hydraulic power tong in china free sample

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Find parts you need to repair or maintain your machines. At Alibaba.com, you can shop for power tong at affordable rates to tackle new obstacles and challenges. In the ever-changing industry, you can find what you need and speak to the supplier directly. Thanks to Alibaba’s collection of wholesale power tong you also get to buy these parts at lower prices, which means you can explore new levels every day more comfortably. From bulldozers to dragline excavators, wheel tractor scrapers to shotcrete machines, any part you need for a heavy-duty mining machinery; you can find it at Alibaba.com.

Looking for purpose-built machine parts? Find them at Alibaba.com. From new components to used parts straight from the manufacturers. Plus, if you need custom-made pieces, you can chat with the supplier, give specifications and wait on delivery. From stone crushers to excavator undercarriage parts, buckets, and even drill bits to get you through the rocks, the power tong from Alibaba offers you the chance to continue operating without a hitch. Whether you are looking to introduce concrete into the rock walls for more consistency and safety during mining, then power tong that goes at wholesale prices at Alibaba will be an excellent addition to your machinery.

Before buying a component, you’d want the equipment to suit your application and offer value. The list of power tong at Alibaba.com lets you dig into earth deposits, and the compare tool checks out other similar parts to give you the information you need to make a purchasing decision. You’ll get wholesale power tong that specializes in mining, with reinforced chassis, and run on more powerful engines. Whether you want to transport minerals or the workers to the mining site, introduce explosives or arms to help you remove materials from your mine pits, Alibaba.com has it all.

Find parts you need to repair or maintain your machines. At Alibaba.com, you can shop for casing hydraulic power tong at affordable rates to tackle new obstacles and challenges. In the ever-changing industry, you can find what you need and speak to the supplier directly. Thanks to Alibaba’s collection of wholesale casing hydraulic power tong you also get to buy these parts at lower prices, which means you can explore new levels every day more comfortably. From bulldozers to dragline excavators, wheel tractor scrapers to shotcrete machines, any part you need for a heavy-duty mining machinery; you can find it at Alibaba.com.

Looking for purpose-built machine parts? Find them at Alibaba.com. From new components to used parts straight from the manufacturers. Plus, if you need custom-made pieces, you can chat with the supplier, give specifications and wait on delivery. From stone crushers to excavator undercarriage parts, buckets, and even drill bits to get you through the rocks, the casing hydraulic power tong from Alibaba offers you the chance to continue operating without a hitch. Whether you are looking to introduce concrete into the rock walls for more consistency and safety during mining, then casing hydraulic power tong that goes at wholesale prices at Alibaba will be an excellent addition to your machinery.

Before buying a component, you’d want the equipment to suit your application and offer value. The list of casing hydraulic power tong at Alibaba.com lets you dig into earth deposits, and the compare tool checks out other similar parts to give you the information you need to make a purchasing decision. You’ll get wholesale casing hydraulic power tong that specializes in mining, with reinforced chassis, and run on more powerful engines. Whether you want to transport minerals or the workers to the mining site, introduce explosives or arms to help you remove materials from your mine pits, Alibaba.com has it all.

The tong should be secured for both make-up or break-out operation, by utilizing the snub line. If this is not done, the tong may be thrown against operator causing physical harm.

To be sure connectors are completely tight, first tighten them until trave lis restricted and the end of the thread travel appears to be reached. Then try to tighten the connector further to be sure first restriction was not a false tightness. Then continue to tighten the fitting until connection is tight.

When using the mechanical shift lever to change speeds, the power tong must first come to a complete stop before shifting. When using tongs hydraulic shift two-speed motor to change speeds, the tong may be shifted "On the Run."

Eckel tongs have proven to basically to last forever with minimal maintenance as all they are manufactured with the highest quality of steel. Using Eckel equipment tells your customer that you have the highest quality equipment on the market.

Tong size is determined by range of tubulars you will run. For example a 5-1/2 Hydra-Shift® is capable of running tubulars 5-1/2-inches and smaller while the 14 UHT is capable of running tubulars 14-inches and smaller. It is important not to use a large range of sizes with just one tong. If you have a 10-3/4 Standard and you regularly run 4-1/2-inch tubing with this tong, you might consider using a smaller tong.

Determine the maximum torque that will be required to break-out the tubular you are running. There are many influences that figure in to break out torque. In some instances it requires up to 2-1/2 times the makeup torque to break out a connection.

Wrap-Around dies offer a much larger surface contact on the tubular. This distributes the force uniformly around the tubular reducing needless pipe damage due to point loading.

PSI pressure determines the maximum torque the tong will safely be able to reach. Eckel rates all their tongs at the industry standard 2500 PSI. A competitor with a similar size tong may show more or the same torque as an Eckel tong due to a higher PSI from the power unit (which is in fine print) in an effort to fool you, thinking there tong is equal to the industry standard (Eckel tong.)

