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. 2026 Jan 16;33:103553. doi: 10.1016/j.fochx.2026.103553

Operational parameters and key characteristics of amino acids and flavor compounds profiles of bovine bone clear and white broths

Qingyang Zhang a,b, Meiyu Zheng a,b, Zhen Lu a,b, Kangyu Zhao a,b, Shu Wang c, Bin Li d, Zhigang Hu d, Sihong Zhang a,b, Dongping He a,b,c, Fenfen Lei a,b,c, Li Zhou a,b,c,⁎
PMCID: PMC12853060  PMID: 41623986

Abstract

Emulsification was found to enhance the nutritional value and flavor of bovine bone broth, although the specific processing conditions and their impact on amino acid profiles and flavor compounds remained unclear. To address this knowledge gap, this study identified critical parameters for producing clear broth (simmering time of 4 h, temperature of 115 °C, bone size of 4.0 cm, and liquid-to-solid ratio of 1:4) and established optimal conditions for white broth (a shear rate of 5000 r/min, emulsification pressure of 18 kPa, temperature of 50 °C, and duration of 90.0 min). Analysis identified 17 amino acids in both broths, while volatile compound profiling revealed 57 and 64 distinct compounds in the clear and white broths, respectively. The findings provide a theoretical foundation for the high-value utilization of bovine bones.

Keywords: Bovine bone, Broth, Emulsification, Amino acids, Volatile substances

Graphical abstract

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Highlights

  • •

    Operational parameters for bovine bone clear and white broths were investigated.

  • •

    Amino acid profiles (17 types) of bovine bone clear and white broth were compared.

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    Types of 57 and 64 volatile compounds were identified in the clear and white broths.

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    Emulsification mechanism in forming white broth's unique quality was explored.

1. Introduction

Within the meat industry, an estimated 40–60% by weight of animal slaughter by-products remain underutilized (Aspevik et al., 2018). Bovine bones, a significant by-product of beef cattle slaughter and processing, are a rich source of nutritional compounds, including fatty acids, amino acids, vitamins, hormones, and minerals such as calcium and phosphorus (Irshad & Sharma, 2015). Furthermore, they contain collagen, which has been demonstrated to promote bone cell proliferation and mitigate osteoporosis (Ye et al., 2019). Influenced regional dietary cultural variations, animal bones are frequently utilized to produce clear broth or for further processing into white broth (Guan et al., 2024). Bovine bone clear broth, characterized by its clear appearance and rich flavor, is commonly utilized as the foundational ingredient for white broth preparation, whereas bovine bone white broth, known for its intense flavor and milky hue, plays a significant role in various delicacies such as hot pot broth and noodle soup (Zhao et al.,2024).

The nutritional value of bone broths was evaluated through total nitrogen content, which serves as a comprehensive indicator of nitrogen-containing compounds (such as proteins, amino acids) (Ding et al., 2023). Additionally, collagen, which accounted for over 90% of the protein present in bone broth, served as an important indicator for the evaluation of broth quality (Ma et al., 2023). From a sensory standpoint, research indicates that high temperature simmering more effectively develops and enhances the flavor profile of bone broth compared to boiling at lower temperatures (Moon et al., 2015). This thermal processing facilitates the degradation of proteins into free amino acids and small peptides, which are key flavor precursors. The significant influence of thermal conditions on the generation of these compounds has been further elucidated in studies on other systems, such as Solen grandis meat (Zheng et al., 2025). Furthermore, the flavor of bone broth is not derived from bones alone; residual meat adhering to the bones also exerts a synergistic effect on the overall flavor. The development of meaty flavors is primarily attributed to the thermal degradation of proteins and lipids, with the resultant volatile compounds contributing distinct sensory notes (Wu et al., 2025).

As a typical oil-in-water emulsion system, bone broth is characterized by its richness in nutrients, as well as the demonstration of good stability and a unique rich taste (Zhang et al., 2024). The formation of the broth system was influenced by various cooking techniques, such as mechanical stirring emulsification, high-pressure homogenization emulsification, ultrasonic emulsification, among others (Qayum et al., 2023). The emulsifying stability index, serving as an indicator of the dispersion degree of emulsions, directly reflected the uniform distribution of dispersed phases such as fat globules in the continuous phase (Celis et al., 2016). Broths with higher emulsifying stability index were associated with enhanced stability and a more pronounced taste richness (Zhang et al., 2024). Presently, research on bovine bone broth primarily centered on the formation mechanism of emulsification and physicochemical properties, while systematic studies on its emulsification process were relatively scarce. Additionally, a deep exploration in the amino acid and volatile substances profile of the bovine bone clear and white broths was lacking. This, to some extent, hindered the quality enhancement of industrial production of bovine bone broth products.

Given the current state of research, a comparative investigation of the preparation methods, nutritional content, and flavor profiles of clear and white bovine bone broths is warranted. This study employed fresh bovine bones to systematically evaluate the impact of simmering time, temperature, and liquid-to-solid ratio on clear broth quality, using total nitrogen and collagen content as key metrics. To overcome the limitations of traditional methods, high-pressure homogenization was utilized to optimize the emulsification and stability of white broth. Furthermore, the amino acid compositions and volatile flavor compounds of both broths were characterized using an amino acid analyzer and headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry. The findings aim to advance the understanding of bovine bone broth and provide a scientific foundation for its industrial production and functional food development.

2. Materials and methods

2.1. Materials

Fresh bovine bone raw materials were purchased from the local Walmart supermarket (Wuhan, China), cut into a designed size, and stored in a refrigerator at −4 °C for downstream utilization. Analytical grade reagents including anhydrous ethanol (≥ 99.7%, CAS: 64–17-5), sodium chloride (≥ 99.8%, CAS: 7647-14-5), sodium hydroxide (≥ 96.0%, CAS: 1310–73-2), concentrated sulfuric acid (95.0–98.0%, CAS: 7664-93-9), magnesium oxide (≥ 98.5%, CAS: 1309-48-4), sodium acetate (≥ 99.0%, CAS: 127–09-3), boric acid (≥ 99.8%, CAS: 10043–35-3), methanol (≥ 99.5%, CAS: 67–56-1), hydrochloric acid (36.0–38.0%, CAS: 7647-01-0), chloramine (T) (≥ 98%, CAS: 127–65-1), glacial acetic acid (≥ 99.8%, CAS: 64–19-7), n-propanol (≥ 99.5%, CAS: 71–23-8), isopropyl alcohol (≥ 99.7%, CAS: 67–63-0), chloroform (≥ 99.9%, CAS: 67–66-3), trichloroacetic acid (≥ 99.7%, CAS: 76–03-9), potassium sulfate (≥ 99.0%, CAS: 7778-80-5), acetic acid (≥ 99.9%, CAS: 64–19-7), copper (II) sulfate pentahydrate (≥ 99.0%, CAS: 7758-99-8), methyl red (92.0% (LC), CAS: 493–52-7), β-Cyclodextrin (≥ 99.0%, CAS: 7585-39-9), and magnesium acetate (≥ 98%, CAS: 142–72-3) were purchased from Sinopharm Group Chemical Reagent Co., Ltd. (Shanghai, China). Standard amino acids, p-dimethylaminobenzaldehyde (≥ 99%, CAS: 100–10-7), bromocresol green (CAS: 76–60-8), citric acid (≥ 99.5%, CAS: 77–92-9), petroleum ether (≥ 99.0%, CAS: 8032-32-4), lanthanum oxide (≥ 99.9%, CAS: 1312-81-8), glucose (≥ 99%, CAS: 50–99-7), phenolphthalein indicator (≥ 98%, CAS: 77–09-8), and sodium L-glutamate (≥ 99%, CAS: 142–47-2) were also procured from the same company. Bovine serum albumin (> 96%, CAS: 9048-46-8) was sourced from Shanghai Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China). Arabic gum (≥ 98%, CAS: 9000-01-5) and starch (≥ 96%, CAS: 9005-84-9) were obtained from Zhengzhou Rongyuan Chemical Products Co., Ltd. (Zhengzhou, Henan, China).

