methods 2.1. Materials Fresh Huping jujubes ( Ziziphus jujuba Mill. 'Huping') were procured from a local market in Taigu District, Jinzhong City, Shanxi Province, China, and stored at 4℃ until use. Lactiplantibacillus plantarum and Lactobacillus acidophilus were obtained from the Microbiology Laboratory, College of Food Science and Engineering, Shanxi Agricultural University. DPPH, ABTS, 3,5-dinitrosalicylic acid, Folin-Ciocalteu reagent, gallic acid, rutin, and other chemicals were of analytical grade (Aladdin Reagent Co., Ltd., Shanghai, China). 2.2. Preparation of fresh jujube juice Huping jujubes were washed, pitted, and homogenized with deionized water (1:2, w/v) using a high-speed tissue homogenizer. The slurry was enzymatically hydrolyzed with pectinase (0.8 g/L) and cellulase (1.6 g/L) at 50℃ for 4 h, followed by enzyme inactivation at 90℃ for 10 min. After cooling to ambient temperature, the mixture was sequentially filtered through 80-, 200-, and 500-mesh sieves. The total soluble solids (TSS) content was standardized by sucrose addition, and the juice was stored at 4℃ prior to use [14] . 2.3. Preparation of FJJ using US-UHP A mixed culture of L. plantarum and L. acidophilus (2:1, v/v) was cultivated anaerobically in MRS broth at 37℃ for 24 h. Cells at the logarithmic phase were harvested by centrifugation (4000 rpm, 10 min), washed twice with sterile saline (0.9% NaCl), and resuspended to 10 8 CFU/mL. Five treatment groups were established: (1) FJJ-C: uninoculated control; (2) FJJ-P: pasteurization (85℃, 15 min) followed by inoculation; (3) FJJ-H: UHP(500 MPa, 10 min) followed by inoculation; (4) FJJ-P-U: pasteurization plus US treatment (300 W, 40 kHz, pulsed mode 5 s on/5 s off, 15 min, ≤35℃) applied 2 h post-inoculation; (5) FJJ-H-U: UHP plus US treatment under identical conditions. US treatment was applied at 2 h post-inoculation, which was considered to correspond to the early exponential growth phase of the mixed starter culture under the present fermentation conditions (37 °C, 1.5% v/v inoculum, ≥10 8 CFU/mL). This timing was selected mainly based on previous studies showing that US intervention during the early logarithmic phase more effectively promotes LAB activity, metabolic transformation, and quality development than treatment applied at other growth stages [19] , [20] , [21] . Each group was inoculated at 1.5% (v/v) and fermented anaerobically at 37℃ for 40 h. Samples were withdrawn at designated intervals for physicochemical analysis (0, 8, 16, 24, 32, and 40 h), antioxidant activity and Zeta potential and particle size (0, 24, and 40 h), viable LAB counts (16 and 40 h), sensory evaluation (24 and 40 h), and volatile compound analysis via GC-IMS, HS-SPME-GC–MS, and electronic nose (24 h). 2.4. Determination of pH The pH was measured using a calibrated pH meter (PHS-3C, Shanghai Sanxin Instrument Factory, China). Aliquots (10 mL) of supernatant were sampled every 8 h, and measurements were performed in triplicate [22] . 2.5. Determination of TSS content TSS was determined using a digital refractometer (SW-32D, Speedwell Electronic Technology Co., Ltd., China) at 25℃. The instrument was calibrated with deionized water before each measurement. Results were expressed as °Brix, and all measurements were performed in triplicate [14] . 2.6. Determination of browning index (BI) Supernatant samples (2 mL) were mixed with an equal volume of 95% (v/v) ethanol, vortexed, and centrifuged at 10,000 rpm for 10 min. Absorbance was measured at 430 nm using a UV–Vis spectrophotometer (Evolution-220, Thermo Fisher Scientific, China) with distilled water as blank. The browning index (BI) was calculated as: BrowningIndex ( B I ) = A 430 × D i l u t i o n F a c t o r 2.7. Determination of color Color parameters were measured using a tristimulus colorimeter (CR-400, Konica Minolta, Japan) calibrated with a standard white plate ( L* = 97.30, a* = −0.23, b* = 0.22). Supernatant samples (10 mL) were transferred to Petri dishes for measurement. Results were expressed as CIE Lab* values, where L* indicates lightness, a* indicates redness (+)/greenness (−), and b* indicates yellowness (+)/blueness (−). Measurements were conducted in triplicate [23] . 2.8. Determination of ascorbic acid content Ascorbic acid content was determined spectrophotometrically at 265 nm (Evolution-220, Thermo Fisher Scientific, China). A standard curve was prepared using ascorbic acid (0–10 μg/mL) dissolved in 2% oxalic acid solution. Samples were appropriately diluted with the same solution, and ascorbic acid content was calculated from the standard curve and expressed as mg/100 g. 2.9. Determination of total phenolic content Total phenolic content (TPC) was determined by the Folin-Ciocalteu method [24] . Briefly, diluted samples (1 mL, 1000-fold dilution) were mixed with Folin-Ciocalteu reagent (0.5 mL), followed by 10% Na 2 CO 3 (1.5 mL) and distilled water (7 mL). After incubation at 25℃ for 60 min in darkness, absorbance was measured at 710 nm. Results were expressed as gallic acid equivalents (mg/mL) based on a standard curve. 2.10. Determination of total flavonoid content Total flavonoid content (TFC) was measured using the aluminum nitrate colorimetric method with modifications [25] . Diluted samples (2.5 mL to 5.0 mL) were sequentially mixed with 5% NaNO 2 (1 mL, 6 min), 10% Al(NO 3 ) 3 (1 mL, 6 min), 4% NaOH (10 mL), and 60% ethanol (8 mL). After incubation at 25℃ for 12 min, absorbance was recorded at 510 nm. Results were expressed as rutin equivalents (mg/mL). 2.11. Determination of reducing sugar content Reducing sugar content was determined by the 3,5-dinitrosalicylic acid (DNS) method. Glucose standard solutions (0.2–1.0 mL of 1.0 mg/mL) were adjusted to 1.0 mL with distilled water, mixed with DNS reagent (2.0 mL), and heated in a boiling water bath for 7 min. After rapid cooling, the mixtures were diluted to 20 mL. Absorbance was measured at 540 nm against a reagent blank, and reducing sugar content was calculated from a glucose standard curve. 