Gallons Per Minute determines the rotational speed of the tong. A low GPM will cause the tong to operate at a lower speed while a high GPM will result in the tong to rotate at a higher speed. Eckel offers an RPM (Revolutions per minute) Control which is a flow divider to decrease the amount of hydraulic fluid that reaches the tong if needed, the remaining fluid is returned to the power unit reservoir. By decreasing the amount of fluid reaching the tong the operator is able to control the maximum RPM of the tong.

Field tests have shown depending on several factors most power units used in above 32 degrees Fahrenheit conditions no matter if your hydraulic oil tank holds 200 gallons of oil, will exceed 150 degrees during a short 8 hour job. Most power units without hydraulic oil coolers exceed 170 degrees which is way past the recommended guide lines.

Global power tong market value is anticipated to grow with an impressive CAGR during the forecast years on the backbone of growing demands for efficient and effective equipment for oil excavation and reservoir drilling activities. Surge in the demand for oil and natural gas from the various end use sectors also drives the growth of the global power tong market in the upcoming five years. Rising concerns regarding the safety of the reserve workers and drilling site workers in case of functional at a complicated site, upsurges the demand for better and efficient drilling equipment and safety equipment also supports the growth of the global power tong market in the next five years.

Increasing consumption of oil & gas in the various end use industries, drive the growth of the global power tong market in the upcoming five years. With higher consumption, consistent increase in the production of the oil and gas through oil wells is also experienced. Growing number of oil reserves and higher drilling activities further facilitates the growth of the global power tong market in the future five years. The demand for the power tongs is also increasing due to drilling for oil in the complicated and difficult terrain. Excavation activities in the difficult terrain makes it impossible for the workers to access the location physically. In those terms, efficient drilling equipment and safety tools are required for the drilling, thereby aiding the growth of the global power tong market in the next five years.

Global oil production in the year 2020 was recorded to be 4165.1 million tonnes and its consumption was about 88477 thousand barrels per day. The United States oil production capacity was 712.7 million tonnes in the year 2020. Demands from the end use industries are exhaustive. Increasing number of passenger cars and commercial vehicles further upsurge the demand for higher production of oil and thus aids the growth of the global power tong market in the forecast years.

Demand for higher performance, efficiency, and cost effectiveness demands for the consistent research and technological advancement in the oil drilling equipment and safety equipment. It is highly mandatory that these equipment are safe switched and are easily manageable by the remote network based control to the equipment. Also, higher production of the oil and gas demands for the advanced and evolved equipment that has better performance as well as can be equipped for longer duration of time and is fail safe. The cost of production, distribution and supply of oil and oil products is very expensive, and still, the demands for this energy produced is increasing rapidly, thus creating further demand for efficient methods of oil excavation, drilling, and supply.

The global power tong market segmentation is based on type, product type, function, location, regional distribution, and competitive landscape. Based on type, the market is differentiated between hydraulic and pneumatic. By product type, the market is further segmented into chisel tong, casing tong, rotary tong, manual tong, and others. Based on function, the market is distinguished between breakout tong and makeup tong. By location, the market is bifurcated between offshore and onshore location. The market analysis also studies the regional segmentation, divided among Asia-Pacific region, North American region, European region, South American region, and Middle East & African region.

Keystone Energy Tools, Texas International Oilfield Tools Limited, Eckel, Weatherford International, ProTorque, Edcon Power Tongs and Oilfield Services Ltd., TNT Power Tongs, Starr Power Tongs, Besdrill Machinery, McCoy Global Inc., are enlisted in a partial list of major market players of the global power tong market.

In this report, global power tong market has been segmented into following categories, in addition to the industry trends which have also been detailed below:

With the given market data, TechSci Research offers customizations according to a company"s specific needs. The following customization options are available for the report:

Monolayer graphene was produced by chemical vapour deposition, as reported previously2 (99.999%) for 30 min. After that, the Cu foil was annealed at 1,000 °C in 4 sccm H2 for another 30 min. Then, 16 sccm CH4 (99.999%) was introduced as the carbon source in the atmosphere of 4 sccm H2. The growth lasted for 15 min. Finally, the tube furnace was immediately cooled to room temperature under H2 gas. The graphene film on the Cu foil was transferred to a SiO2/Si substrate by the electrochemical bubbling method

The g-FET device was fabricated via a thermally assisted bilayer lift-off process2/Si wafer. After photolithography (Microwriter ML3, Durham Magneto Optics), 5/45 nm Cr/Au was deposited by using a thermal evaporator (Angstrom Engineering). The sample was heat treated on a hot plate at 170 °C for 1 min to allow photoresist reflow. Due to the higher reflow temperature of LOR (MicroChem LOR series reflow temperature >250 °C, DOW MicroPosit S1800 series reflow temperature <200 °C), the bottom photoresist undercut remains un-deformed while the top layer deformed. The introduced gap allowed the Remover PG stripper solution to access the photoresist, and improved the efficiency of the lift-off process considerably. The sample was treated by oxygen plasma to remove the photoresist residues. Graphene synthesized by chemical vapour deposition was then transferred onto the wafer using poly(methyl methacrylate) (PMMA) carrier layer to connect the drain and the source electrodes2 plasma etching techniques to define the sensing region. Typically, the width and length of the graphene channel were 60 μm and 30 μm, respectively.