2.2. Preparation of bovine bone clear broth

Fresh bovine bone chunks were soaked in water (25 ± 1 °C) for 15 min and pre-cooked in boiling water for 5 min using an MC-22MC01 induction cooker (Midea Group Co., Ltd., Foshan, Guangdong, China) to eliminate blood and surface impurities. Subsequently, 500 g of these bones were transferred to a Y-80YC9510C pressure cooker (Supor Home Appliances Manufacturing Co., Ltd., Hangzhou, Zhejiang, China) with a working pressure of 50 kPa, and a specific quantity of distilled water and NaCl (0.25 wt%) were added. The mixture was brought to a boil at 100 °C for 5 min and then simmered under controlled conditions. After, the bone broth was cooled down and filtered through a linen cloth to eliminate the solid lipid and bone residue. The bovine bone clear broth was then evaporated and concentrated to one-fifth of its original volume using an XD-5000DQ rotary evaporator (Shanghai Xian De Experimental Instrument Co., Ltd., Shanghai, China) at 80.0 °C, 90 r/min, and a pressure range of −0.08 to −0.1 MPa. Finally, the obtained bovine bone clear broth samples were placed in polyethylene bags and stored in a refrigerator at −18 °C, and the separated fat was also stored in the refrigerator at −4 °C. In this study, the size of bovine bone chunk (2.0, 4.0, 6.0, 8.0, 10.0 cm), bone-to-water ratio (1:1, 1:2, 1:3, 1:4, 1:5, m/m), simmering temperature (105, 110, 115, 120 °C), and simmering time (1.0, 2.0, 3.0, 4.0, 5.0, 6.0 h) were utilized as independent variables to investigate the optimal brewing process for bovine bone clear broth.

2.3. Preparation of bovine bone white broth

The bovine bone clear broth, as detailed in section 2.2, underwent the addition of additives following the method described by Chranioti et al. (2015) with slight modification, shear emulsification, and high-pressure homogenization to achieve bovine bone white broth. Specifically, 500 mL of the bovine bone broth samples were thawed using a HH-2 digital constant temperature water bath pot (Guohua Electrical Appliances Co., Ltd. Changzhou, Jiangsu, China) and heated to 55 °C. To emulsify the mixture, 2.5 wt% Arabic gum and 1.0 wt% modified starch were used. These emulsifiers, along with 15.0 wt% beef tallow lipid (from section 2.2) and 0.5 wt% β-Cyclodextrin (as a stabilizer), were combined using a D2004W/JB50-S high-speed mixer (SaiDe Instrument Co., Ltd. Hangzhou, Zhejiang, China) at a shear rate of 4000–6000 r/min to form a stable emulsion. Subsequently, a high-pressure GYB1001 homogenizer (Shanghai Donghua high pressure homogenizer Co., Ltd., Shanghai, China) was utilized for the further emulsification process of bovine bone white broth for the preparation of white broth, the impacts of independent variables including emulsification temperature (45, 50, 55, 60, 65 °C), emulsification pressure (16.0, 17.0, 18.0, 19.0, 20.0 KPa), and emulsification time (60, 70, 80, 90, 100 mins) on its emulsification effect were investigated.

2.4. Physicochemical properties determinations

2.4.1. Meat-bone mass ratio

The meat attached to the surface of the fresh bovine bones was carefully removed using a knife, with care being taken to avoid the mixing of bone fragments into the meat. The bovine bones and the removed meat were weighed separately using a JA31002 electronic balance (Shanghai Hengping Instrument Co., Ltd. Shanghai, China). The calculation for the meat-to-bone ratio was conducted following Eq. (1).

Meat−bone mass ratio%=m2m1−m2×100 (1)

Where, m1: mass of bovine bone with meat attached (g), m2: mass of the meat removed from the bovine bone (g).

2.4.2. Moisture content

The moisture content of the bovine bone samples was determined following the China National Standard GB 5009.3–2016. The bones were first crushed into particles (diameter ≤ 2 mm) using a dedicated bone crusher (PG-180, Zhengzhou Huazhu Environmental Protection Equipment Co., Ltd. Zhengzhou, China). A 20 g ± 0.0001 g aliquot of the crushed sample was evenly spread in a pre-dried glass Petri dish, ensuring a thickness not exceeding 3 mm. The samples were then dried in an electric blast drying oven (GZX-9070MBE, Shanghai Boxun Industrial Co., Ltd. Shanghai, China) at 105 °C for 4 h until a constant weight was achieved. After drying, the samples were cooled for 1 h in a vacuum desiccator (5211-0250PK, Thermo Scientific™ Nalgene™) and weighed. Moisture content was calculated as the ratio of weight loss during drying to the original sample mass, as per Eq. (2).

Moisture content%=m1−m2m1−m3×100 (2)

Where, m1: total mass of the Petri dish and the bovine bone sample before drying (g); m2: total mass of the Petri dish and the bovine bone sample after drying (g); m3: mass of the Petri dish (g).

2.4.3. Fat content determination

The fat content of the bovine bone samples was determined according to the China National Standard GB/T 5009.6–2016. Briefly, a 5.0 g aliquot of the pre-treated bovine bone powder (from Section 2.4.2) was enclosed in filter paper and placed into a SOX606 Soxhlet extractor (Shanghai Lijing Scientific Instrument Co., Ltd. Shanghai, China). A 50 mL dried flat-bottom flask was used as the receiving vessel, and 20 mL of anhydrous ether was added to the condenser tube, filling approximately two-thirds of the flask's volume. Continuous reflux extraction was performed for 10 h using a HH-2 constant temperature water bath (Guohua Instrument Manufacturing Co., Ltd. Changzhou, China) set at 70 °C. Following extraction, the ether-fat mixture was transferred to a RE52C rotary evaporator (Shanghai Yarong Biochemical Instrument Co., Ltd. Shanghai, China) and concentrated at 40 °C until a constant weight was achieved. The fat content was then calculated based on the mass of the residual material, as defined in Eq. (3).

Fatcontent%=m1−m2m×100 (3)

Where, m: mass of the bovine bone powder sample; m1: mass of the bottle and the fat after evaporation; m2: mass of the empty bottle.

2.4.4. Ash content determination

The ash content of bovine bones was determined according to the China National Standard GB 5009.4–2016. A 2.0 g sample (prepared per method 2.4.2) was placed in a porcelain crucible and treated with 3.0 mL of magnesium acetate solution (80.0 g/L) for 10 min. After drying on a 100 °C water bath, the sample was carbonized by gradual heating to 250–300 °C on a hot plate (HTL-500EX, Shenzhen Bodar Life Science and Technology Co., Ltd. Shenzhen, China) until smokeless. The crucible was then transferred to a muffle furnace (SGM·M6/10, Sigma High Temperature Electric Furnace Co., Ltd. Shanghai, China) and ashed at 550 ± 10 °C for 4 h. This ashing process was repeated until consecutive mass differences were ≤ 0.3 mg. A blank test using 3.00 mL of magnesium acetate solution was processed identically. The ash content was calculated using Eq. (4).

