Determination of insecticide residues in beverages based on MIL-100(Fe) dispersive solid-phase microextraction in combination with dispersive liquid-liquid microextraction followed by HPLC-MS/MS.

A novel dispersive solid-phase microextraction method based on a metal-organic framework (MIL-100(Fe)) combined with a dispersive liquid-liquid microextraction technique was proposed for the extraction and enrichment of four insecticides in beverages. The qualitative and quantitative analysis of these insecticides was conducted using HPLC-MS/MS. To optimize the extraction process, several parameters were investigated, and the main variables were optimized using CCD-based RSM. The developed method displayed a wide linear range of 1.000-1000 ng/L and R2 values >0.993 for all four calibration curves. The method demonstrated high sensitivity, with LODs and LOQs of 0.3-0.6 ng/L and 0.8-1.0 ng/L, respectively. In addition, the greenness of the proposed method was assessed using the Complex GAPI tool, and the results showed that the proposed method exhibits benefits, such as minimal usage of organic solvents and negligible matrix influence, making it a suitable method for the detection of insecticide residues in beverages.

Self-powered molecularly imprinted photoelectrochemical sensor based on Ppy/QD/HOF heterojunction for the detection of bisphenol A.

As an emerging porous material, hydrogen-bonded organic framework materials (HOFs) still pose application challenges. In this work, the designed type "I + II" heterojunction extracted hot electrons from HOFs using quantum dots (QDs) and polypyrrole (Ppy), improving the stability and photoelectrochemical performance of materials. In addition to serving as a potential well, electropolymerized Ppy was used as a recognition element for bisphenol A (BPA), and a novel self-powered molecularly imprinted photoelectrochemical (MIP-PEC) sensor was designed. The sensing platform showed a linear relationship from 1 × 10-10 to 1 × 10-7 mol∙L-1 and from 1 × 10-7 to 1 mol∙L-1 with an acceptable detection limit of 4.2 × 10-11 mol∙L-1. This is the first application of HOFs in constructing MIP-PEC sensors and a new attempt to improve the stability of HOFs for the application of porous crystal materials in the sensing field.

Study on Purification, Identification and Antioxidant of Flavonoids Extracted from Perilla leaves.

The flavonoids from Perilla leaves were extracted using flash extraction assisted by ultrasonic extraction with ethanol. Subsequently, macroporous resin was employed for the isolation and purification of these flavonoids, followed by an investigation into their antioxidant activity. The process conditions for the extraction of flavonoids from Perilla leaves were designed and optimized using a one-way experiment combined with a response surface methodology. The optimal extraction conditions were determined as follows: the liquid-solid ratio was 20:1, ethanol volume fraction of 60%, ultrasound temperature of 60 °C, ultrasound time of 10 min and flash evaporation time of 60 s. The optimal extraction rate of flavonoids is 9.8 mg/g. In terms of separation and purification, a high-performance macroporous resin (HPD450 resin) with high purification efficiency was selected through static analysis and adsorption experiments. The optimal enrichment conditions were as follows: loading concentration of 0.06 mg/mL, optimal loading concentration of 20 mL, elution concentration of 70% and 76 mL, providing a reference for the further development and utilization of Perilla leaf flavonoids.

Correlation Analysis of Microbial Community Changes and Physicochemical Characteristics in Aged Vinegar Brewing.

This study aimed to explore key physicochemical characteristics and evolutionary patterns of microbial community structure during the fermentation of aged vinegar. The correlation between microorganisms and physicochemical characteristics during fermentation was examined. The results revealed significant differences in genera at different stages of fermentation. The dominant bacteria in R1 were Bacillus, Lactobacillus, Aspergillus, and Issatchenkia. During the R2 fermentation stage, Lactobacillus, Acetobacter, and Saccharomyces exhibited an upward trend and finally became the dominant bacteria. Aspergillus was the main bacterial genus at the end of overall fermentation. The correlation analysis showed that the bacterial genera significantly positively and negatively correlated with reducing sugars and amino acid nitrogen were the same in Cuqu. Similarly, the bacterial genera significantly positively and negatively correlated with pH and saccharification power were the same. pH, reducing sugar, and saccharification ability were mainly positively correlated with bacterial genera during fermentation. Further, studies found that the overall correlation between fungal communities and physicochemical characteristics was weaker than the correlation with bacteria during fermentation.