MolEMS was obtained by self-assembling four oligonucleotides. The sequences of the oligonucleotides were designed as listed in Supplementary Table 1, and then were prepared and purified by Sangon Biotechnology. Once received, we centrifuged the tubes at 10,200 rpm at 4 °C for 5 min, then dissolved the DNA powder in ultrapure water (18.2 MΩ, Milli-Q system). The concentrations of ss-DNA stock solutions were calculated on the basis of their absorbance at 260 nm using ultraviolet-visible spectroscopy (Lambda750, Perkin-Elmer) through the Beer–Lambert lawA = εbc, where A is the absorbance, ε is the molar absorptivity, b is the optical path length and c is the molar concentration. Equimolar quantities at a final concentration (1 μM) of four strands for the assemblage of the MolEMSs were mixed in 1× TM buffer at 95 °C for 10 min, and were cooled to 4 °C immediately using a thermal cycler (SimpliAmp, Thermo Fisher Scientific)2+, respectively, or complementary oligonucleotide for detecting ss-DNA-T, cDNA and RNA. Finally, the MolEMSs were stored in 1× TM buffer at 4 °C for later use.

The immobilization of MolEMS involves non-covalently functionalizing 1-pyrenebutanoic acid succinimidyl ester (PASE, Sigma-Aldrich) on monolayer chemical-vapour-deposition graphene (Supplementary Fig. 3) via π–π stacking, and covalently linking PASE to the amino groups at the bottom of the base of MolEMS. The g-FET device was immersed in a dimethylformamide (Sigma-Aldrich) solution of 5 mM PASE for 1.5 h at room temperatureπ–π interactions between graphene and the pyrene groups of PASE. A PDMS well was stamped above the graphene channel to hold the solution. After rinsing thoroughly with ethanol and ultrapure water, 50 μl 1× TM buffer with 100 nM MolEMS was added in the PDMS well for 12 h at room temperature. Following this incubation, the solution in PDMS well was changed to 100 mM ethanolamine (Sigma-Aldrich) in 1× TM buffer solution for 1 h, which deactivated and blocked the excess reactive groups remaining on the graphene surface (Supplementary Fig. 4). Then, the solution in the PDMS well was changed to 1× TM buffer, and the device was rinsed at least three times. Finally, 80 μl buffer or VTM was added to the PDMS well, and it was sealed by a piece of glass or wafer. The device was stored at 4 °C in the dark.

The morphologies of g-FET devices modified with MolEMSs and ss-DNA aptamers were analysed in fluids (1× TM buffer) by AFM (Fastscan, Bruke) operated in ScanAsyst mode using an ultrasharp tip (Fluid + , Bruke) of ~2–3 nm radius. In the AFM images, we determined the number of MolEMSs in a certain area on the graphene surface, and calculated the density via dividing the number by the area. The average density was obtained by measuring different locations. The g-FET devices after serum incubation were rinsed by ultrapure water and dried by nitrogen, then were analysed in air by AFM operated in ScanAsyst mode using a ~20–25 nm radius tip (Scanasyst air).

The graphene samples were measured by Raman spectrometer (LabRam HR Evolution, Horiba Jobin Yvon, 532 nm Ar ion laser) and high-resolution transmission electron microscope (Tecnai G2 F20 S-Twin, acceleration voltage 200 kV). The g-FET devices modified with MolEMSs or ss-DNA aptamers were washed thoroughly by ultrapure water, dried by nitrogen and then scanned by scanning electron microscope (Gemini SEM500, Zeiss) at 0.8 keV. MolEMSs were analysed using 3% agarose gel electrophoresis in 1× TBE Mg2+ buffer (89 mM Tris, 89 mM boric acid, 2 mM EDTA, 12.5 mM Mg2+, pH ~8.0) at 95 V for 40 min, and then the gels were stained in Sybr safe (Thermo Fisher) for 30 min followed by imaging under ultraviolet exposure (Gel-doc XR + , Bio-Rad). Before and after modifying with PASE and MolEMS, the graphene was measured by X-ray photoelectron spectroscopy (Thermo Fisher Scientific K-Alpha instrument) with an Al X-ray source (hν = 1,486.6 eV).

FRET analysis was performed as follows−1 scan rate; (3) the FRET efficiency (EFRET) was determined from the decrease in donor fluorescence intensity via:

where IDA and ID denote the measured maximum fluorescence intensity of the donor (Cy3) for the DNA nanostructures with or without the acceptor (Cy5), respectively. The sequences (5′-3′) of DNA strands used in the FRET experiments are presented in the Supplementary Table 1.