2.12. Determination of antioxidant activity 2.12.1. ABTS free radical scavenging rate The ABTS + working solution was prepared by diluting the stock solution with PBS (pH 7.4) to an absorbance of 0.70 ± 0.02 at 734 nm. Aliquots of sample (0.5 mL) were combined with the ABTS + reagent (5 mL) and allowed to react in the dark for 6 min at room temperature. Absorbance (A i ) was measured at 734 nm. A blank (A 734 ) using distilled water and a sample control (A j ) using PBS instead of ABTS + were prepared. The scavenging rate was calculated as [26] : A B T S s c a v e n g i n g r a t e % = ( 1 - A i - A j A 734 ) × 100 % 2.12.2. DPPH free radical scavenging rate Samples (2 mL) were mixed with DPPH solution (2 mL, 0.1 mmol/L in ethanol) and incubated in darkness for 30 min. Absorbance (A 1 ) was measured at 517 nm. The sample blank (A 2 , ethanol replacing DPPH) and reagent blank (A 0 , ethanol replacing sample) were measured. The scavenging rate was calculated as [23] : D P P H s c a v e n g i n g r a t e % = ( 1 - A 1 - A 2 A 0 ) × 100 % 2.13. Determination of LAB count Viable LAB counts were determined by pour plate method on MRS agar. Samples (1 mL) were serially diluted (10 -1 to 10 -8 ) in sterile saline (0.9% NaCl), and appropriate dilutions (1 mL) were inoculated in duplicate. Plates were incubated anaerobically at 37℃ for 48 h. Colonies (30–300 per plate) were enumerated, and results were expressed as CFU/mL. 2.14. Determination of zeta potential and particle size Samples were filtered (0.45 μm membrane), vortexed for 1 min, and analyzed using a nanoparticle analyzer (NS-90Z Plus, Zhuhai Omec Instruments, China) at 25℃ The Z-average particle size, polydispersity index (PdI), and zeta potential were recorded in triplicate [27] . 2.15. GC-IMS analysis Volatile compounds were analyzed by GC-IMS (FlavourSpec®, G.A.S., Germany). Samples (1 mL) with NaCl (3 g) were incubated in 20 mL headspace vials at 60℃ for 15 min. Separation was performed on an MXT-200 column (30 m × 0.53 mm × 1 μm) at 70℃ using nitrogen carrier gas (≥99.999%). The carrier gas flow rate program: 2 mL/min (held 2 min), ramped to 100 mL/min over 20 min (held 20 min); drift gas: 150 mL/min. Other conditions: injection needle 80℃, ionization chamber 45℃, injection volume 500 μL. Compounds were identified by comparing retention time, drift time, and mass spectra with the NIST library and C4-C9 n-alkane retention indices [28] . 2.16. HS-SPME-GC–MS analysis Volatile flavor compounds were analyzed by HS-SPME-GC–MS [29] .Samples (5 mL) with NaCl (1.5 g) were sealed in 20 mL headspace vials with PTFE septa and equilibrated at 50℃ for 15 min. A preconditioned SPME fiber (65 μm CAR/PDMS, Supelco, USA; 270℃, 30 min) was exposed to the headspace at 50℃ for 40 min, then desorbed in the GC inlet at 250℃ for 5 min. Separation was performed on a TG-WAX MS column (30 m × 0.25 mm × 0.25 μm, Thermo Scientific, USA) with helium carrier gas (1.0 mL/min, splitless mode). Temperature program: 40℃ (2 min), 5℃/min to 100℃, 8℃/min to 250℃ (3 min). MS conditions: EI mode (70 eV), ion source 230℃, transfer line 250℃, scan range m / z 35–450. Compounds were identified by comparing retention indices (calculated using C7-C40 n-alkanes) and mass spectra with the NIST 20 library (match score > 80%). Relative contents were calculated by peak area normalization. 2.17. Electronic nose analysis Volatile profiles were analyzed using an electronic nose (PEN3, Airsense Analytics, Germany) equipped with a metal oxide semiconductor sensor array. Samples (5 mL) were equilibrated in headspace vials at 25℃ for 20 min. Prior to measurement, sensors were flushed with clean air (300 mL/min, 300 s). Headspace gas was collected for 60 s with sensor response recorded for 120 s. Stable response signals were extracted for statistical analysis [28] . 2.18. Sensory evaluation analysis Sensory evaluation was performed by a trained panel (n = 10; 5 males, 5 females) using a double-blind protocol [30] . This sensory study was exempt from formal ethics committee review as per institutional guidelines at Shanxi Agricultural University, given the minimal risk associated with evaluating food products prepared from safe ingredients. Prior to participation, all panelists received detailed information about the study and provided written informed consent. Participants volunteered freely, with the explicit right to withdraw at any time. Confidentiality of personal data was maintained throughout the study. Randomly coded samples were evaluated for color (10 points), aroma (30 points), taste (40 points), texture (20 points), and overall acceptability. Graded descriptive indicators were established for each attribute. Scores were expressed as mean values for statistical analysis. 2.19. Statistical analysis All experiments were performed in triplicate. Data were analyzed using one-way ANOVA followed by Duncan's multiple range test (IBM SPSS Statistics 25.0), with significance set at p < 0.05. PCA was performed using SIMCA 14.1. Data visualization was conducted using Origin 2021. 3. Results and Discussion 3.1. Synergistic effects of US-UHP on the pH of FJJ pH serves as a critical indicator for monitoring fermentation progression, directly reflecting the acid-producing capacity of lactic acid bacteria and metabolic activity [14] . As shown in Fig. 1 A, the pH in the FJJ-C group remained relatively stable throughout fermentation, whereas all treatment groups exhibited a consistent decreasing trend. Initially (0 h), pH values across treatment groups ranged from 4.2 to 4.3. A substantial pH decline was observed during the 8–16 h period, followed by a decelerated rate of decrease from 16 to 24 h. During the late fermentation stage (24–40 h), pH values stabilized at 3.4–3.5, significantly lower than the FJJ-C group. This indicates vigorous LAB metabolic activity during the initial stage, characterized by abundant organic acid production. The subsequent stabilization indicates nutrient depletion and organic acid accumulation, which attenuate bacterial growth [31] . Notably, the FJJ-P-U (pH 3.41) and FJJ-H-U (pH 3.40) groups achieved the lowest terminal pH values, consistent with enhanced LAB proliferation under US-assisted conditions. This phenomenon can be attributed to the cavitation effect of US, which enhances cell membrane permeability and promotes nutrient uptake, thereby accelerating LAB metabolic activity and acid production [29] . Fig. 1. Open in a new tab Effects of US-UHP treatment on the quality parameters of fermented jujube juice. (A) pH value; (B) total soluble solids content; (C) browning index; (D) visual appearance. 