The Separation, Purification, Structure Identification, and Antioxidant Activity of Elaeagnus umbellata Polysaccharides.

In order to investigate the antioxidant activity of Elaeagnus umbellata polysaccharides, the physicochemical characteristics of purified Elaeagnus umbellata polysaccharides (EUP, consisting of two fractions, EUP1 and EUP2) were investigated using UV spectrophotometry, high-performance liquid chromatography (HPLC), high-performance gel permeation chromatography (HPGPC), and Fourier transform infrared spectroscopy (FT-IR). This revealed that EUP1 and EUP2 were acidic polysaccharides with an average molecular weight (MW) of 63 and 38 kDa, respectively. EUP1 mainly consisted of L-rhamnose and D-galactose in a molar ratio of 2.05:1, and EUP2 consisted of D-mannose, L-rhamnose, D-galactose, and D-arabinose in a molar ratio of 2.06:1:2.78:1. Furthermore, EUP exhibited considerable antioxidant potential for scavenging hydroxyl, superoxide anion, DPPH, and ABTS radicals. Therefore, EUP can be developed as a potential antioxidant for the functional food or pharmaceutical field.

Determination of Flavonoid Compounds in Shanxi Aged Vinegars Based on Hydrophobic Deep Eutectic Solvent VALLME-HPLC Method: Assessment of the Environmental Impact of the Developed Method.

This research presents a novel, eco-friendly, vortex-assisted liquid-liquid microextraction (VALLME) approach, integrating hydrophobic deep eutectic solvents (DESs) with HPLC for the identification and quantification of nine specific flavonoids in Shanxi aged vinegar (SAV). The parameters of DES-VALLME, including the ratio of trioctylmethylammonium chloride to 1,4-butanediol (1:6), DES volume (150 µL), vortex duration (5 min), the concentration of NaCl (0.40 g), and centrifugation time (10 min), were optimized to achieve the maximum extraction efficiency of target substances. Under these optimal conditions, quantitative analyses performed via HPLC demonstrated a broad linear range of 0.20-50.00 µg/mL and correlation coefficients (r2) greater than 0.9944 for all nine calibration curves. The limits of detection (LOD) and limits of quantitation (LOQ) were 0.09-0.18 µg/mL and 0.30-0.60 µg/mL, respectively, ensuring high sensitivity. The relative standard deviations for intra-day and inter-day variability were within the acceptable range, 2.34-3.77% and 3.04-4.96%, respectively, demonstrating the method's reliability. The recovery rates ranged from 85.97% to 108.11%, underscoring the method's precision. This technique exhibited a significant enrichment effect (enrichment factor: 43 to 296) on SAV flavonoids. Notably, the eco-friendliness of this procedure was evaluated using the Analytical Eco-Scale, Green Analytical Procedure Index, and Analytical Greenness Metric. The results suggested that this technique is a viable green alternative to traditional flavonoid determination methods in SAV. In summary, this novel method provides a theoretical basis for assessing flavonoid content in SAV samples and tracing SAV products. This contribution has significant implications for enhancing analytical techniques in food chemistry and environmental science and the sustainable development of the food industry.

A universal design of turn-on fluorescent aptasensor based on luminescent MOFs: Application for the detection of bisphenol A in water, milk and chicken samples.

A universal design of turn-on fluorescent aptasensor based on aptamer functionalized gold nanoparticles (AuNPs) and luminescent metal-organic frameworks (LMOFs) complex (AuNPs-Apt/NH2-MIL-125(Ti)) was realized for bisphenol A (BPA) detection. LMOF NH2-MIL-125(Ti) was prepared using facial hydrothermal method. BPA aptamer functionalized AuNPs were prepared and adsorbed on the surface of NH2-MIL-125(Ti) to obtain platform of the fluorescent aptasensor. The fabrication process, sensing performance and applicability of the proposed aptasensor were characterized and investigated carefully. Linear detection range of the constructed aptasensor was from 1 × 10-9 mol L-1 to 1 × 10-4 mol L-1 with good selectivity, repeatability, stability and reproducibility under optimal experimental conditions. Meanwhile, the fluorescent aptasensor was successfully utilized for BPA detection in real samples with the recoveries of 95.80%-103.12%. The proposed aptasensor based on AuNPs-Apt/NH2-MIL-125(Ti) holds significant potential for BPA detection in environmental and food samples, promoting the construction and application of LMOFs-based aptasensor.