Graphene was transferred by PMMA onto a quartz substrate with pre-patterned gold electrodes and patterns. The graphene surface was immobilized with Cy3-conjugated 17bp-15T MolEMSs, Cy3-conjugated 7 bp DNA nanostructures or Cy3-conjugated 17 bp DNA nanostructures. A PDMS well was stamped on the top of graphene and filled with 1× TM buffer. To measure the electrical-field-driven fluorescence quenching (Supplementary Note 2), a Ag/AgCl electrode was inserted into the solution to apply a Vlg, which generated an electrical field and drove Cy3 on 17bp-15T MolEMS downwards. The fluorescence intensities were imaged by using a confocal fluorescence microscope (C2+, Nikon)

Human kidney cells HEK293 and African green monkey kidney cell line Vero E6 (Cell bank of Chinese Academy of Sciences) were cultured at 37 °C with 5% CO2 in Dulbecco’s modified Eagle medium (Gibco) containing 2 mM l-glutamine, 50 U ml−1 penicillin, 100 mg ml−1 streptomycin and 10% (vol./vol.) foetal bovine serum (Gibco). SARS-CoV-2 strain nCoV-SH01 (GenBank, MT121215.1), from a laboratory-confirmed COVID-19 severe pneumonia case provided from CDC Shanghai, was propagated in Vero E6 cells in a biosafety level 3 laboratory

Sample 1 and sample 5 were extracted from African green monkey kidney cell line Vero E6 and human kidney cells HEK293, respectively, by using TRIzol reagent (Invitrogen) following the manufacturer’s instructions. Viral RNA or human RNA was used for synthesizing first-strand cDNA by SuperScript III CellsDirect cDNA Synthesis Kit (Invitrogen). Viral cDNA was then quantified by qRT–PCR (qRT–PCR detection kit Tiangen). For MolEMS g-FET testing, the vials containing 108 copies per μl viral cDNA were centrifuged at 3,000 rpm at 4 °C for 5 min and diluted into 5 × 107 copies per μl first in full artificial saliva (Solarbio) and 2% of RNase inhibitor (Thermo Fisher), then were serially diluted in full artificial saliva to concentrations of 5,000,000, 500,000, 50,000, 5,000, 500, 50, 5, 0.5, 0.1 and 0.02 copies per μl.

Sample 2, sample 3 and sample 4 were SARS-CoV-2, SARS-CoV and MERS-CoV IVT RNA reference materials (nt 13321–15540, GenBank no. MT027064.1, no. NC004718.3 and no. NC019843.3), respectively, with a titre of 105 copies per μl, provided by Shanghai Institute of Measurement and Testing Technology. For MolEMS g-FET testing, the samples were centrifuged at 3,000 rpm at 4 °C for 5 min and diluted into 5,000 copies per μl first in full artificial saliva (Solarbio) and 2% of RNase inhibitor (Thermo Fisher), and then were serially diluted in full artificial saliva to a concentration of 500, 50, 5, 0.5 and 0.1 copies per μl.

The clinical samples used were collected from Department of Laboratory Medicine, Shanghai Public Health Clinical Center. Thirty-three nasopharyngeal swab samples were from patients with qRT–PCR-positive COVID-19. Twenty-three nasopharyngeal swab samples were from qRT–PCR-negative patients in fever clinic. Four nasopharyngeal swab samples were from patients with influenza A. Two nasopharyngeal swab samples were from patients with influenza B. Other 25 nasopharyngeal swab samples were from healthy volunteers. To release the RNA, 200 μl VTM (Yocon) used to store the swab was heated at 56 °C for 30 min. Then, the medium was directly used for SARS-CoV-2 nucleic acid testing by MolEMS g-FETs without requirement of the extraction procedure. This research was approved by the Shanghai Public Health Clinical Center Ethics Committee (approval ID number 2020-Y114-01) with informed consent from participants.

Sample 1 was extracted by TRIzol reagent (Invitrogen) following the manufacturer’s instructions. Viral RNA was used for synthesizing first-strand cDNA by using SuperScript III CellsDirect cDNA Synthesis Kit (Invitrogen). To quantify viral cDNA in sample 1, qRT–PCR was performed in a 20 μl reaction (containing 2 μl cDNA sample) using qRT–PCR detection kit (Tiangen) on an MXP3000 cycler (Stratagene) as follows: 95 °C 5 min; 40 cycles of 95 °C 10 s, 50 °C 30 s and 72 °C 30 s. PCR primers (Genewiz) targeting SARS-CoV-2 RdRp gene (nt 1113–1326) were (forward/reverse) 5′-CTTGTGTATGCTGCTGACCC-3′/5′-GCAGCATTACCATCCTGAGC-3′. The copies of cDNA in sample 1 were calculated based on the standard curve (Supplementary Fig. 29).

To quantify viral RNA in clinical samples, 200 μl swab samples were extracted following instructions as described in the protocol (magnetic bead nucleic acid extraction kit, SDK60165) from Jiangsu Shuoshi Biological Technology. qRT–PCR was performed in a 25 μl reaction system by using 2019-nCoV nucleic acid detection kit (DaAn Gene). Five microlitres of the extracted nucleic acid elution sample was taken as the amplification template for amplification on Applied Biosystems 7500 (BJ001271, ABI), according to the protocol (2019-nCoV nucleic acid detection kit, DA0931). The copies of RNA in the clinical samples were calculated based on the standard curve (Supplementary Fig. 39).