3.2. Synergistic effects of US-UHP on the TSS of FJJ TSS content is an important indicator reflecting sugar consumption and fermentation progress in juice [32] . As shown in Fig. 1 B, the TSS content in the FJJ-C group remained essentially unchanged during fermentation, whereas all treatment groups exhibited a decreasing trend over time. The FJJ-H-U group demonstrated significantly lower TSS content than other groups at 24 h ( p < 0.05), reaching its minimum value (11.12°Brix) at 40 h. This reduction reflects the metabolic characteristics of LAB utilizing sugars as the primary carbon source for lactic acid fermentation. As LAB continuously metabolize soluble sugars for growth and reproduction, converting them into lactic acid and other metabolites, TSS progressively decreases with increasing biomass [33] . Furthermore, the cavitation effect of US disrupts macromolecular soluble solids such as polysaccharides and pectin, promoting their degradation or precipitation and achieving synergistic enhancement [34] . 3.3. Synergistic effects of US-UHP on the BI of FJJ BI is a crucial indicator for assessing the degree of browning during the processing of fruit juice [35] . As shown in Fig. 1 C, the browning index of all treatment groups increased within 40 h of fermentation. The FJJ-C group exhibited a significant increase, reaching 0.26 at 40 h, which was significantly higher than other treatment groups ( p < 0.05). Within the 40 h fermentation period, the BI of both the FJJ-H-U and FJJ-H groups remained stable, ranging between 0.11 and 0.15. These results confirm that jujube juice without pretreatment is prone to Maillard reactions and enzymatic browning during fermentation, whereas UHP effectively inhibits browning [36] . 3.4. Synergistic effects of US-UHP on the color of FJJ As shown in Fig. 1 D, fresh jujube juice exhibited an overall browning trend during fermentation, with the color gradually deepening from light yellow over time, consistent with the color data presented in Table 1 . The L* value of the FJJ-C group decreased significantly from 17.76 to 11.96 during 0–40 h, while the L* value of the FJJ-H group remained between 19.20 and 19.70 during the late fermentation stage (24–40 h). The a* value of the FJJ-H-U group showed minimal variation and remained relatively stable throughout fermentation. The b* value of the FJJ-H group was significantly higher than that of other groups during the middle and late fermentation stages (8–40 h) ( p < 0.05). These data analyses indicate that UHP has minimal impact on fruit pigments such as chlorophyll, carotenoids, and anthocyanins [37] . By effectively inhibiting polyphenol oxidase activity and Maillard reactions through low-temperature non-thermal mechanisms, UHP better preserves color after processing [38] . Table 1. Color difference values ( L *, a *, and b *) of fresh jujubes under different processing treatments during fermentation. Group Parameter FJJ-C FJJ-P FJJ-P-U FJJ-H FJJ-H-U 0 h L* 17.76 ± 0.54 Aa 15.42 ± 0.26 Bb 14.44 ± 0.77 Cbc 13.63 ± 0.36 Cc 17.25 ± 0.94 Aa a* −0.92 ± 0.07 Bab −1.03 ± 0.06 Ab −1.02 ± 0.08 Aab −1.03 ± 0.06 Ab −0.90 ± 0.03 Aa b* −0.28 ± 0.05 Ab 0.98 ± 0.04 Aa −2.61 ± 0.11 Ed −0.38 ± 0.05 Bb −1.79 ± 0.11 Ec 8 h L* 15.64 ± 0.52 Ba 14.01 ± 0.23 Cb 12.36 ± 0.32 Dc 13.97 ± 0.44 Cb 12.36 ± 0.32 Cc a* −1.07 ± 0.03 Ca −1.43 ± 0.07 Bcd −1.47 ± 0.09 Cd −1.23 ± 0.17 Bab −1.28 ± 0.03 Cbc b* −1.74 ± 0.11 Bd −0.47 ± 0.08 Bab −0.57 ± 0.07 Ab −0.34 ± 0.05 Ba −0.97 ± 0.06 Cc 16 h L* 14.79 ± 1.02 Bb 19.59 ± 1.21 Aa 20.54 ± 0.31 Aa 15.83 ± 0.28 Bb 15.26 ± 0.79 Bb a* −0.95 ± 0.05 Ba −1.42 ± 0.13 Bc −1.11 ± 0.04 Ab −1.08 ± 0.05 ABab −1.03 ± 0.09 Bab b* −2.33 ± 0.30 Cc −0.77 ± 0.06 Cb −0.93 ± 0.17 Bb 0.16 ± 0.02 Aa 0.37 ± 0.05 Aa 24 h L* 12.60 ± 0.38 Cd 16.12 ± 1.10 Bb 14.07 ± 0.10 Cc 19.21 ± 0.78 Aa 14.68 ± 0.19 Bc a* −0.82 ± 0.04 Aa −1.09 ± 0.15 Ab −1.31 ± 0.09 Bc −1.10 ± 0.07 ABb −1.04 ± 0.08 Bb b* −3.00 ± 0.14 Ec −3.11 ± 0.19 Fc −2.20 ± 0.12 Db −1.49 ± 0.13 Ea −1.22 ± 0.20 Da 32 h L* 11.31 ± 0.78 Cc 15.75 ± 0.70 Bb 15.87 ± 0.24 Bb 19.70 ± 0.51 Aa 15.66 ± 0.29 Bb a* −0.73 ± 0.02 Aa −1.38 ± 0.06 Bc −1.05 ± 0.04 Ab −1.04 ± 0.04 Ab −0.98 ± 0.09 ABb b* −3.02 ± 0.12 Ed −2.75 ± 0.19 Ec −1.3 ± 0.07 Cb −1.26 ± 0.06 Db −0.63 ± 0.07 Ba 40 h L* 11.96 ± 0.89 Cd 15.84 ± 0.27 Bb 15.52 ± 0.35 Bbc 19.37 ± 0.18 Aa 14.69 ± 0.26 Bc a* −1.11 ± 0.06 Cb −1.02 ± 0.08 Aab −1.11 ± 0.06 Ab −0.95 ± 0.06 Aa −0.94 ± 0.02 ABa b* −2.71 ± 0.07 Dd −2.28 ± 0.25 Dc −1.33 ± 0.13 Cb −0.83 ± 0.06 Ca −1.25 ± 0.07 Db Open in a new tab Note: Values are expressed as mean ± SD (n = 3). Different uppercase letters (A-F) within the same column indicate significant differences among fermentation times at p < 0.05. Different lowercase letters (a-d) within the same row indicate significant differences among treatments at p < 0.05. 