Ashcontentg/100g=m1−m2−m0m3−m2×100 (4)

Where, m1: total mass of the porcelain crucible and ash (g); m2: mass of the porcelain crucible (g); m0: mass of magnesium chloride (product generated after burning magnesium acetate) (g); m3: total mass of the porcelain crucible and bovine bone sample (g).

2.5. Nitrogen and crude protein content determination

The total nitrogen content was determined by the Kjeldahl method according to China National Standard GB/T 6432–2018, with crude protein content expressed as a percentage of dry weight. Precisely 1.0 g of bovine bone powder or broth sample was weighed into a 250 mL digestion tube. A mixture of 6.0 g potassium sulfate and 0.4 g anhydrous copper sulfate was added as catalyst, followed by 12 mL concentrated sulfuric acid and two glass beads. After thorough mixing, digestion was initiated at 200 °C until carbonization occurred, then continued at 420 °C for 2 h until the solution turned clear. The cooled digest was diluted to 50 mL and transferred to a distillation apparatus. Upon adding 50 mL of 40.0 wt% sodium hydroxide to alkalize the solution, the liberated ammonia was distilled into 25 mL of 4.0 wt% boric acid solution with the condenser outlet immersed. The receiver solution was titrated with 0.1 mol/L hydrochloric acid using a mixed methyl red-bromothymol blue indicator (10 drops), and nitrogen content was calculated using Eq. (5).

w=V2−V1×c×141000m×V′V×100 (5)

Where, V1: the volume of standard hydrochloric acid solution consumed by the titrated sample (mL); V2: the volume of the standard hydrochloric acid solution consumed by the titrated blank (mL); c: the concentration of the standard hydrochloric acid titration solution (mmol/L); 14 represents the molar mass of nitrogen element (g/mol); m: weight of the sample; V′: distillation digestion solution volume (mL); V: sample digestion solution volume (mL).

The determination of crude protein content in the bovine bone was conducted following the same method as described above, where the measured nitrogen content was multiplied by a factor of 6.25 to convert it to crude protein content.

2.6. Collagen content determination

The collagen content in bovine bone broth was quantified according to the China Industrial Standard NY/T 3608–2020. Specifically, a hydroxyproline standard stock solution (0.500 g/L) was prepared by dissolving 50.00 mg of hydroxyproline standard in deionized water, adding one drop of 3.0 mol/L HCl, and diluting to 100 mL. For sample preparation, 2.0 mL of bone broth was hydrolyzed with 10.0 mL of 3.0 mol/L HCl at 105 °C for 18 h. The hydrolysate was filtered (0.45 μm), diluted to 100.0 mL, and further diluted to a hydroxyproline concentration within the linear range of 0.5–5.0 μg/mL. For color development, 2.0 mL of the diluted sample was mixed with 2.0 mL of chloramine T reagent (0.05 mol/L, pH 6.0) and reacted at room temperature for 20 min. Then, 2.0 mL of dimethylaminobenzaldehyde reagent was added, and the mixture was incubated at 60 °C for 20 min. Absorbance was measured at 600 nm using a UV-900 spectrophotometer (Shanghai DaPu Instrument Co., Ltd.) with deionized water as the blank. A standard curve was constructed by plotting absorbance against hydroxyproline concentration (Fig. S1), and collagen content was calculated using Eq. (6).

Collagen content%=X×F=c×100×nm×1000×100×F (6)

Where, X: Content of hydroxyproline in the sample (%); c: Concentration of hydroxyproline in the sample solution obtained from the standard curve (μg/mL); m: Sample mass (g); n: Dilution factor of the hydrolyzed sample; F: Conversion factor for collagen (7.9 for bovine bone); 50: Volume after digestion of the sample (mL); 1000- Conversion factor.

2.7. Emulsifying stability index determination

The determination of the emulsifying stability index of the bovine bone broth samples was conducted using a turbidimetric method outlined by Yu, Li, Li and Chen, (2022b) according to the principle that the emulsifying stability index of water-in-oil emulsion is positively correlated with the absorbance measured using a spectrophotometer at a wavelength of 500 nm. Specifically, the bovine bone broth samples obtained through the investigated parameters of shear rate, emulsification temperature, emulsification pressure, and emulsification time were sampled in 30 μL aliquots and placed in 10-mL plastic tubes, and then diluted to 6 mL (200-fold) with 0.1 wt% SDS and thoroughly mixed with a VM-96 A vortexer (Shanghai Chenhui Instrument Co., Ltd. Shanghai, China). Subsequently, the absorbance of the liquid was immediately measured using a UV-VIS spectrophotometer (UV-3100PC, VWR International, Radnor, Pennsylvania, USA) equipped with a 10 mm × 10 mm × 30 mm quartz cuvette at a wavelength of 500 nm. The emulsifying stability index of the bovine bone white broth with the change of shear rate was determined at the emulsification temperature of 50 °C, emulsification pressure of 18.0 kPa, and emulsification time of 80 min. Meanwhile, the emulsification parameters including temperature (45–65 °C), pressure (16.0–20.0 kPa), and time (60–100 min) were investigated.

2.8. Amino acid profile determination

The amino acid profile of bovine bone broth was determined according to China National Standard GB 5009.124–2016. For total amino acids, 4.0 mL of sample was hydrolyzed with 10 mL of 6 mol/L HCl and 4 drops of phenol in a sealed tube under nitrogen atmosphere at 110 °C ± 1 °C for 22 h. The hydrolysate was filtered, diluted to 50 mL, and 1.0 mL was dried under reduced pressure at 40–50 °C. The residue was reconstituted in 1.0 mL sodium citrate buffer, filtered (0.22 μm), and analyzed. For free amino acids, 400 μL of sample was mixed with 100 μL of 10% sulfosalicylic acid, held at 4–8 °C for 60 min, and centrifuged twice at 14,000–15,000 rpm. The supernatant was diluted 1:200 and filtered (0.22 μm). Analysis was performed using an L-8900 amino acid analyzer (Hitachi High Tech) with a sulfonic acid cation exchange column at detection wavelengths of 570 nm and 440 nm. Amino acid concentrations were quantified by peak area using EZChrom Elite software according to Eq. (7).

Xi=ci×F×V×Mm×109×100 (7)

Where, Xi: Content of amino acid i in the sample (g/100 g); ci: Content of amino acid i in the sample determination solution (nmol/mL); F: Dilution factor; V: Volume of sample hydrolysate transferred to a volumetric flask (mL); M: Molar mass of amino acid i; m: Weight of sample (g); 109: Coefficient for converting sample content from nanogram (ng) to gram (g); 100: Conversion factor.