The standard curves of the detection kits were established by using plasmid COVID-19 containing cloned target sequence as the standard. The stock of plasmid DNA was determined by spectrophotometric analysis. The mass of one copy DNA was calculated as:

where n is the plasmid size (bp) and m is the mass (g). A series of serial dilutions were performed to achieve a working stock of plasmid DNA. The standard curves for quantitation were obtained by measuring the serially diluted plasmid DNA (Supplementary Figs. 29 and 39).

Source–drain electrodes were wire bonded using Al wires. Then, 80 μl or 100 μl buffer solution (Supplementary Note 3) or biofluids were added into the PDMS well as electrolyte throughout the measurement, and a Ag/AgCl reference electrode was inserted into the solution as the liquid gate electrodeIds stabilized, Ids versus Vlg curves were obtained by sweeping Vlg in a certain range, and the Dirac point was obtained at Vlg where Ids reaches its minimum.

To measure targets by using MolEMS g-FETs, the device was first immersed in 80 μl or 100 μl buffer solution or biofluids, and then a stable Vds bias (Vds = 50 mV) was applied between the drain and source electrodes. After the Ids stabilized, time-resolved Ids measurement was performed. To add the testing solutions, 50% or 100% volume solution was taken out from the PDMS well and then was replaced by pre-mixed samples of the same volume. The solutions with a certain concentration of analytes (thrombin, casein, BSA, Hg2+, Fe3+, Cd2+, Zn2+, Ca2+, Cu2+, ATP, CTP, GTP, ss-DNA-T, ss-DNA-R1, ss-DNA-R2, ss-DNA-R3, ss-DNA-mis-3′, ss-DNA-mis-m, ss-DNA-mis-5′, SARS-CoV-2 cDNA, human cDNA, SARS-CoV-2 IVT RNA, SARS-CoV IVT RNA, MERS-CoV IVT RNA) or clinical samples (P1–P33, F1–F23, A1–A4, B1, B2, H1–H25) were added, and at the same time Ids versus t curves were recorded. To improve the sensitivity, negative Vlg (that is, −0.5 V) was applied to drive MolEMS during the measurements. As a comparison, Ids versus t curves were also measured at Vlg = 0 V. To conduct measurement in full serum, the MolEMS g-FET devices were immersed in 100% foetal bovine serum (Zhejiang Tianhang Biotechnology) for 2 h before usage.

To conduct measurements in full artificial saliva, the MolEMS g-FET devices were immersed in 100% artificial saliva (Solarbio) for 1 h before usage. To conduct measurements of the clinical samples, the MolEMS g-FET devices were immersed in VTM (Yocon) for ~1–2 h before usage. To conduct measurements in full serum, thrombin, ATP and ss-DNAs were spiked to 100% foetal bovine serum and diluted to different concentrations by adding full serum. To examine the long-term stability, the MolEMS g-FET devices were immersed in 100% foetal bovine serum for 15 days or in VTM for 3, 10, 15 and 21 days, and then electrical measurements were carried out under the same conditions as described above.

The threshold value to diagnose COVID-19 by MolEMS g-FETs was taken from three times the largest response of the negative samples. The diagnosis time was read from the interception of the threshold value and the real-time ∆Ids/Ids0 responses. As an example, most ∆Ids/Ids0 values of negative samples were below 0.08%; thus, the threshold value was set to 0.24%. The response time of P4, P7, P18, and P21 was therefore ~60 s, ~22 s, ~90 s and ~20 s according to the time when the ∆Ids/Ids0 response reached the threshold value (Extended Data Fig. 7), respectively.

For on-site and point-of-care detection, a prototype of the COVID-19 testing system was assembled by using MolEMS g-FETs (Supplementary Fig. 41). The testing system was composed of two parts. The first was a MolEMS g-FET-integrated testing module, which served as the consumable material in the test. To avoid cross interference between different samples, one module was used for testing one sample. The second was the main system with size of 11.5 cm × 9 cm × 5.5 cm, which included a controller, digital converters, a timer, a rechargeable lithium battery, a signal processing module, a signal amplifier regulator module and a signal output module that could connect with smartphone or computer by USB, WiFi or Bluetooth. The main system was reused for multiple tests by simply replacing the MolEMS testing module. The operation of this system took three steps (Supplementary Fig. 42). The first step was to connect the testing module with the main system and connect the computer or the smartphone with the testing system by USB, Wifi or Bluetooth. The second step was to open the seal which covered the PDMS well of the testing module. The third step was to add the testing sample into the PDMS well and to read the results from the computer or the smartphone.