3.5. Synergistic effects of US-UHP on the ascorbic acid content of FJJ As shown in Fig. 2 A, the ascorbic acid content in all groups exhibited a decreasing trend during fermentation, with the most pronounced degradation observed in the FJJ-C group, which retained only 0.57 mg/100 g at 40 h. The FJJ-H and FJJ-H-U groups showed the gentlest downward trends, with ascorbic acid contents significantly higher than those of other groups at 24 h and 40 h ( p < 0.05). After 40 h of fermentation, the ascorbic acid content in the FJJ-P-U group (0.81 mg/mL) was higher than that in the FJJ-P group (0.44 mg/mL). Since ascorbic acid is highly sensitive to thermal treatment, UHP (500 MPa), as a non-thermal technology, favors the retention of bioactive components including ascorbic acid, whereas conventional thermal sterilization readily induces ascorbic acid degradation [39] . Additionally, UHP can slow down the enzymatic and non-enzymatic oxidation of ascorbic acid by inactivating the ascorbate oxidase system and regulating dissolved oxygen levels [40] . US treatment promotes the release of antioxidants such as polyphenols, enhances the overall antioxidant capacity of the system, and indirectly protects ascorbic acid from oxidative degradation. Fig. 2. Open in a new tab Effects of US-UHP treatment on bioactive compounds and functional properties of fermented jujube juice. (A) Ascorbic acid content during fermentation (0–40 h); (B) ascorbic acid content at 24 h and 40 h; (C) total phenolic content during fermentation (0–40 h); (D) total phenolic content at 24 h and 40 h; (E) total flavonoid content during fermentation (0–40 h); (F) total flavonoid content at 24 h and 40 h; (G) reducing sugar content during fermentation (0–40 h); (H) reducing sugar content at 24 h and 40 h; (I) ABTS radical scavenging activity; (J) DPPH radical scavenging activity; (K) lactic acid bacteria (LAB) viable counts; (L) representative colony morphology of LAB. Different lowercase letters indicate significant differences ( p < 0.05). Asterisks (*) denote significant differences between groups at 24 h and 40 h as determined by independent samples t -test (* p < 0.05; ** p < 0.01; *** p < 0.001). 3.6. Synergistic effects of US-UHP on the TPC of FJJ As shown in Fig. 2 C, the TPC in all treatment groups exhibited an initial increase followed by a decrease during fermentation. The FJJ-C group showed a relatively low overall level of total phenolic content. Although the total phenolic content in the FJJ-P group was higher than that in the control group, reaching a peak of 2.52 mg/mL at 16 h, it decreased markedly in the late fermentation stage to 1.79 mg/mL at 40 h, indicating a weak protective effect. The FJJ-H and FJJ-H-U groups displayed optimal retention of total phenolic content, maintaining relatively high levels throughout the fermentation period. Conventional pasteurization cannot achieve complete sterility, resulting in products requiring refrigeration with a short shelf life, while causing losses of certain nutritional and flavor components [41] . After UHP treatment, the total phenolic content in samples was significantly higher than that in the untreated control group ( p < 0.05), attributed to improved extraction efficiency of phenolic compounds from cellular tissues and activation of their biosynthetic pathways [42] . UHP at 500–600 MPa can disrupt cell walls to release phenolic compounds, thereby enhancing their extraction efficiency and bioaccessibility, while inactivating oxidases and reducing phenolic oxidation [43] . US treatment further enhances cell wall disruption and substrate release via cavitation effects, thereby achieving synergistic enhancement [44] . 3.7. Synergistic effects of US-UHP on the TFC of FJJ As shown in Fig. 2 E, the TFC in the FJJ-C group decreased from 0.09 mg/mL to 0.07 mg/mL during 0–40 h, exhibiting a downward trend. In contrast, the TFC in all treatment groups exhibited an overall upward trend. At 40 h, the FJJ-H-U group exhibited the highest TFC (0.16 mg/mL), while the value in the FJJ-P-U group (0.14 mg/mL) was significantly higher than that in the FJJ-P group (0.13 mg/mL) ( p < 0.05). These results indicate that flavonoids underwent a certain degree of degradation during spontaneous fermentation. The cavitation effect of US promoted the dissolution and metabolic conversion of flavonoid precursors [44] . The combined UHP and US treatment exerted the optimal protective effect on flavonoids. Since UHP has a limited effect on the covalent bonds of low-molecular-weight compounds, it effectively preserves the nutritional value and sensory characteristics of fruits and vegetables [45] . The cavitation effect of US treatment complements UHP by forming a synergistic protective mechanism. 3.8. Synergistic effects of US-UHP on the reducing sugar content of FJJ As shown in Fig. 2 G, the reducing sugar contents in all treatment groups at the initial fermentation stage ranged from 72.30 to 74.00 mg/mL, showing a downward trend with extended fermentation time. The FJJ-C group exhibited a gentle decrease, while the FJJ-H-U group showed the most significant reduction with a consumption rate of 54.7%. At 40 h of fermentation, the reducing sugar content in the FJJ-H-U group was 33.47 mg/mL, significantly the lowest among all groups ( p < 0.05). The value in the FJJ-P-U group was 35.67 mg/mL, obviously lower than those in the FJJ-P and FJJ-U groups. The rapid consumption of reducing sugars is highly consistent with the pH decline and viable LAB count increase during fermentation, confirming that US-UHP combined treatment can accelerate fermentation by improving the accessibility of fermentation substrates [46] . 3.9. Synergistic effects of US-UHP on the antioxidant activity of FJJ As shown in Fig. 2 I and 2 J, the free radical scavenging rates of almost treatment groups were higher than those of the control group, with the most pronounced enhancements observed in the FJJ-P-U and FJJ-H-U groups. In terms of ABTS scavenging rate, the FJJ-P, FJJ-P-U, and FJJ-H-U groups exhibited significantly higher values than the FJJ-C group at 24 h ( p < 0.05). At 40 h, the ABTS scavenging rates of the FJJ-P-U and FJJ-H-U groups remained at a high level of approximately 85%. For DPPH scavenging rate, all treatment groups showed significantly higher values at 24 h than at 40 h ( p < 0.001). The FJJ-P-U and FJJ-H-U groups presented markedly higher DPPH scavenging rates than the FJJ-C group at 24 h ( p < 0.05), reaching 74.97% and 74.86%, respectively. Even at 40 h, their DPPH scavenging rates remained above 50%, significantly superior to the FJJ-C group. These results indicate that DPPH free radical scavenging activity decreases with prolonged fermentation time, while US-UHP treatment can effectively improve DPPH free radical scavenging activity. UHP effectively eliminates undesirable microorganisms and moderately disrupts cellular structures, creating favorable conditions for the release of antioxidant substances [44] . US-assisted fermentation further promotes cell wall rupture through cavitation effects and mechanical shearing actions, improving the extraction efficiency of antioxidant components including polyphenols and flavonoids. This combined treatment enhances microbial metabolic activity and facilitates the transformation and accumulation of bioactive components [47] . 