2.9. Volatile substances profile determination

The volatile compounds in bone broth sample were analyzed by headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME-GC–MS) at the Wuhan Institute for Food and Cosmetic Control (Wuhan, China), following a method adapted from Zhou et al. (2025). Specifically, a 2.0 mL broth sample, mixed with 5 mL of boiling water in a 20 mL headspace vial, was subjected to automated pretreatment using a CTC PAL autosampler (Agilent Technologies, Inc. Santa Clara, CA, USA). The sample was equilibrated at 60 °C for 10 min, followed by extraction for 30 min at the same temperature using a 65 μm polydimethylsiloxane/divinylbenzene fiber at a stirring rate of 250 r/min. Thermal desorption was performed in the injection port of a 7890B-5977 A GC–MS system (Agilent Technologies, Inc. Santa Clara, CA, USA) at 230 °C for 10 min. Chromatographic separation was achieved on an Agilent DB-WAX capillary column (30 m × 0.25 mm × 0.25 μm) with high-purity helium as the carrier gas at a constant flow of 2 mL/min. The oven temperature was programmed as follows: 55 °C for 3 min, increased to 115 °C at 10 °C/min and held for 10 min, then raised to 255 °C at 5 °C/min and held for 10 min. High-purity helium gas was employed as the carrier gas at a flow rate of 2 mL/min, with a column head pressure of 61.80 kPa, injection port temperature of 255 °C, and splitless injection mode. The mass spectrometry parameters included the use of an electron ionization source (EI) at a source temperature of 230 °C, gas interface temperature of 280 °C, and quadrupole temperature of 150 °C. The range of scanning was established from 40 to 500 m/z. Volatile substances displaying chromatographic peaks surpassing a signal-to-noise ratio of 300 were recognized using the Canvas workstation software (version 2.5.0, Xuejing Technology Co., Ltd. Beijing, China), with compounds demonstrating a matching level of ≥80 in the NIST20 library (NIST, Gaithersburg, MD, USA).

2.10. Statistical analysis

All experiments using independent samples were conducted in triplicate, and the results were reported as mean ± standard deviation (SD). Statistical analyses were conducted using the Statistical Package for the Social Science (SPSS) version 25.0 (IBM Crop., Armonk, NY, USA). Independent t-tests were employed to compare differences in the content of individual total nitrogen content and collagen content. Meanwhile, One-way ANOVA and Post-hoc Tukey HSD tests were used to determine significant differences in the emulsifying stability index, aminoacid content, and volatile substance content. Figures were created using GraphPad Prism 10.1.2 (GraphPad Software, Boston, MA, USA). Significant differences were reported at the 95% confidence interval (p-value <0.05).

3. Results and discussion

3.1. Main components of fresh bovine bones

The moisture, protein, fat, ash in the bovine bone samples were determined to be 23.87 ± 1.26%, 17.65 ± 0.85%, 28.37 ± 1.32%, and 29.21 ± 1.82%, respectively. Meanwhile, the meat-bone ratio was tested to be 23.94 ± 1.76%. Determining the basic chemical composition of fresh bovine bones serves as the foundation for subsequently optimizing the broth preparation process and analyzing the nutritional components of the final product, as the composition of the raw bone material directly determines the potential content of nutritional and flavor compounds in the broth (Meng et al., 2022). Fresh bovine bones are rich in various components, among which protein (17.65%) and fat (28.37%) levels are particularly notable. Additionally, a higher meat-to-bone ratio (23.94%) was observed to result in an increase in fat content in the raw materials. The quality evaluation of bone broth typically involves its sensory characteristics, physicochemical properties, and nutritional value. Protein and fat, as the primary macronutrients in bovine bones, significantly influence the flavor, mouthfeel, nutritional composition, and overall acceptability of the resulting broth (Li et al., 2022,Ma et al., 2023). A series of amino acids were generated through heat-induced protein hydrolysis in beef bones during the broth-making process (Yu et al., 2017). These amino acids underwent Maillard reactions with reducing sugars, leading to the formation of characteristic flavor compounds, such as pyrazines and furans, which are considered the primary contributors to flavor development in beef bone soup (Sun et al., 2022). During the cooking process, fats, serving as a significant reservoir for aroma compounds, underwent simultaneous hydrolysis and oxidation reactions, leading to the production of fatty acids, which further degraded into volatile compounds such as aldehydes and ketones (Qi et al., 2020).

3.2. Processing parameters of bovine bone clear broth

The impacts of processing parameters including bone size, bone-water mass ratio, simmering temperature, and simmering time on the quality of the bovine bone clear broth were investigated. The total nitrogen content and collagen content were selected as dependent variables.

3.2.1. Bone size and bone-water mass ratio

Both total nitrogen and collagen content in bovine bone clear broth exhibited a significant inverse relationship with bone size (Fig. 1a). A marked decrease occurred as bone size increased from 2.0 cm to 6.0 cm. Subsequently, total nitrogen content stabilized within the 6.0–10.0 cm range, whereas collagen content continued to decline gradually (Fig. 1a). At smaller bone sizes (2.0 cm, 4.0 cm), total nitrogen content reached 1900 mg/L and 1870 mg/L, respectively. However, when bone size exceeded 4.0 cm, a significant reduction was observed, with values falling to 1625 mg/L at 8.0 cm. In contrast, collagen content demonstrated a non-linear trend, characterized by a slight initial increase followed by a continuous decrease (Fig. 1a). While bone sizes ≤4.0 cm had a minimal impact, a consistent decline in collagen content was noted beyond this threshold, decreasing from 50.0 mg/mL at 4.0 cm to 46.5 mg/mL at 10.0 cm (Fig. 1a). These findings align with Zeng QingQing and Zhang LiYan (2014), who reported that smaller chicken bones facilitate more efficient nutrient leaching. Conversely, larger bones increase internal heat and mass transfer resistance, thereby prolonging the extraction of proteins and reducing leach efficiency (Zhang et al., 2023). Consequently, a bone size of 4.0 cm is recommended to achieve an optimal simultaneous release of total nitrogen (1870 mg/L) and collagen (50.0 mg/mL).

Fig. 1.

Fig. 1

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Effect of bone size (a) and solid-to-liquid (bone-to-water) ratio (b) on total nitrogen content (left) and collagen content (right) in bovine bone clear broth.

Both total nitrogen and collagen content in bovine bone clear broth demonstrated a significant positive correlation with the liquid-to-solid ratio (m/v), increasing substantially as the ratio widened (Fig. 1b). At a 1:1 ratio, values were relatively low (1545.3 mg/L and 44.8 mg/mL, respectively), but rose progressively with increasing solvent volume, peaking at 1909.6 mg/L and 60.1 mg/mL at a 1:5 ratio (Fig. 1b). This trend is consistent with dissolution kinetics, whereby a higher ratio increases the bone-solvent contact area, thereby enhancing the extraction efficiency of nitrogenous and collagenous compounds (Yildirim et al., 2011). A marked increase in total nitrogen content occurred between the 1:3 and 1:4 ratios, while collagen content rose most significantly between 1:2 and 1:3. To optimize mass transfer efficiency against production costs, a liquid-to-solid ratio of 1:4 was identified as optimal.

3.2.2. Simmering operations on the bovine bone clear broth

During the 6.0-h simmering process, the total nitrogen content and collagen leaching in bovine bone clear broth were significantly influenced by the interaction between temperature and time (Fig. 2a, b). At any given time, the effect of temperature on both parameters followed the order: 115 °C > 120 °C > 110 °C > 105 °C. In the initial stage (1.0 h), the total nitrogen content reached 1160 mg/L at 105 °C and 1344 mg/L at 115 °C, with corresponding collagen contents of 24.1 mg/mL and 31.8 mg/mL, respectively (Fig. 2a, b). Temperature served as the dominant factor, as elevated temperatures induce contraction of connective tissues (e.g., periosteum and fascia), promoting protein release and breakdown, thereby increasing nitrogen and myofibrillar protein content (Meng et al., 2022). Furthermore, high temperatures alter collagen structure, facilitating partial hydrolysis into water-soluble gelatin, with leaching amounts positively correlated with thermal intensity (Gómez-Guillén et al., 2002). However, excessively high temperatures (>115 °C) can disrupt amino acid structures and intensify the Maillard reaction, leading to the formation of melanoidins from nitrogen-containing compounds and reducing sugars, ultimately reducing detectable nitrogen content (Starowicz & Zieliński, 2019).