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This present work underlines the effect of pH-shifting at pH 2 and pH 12 individually or combined with ultrasound treatment to modify the molecular structure of β-conglycinin (7S) on its emulsifying properties and stability. Fourier transform infrared (FTIR) spectroscopy and intrinsic fluorescence spectroscopy showed that pH-shifting improves the molecular structure of 7S, while ultrasound further promotes structural changes. In particular, the pH-shifting at pH 12 combined with ultrasound treatment (U-7S-12) resulted in more significant changes than the pH-shifting at pH 2 combined with ultrasound (U-7S-2). U-7S-12 showed a significant reduction in protein particle size from 152 to 34.77 nm and a relatively smooth protein surface compared to 7S. The protein had the highest surface hydrophobicity and flexibility at 81,560.0 and 0.45, respectively, and the free sulfhydryl content from 1.57 to 2.02 μmol/g. In addition, we characterized the emulsions prepared after 7S treatment. The single or combined treatment increased the interfacial protein adsorption of the samples, which showed lower viscosity and shear stress compared to 7S. The U-7S-12 emulsion exhibited the highest emulsifying properties and was more stable than other emulsions under creaming, heating, and freeze–thaw conditions. In summary, the concerted action of pH-shifting and ultrasound can modify the structure, and combined alkaline pH-shifting and ultrasound treatment can further improve the emulsifying properties and stability of 7S.

β-Conglycinin (7S), a glycoprotein, is one of the major components of soy protein isolate (SPI). 7S is a trimer that consists of three main subunits, α (68 kDa), α" (72 kDa), and β (52 kDa) [1]. The functional properties of 7S are closely related to its molecular structure, and its emulsifying properties change in response to structural changes [2]. However, the molecular structure of 7S is affected by the processing conditions or external environment, such as acidity, ionic strength, and temperature [3], affecting its emulsifying properties. This limits the further application of 7S as a natural emulsifier in foods such as ice cream and mayonnaise.

Nowadays, several techniques, such as hydrolysis, ultrasound, extreme pH treatment, and heating, are used to alter the conformation of 7S to enhance its emulsifying properties and expand its application [4], [5], [6], [7]. pH-shifting is a relatively simple method of chemically modifying proteins. Under extremely acidic or alkaline conditions, the structure of proteins partially unfolds. When the pH returns to neutral (7.0), the structure refolds but does not fully return to its natural conformation. This modified structure, somewhere between denatured and nondenatured, is called a “molten globule” [8]. Jiang et al. [9] found that the structure of SPI unfolds at acidic or alkaline pH and then refolds at neutral pH, thus improving the emulsification activity and emulsion stability of SPI. Chen et al. [10] demonstrated that secondary and tertiary conformations in whey isolate changed after pH-shifting, with improved protein emulsifying properties.

However, the improvement in protein emulsifying properties through pH-shifting is limited [11], therefore, there is a need to use other approaches to further modify proteins. Being a green, simple, and promising modification modality, ultrasound is widely used to change the functional properties of proteins [12]. Ultrasound cavitation–induced activity can undermine intramolecular protein bonds, altering the protein conformation [13]. Research has shown that the combined pH-shifting and ultrasound treatment can maximize the nutritional values of proteins, as unfolded proteins are more susceptible to ultrasound treatment under extreme pH conditions. Alavi et al. [14] showed that ultrasound combined with alkali treatment can alter the structure of faba bean protein, enhancing its foaming capacity and stability. Huang et al. [15] revealed that combined acidic pH-shifting and ultrasound treatment remarkably improves the emulsifying properties of SPI. Samaneh Pezeshk et al. [16] found that the combination of pH shifting and ultrasound treatment (400 W, 10 min) significantly improved the emulsification and foaming properties of proteins recovered from rainbow trout by-products compared with pH shifting alone. Yan et al. [17] found that the best functional properties were obtained for SPI treated with 400 W of ultrasound power. However, the effect of combined pH-shifting and ultrasound treatment on the structure and functional characteristics of 7S has been less studied.

Therefore, this paper aims to evaluate the effect of pH-shifting alone or combined with ultrasound on the structure and emulsifying properties as well as 7S stability. The findings could provide solutions for altering and modifying the structure of 7S, developing as a promising emulsifier to enhance its application in the food industry.

Defatted soybean powder was purchased from Yihai Kerry Cereals, Oils and Foodstuffs Co., ltd. (Harbin, China), corn oil from a store in Harbin (China), Nile red and Nile blue from Sigma-Aldrich (St. Louis, MO, USA), and trypsin (porcine pancreas, 2,500 u/mg) were supplied by Yuanye Bio-Technology Co., ltd. (Shanghai, China). All chemicals were purchased from supply shops.

7S was prepared, as described previously by Ren et al. [18]. Briefly, defatted soybean flour was dissolved in deionized (DI) water (1:10, w/v) and stirred with 2 mol/L NaOH to pH 7.5 for 1 h, then centrifuged at 10,000 g for 30 min. Dry sodium bisulfite (0.98 g/L) was added to the supernatant and the pH was adjusted to 6.4. The suspension was kept overnight at 4 °C and then centrifuged at 6500 g for 20 min. The NaCl (14.61 g/L) was added to the supernatant and adjusted to pH 5.0. The mixture was centrifuged at 10,000 g for 30 min, and the supernatant was diluted with DI (1:1, v/v), then adjusted to pH 4.8 and centrifuged at 6500 g for 20 min. The precipitates were washed three times with DI and redispersed, pH adjusted to 7.0. The solution was dialyzed at 4 °C for 48 h and then lyophilized for further use. The protein content of 7S measured by the Dumas combustion method was 93.32 ± 0.28 %.