3.10. Synergistic effects of US-UHP on the LAB counts of FJJ As shown in Fig. 2 K, the viable LAB counts in the FJJ-C group reached 2.86 × 10 7 CFU/mL after 16 h of fermentation, significantly the lowest among all groups ( p < 0.05). The value in the FJJ-C group dropped sharply to 1.17 × 10 5 CFU/mL at 40 h of fermentation ( p < 0.005). The viable bacterial counts in all other treatment groups remained above 1.0 × 10 8 CFU/mL. The FJJ-H group showed the highest count of 4.77 × 10 8 CFU/mL at 40 h of fermentation ( p < 0.05). These results suggest that different sterilization treatments can inactivate competitive indigenous microorganisms and provide more favorable ecological niches for target LAB. UHP treatment maintains a relatively high level of viable bacterial count during the entire fermentation period, indicating that UHP disrupts the cellular structure of jujube tissues via isostatic pressure, releases more available nutrients, and improves the nutrient accessibility of the fermentation medium [48] . Emerging technologies applied at sublethal levels to LAB before or during fermentation can modify their growth rate and induce metabolic pathways for varied production yields [49] . The cavitation effect of US enhances cell membrane permeability of LAB and promotes nutrient uptake [50] . 3.11. Synergistic effects of US-UHP on the zeta potential and particle size distribution of FJJ As shown in Fig. 3 A, the zeta potential of the FJJ-C group remained around − 7 mV within 40 h, with minor fluctuations. Compared to the control group, the absolute zeta potential values of all treatment groups significantly decreased over fermentation time and stabilized after 16 h, indicating that fermentation reduces the surface charge density of particles, thereby weakening electrostatic repulsion between them. Regarding particle size, the Z-average data in Fig. 3 C reveal that, except for the FJJ-C group, the Z-average of all treatment groups at 24 h was significantly lower than at 0 h ( p < 0.05). At 40 h of fermentation, the particle size of the FJJ-C group rebounded substantially to 1597.67 d.nm, while other fermentation groups maintained low levels (33.41–53.21 d.nm). Regarding the polydispersity index (PdI), Fig. 3 D shows that PdI values of the FJJ-H and FJJ-H-U groups at 0 h were the lowest. Over fermentation time, PdI values in all treatment groups exhibited an upward trend, indicating a decrease in system uniformity. At 40 h, the PdI values of the FJJ-P-U and FJJ-H-U groups were lower than those of the FJJ-H and FJJ-P groups. The particle size distribution curves in Fig. 3 E indicate that all treatment groups at 0 h displayed either unimodal or bimodal distributions. Fig. 3 F and G reveal multimodal distributions at 24 h and 40 h of fermentation. The average particle size data suggest that LAB addition initially increases system particle size; however, metabolites such as pectinase and cellulase effectively degrade macromolecular polysaccharides. The continuous metabolic activity of LAB contributes to maintaining the colloidal stability of the system [51] . The superior dispersibility observed in UHP-treated groups at 0 h is closely related to the fact that UHP processing at low temperatures avoids irregular thermal aggregation of proteins and polysaccharides. UHP can affect the stability of pressure-sensitive macromolecules such as proteins, promoting conformational changes and moderate depolymerization, leading to particle homogenization and improved enzymatic hydrolysis efficiency [52] . Fig. 3. Open in a new tab Effects of US-UHP treatment on the colloidal stability of fermented jujube juice. (A) Zeta potential during fermentation (0–40 h); (B) Zeta potential at 24 h and 40 h; (C) Z-average particle size; (D) polydispersity index (PDI); (E) particle size distribution at 0 h; (F) particle size distribution at 24 h; (G) particle size distribution at 40 h. Different lowercase letters indicate significant differences ( p < 0.05). Asterisks (*) denote significant differences between groups at 24 h and 40 h as determined by independent samples t -test (* p < 0.05; ** p < 0.01; *** p < 0.001). 3.12. Synergistic effects of US-UHP on volatile flavor compounds of FJJ based on GC-IMS As presented in Fig. 4 A and 4B, the FJJ-C group exhibited dense signals in the low retention time region, indicating abundant volatile compounds in unsterilized jujube juice. The fingerprint analysis ( Fig. 4 C) revealed significant differences among treatment groups. In Region a, signal intensities of isobutanol, isoamyl alcohol, and ethyl acetate in the control group significantly exceeded those in sterilization groups ( p < 0.05). The enhanced alcohol and ester signals in Region A resulted from residual microorganisms (yeast, LAB, etc.) in unsterilized juice, which metabolized higher alcohols and esters via the Ehrlich pathway and esterase catalysis [50] . In Region b, 3-octanone content in UHP-treated groups was notably higher than in thermal treatment groups. thermal processing, while achieving sterilization through high temperatures, simultaneously degrade aroma compounds (such as 3-octanone) and their precursors, resulting in low final retention levels [53] . Importantly, Region c demonstrated that undesirable flavor compounds (butyric acid, 2-hexanone, hexanal) were significantly lower in the FJJ-H-U group compared to the FJJ-H group. The cavitation effect of US may disrupt oxidative enzyme structures or promote antioxidant release, synergistically inhibiting lipid oxidation [54] . In Region d, the contents of 1-heptanol and trans -2-hexenal