Fig. 2.

Fig. 2

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Effects of simmering time (a) and temperature (b) on total nitrogen content and collagen content in the bovine bone clear broth.

Extending the simmering time from 1.0 h to 4.0 h resulted in increased total nitrogen and collagen content across all temperatures, with maximum values attained at the 4.0-h mark (Fig. 2a, b). Specifically, at 115 °C, total nitrogen content rose from 1344 mg/L to 2095 mg/L (a 35.85% increase), while collagen content increased from 31.8 mg/mL to 60 mg/mL (a 47% increase) (Fig. 2a, b). This rise is likely due to thermally promoted protein degradation in meat and bone, which accelerates the release of amino acids and peptides (Ma et al., 2023). However, a further extension of the simmering time to 6.0 h led to a gradual decline in both total nitrogen and collagen content, with values at 115 °C decreasing to 1958 mg/L and 50.9 mg/mL, respectively (Fig. 2a, b). This subsequent decrease may be attributed to the prolonged thermal hydrolysis of collagen into low molecular weight peptides, thereby reducing the detectable content (He et al., 2021). Conversely, Meng et al. (2022) observed that volatile flavor compounds are most abundant at 4.0 h and 5.0 h of cooking. Therefore, to balance extraction efficiency with flavor retention, a simmering time of 4.0 h at 115 °C is recommended.

3.3. Emulsification of bovine bone white broth

3.3.1. Effects of shear rate on the emulsifying stability index

During the emulsification of bovine bone clear broth, emulsifying stability index exhibited a trend of initial increase followed by a decrease with rising shear rate (Fig. 3). As the shear rate increased from 4000 r/min to 5000 r/min, emulsifying stability rose rapidly from 0.61% to 0.67%, peaking at 0.69% at 5000 r/min. Beyond this critical rate, stability declined, falling to 0.69% at 6000 r/min (Fig. 3). At 5000 r/min, increased turbulent kinetic energy promotes a dynamic equilibrium of collision and aggregation among dispersed phase particles, yielding a more uniform emulsion (Ji et al., 2020). However, when shear rate or pressure exceeds this critical threshold, excessive mechanical energy can disrupt the emulsifier film at the droplet interface, significantly increasing coalescence probability, reducing interfacial film elasticity and stability, and consequently diminishing the emulsifying stability index (Jafari et al., 2008).

Fig. 3.

Fig. 3

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Effects of shear rate on the emulsifying stability index of bovine bone white broth.

3.3.2. Effects of emulsification parameters on the emulsifying stability index

The emulsifying stability index (ESI) of bovine bone broth is significantly influenced by emulsification temperature. An increase in temperature to 50 °C elevated the ESI from 0.58% to 0.68%, representing a 15.5% increase (Fig. 4a). However, further heating to 65 °C reduced the ESI to 0.62%. This pattern can be attributed to the reduction of oil-water interfacial tension within a specific thermal range, promoting droplet refinement and enhancing emulsion stability (Ravera et al., 2021). Additionally, natural surfactants in bone broth, such as bile salts and lysolecithin, transition from micellar to monomeric states with rising temperature, increasing the concentration of free monomers available for rapid adsorption and interfacial film repair (Gutiérrez-Méndez et al., 2022). Emulsification pressure similarly affected ESI in a non-linear manner, with a maximum value of 0.69% observed at 18.0 kPa (Fig. 4b). Emulsification time was also a critical parameter: extending the duration from 60.0 min to 80.0 min raised the ESI to 0.68%, peaking at 0.69% after 90.0 min (Fig. 4c). Prolonged mechanical input initially promotes oil droplet fragmentation, reducing average particle size and enhancing dispersion stability through improved adsorption of proteins and emulsifiers at the interface (Sousa et al., 2022). However, exceeding 90.0 min diminished the ESI to 0.68% at 100 min (Fig. 4c), likely due to protein denaturation and impaired interfacial film integrity under prolonged high-shear conditions (Yu, Li, Sun, Yan, & Zou, 2022a). Altogether, to achieve a stable and uniform bovine bone broth emulsion with a delicate texture, the optimal emulsification parameters are a shear rate of 5000 r/min, pressure of 18 kPa, temperature of 50 °C, and time of 90.0 min.

Fig. 4.

Fig. 4

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Effects of emulsification parameters, temperature (a), pressure (b), and time (c), on the emulsifying stability index.

3.4. Amino acids profile analysis

Analysis of the amino acid composition in bovine bone clear broth and white broth revealed significant differences in the distribution of total and free amino acids (Table 1). Seventeen amino acids were identified in both the clear and white broths. Among which, glycine, alanine, proline, aspartic acid, and glutamic acid were determined to be the primary amino acids (Table 1), consistent with the amino acid profile characteristics of bone broth documented by Gauza-Włodarczyk et al. (2017). It is noteworthy that umami amino acids (12.85% ∼ 16.95%) and sweet amino acids (29.29% ∼ 63.11%) constitute a significant proportion, forming the main components of the broth's unique taste (Table 1). Free amino acids accounted for only 3.36% to 9.62% of the total amino acids (Table 1), indicating that amino acids in the broth primarily exist in a bound state.

Table 1.

Amino acids profile of bovine bone clear broth and white broth.

Amino acid type Total aminoacid content(mg/100 mL)
 Free aminoacid content(mg/100 mL)
Clear broth White broth Clear broth White broth
Aspartic acid 208.12 ± 13.26 142.31 ± 7.32 8.38 ± 1.52 16.47 ± 2.13
Threonine* 68.31 ± 3.63 43.71 ± 3.36 5.74 ± 0.82 5.18 ± 0.75
Serine 79.07 ± 3.28 45.14 ± 2.53 0.81 ± 0.05 5.75 ± 0.72
Glutamic acid 208.28 ± 9.83 131.76 ± 7.26 8.78 ± 1.73 12.11 ± 1.65
Glycine 638.31 ± 35.71 540.36 ± 24.83 6.19 ± 0.82 50.34 ± 2.62
Alanine 558.26 ± 27.16 371.22 ± 17.61 5.45 ± 0.74 35.82 ± 2.83
Cystine 183.87 ± 9.53 147.37 ± 6.35 6.28 ± 0.81 13.77 ± 1.62
Valine * 128.45 ± 7.34 96.06 ± 7.58 10.26 ± 1.15 8.06 ± 1.34
Methionine * 16.14 ± 1.32 12.19 ± 1.83 3.14 ± 0.62 2.09 ± 0.47
Isoleucine* 35.04 ± 2.67 23.15 ± 1.95 2.97 ± 0.55 1.91 ± 0.35
Leucine* 66.08 ± 3.43 42.64 ± 2.86 5.09 ± 0.72 3.44 ± 0.63
Tyrosine 15.17 ± 1.56 9.18 ± 1.21 1.04 ± 0.16 1.33 ± 0.16
Phenylalanine* 29.34 ± 2.63 20.07 ± 2.36 6.42 ± 1.37 1.67 ± 0.23
Histidine 48.31 ± 3.17 41.46 ± 3.52 2.13 ± 0.34 3.52 ± 0.81
Lysine* 130.29 ± 6.62 89.32 ± 4.53 11.36 ± 1.58 7.62 ± 1.42
Arginine 114.03 ± 6.53 77.24 ± 4.76 7.87 ± 1.27 7.25 ± 1.22
Proline 483.24 ± 23.51 300.14 ± 21.62 9.33 ± 1.31 28.94 ± 2.39
Total umami amino acid content(%) 13.83 12.85 16.95 13.92
Total sweet amino acid content(%) 62.30 62.91 29.29 63.11
Sulfur-containing amino acid content(%) 0.54 0.57 3.10 1.02
Total amino acid 3010.31 2133.32 101.24 205.27
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Note: The* labeled amino acids are essential for the human body. Umami amino acids are glutamic acid and aspartic acid; sweet amino acids are serine, threonine, glycine, alanine, histidine, and proline, and sulfur-containing amino acids refer to methionine.