MOF/COF heterostructure hybrid composite-based molecularly imprinted photoelectrochemical sensing platform for determination of dibutyl phthalate: A further expansion for MOF/COF application.

A novel metal-organic framework (MOF)/covalent-organic framework (COF) heterostructure hybrid composite (NH2-UiO-66/TpPa-1-COF) with excellent photoactivity was developed, which further acted as the photoelectrochemical sensitized layer of a molecularly imprinted photoelectrochemical (MIP-PEC) sensor for extremely sensitive and selective determination of dibutyl phthalate (DBP). The NH2-UiO-66/TpPa-1-COF was synthesized using a simple one-step solvothermal method, which showed improved photocurrent response owing to heterojunction formation, favorable energy-band configuration and strong light absorption capacity. To improve the sensing performance, molecularly imprinted polymer (MIP) was developed by sol-gel polymerization method as the recognition component of PEC sensor. The specific binding of imprinting sites towards DBP could block the electron transfer, causing decreased photocurrent response of the MIP-PEC sensor. The MIP-PEC sensor showed a wide detection range from 0.1 nmol L-1 to 100 µmol L-1 with a limit of detection of 3.0 × 10-11 mol L-1 under optimal conditions. Meanwhile, the proposed MIP-PEC sensor showed good stability, selectivity, reproducibility, and applicability in real samples. This is the first attempt to apply MOF/COF heterostructure hybrid composite for MIP-PEC sensor construction, providing new insight into the potential applications of microporous crystalline framework heterostructure hybrid composite in the sensing field.

Structure, physicochemical properties, antioxidant, and hypoglycemic activities of water-soluble polysaccharides from millet bran.

This study aimed to investigate the composition, structure, in vitro antioxidant, and hypoglycemic activities of a novel water-soluble MBP-1, an extract from millet bran, isolated by DEAE-52 cellulose and purified by Sephadex G-100. The results showed that MBP-1 was mainly composed of xylose, mannose, galactose, rhamnose, and arabinose with a molar ratio of 0.72:0.59:76.26:1.04:0.83 and a molecular weight of 6.6×104  Da, and its purity was 98%, and the yield was 3.76%. MBP-1 has an irregular granular structure by atomic force microscopy and scanning electron microscopy, and the anomeric carbon in MBP-1 molecule has α-configuration and ß-configuration by NMR and FTIR. The in vitro scavenging abilities of MBP-1 for·OH, DPPH, O2·- , and ABTS+ were 73.5%, 80%, 69.8%, and 75.2%, respectively, and the chelating activity for Fe2+ was 50%, and the inhibition rates of α-glucosidase and α-amylase were 78.5% and 74.6%, respectively, which indicated that MBP-1 possessed strong antioxidant and hypoglycemic activities. The results indicated that MBP-1 have certain application prospects in food-related fields.

Antidiabetic Effect of Millet Bran Polysaccharides Partially Mediated via Changes in Gut Microbiome.

Diabetes is a type of metabolic disease associated with changes in the intestinal flora. In this study, the regulatory effect of millet bran on intestinal microbiota in a model of type 2 diabetes (T2DM) was investigated in an effort to develop new approaches to prevent and treat diabetes and its complications in patients. The effect of purified millet bran polysaccharide (MBP) with three different intragastric doses (400 mg/kg, 200 mg/kg, and 100 mg/kg) combined with a high-fat diet was determined in a streptozotocin (STZ)-induced model of T2DM. By analyzing the changes in indicators, weight, fasting blood sugar, and other bio-physiological parameters, the changes in gut microbiota were analyzed via high-throughput sequencing to establish the effect of MBP on the intestinal flora. The results showed that MBP alleviated symptoms of high-fat diet-induced T2DM. A high dosage of MBP enhanced the hypoglycemic effects compared with low and medium dosages. During gavage, the fasting blood glucose (FBG) levels of rats in the MBP group were significantly reduced (p < 0.05). The glucose tolerance of rats in the MBP group was significantly improved (p < 0.05). In diabetic mice, MBP significantly increased the activities of CAT, SOD, and GSH-Px. The inflammatory symptoms of liver cells and islet cells in the MBP group were alleviated, and the anti-inflammatory effect was partially correlated with the dose of MBP. After 4 weeks of treatment with MBP, the indices of blood lipid in the MBP group were significantly improved compared with those of the DM group (p < 0.05). Treatment with MBP (400 mg/kg) increases the levels of beneficial bacteria and decreases harmful bacteria in the intestinal tract of rats, thus altering the intestinal microbial community and antidiabetic effect on mice with T2DM by modulating gut microbiota. The findings suggest that MBP is a potential pharmaceutical supplement for preventing and treating diabetes.