were significantly lower in the FJJ-P-U group compared to the FJJ-H group. This may be attributed to HPP maintaining enzymatic activity to preserve the dynamic balance of flavor compounds, whereas thermal treatment inactivates enzymes and disrupts compound structures, achieving sterilization at the expense of certain flavor components. In Region e, the higher methyl butyrate content in UHP-treated groups was attributed to UHP disrupting cell membrane permeability, facilitating contact between substrates and synthases, and promoting ester accumulation [52] . Fig. 4. Open in a new tab GC-IMS analysis of volatile compounds in fermented jujube juice treated with US-UHP. (A) Three-dimensional topographic spectrum; (B) two-dimensional difference spectrum; (C) fingerprint gallery of volatile compounds; (D) principal component analysis (PCA) score plot; (E) PCA loading plot; (F) Sankey diagram of decreased flavor compounds; (G) Sankey diagram of increased flavor compounds. PCA analysis ( Fig. 4 D) demonstrated that the cumulative contribution rate of the first two principal components reached 96.39% (R 2 = 0.9357, P = 0.0010). The control and sterilization groups were distinctly separated along the PC1 axis, while thermal treatment groups occupied the positive PC2 region and UHP groups the negative region. The loading plot ( Fig. 4 E) revealed that ethanol was strongly correlated with the control group, whereas 2-pentanone, acetone, and acetic acid were closely associated with thermal treatment groups. Combining Fig. 4 F and 4G, the US groups (FJJ-P-U, FJJ-H-U) showed significant reductions in alcohol and fusel odors, potentially due to US's regulation of microbial metabolism, which decreases ethanol and related fusel alcohol production [55] , [38] . The FJJ-P-U treatment was most prominent in enhancing floral and sweet notes. Although thermal treatment increased creamy aroma, it resulted in significant loss of fruity notes. Overall, the combination of UHP and US treatments effectively inhibited undesirable flavor compound formation while promoting the accumulation of floral, fruity, and sweet aroma substances, offering a viable technological approach to improving the flavor quality of FJJ. 3.13. Synergistic effects of US-UHP on volatile flavor compounds of FJJ based on HS-SPME-GC–MS For the composition and classification of volatile compounds ( Fig. 5 A), various classes were identified, with organic acids being the most abundant (7 species), followed by alcohols (6 species) and aldehydes (5 species). Ketones (4 species) and aromatic alcohols (3 species) also accounted for considerable proportions. As shown in Fig. 5 B, Phenolic ethers were overwhelmingly dominant in the FJJ-C group. After fermentation, organic acids became the predominant class, however, following US treatments (FJJ-P-U, FJJ-H-U), the relative proportion of organic acids decreased significantly, while other flavor classes increased noticeably, resulting in a more complex and harmonious flavor profile [56] . PCA results ( Fig. 5 C) showed that the cumulative contribution rate of PC1 and PC2 reached 73.46% (R 2 = 0.9357, P = 0.0010). The FJJ-C group was significantly separated from all treatment groups along the PC1 axis, confirming that LAB fermentation is the dominant factor driving flavor transformation. US groups exhibited higher PC1 scores compared to their non-US counterparts. The FJJ-H-U group showed a significantly negative score on PC2 (−2.5), clearly distinguishing it from other groups, indicating that US-UHP exerted a unique synergistic regulatory effect. Loading plot analysis ( Fig. 5 D) revealed that compounds characteristic of floral and fruity notes, including 1-hexanol, ethyl hexanoate, benzaldehyde, and decanal, contributed positively to the PC1 axis. As a non-thermal technology, UHP better preserves aromatic and nutritional compounds in juice. The cavitation effect of US releases more reaction precursors by disrupting cell structures and accelerates microbial metabolism, promoting the synthesis of key aroma compounds such as esters and aldehydes [38] . The distinctiveness of the FJJ-H-U group in the negative PC2 direction may result from synergistic effects further modulating Maillard reactions or specific metabolic pathways. Fig. 5. Open in a new tab HS-SPME-GC–MS analysis of volatile compounds in fermented jujube juice treated with US-UHP. (A) Classification of volatile compound categories; (B) stacked bar chart of relative content of flavor substances; (C) PCA score plot; (D) PCA loading plot; (E) circular stacked bar chart of volatile compounds distribution; (F) variable importance in projection (VIP) score plot of differential markers; (G) relative content of acetic acid; (H) relative content of 3-methyl-1-butanol; (I) relative content of 3-methyl-2-butanol. Different letters represent significant differences at p < 0.05. Further analysis using the volatile compound heatmap fingerprint ( Fig. 5 E) revealed that hexanol and 2-methyl-1-butanol were significantly produced in the fermentation groups, with enhanced signals in US groups. Benzaldehyde and decanal were most prominent in the FJJ-H-U group. Ethyl hexanoate was predominantly present in fermentation groups, exhibiting stronger signals in US-treated samples. Multiple furan derivatives were detected or showed enhanced signals in US-treated groups, suggesting that US may induce or accelerate non-enzymatic browning reactions [57] . The prominent 2-phenylacetaldehyde signal in the FJJ-H-U group further confirmed the specific enhancing effect of US-UHP on aromatic compound metabolic pathways. Based on VIP analysis, key discriminatory factors with VIP values greater than 1 were identified, including acetic acid, 3-methyl-1-butanol, and 3-methyl-2-butanol ( Fig. 5 F). Acetic acid exhibited the highest VIP value in the FJJ-H-U group, positioning it as a characteristic flavor marker of US-enhanced fermentation processes. Further analysis of individual volatile compounds revealed treatment-specific accumulation patterns. The relative content of acetic acid was