In-depth analysis revealed that the total amino acid content of the clear broth was significantly higher than that of the white broth. This is attributed to the fact that the clear broth is directly filtered after stewing, and its aqueous phase contains all the thermally dissolved proteins and peptides. In contrast, the white broth undergoes a high-speed shearing and high-pressure homogenization emulsification process after equivalent stewing. The intense mechanical energy input and the formation of new oil-water interfaces during this process may lead to the integration of some of the dissolved proteins and macromolecular peptides into the emulsified oil phase or the interfacial membrane through interfacial adsorption or interaction with added emulsifiers, thereby reducing their measurable concentration in the aqueous phase and resulting in a relative decrease in total amino acid content (Frempong et al., 2024,Li et al., 2022).

Simultaneously, the total free amino acid content in the white broth (205.27 mg/100 mL) was approximately twice that of the clear broth (101.24 mg/100 mL). This could be resulted from the powerful shear energy provided by emulsification and homogenization, which is utilized to construct the emulsion, also acts as a physical means that may further break peptide bonds, promoting the continuous degradation of proteins and larger peptides into smaller peptides and free amino acids (Hu et al., 2023). This process results in the white broth having a lower total amino acid content but a richer pool of flavor precursors (free amino acids), which aligns with its more intense flavor perception (Fig. 5, Fig. 6). Furthermore, the amino acid profile in bovine bone broth is characterized by equilibrium, showcasing abundant levels of essential amino acids such as lysine, valine, threonine, and leucine. Serving as fundamental constituents of proteins, these essential amino acids are recognized to contribute to muscle tissue repair and the regulation of the synthesis of bioactive compounds, thus playing a role in the metabolism and immune response of organisms (Di & Jia, 2024).

Fig. 5.

Fig. 5

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Radial stacked bar chart of type and relative content of volatile substances in the bovine bone clear (green color bar) and white (yellow color bar) broth. Note: a detailedTable. 1S regarding all the volatile substances was provided as a supplementary file. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Fig. 6.

Fig. 6

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Number of types of volatile substances in bovine bone clear and white broth.

3.5. Volatile compounds analysis

A total of 57 and 64 volatile compounds were identified in bovine bone clear broth and white broth, respectively, encompassing hydrocarbons, alcohols, aldehydes, ketones, esters, acids, sulfur-, oxygen-, and nitrogen-containing compounds (Fig. 5, Fig. 6; Supplementary Table S1, Fig. S2, Fig. S3). The predominant flavor compounds in white and clear broth were hydrocarbons (46.9%; 35.51%), alcohol (21.56%; 30.27%), aldehydes (8.93%; 11.96%), and esters (13.33%; 7.23%). Principal component analysis (PCA) of the 72 shared flavor compounds revealed that the first two principal components (PCs) accounted for 78.9% and 20.1% of the variance, respectively, cumulatively explaining 99% of the total variance and capturing most flavor-related information (Fig. 7a, b). As shown in Fig. 7a, clear and white broth samples were clearly separated along PC1, which contributed 78.87% of the variance and served as the primary discriminant axis. However, the PCA loading plot (Fig. 7b) indicated that no single compound dominated this separation; rather, loadings were widely distributed in a semicircular pattern, suggesting that flavor differences arise from collective variation among multiple compounds rather than individual “key aromas”.

Fig. 7.

Fig. 7

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PCA score plot (a) and loading plot (b) of flavor compounds in the two types of bone broth.

Volatile profiles were closely linked to processing techniques. The higher ester content in clear broth likely results from stewing-promoted esterification (Wang et al., 2024), whereas the elevated aldehyde levels in white broth are attributed to emulsification-induced lipid oxidation—intense shearing increases the fat‑oxygen interface, accelerating oxidative degradation of unsaturated fatty acids (Mahmood & Nuru Garba, 2024; Aslam & Schroën, 2023). These aldehydes impart grassy, fatty, and citrus notes that underpin white broth's flavor (Adams et al., 2011; Zhao et al., 2024). Hydrocarbons were most abundant in both broths. While many straight-chain alkanes have high odor thresholds and act mainly as carriers or intermediates (He et al., 2022), certain unsaturated or branched hydrocarbons contribute fatty, sweet, or waxy aromas and enhance overall complexity (Wang et al., 2022). Notably, certain key compounds significantly shape each broth's characteristic aroma. Dimethyl disulfide (7.58% in white broth), derived from sulfur-containing amino acids via Strecker degradation, contributes onion-like and meaty notes essential to white broth's profile (Stöppelmann et al., 2023). In contrast, clear broth contains more pyrazines formed in advanced Maillard stages, providing nutty and roasted aromas. Overall, although both broths share similar volatile types, distinct concentrations and key active compounds yield differentiated flavor profiles: clear broth emphasizes roasted and meaty notes from thermal reactions, while white broth exhibits stronger fatty, green, and sulfurous meaty aromas from lipid oxidation and sulfur compound release.

4. Conclusion

In conclusion, this study systematically elucidated the preparation processes, amino acid profiles, and volatile compounds of bovine bone clear broth and white broth. Key findings demonstrate that bone size, liquid-solid ratio, cooking temperature and time significantly influenced clear broth quality, while optimal white broth emulsification parameters were established (5000 r/min, 50 °C, 18 kPa, 90 min). Clear broth exhibited higher total amino acids, whereas white broth contained doubled free amino acids. Volatile analysis revealed 57 and 64 compounds in clear and white broths respectively, with hydrocarbons, alcohols, aldehydes and esters as predominant flavor compounds derived from lipid oxidation, Maillard reaction and protein degradation.

Future studies should systematically optimize emulsifier combinations to enhance emulsion stability in bovine bone broth and explore spice incorporation for flavor modification. Further investigation is warranted into the temporal evolution of flavor compounds during cooking, along with storage kinetics and microbial safety assessment, to support industrial development of high-quality bovine bone broth.

CRediT authorship contribution statement

Qingyang Zhang: Writing – original draft, Methodology, Investigation, Conceptualization. Meiyu Zheng: Writing – original draft, Methodology, Investigation. Zhen Lu: Methodology, Investigation. Kangyu Zhao: Methodology, Data curation. Shu Wang: Methodology, Investigation, Data curation. Bin Li: Methodology, Investigation. Zhigang Hu: Methodology, Conceptualization. Sihong Zhang: Resources, Methodology, Conceptualization. Dongping He: Resources. Fenfen Lei: Supervision, Funding acquisition. Li Zhou: Supervision, Project administration, Methodology, 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.

Acknowledgements

This work was supported by the Department of Science and Technology of Hubei Province (2025DJA027), the Open Research Fund of the Key Laboratory of Edible Oil Quality and Safety, State Administration for Market Supervision (SYYKF202306), the Young Elite Scientist sponsorship program by the China Association for Science, and Technology (YESS20200380), and the Guangxi Key Research and Development Program (AB23026140).

Footnotes

Appendix A

Supplementary data to this article can be found online at https://doi.org/10.1016/j.fochx.2026.103553.