Probiotic Properties Including the Antioxidant and Hypoglycemic Ability of Lactic Acid Bacteria from Fermented Grains of Chinese Baijiu.

In this study, lactic acid bacteria (LAB) strains were isolated from fermented grains of traditional Chinese Baijiu, and their probiotic properties were characterized. Eleven out of 29 LAB strains showed good tolerance to the gastrointestinal tract and bile salts. The surface characteristics (auto-aggregation, co-aggregation, hydrophobicity), safety (hemolytic and antibiotic sensitivity), antibacterial activity against three foodborne pathogens, and antioxidant and hypoglycemic properties of the 11 LAB strains were investigated. Principal component analysis (PCA) was used to comprehensively evaluate LAB strains and their probiotic properties. It was found that Weissella cibaria (OP288150), Pediococcus acidilactici (OP288151), Pediococcus pentosaceus (OP288154), Pediococcus pentosaceus (OP288156) and Levilactobacillus brevis (OP288158) showed high probiotic properties, with potential for commercial development. The results also demonstrated that fermented grains of Chinese Baijiu can be used as a source of high-quality probiotics.

Portable and on-site electrochemical sensor based on surface molecularly imprinted magnetic covalent organic framework for the rapid detection of tetracycline in food.

In this study, for the first time, surface molecularly imprinted magnetic covalent organic frameworks (Fe3O4@COFs@MIPs) were combined with disposable screen-printed electrode (SPE) to construct a portable and on-site electrochemical sensor for the rapid detection of tetracycline (TC). The Fe3O4@COFs@MIPs, which was prepared by layer-by-layer modification method, had good magnetism and excellent adsorption ability. With the help of disposable SPE, equipped with a magnet, the electrode modification process was simplified and the detection efficiency was improved. Under optimal conditions, the fabricated electrochemical sensor exhibited linearity ranging from 1 × 10-10 to 1 × 10-4 g mL-1. It had good selectivity, excellent reproducibility, desirable stability and remarkable applicability. The fabricated sensor was successfully applied to detect TC in real samples with satisfactory recoveries (96.15-106.20%). The detection strategy separated the recognition and adsorption process from the electrochemical detection process, providing a design idea for the application of COFs in the construction of high-efficiency molecularly imprinted electrochemical sensors.

A novel metal-organic frameworks composite-based label-free point-of-care quartz crystal microbalance aptasensing platform for tetracycline detection.

A novel label-free point-of-care quartz crystal microbalance (QCM) aptasensing platform based on metal-organic frameworks (MOFs) and gold nanoparticles (AuNPs) was fabricated for tetracycline (TC) detection. MOFs (HKUST-1) and AuNPs were modified onto the sensing interface of QCM sensor to enhance the sensing performance of the QCM aptasensor. TC aptamer with sulfhydryl group was fixed through Au-S bond. The recognition performance of the aptasensor was predicted and verified by the computer simulation. At the optimal conditions, the frequency change of the sensor was adopted for quantitative detection of TC. The prepared QCM aptasensor exhibited a wide linear range from 1 × 10-10 g mL-1 to 1 × 10-5 g mL-1 with low limit of detection (0.8 × 10-11 g mL-1). High sensitivity, good selectivity, acceptable recoveries (87.6-91.4%) in real samples were obtained. For the first time, MOFs were utilized in the construction of QCM aptasensing platform, providing a promising application way of MOFs in the QCM sensing.

The antibacterial mechanism of perilla rosmarinic acid.