The structure of 7S was altered, as described previously by Jiang et al. [19] with slight modifications. A 7S solution (10 mg/mL) was stirred at 21 °C for complete hydration and used as a control solution (labeled 7S). For pH-shifting alone, the pH of 7S solutions was adjusted to 2.0 or 12.0 using 2 mol/L HCl or NaOH, respectively, stirring for 1 h and then adjusting the pH to neutral. These samples were labeled 7S-2 and 7S-12, respectively. For ultrasound treatment alone, the ultrasound power and time were determined with slight modifications according to the method of et al [16] and Yan et.al [17]. The 7S solution was processed using a JY 92-IID ultrasound device (Xinzhi Biotechnology Co., ltd., Ningbo, China) at 400 W for 10 min and labeled U-7S. For combined pH-shifting and ultrasound treatment, the pH of 7S solutions was set to 2.0 or 12.0 and the solutions were stirred for 1 h and ultrasound at 400 W for 10 min; the samples were labeled U-7S-2 and U-7S-12.

The fluorescence spectra of the 7S samples were measured, as described previously by Tong et al. [20]. The protein content of all samples was diluted to a final protein concentration of 0.1 mg/mL. The excitation wavelength was 280 nm, the emission wavelength was 300–400 nm, the slit width was 5 nm, and a data interval of 0.5 nm.

The 7S samples were freeze-dried after different treatments and characterized using a Nicolet IS50 Fourier transform infrared (FTIR) spectrometer (Thermo Fisher, Madison, USA). Pure potassium bromide (KBr) was mixed with the samples and pressed into tablets. The scanning waveband range was set to 4000–400 cm−1, and the resolution was set to 4 cm−1 with 32 scans [21].

Before measurement, the sample concentration was adjusted to 1 mg/mL. The ζ-potential and particle size of different samples were analyzed using Zetasizer Nano ZS90 (Malvern, England), as described previously by Li et al. [22].

The free sulfhydryl (SH) content of 7S was measured, as described previously by Zhu et al. [23]. All samples were diluted to a concentration of 1 mg/mL using Tris-glycine solution (0.09 mol/L glycine, 0.086 mol/L Tris, 4 mmol/L ethylenediaminetetraacetic acid, and 8 mol/L urea). Next, 20 μL of 5,5′-dithiobis-bis-(2-nitrobenzoic acid) (Elman"s reagent, in Tris-glycine solution, 4 mg/mL) was added, and the reaction was carried out in the dark for 60 min. Absorbance was measured at 412 nm using the UV-2600 spectrophotometer (Shimadzu, Japan). The free sulfhydryl content was expressed as follows:

To determine the surface hydrophobicity (H0) of 7S, 50 μL of 1-anilinonaphthalene-8-sulfonic acid (ANS) (8.0 mmol/L) was added to 5 mL of 7S samples with different protein concentrations (0.001, 0.01, 0.05, 0.1, 0.15, and 0.20 mg/mL), and the reaction was carried out for 15 min protected from light. Fluorescence intensity was measured using the RF-6000 spectrofluorophotometer (Hitachi Co., ltd., Kyoto, Japan) with a slit width of 5 nm at an excitation wavelength of 390 nm and an emission wavelength of 470 nm. H0 was expressed as the slope of the protein concentration versus the fluorescence intensity curve [19].

Molecular flexibility (F0) was determined, as described previously by Zhu et al. [23]. The sample (1 mg/mL, w/v) was dispersed in Tris-HCl (0.05 mol/L) and mixed with trypsin at 16:1 (v/v). The reaction was terminated with 5 mg/mL trichloroacetic acid (TCA) after incubation at 37 °C for 5 min. After centrifugation at 5,000 × g for 30 min, the absorbance of the supernatant was measured at 280 nm to determine F0.

All 7S samples were lyophilized and then characterized using scanning electron microscopy (S-480 SEM, Hitachi High-Technologies Corporation, Japan) at 5000 × and 10,000 × magnifications and 15 kV accelerating voltage [24].

Oil-in-water (O/W) emulsions were obtained from corn oil and 1 % (w/v) 7S dispersion at 1:9 (v/v). Briefly, the corn oil and 7S mixture were homogenized at 10,000 rpm/min for 2 min (ULTRA-TURRAX, T18, IKA, Germany) and then homogenized thrice at 80 MPa to obtain an emulsion (Microfluidizer, M−110P, Microfluidics, Newton, Britain).

The microstructure of the emulsions was analyzed by ultrahigh-resolution microscopy (URM), as described previously by Bai et al. [25]. Briefly, 1 mL of each sample was stained by adding 40 μL of Nile blue and 40 μL of Nile red. After 30 min in the dark, 5 μL of the emulsion was placed on a coverslip, covered with a microscope slide, and sealed with nail polish to prevent flowing. Finally, the structure was observed at excitation wavelengths of 488 and 633 nm using OMX Delta Vision URM equipment (General Electric Co., Boston, MA, USA).