Appendix A. Supplementary data

Supplementary material

mmc1.docx (510.9KB, docx)

Data availability

Data will be made available on request.

References

  1. Adams A., Kitryte V., Venskutonis R., De Kimpe N. Model studies on the pattern of volatiles generated in mixtures of amino acids, lipid-oxidation-derived aldehydes, and glucose. Journal of Agricultural and Food Chemistry. 2011;59(4):1449–1456. doi: 10.1021/jf104091p. [DOI] [PubMed] [Google Scholar]
  2. Aslam A., Schroën K. Lipid oxidation in food emulsions: A review dedicated to the role of the interfacial area. Current Opinion in Food Science. 2023;51 doi: 10.1016/j.cofs.2023.101009. [DOI] [Google Scholar]
  3. Aspevik T., Oterhals Å., Rønning S.B., Altintzoglou T., Wubshet S.G., Gildberg A., Lindberg D. Valorization of proteins from co-and by-products from the fish and meat industry. Chemistry and chemical technologies in waste valorization. 2018;123-150 doi: 10.1007/978-3-319-90653-9_5. [DOI] [Google Scholar]
  4. Celis M.T., Contreras B., Forgiarini A., Rosenzweig L., P., & Garcia-Rubio, L. H. Effect of emulsifier type on the characterization of O/W emulsions using a spectroscopy technique. Journal of Dispersion Science and Technology. 2016;37(4):512–518. doi: 10.1080/01932691.2015.1048459. [DOI] [Google Scholar]
  5. Chranioti C., Nikoloudaki A., Tzia C. Saffron and beetroot extracts encapsulated in maltodextrin, gum Arabic, modified starch and chitosan: Incorporation in a chewing gum system. Carbohydrate Polymers. 2015;127:252–263. doi: 10.1016/j.carbpol.2015.03.049. [DOI] [PubMed] [Google Scholar]
  6. Di C., Jia W. Food-derived bioactive peptides as momentous food components: Can functional peptides passed through the PI3K/Akt/mTOR pathway and NF-κB pathway to repair and protect the skeletal muscle injury? Critical Reviews in Food Science and Nutrition. 2024;64(25):9210–9227. doi: 10.1080/10408398.2023.2209192. [DOI] [PubMed] [Google Scholar]
  7. Ding Y., Yan C., Dai W., Wang Y., Liu S., Zheng R., Zhou X. Flavor improving effects of cysteine in xylose–glycine–fish waste protein hydrolysates (FPHs) Maillard reaction system. Bioresources and Bioprocessing. 2023;10(1):95. doi: 10.1186/s40643-023-00714-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frempong K.E.B., He G., Kuang M., Xue M., Wang J., Wei Y., Zhou J. Sulfonated cottonseed hydrolysates with adjustable amphiphilicity as environmental -stress stable emulsifiers. Food Chemistry. 2024;454 doi: 10.1016/j.foodchem.2024.139787. [DOI] [PubMed] [Google Scholar]
  9. Gauza-Włodarczyk M., Kubisz L., Włodarczyk D. Amino acid composition in determination of collagen origin and assessment of physical factors effects. International Journal of Biological Macromolecules. 2017;104:987–991. doi: 10.1016/j.ijbiomac.2017.07.013. [DOI] [PubMed] [Google Scholar]
  10. Gómez-Guillén M.C., Turnay J., Fernández-Dıaz M.D., Ulmo N., Lizarbe M.A., Montero P. Structural and physical properties of gelatin extracted from different marine species: A comparative study. Food Hydrocolloids. 2002;16(1):25–34. doi: 10.1016/S0268-005X(01)00035-2. [DOI] [Google Scholar]
  11. Guan H., Tian Y., Feng C., Leng S., Zhao S., Liu D., Diao X. Migration of nutrient substances and characteristic changes of chicken white soup emulsion from chicken skeleton during cooking. Foods. 2024;13(3):410. doi: 10.3390/foods13030410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gutiérrez-Méndez N., Chavez-Garay D.R., Leal-Ramos M.Y. Lecithins: A comprehensive review of their properties and their use in formulating microemulsions. Journal of Food Biochemistry. 2022;46(7) doi: 10.1111/jfbc.14157. [DOI] [PubMed] [Google Scholar]
  13. He L., Gao Y., Wang X., Han L., Yu Q., Shi H., Song R. Ultrasonication promotes extraction of antioxidant peptides from oxhide gelatin by modifying collagen molecule structure. Ultrasonics Sonochemistry. 2021;78 doi: 10.1016/j.ultsonch.2021.105738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. He X., Li B., Yu X., Zhuang Y., Li C., Dong L., Wang S. Inhibiting effects of ginger and rosemary on the formation of heterocyclic amines, polycyclic aromatic hydrocarbons, and trans fatty acids in fried pork balls. Foods. 2022;11(23):3767. doi: 10.3390/foods11233767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hu J., Yu B., Yuan C., Tao H., Wu Z., Dong D., Lu Y., Zhang Z., Cao Y., Zhao H., Cheng Y., Cui B. Influence of heat treatment before and/or after high-pressure homogenization on the structure and emulsification properties of soybean protein isolate. International Journal of Biological Macromolecules. 2023;253 doi: 10.1016/j.ijbiomac.2023.127411. [DOI] [PubMed] [Google Scholar]
  16. Irshad A., Sharma B.D. Abattoir by-product utilization for sustainable meat industry: A review. Journal of Animal Production Advances. 2015;5(6):681–696. doi: 10.5455/japa.20150626043918. [DOI] [Google Scholar]
  17. Jafari S.M., Assadpoor E., He Y., Bhandari B. Re-coalescence of emulsion droplets during high-energy emulsification. Food Hydrocolloids. 2008;22(7):1191–1202. doi: 10.1016/j.foodhyd.2007.09.006. [DOI] [Google Scholar]
  18. Ji Y., Bellettre J., Montillet A., Massoli P. Fast oil-in-water emulsification in microchannel using head-on impinging configuration: Effect of swirl motion. International Journal of Multiphase Flow. 2020;131 doi: 10.1016/j.ijmultiphaseflow.2020.103402. [DOI] [Google Scholar]
  19. Li X., Meng H., Zhu Y., Shu S., Bao Y., Jiang S., Liu Y. Correlation analysis on sensory characteristics and physicochemical indices of bone broth under different processing methods. Food Chemistry Advances. 2022;1 doi: 10.1016/j.focha.2022.100036. [DOI] [Google Scholar]
  20. Ma C., Tian X., Li Y., Guo J., Wang X., Chen S., Wang W. Using high-temperature cooking for different times for bone soup: Physicochemical properties, protein oxidation and nutritional value. Journal of Food Composition and Analysis. 2023;122 doi: 10.1016/j.jfca.2023.105467. [DOI] [Google Scholar]
  21. Mahmood R.M., Nuru Garba N.G. Experimental analysis of the effect of shear forces on oil-water separation, using a high shear mixer. International Journal of Science for Global Sustainability. 2024;10(2):182–189. doi: 10.57233/ijsgs.v10i2.664. [DOI] [Google Scholar]
  22. Meng Q., Zhou J., Gao D., Xu E., Guo M., Liu D. Desorption of nutrients and flavor compounds formation during the cooking of bone soup. Food Control. 2022;132 doi: 10.1016/j.foodcont.2021.108408. [DOI] [Google Scholar]
