Influence of pH and Heat Treatment on the Physicochemical, Interfacial, and Emulsifying Properties of Hemp Seed Protein Dispersions

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This manuscript investigates the effects of pH and heat treatment on the physicochemical, interfacial, and emulsifying properties of hemp seed protein isolate (HPI) dispersions. The research topic is highly relevant to food colloid science and industrial applications of plant-based emulsifiers, which aligns well with the scope of Foods. Comprehensive experiments (including diffusion kinetics, interfacial rheology, CLSM observation, and emulsion stability evaluation) are carried to investigate the emulsifying properties of HPI and HHPI. However, several critical issues regarding experimental details, data interpretation, and academic rigor need to be addressed to meet the publication standards of Foods.

• The rationale for selecting pH 2, 7, and 10 is not adequately explained, what is the scientific basis for specific pH value (2, 7, 10) were investigated in this work. Are these conditions representative of actual food processing environments?
• This study states that both pH and thermal treatment were employed in the treatment of HPI, but it lacks a clear, concise summary of the key findings specifically related to thermal treatment in abstract.
• Language Error (Line 80): The phrase "remain...still" is redundant.
• Surface Hydrophobicity (Line 149): The method describes diluting protein suspensions with Milli-Q water prior to surface hydrophobicity measurement. Will this dilution change the actual pH value of the sample and causing a deviation in the test results?
• The statement that heating did not affect protein solubility is highly unusual and contradicts well-established principles of protein chemistry. Thermal treatment typically causes denaturation, aggregation, and changes in solubility. The authors must provide robust evidence and cite relevant literature to support this exceptional claim. A more thorough discussion of the underlying mechanisms is required.
• Figure 2 shown that HPI7 and HHPI7 has no obvious difference in proportion of aggregated β-sheet, yet the particle size is reported to increase significantly in Figure 3 and Line 550-551. This apparent contradiction needs to be resolved.
• The conclusion drawn in Line 375-380 appears to be speculative rather than data-driven. It requires stronger support from cited literature.
• Reference 30 does not list a "Chang" author (line 372), all reference should be carefully revised.
• Line 381-382, comparing only the equilibrium interface pressure is insufficient and scientifically misleading. As can be seen from Figure 4, HHPI2 and HHPI7 reach the equilibrium interfacial pressure more quickly than that of HPI2 and HPI7.
• The quality of microscopy images provided in Figure 8 is inadequate. The oil droplets are not clearly visible, making it impossible to reliably identify or discuss protein shells around them.
• The legend for Figure 8 is insufficient. It must explicitly state which color/shade corresponds to the oil phase and which corresponds to the protein phase to allow for correct interpretation by the reader.

Author Response

Please find our reply in the following word document attached here

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Check line 61

Line 63 – “previous studies”, however cite only one.

Mention the replicate of each test

It is not so clear the code of each sample

Lines 319, 345, 413, 440, 442, 514, 519  cite the authors

References should follow the journal indications

Please, note some troubles in references citation along the text (mark in red)

The samples produced should be presented with their respective codes in a table in M&M session

Comments for author File: Comments.pdf

Comments on the Quality of English Language

No comments

Author Response

Please find our reply in the following attached word document

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Please carefully revise the manuscript, paying particular attention to ensuring that all references are properly cited.

Line 344 and Line 766: Citation "Wang" does not match reference 36

Line 196 and Line 731: Citation "Kramer" does not match reference 21

Line 481 and Line 787: Citation "Zhang" does not match reference 45

Line 478 and Line 785: Citation "Yu" does not match reference 44

……

Author Response

We agree with the reviewer that a careful revise of the manuscript was needed to ensure that all the references were cited correctly. We then double-checked all the references cited and adopted in the manuscript as well as its bibliography. We corrected all of the mistakes of the previous version in this final one. We also improved the academic english level of the overall manuscript in order to be clear for the reader.