Rosmarinic acid (RosA) is a phenolic acid compound extracted from perilla. In this experiment, the Oxford cup method was used to verify the antibacterial activity of PerillaRosA against Escherichia coli, Staphylococcus aureus, Salmonella, and Bacillus subtilis. By polyacrylamide gel electrophoresis, the effect of RosA on bacterial nucleic acid and bacterial Na+ /K+ -ATP-ase activity, and scanning electron microscope to exploration of its antibacterial mechanism preliminarily. The results showed that RosA had antibacterial properties against all four bacteria. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of E. coli were 0.8 and 0.9 mg/ml, respectively. The MIC and MBC of Salmonella were 0.9 and 1.0 mg/ml, respectively. The MIC and MBC of S. aureus and B. subtilis were both 1.0 and 1.1 mg/ml. RosA has the bacteriostasis function, which can destroy bacterial cells and cell proteins and inhibit the activity of Na+ /K+ -ATP-ase in cells.

Surface molecularly imprinted magnetic MOFs: A novel platform coupled with magneto electrode for high throughput electrochemical sensing analysis of oxytetracycline in foods.

In order to solve inherent problems of traditional molecularly imprinted electrochemical sensors (MIECS), a novel platform of surface molecularly imprinted magnetic metal-organic frameworks (mMOFs@MIPs) was coupled with magneto electrode to establish magnetic MIECS for the recognition of oxytetracycline (OTC). mMOFs@MIPs were synthesized using layer-by-layer modification method for the recognition of OTC. With the help of magneto electrodes, mMOFs@MIPs can be magnetically modified on the electrode surface, forming the electrochemical sensing interface. The imprinted cavities of mMOFs@MIPs can act as the electron channel of the probe to realize label-free detection of OTC. A linear response was obtained within the OTC concentration range of 1.0 × 10-9 g mL-1-1.0 × 10-4 g mL-1. The applicability of the sensor was estimated using the spiking and recovery method in milk samples with the recoveries ranging from 89.0% to 103.1%. It has potential applications in food safety analysis with high throughput detection capability, high specificity and good stability.

Studies on Adsorption Kinetics and Thermodynamics of Macroporous Resin for Rosmarinic Acid.

This experiment treated perilla seeds with different concentrations of NaCl solution to enrich and purify their rosmarinic acid (RosA). The results showed that low concentrations of salt (0-20 mmol/L) promoted seed germination, while high concentrations (> 20 mmol/L) inhibited germination. When the salt concentration was 20 mmol/L, the germination rate was the highest. The content of RosA in germinated perilla seeds was 3.5 mg/g, which was 3.5 times as much as that in the seeds without germination. The RosA was purified using NK-109 macroporous resin and its adsorption kinetics, isotherms and thermodynamics were determined. The adsorption kinetics showed that the adsorption behavior of RosA in NK-109 resin conformed to the pseudo-second-order kinetic model. The model for RosA in the NK-109 resin exhibited Langmuir adsorption based on a spontaneous exothermic process according to its adsorption thermodynamics, which included both physical and chemical adsorption. The optimized process conditions were as follows: the loading concentration of 0.04 mg/mL, loading volume of 40 mL, 70% methanol as the eluent with the volume of 60 mL, and the purity of RosA was 42.1%.

Development of a molecularly imprinted photoelectrochemical sensing platform based on NH2-MIL-125(Ti)-TiO2 composite for the sensitive and selective determination of oxtetracycline.

In this work, a molecularly imprinted photoelectrochemical (MIP-PEC) sensor based on a novel PEC composite of metal-organic frameworks (MOFs) and TiO2 (NH2-MIL-125(Ti)-TiO2) was established for the ultrasensitive and selective detection of oxytetracycline (OTC). This is the first attempt of applying MOFs in the construction of MIP-PEC sensor. The NH2-MIL-125(Ti)-TiO2 was synthesized by a simple one-step solvothermal method and modified onto the surface of indium tin oxide (ITO) electrode as the photosensitive layer. Subsequently, molecularly imprinted polymer (MIP) was modified as recognition element by electropolymerization. The NH2-MIL-125(Ti)-TiO2 showed an enhanced photocurrent response due to stronger light absorption capacity and matched energy band. Furthermore, MIP greatly improved the selectivity and sensitivity of the constructed PEC sensor. The photocurrent response of the MIP-PEC sensor was reduced after OTC recognition because the specific binding of OTC to the imprinted cavities blocked the electron transfer of the electrode. Under optimal experimental conditions, the MIP-PEC sensor exhibited a wide detection range from 0.1 nM to 10 µM with a low limit of detection (LOD) of 60 pM, as well as certain reproducibility, stability and good applicability in real samples. The proposed sensor provides ideas for the application of MOFs in the construction of PEC sensors and will offer an alternative method for the detection of other pollutants in the field of food safety.