  23. Moon S.H., Kim J., Hwang K.T., Cha Y.B. Physicochemical and sensory characteristics of beef-bone broths prepared under atmospheric pressure and overpressure. Korean Journal of Food Science and Technology. 2015;47(6):725–732. doi: 10.9721/KJFST.2015.47.6.725. [DOI] [Google Scholar]
  24. Qayum A., Rashid A., Liang Q., Wu Y., Cheng Y., Kang L., Ma H. Ultrasonic and homogenization: An overview of the preparation of an edible protein–polysaccharide complex emulsion. Comprehensive Reviews in Food Science and Food Safety. 2023;22(6):4242–4281. doi: 10.1111/1541-4337.13221. [DOI] [PubMed] [Google Scholar]
  25. Qi J., Xu Y., Xie X.F., Zhang W.W., Wang H.H., Xu X.L., Xiong G.Y. Gelatin enhances the flavor of chicken broth: A perspective on the ability of emulsions to bind volatile compounds. Food Chemistry. 2020;333 doi: 10.1016/j.lwt.2024.116905. [DOI] [PubMed] [Google Scholar]
  26. Ravera F., Dziza K., Santini E., Cristofolini L., Liggieri L. Emulsification and emulsion stability: The role of the interfacial properties. Advances in Colloid and Interface Science. 2021;288 doi: 10.1016/j.cis.2020.102344. [DOI] [PubMed] [Google Scholar]
  27. Sousa A.M., Pereira M.J., Matos H.A. Oil-in-water and water-in-oil emulsions formation and demulsification. Journal of Petroleum Science and Engineering. 2022;210 doi: 10.1016/j.petrol.2021.110041. [DOI] [Google Scholar]
  28. Starowicz M., Zieliński H. How Maillard reaction influences sensorial properties (color, flavor and texture) of food products? Food Reviews International. 2019;35(8):707–725. doi: 10.1021/acs.jafc.3c03153. [DOI] [Google Scholar]
  29. Stöppelmann F., Chan L.F., Liang J., Greiß M., Lehnert A.S., Pfaff C., Zhang Y. Generation of meaty aroma from onion (Allium cepa L.) with Polyporus umbellatus: Fermentation system, sensory profile, and aroma characterization. Journal of Agricultural and Food Chemistry. 2023;71(35):13054–13065. doi: 10.1021/acs.jafc.3c03153. [DOI] [PubMed] [Google Scholar]
  30. Sun A., Wu W., Soladoye O.P., Aluko R.E., Bak K.H., Fu Y., Zhang Y. Maillard reaction of food-derived peptides as a potential route to generate meat flavor compounds: A review. Food Research International. 2022;151 doi: 10.1016/j.foodres.2021.110823. [DOI] [PubMed] [Google Scholar]
  31. Wang L., Li C., Al-Dalali S., Liu Y., Zhou H., Chen C., Wang Y. Characterization of key aroma compounds in traditional beef soup. Journal of Food Composition and Analysis. 2022;114 doi: 10.1016/j.jfca.2022.104839. [DOI] [Google Scholar]
  32. Wang Q., Xie J., Wang L., Jiang Y., Deng Y., Zhu J.…Yang Y. Comprehensive investigation on the dynamic changes of volatile metabolites in fresh scent green tea during processing by GC-E-nose, GC–MS, and GC × GC-TOFMS. Food Research International. 2024;187 doi: 10.1016/j.foodres.2024.114330. [DOI] [PubMed] [Google Scholar]
  33. Wu S., Bai Y., Xu B., Li X., Yao Z., Li J., Sun Y. The formation and change of volatile flavor compounds during the cooking of sheep bone soup. Foods. 2025;14(6):949. doi: 10.3390/foods14060949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ye M., Jia W., Zhang C., Shen Q., Zhu L., Wang L. Preparation, identification and molecular docking study of novel osteoblast proliferation-promoting peptides from yak (Bos grunniens) bones. RSC Advances. 2019;9(26):14627–14637. doi: 10.1039/C9RA00945K. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yildirim A., Öner M.D., Bayram M. Fitting Fick’s model to analyze water diffusion into chickpeas during soaking with ultrasound treatment. Journal of Food Engineering. 2011;104(1):134–142. doi: 10.1016/j.jfoodeng.2010.12.005. [DOI] [Google Scholar]
  36. Yu C., Li S., Sun S., Yan H., Zou H. Modification of emulsifying properties of mussel myofibrillar proteins by high-intensity ultrasonication treatment and the stability of O/W emulsion. Colloids and Surfaces A: Physicochemical and Engineering Aspects. 2022;641 doi: 10.1016/j.colsurfa.2022.128511. [DOI] [Google Scholar]
  37. Yu J.J., Li N., Li S., Chen Y. Solubilization mechanism and structural properties of high-denatured peanut protein treated by shearing. Journal of Food Process Engineering. 2022;45(2) doi: 10.1111/jfpe.13951. [DOI] [Google Scholar]
  38. Yu T.Y., Morton J.D., Clerens S., Dyer J.M. Cooking-induced protein modifications in meat. Comprehensive Reviews in Food Science and Food Safety. 2017;16(1):141–159. doi: 10.1111/1541-4337.12243. [DOI] [PubMed] [Google Scholar]
  39. Zeng QingQing Z.Q., Zhang LiYan Z.L. Study on emulsification conditions and emulsion stability of chicken bone stock. China Condiment. 2014;1–6 https://www.cabidigitallibrary.org/doi/full/10.5555/20143172460. [Google Scholar]
  40. Zhang H., Liu H., Qi L., Xv X., Li X., Guo Y., Richel A. Application of steam explosion treatment on the collagen peptides extraction from cattle bone. Innovative Food Science & Emerging Technologies. 2023;85 doi: 10.1016/j.ifset.2023.103336. [DOI] [Google Scholar]
  41. Zhang L., Cao X., Zhu B., Wang H., Zhong K., Shi B., Zhao L. Enhancing saltiness perception in bone broth: The additive effect of oil and optimization of sodium-reduction formula for consumer acceptability. Journal of the Science of Food and Agriculture. 2024;104(10):6108–6117. doi: 10.1002/jsfa.13438. [DOI] [PubMed] [Google Scholar]
  42. Zhao H., Zhang S., Ma D., Liu Z., Qi P., Wang Z., Di S., Wang X. Review of fruits flavor deterioration in postharvest storage: Odorants, formation mechanism and quality control. Food Research International. 2024;182 doi: 10.1016/j.foodres.2024.114077. [DOI] [PubMed] [Google Scholar]
  43. Zhao L., Gao H., Xiao Y., Sun L., Li S., Zhu Y., Li M. Preparation and characteristics of tea water-insoluble protein/chitosan composite particles. LWT. 2024;212 doi: 10.1016/j.lwt.2024.117009. [DOI] [Google Scholar]
  44. Zheng J., Li J., Song Z., Liu Y., Yu D., Wang Q., Jiang P. Effects of different thermal cooking methods on the characteristic taste and flavor compounds of Solen grandis meat. LWT. 2025;117439 doi: 10.1016/j.lwt.2025.117439. [DOI] [Google Scholar]
  45. Zhou L., Zheng M., Zhang T., Wang S., Jiang X., Li B., Zhang Q. Chemical property, bioactive constituent and volatile compound changes of beef tallow during an improved flavor-retaining refining approach. Food Chemistry. 2025;477 doi: 10.1016/j.foodchem.2025.143491. [DOI] [PubMed] [Google Scholar]

Associated Data

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Supplementary Materials

Supplementary material

mmc1.docx (510.9KB, docx)

Data Availability Statement

Data will be made available on request.

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