significantly elevated in the FJJ-H-U group compared with the FJJ-H group ( p < 0.05), suggesting that US pretreatment synergistically promoted acetic acid biosynthesis during fermentation ( Fig. 5 G). This enhancement may be attributed to US-induced cell membrane permeabilization, which facilitates substrate accessibility for microbial metabolic pathways. Notably, 3-methyl-1-butanol, a branched-chain higher alcohol derived from leucine catabolism via the Ehrlich pathway, exhibited the highest relative content in the FJJ-C group among all treatments ( Fig. 5 H). In contrast, 3-methyl-2-butanol accumulated preferentially in the FJJ-P-U group ( Fig. 5 I), indicating that the combined US-UHP pretreatment modulated the metabolic flux toward distinct volatile alcohol profiles. Simultaneously, US releases substrates such as polyphenols, sugars, and amino acids. This promotes the formation of pleasant aroma compounds including esters and aldehydes, while inhibiting the accumulation of certain off-odor compounds through selective modulation of microbial community structure or enzyme activities, thereby optimizing the overall flavor quality of fermented fresh jujube juice [58] . 3.14. Synergistic effects of US-UHP on the flavor of FJJ based on electronic nose technology Based on the electronic nose radar plot ( Fig. 6 A), the FJJ-C group exhibited significantly higher response values for methyl-sensitive sensors and alcohol-, aldehyde-, and ketone-sensitive sensors compared to all sterilization treatment groups. While the response profiles among different sterilization groups were relatively similar, they differed markedly from the FJJ-C group. Notably, the FJJ-P-U group showed slightly higher responses for sulfur compound sensors and alcohol-, aldehyde-, and ketone-sensitive sensors compared to the FJJ-P group, indicating that vigorous metabolic activity of microorganisms in unsterilized juice results in higher concentrations of alcohols, aldehydes, ketones, and methyl-containing compounds, while US treatment enhances the responses of sulfur compounds and certain carbonyl compounds [59] . The PCA scatter plot ( Fig. 6 B) revealed that the cumulative contribution rate of PC1 and PC2 reached 93.76% (R 2 = 0.9253, P = 0.0010). The FJJ-C group was independently distributed in the positive PC1 region, completely separated from all sterilization treatment groups. US-treated groups (FJJ-P-U, FJJ-H-U) and non-US groups (FJJ-P, FJJ-H) were located in positive and negative PC2 regions, respectively, with significant separation between these clusters. These results confirm that sterilization treatments effectively alter the volatile flavor composition of samples, with US treatment shifting the flavor attributes of both UHP and thermal treatment groups, and thermal treatment groups exhibiting more pronounced changes. The loading plot ( Fig. 6 C) further revealed that positive contributions to PC1 primarily originated from sulfur compounds, terpenes, and alcohol-, aldehyde-, and ketone-containing compounds, while negative contributions were mainly associated with nitrogen oxides, aromatic compounds, and long-chain alkanes. Integrating these findings with sample distribution patterns, sterilization treatments significantly reduced the contents of alcohols, aldehydes, ketones, and sulfur compounds, while relatively increasing the proportions of nitrogen oxides and aromatic compounds, thereby fundamentally reshaping the overall odor profile of the samples. Fig. 6. Open in a new tab Sensory analysis of fermented jujube juice treated with US-UHP. (A) Radar chart of electronic nose responses; (B) PCA score plot of electronic nose analysis; (C) PCA loading plot of electronic nose analysis; (D) sensory evaluation radar chart at 24 h; (E) sensory evaluation radar chart at 40 h. 3.15. Synergistic effects of US-UHP on the sensory quality of FJJ According to the sensory evaluation radar plots ( Fig. 6 D and 6E), all treatment groups exhibited decreased scores across various sensory indicators at 40 h compared to 24 h. At both time points, the FJJ-H-U group demonstrated the best performance in color＆glossiness, taste, odor, tissue state, and overall acceptability. In contrast, the FJJ-C group received the lowest overall acceptability scores at both fermentation time points. UHP-treated groups (FJJ-H, FJJ-H-U) showed higher scores for most sensory indicators compared to non-UHP groups (FJJ-P, FJJ-P-U) at both 24 h and 40 h. Furthermore, US-assisted groups (FJJ-H-U, FJJ-P-U) achieved higher scores across all sensory indicators compared to their non-US counterparts. These results indicate that UHP, as a non-thermal technology, positively affects color retention, flavor preservation, and taste enhancement of FJJ. The FJJ-H and FJJ-H-U groups maintained bright color and harmonious jujube aroma during the initial fermentation stage and sustained stable sensory quality throughout the later fermentation perio [60] . In contrast, the FJJ-C group and pasteurization groups exhibited significant color darkening and aroma attenuation during the later stages, further confirming that UHP provides significant protection for color-related compounds and flavor precursors [52] . 