Optimization of Ultrasound-Assisted Extraction Using Response Surface Methodology for Simultaneous Quantitation of Six Flavonoids in Flos Sophorae Immaturus and Antioxidant Activity.

Ultrasound-assisted extraction (UAE) was applied to extract rutin (RU), nicotiflorin (NI), narcissoside (NA), kaempferol (KA), isorhamnetin (IS), quercetin (QU), and total flavonoids of Flos Sophorae Immaturus (TFFSI) from Flos Sophorae Immaturus (FSI). Through single factor test and response surface methodology (RSM), the optimal extraction conditions were concluded as follows: ethanol concentration 70%, time 30 min, temperature 61 °C, and liquid/solid ratio 15.30 mL/g, respectively. The actual extraction rates of RU, NI, NA, KA, IS, QU, and TFFSI were 14.6101%, 2.9310%, 7.1987%, 0.1041%, 0.4920%, 2.7998%, and 26.4260%, respectively. The experimental results demonstrated that the extraction method with accuracy and efficiency could be used for the comprehensive evaluation quality control of extracts from FSI. The antioxidant activities of hydroalcoholic extraction from FSI on 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), superoxide anion (•O2-) free radicals, and ferric reducing/antioxidant power (FRAP) were assessed. The results showed that the antioxidation activities of extracts on DPPH, ABTS•+, and •O2- free radicals were reached 89.29%, 97.86%, and 56.61%, and 81.4% in FRAP at 1.0 mg/mL, respectively. The antioxidant capacity of FSI extract was positively correlated with the amount of total flavonoids.

Purification, structural elucidation and in vivo immunity-enhancing activity of polysaccharides from quinoa (Chenopodium quinoa Willd.) seeds.

Quinoa crude polysaccharides (QPS) were extracted from Chenopodium quinoa Willd. The soluble non-starch polysaccharide fraction (QPS1) was subsequently purified by DEAE-52 cellulose and Sephadex G-50 gel chromatography, using QPS as raw materials. Its chemical structure was identified using FT-IR, NMR, AFM, SEM and Congo red staining. High performance gel permeation chromatography (HPGPC) was used to determine molecular weight, and composition by HPLC. QPS1, with a molecular weight of 34.0 kDa, was mainly composed of mannose, rhamnose, galacturonic acid, glucose, galactose, xylose and arabinose at a molar ratio of 2.63:2.40:1.64:6.28:1.95:2.48:5.01. In addition, we evaluated the ameliorative effects of QPS1 on the improvement of anti-cyclophosphamide (CTX)-induced immunosuppression in ICR mice. The result exhibited significantly immune-enhancing activity: QPS1 successfully improved the content of IFN-γ, IL-6, IFN-ɑ, IgM and lysozyme (LYSO) in serum for three weeks, enhanced the phagocytic function of mononuclear macrophages and ameliorated delayed allergy in mice.

Isolation and screening for limonin-producing endophytic bacteria from Citrus maxima (Burm.) Merr. cv. Shatian Yu.

Limonin, a compound of highly oxidized triterpenoids, has potential functions in preventing or slowing the occurrences of many diseases. In this study, five different bacterial strains were isolated and identified from Citrus maxima (Burm.) Merr. cv. Shatian Yu. Morphological characteristics and 16S rRNA gene sequencing identified them as Bacillus spp, in which two limonin-producing endophytes named P and P9 were discovered by high-performance liquid chromatography and mass spectrometry using an inorganic salt medium and two natural media; also the production was greater in natural medium 1 (4.377 and 0.299 mg/L, respectively) than in natural medium 2 (0.159 and 0.025 mg/L, respectively). The growth and fermentation characteristics of strain P were studied, and during the liquid cultivation of Bacillus sp. P, limonin began to accumulate at the eighth hour in the inorganic salt medium, peaked at the 16th hour, and then decreased sharply. Single-factor experiments revealed that the optimum fermentation conditions for limonin production included 14-H-old cells, 15% inoculum, and 3 g/L glucose.