The adsorbed protein (AP) content of the emulsions was measured by diluting 8 mL of freshly prepared emulsion 10 times and centrifuging it at 25 °C at 10,000 × g for 30 min. The unabsorbed protein content was measured using the BCA method, as described previously Wang et al. [24]. The AP content (%) was calculated as follows:

The flocculation index (FI, %) of fresh emulsions was determined using Malvern MasterSizer 6000 (Malvern Instruments ltd, Malvern, Worcestershire, UK) and calculated as follows [26]:

Emulsion viscosity and shear stress were measured using the Discovery Hybrid (DHR)-1 rheometer (TA Instruments, New Castle, DE, USA). Using parallel plate clamps (60 mm) at 25 °C and a 500 µm distance between slits, the viscosity of the emulsions and the relationship between shear stress and shear rate for 0.1–100 s−1 were determined after equilibrating the samples for 10 min [27].

The emulsifying activity (EA) and emulsion stability (ES) of the emulsions were determined. Briefly, 50 μL of a freshly prepared emulsion was obtained in the bottom at 0 and 30 min, 5 mL of SDS (1 mg/mL, pH 7) was added, and the mixture was thoroughly mixed. Absorbance was measured at 500 nm. The absorption value A0 represents the EA, and the ES was calculated as follows [17]:

Fresh emulsions were placed in 10 mL sample bottles, heat-treated in a water bath at 90 °C for 30 min, and then cooled to 25 °C in an ice bath [28]. After storage at 4 °C overnight, the samples were analyzed for stability.

First, fresh emulsions were placed in sample bottles and frozen in a refrigerator at –20 °C for 24 h. Next, the samples were thawed and their particle size and macrostructure were analyzed as previously described by Palazolo et al. [29].

Fresh emulsions (8 mL) after different treatments were placed in 10 mL sample bottles. The emulsions were analyzed for creaming stability by measuring the height change in the serum bottom phase (HC) after 15 d of storage at room temperature [30]. The creaming index (CI) was determined as follows:

SPSS Statistics V17.0 (IBM Corporation, Armonk, NY, USA) was used to analyze the data and Origin 2019 (OriginLab Corporation, Northampton, MA, USA), where p ≤ 0.05 was considered statistically significant.

FTIR spectroscopy of the amide I band (1700–1600 cm−1) mostly from CCreated by potrace 1.16, written by Peter Selinger 2001-2019O stretching vibrations, the amide II band (1550–1500 cm−1) mostly from in-plane N—H bending, and the amide III band (1300–1200 cm−1) mainly from N—H bending and C—N stretching vibrations [31]. From Fig. 1A, the peak at 3285.14 cm−1 is linked to the stretching vibration of the intermolecular hydrogen bond between O—H and N—H. The change in the peak position indicates that ultrasound and pH shifting caused a change in the number of intramolecular hydrogen bonds [11]. The peak value of the amide I band moved from 1653.14 to 1668.18 cm−1 after combined pH-shifting and ultrasound treatment, indicating rearrangement of the protein–peptide chain or a change in the secondary structure, which altered the peak amide I band in 7S [32].

The content of the secondary structures of 7S was analyzed by the amide I band, as shown in Fig. 1B. The amide I band was composed of α-helix (1648–1664 cm−1), β-sheet (1615–1637 cm−1, 1680–1700 cm−1), β-turn (1664–1681 cm−1), and random coil (1637–1648 cm−1), as reported by Zhao et al. [33]. The α-helix of U-7S was transformed into random coils due to expansion of the α-helix area caused by ultrasonic cavitation, resulting in increased random coil content [34]. After combined pH-shifting and ultrasound treatment, all samples showed a decrease in the α-helix and β-sheet content and an increasingly disordered structure. The change in the secondary structure was more pronounced for U-7S-12 compared to U-7S-2, due to disruption of the α-helix of the protein during alkaline pH-shifting, where protein unfolding and rearrangement occurred and molecular flexibility increased. Instead, acidic pH Shifting occurred near the isoelectric point of the protein, forming soluble aggregates and inhibiting the unfolding of the protein structure. Instead, acidic pH-shifting occurred near the isoelectric point of the protein, forming soluble aggregates and inhibiting the unfolding of the protein structure. [35].

Intrinsic fluorescence spectroscopy is a method for characterizing tertiary conformation changes in proteins, where changes in the polarity of chromophores can cause changes in the endogenous fluorescence spectrum [36]. From Fig. 2A, the maximum emission wavelength (λmax) and fluorescence intensity of U-7S slightly increased compared to 7S. Ultrasound treatment of 7S alone probably exposed its hydrophobic groups outside the molecular chain [37]. In addition, 7S-12 showed significantly higher fluorescence intensity compared to 7S-2. This was associated with increased electrostatic repulsion of the protein under extreme alkaline conditions, with more hydrophobic groups exposed [38]. The λmax and fluorescence intensity of U-7S-12 significantly increased, indicating the strongest structural changes. These findings show that combined alkaline pH-shifting and ultrasound treatment can significantly alter the molecular structure of 7S.

Intrinsic fluorescence spectroscopy (A), particle size distr