3.16. Correlation analysis and involved mechanism The Pearson correlation heatmap ( Fig. 7 A) visually depicts the synergistic relationships among physicochemical properties, antioxidant activities, and sensory attributes through color intensity and circle size. pH exhibited a significant positive correlation with soluble solids content (r = 0.92), while both parameters showed significant negative correlations with turbidity (r = -0.95). These findings suggest that LAB fermentation of jujube juice degrades soluble substances, thereby promoting organic acid generation while affecting colloidal stability [51] . Browning index demonstrated a strong positive correlation with soluble solids content (r = 0.96), indicating that increased Maillard reaction substrates may exacerbate non-enzymatic browning. Notably, total phenolic content, total flavonoid content, and reducing sugars exhibited a significant positive correlation (r > 0.94), and all three were positively correlated with the DPPH scavenging rate. Lactic acid fermentation-based biotransformation results in changes in the profile and nature of bioactive compounds and improves the organoleptic properties, shelf life, and bioavailability of vitamins and minerals in fermented juices [46] . A greater synthesis of phenolic compounds is associated with stronger antioxidant activity. Meanwhile, the combined physical effects of US cavitation and UHP disrupt the cell wall and vacuole membranes of jujube tissue, releasing originally bound phenolic and flavonoid compounds [61] . This “cell-disruption-induced release” mechanism directly explains the significant association between increased extractable bioactive components and enhanced radical-scavenging capacity, in agreement with previous reports on UHP-assisted phenolic liberation in fruit matrices. Furthermore, the significant positive correlation between zeta potential and total phenolic and total flavonoid contents (r > 0.89) further indicates that the synergistic effect of US-UHP treatment disrupts the cell wall, and the released polyphenolic compounds, many of which carry charged hydroxyl groups, adsorb onto colloidal particle surfaces [62] , potentially altering the charge distribution of colloids and thereby affecting the physical stability of jujube juice. From the perspective of sensory evaluation, color exhibited significant positive correlations with glossiness, taste, aroma, and texture (r > 0.86), and was highly correlated with overall acceptability (r = 0.98), can be attributed to the fact that US-UHP selectively reduced off-flavors (e.g., hexanal, (Z)-3-hexen-1-ol; Fig. 4 C, region a) while promoting fruity esters (e.g., methyl butyrate; Fig. 4 C, region e) [54] . These findings suggest that US-UHP processing effectively maintains the sensory characteristics of FJJ while preserving favorable physicochemical quality, providing a theoretical basis for process optimization. Fig. 7. Open in a new tab Correlation analysis and proposed mechanism of US-UHP treatment on fermented jujube juice. (A) Pearson correlation heatmap among quality parameters; (B) schematic diagram illustrating the proposed mechanism of US-UHP treatment on fermented jujube juice quality. The proposed mechanism by which US-UHP enhances the quality of FJJ is schematically illustrated in Fig. 7 B. UHP effectively induces the depolymerization of pectin and polysaccharide macromolecules by disrupting the colloidal molecular architecture, thereby reducing juice turbidity and enhancing colloidal stability, consistent with the observed changes in zeta potential and particle size [51] . US treatment, through acoustic cavitation, generates microbubbles that grow during cycles of high and low pressure until they become unstable and burst, releasing substantial energy. This cavitation-induced mechanical force disrupts cell walls, promoting the release of polysaccharides, phenolic, and flavonoid compounds [44] . The synergistic combination of US and UHP has been widely applied for preservation and valorization of nutrients with high efficiency, markedly enhancing both ABTS and DPPH radical scavenging capacities. With respect to flavor modulation, US-UHP synergy significantly attenuates undesirable volatile compounds, including hexanal, heptanal, decanal (oxidative off-flavors), (Z)-3-hexen-1-ol (green/fatty notes), and fusel alcohols such as 3-methyl-1-butanol and 1-hexanol. Concurrently, it promotes the biosynthesis of desirable aroma compounds, particularly fruity esters such as methyl butyrate. Depolymerization is the main change in polysaccharides during the high-pressure processing. Collectively, US-UHP pretreatment achieves comprehensive quality enhancement through macromolecular depolymerization, bioactive compound liberation, and volatile profile optimization, thereby enabling precise sensory modulation while conferring superior antioxidant functionality. 4. Conclusion This study demonstrated that US-UHP pretreatment combined with mixed LAB fermentation is an effective approach for improving fresh jujube juice quality, with FJJ-H-U at 24 h showing the best overall performance. The combined treatment regulated flavor by suppressing undesirable volatiles, including butyric acid, hexanal, and acetaldehyde, while promoting more desirable fruity and floral aroma compounds. It also enhanced antioxidant activity, as evidenced by higher total phenolic content and stronger ABTS and DPPH radical-scavenging capacity. In addition, the substantial decrease in particle size and PDI indicated improved colloidal stability, which was consistent with the superior sensory acceptability. Overall, the main technological advantage of this integrated process lies in its ability to simultaneously achieve flavor regulation, antioxidant enhancement, and colloidal stabilization. These findings support the potential of US-UHP-assisted LAB fermentation as a practical strategy for the high-value processing of fresh jujube into stable and appealing fermented beverages, and may also provide useful insight for other polyphenol-rich fruit matrices. CRediT authorship contribution statement Yu Zhang: Writing – original draft, Supervision, Methodology, Conceptualization. Qinjie Xue: Writing – original draft, Formal analysis, Data curation. Yao Yao: Writing – original draft, Investigation. Jiachen Xie: Writing – original draft, Validation. Jiaxin Yan: Resources. Ruier Yao: Software. Miao Yang: Visualization. Xiaoyan Sun: Project administration. Zhihui Yu: Writing – review & editing, Conceptualization. Ling Ma: Supervision, Funding acquisition. Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Acknowledgment This work was financially supported by the Science and Technology Innovation Project of Jinzhong National Agricultural High-Tech Zone (Grant No. JZGJNGQKJCX202304). Contributor Information Zhihui Yu, Email: zhihuisxau@163.com. Ling Ma, Email: ml0359@163.com. References 1. 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