Analysis of Seed Amino Acids in Vegetable Soybeans Dried by Freeze and Thermal Drying
Round 1
Reviewer 1 Report
The author used three different methods of drying the edamame seed to establish which methods would have a lesser effect on the amino acid content of the plant cultivar with high repeatability.
The introduction is informative, and it also outlines the knowledge gaps well. The methods section is well written, however, the authors should explain how NIR technology distinguishes between individual amino acids (because this study focuses on method development).
The results are analysed well and presented logically. The discussion demonstrates a deeper understanding of the work by referencing relevant studies. The conclusions were supported by the findings of the study.
Author Response
Thank you so much for your positive comments and helpful suggestions. An explanation has been added on how NIR method distinguishes between individual amino acids in the Materials and Methods section. As descripted by the manufacturer (Perten Instruments, Inc.), all individual amino acids were analyzed simultaneously in the machine but separately based on different calibrations each established for a specific amino acid. As the accuracy of NIR method is dependent on calibrations as well as samples types, three sets of calibrations, referred to as whole seed, ground and combined calibration, were installed in the DA 7250 NIR analyzer to analyze different types of samples. The combined calibrations are applicable to both whole seed and ground samples. In addition, the calibrations have been updated periodically by the manufacturer to improve the prediction accuracy.
Reviewer 2 Report
1. In the introduction, as we know that vegetable soybean is more popular in Asian, especially in China and Japan. As author mention in the manuscript, vegetable soybean, known as edamame in Japan and maodou in China. However, most of research references are from western country, less references related to China or Japan. It is better to introduce some research progress in China or Japan.
2. As we know that the precise of NIRs method is depending on calibration and sample types. In the materials and methods, seed amino acid analysis, two set of data were generated including powder and whole seed. Which kinds of amino acid calibration formula were used each other? For the whole seed, the shape of dry vegetable soybean will be changed significantly comparing to mature soybean seed. How to avoid the different when you used the same calibration formula? As we know that powder maybe can avoid these kind of problem, however, in the later data analysis authors only used the data from the whole seed although they are significant different both calibrations of powder and whole seed. We know that whole seed method is simple and easy to conduct in this study. As scientific data, their accuracy is more important than convenience.
3. In soybean seed amino acid components, some of components are less content in soybean seeds, such as cysteine and methionine, which are more important for soybean amino acid component. How to avoid the large variation or SD based on NIRs method in this study?
4. In order to validate the repeatability of amino acid determination in soybean seed by NIRs method, it is better to analyze amino acid content based on both NIRs and analytical chemical methods to clarify NIRs method is not significant different with analytical chemical methods.
Author Response
- In the introduction, as we know that vegetable soybean is more popular in Asian, especially in China and Japan. As author mention in the manuscript, vegetable soybean, known as edamame in Japan and maodou in China. However, most of research references are from western country, less references related to China or Japan. It is better to introduce some research progress in China or Japan.
Response: It is a great idea. We appreciate your suggestion and have added such information and provided related references.
- As we know that the precise of NIRs method is depending on calibration and sample types. In the materials and methods, seed amino acid analysis, two set of data were generated including powder and whole seed. Which kinds of amino acid calibration formula were used each other? For the whole seed, the shape of dry vegetable soybean will be changed significantly comparing to mature soybean seed. How to avoid the different when you used the same calibration formula? As we know that powder maybe can avoid these kind of problem, however, in the later data analysis authors only used the data from the whole seed although they are significant different both calibrations of powder and whole seed. We know that whole seed method is simple and easy to conduct in this study. As scientific data, their accuracy is more important than convenience.
Response: Thank you for your comments. To address your concerns, we have provided some explanations on the calibrations and uses. We also added the data of ground samples to Table 5 and 6 for full information and a better comparison. Considering page issues, we did not add the data of ground samples to Table 7 directly. Instead, we provided a Supplementary Material, including three tables to present the results of ground samples under low-heat drying (Table S1) and freeze drying (Table S2), as well as the data of whole seed samples under freeze drying (Table S3). Some text changes or additions have been made accordingly.
- In soybean seed amino acid components, some of components are less content in soybean seeds, such as cysteine and methionine, which are more important for soybean amino acid component. How to avoid the large variation or SD based on NIRs method in this study?
Response: Thanks for your comments again. The related information and discussion have been added as suggested.
- In order to validate the repeatability of amino acid determination in soybean seed by NIRs method, it is better to analyze amino acid content based on both NIRs and analytical chemical methods to clarify NIRs method is not significant different with analytical chemical methods.
Response: Yes, analytical chemical method is a standard to evaluate seed composition. As discussed in the manuscript, the best way is to evaluate the fresh samples directly using traditional chemical method. But it’s almost impossible to timely treat many samples in this way, in addition to other issues. NIR is an easy and cost-effective method, but the concern is the calibrations used. To date, no calibrations has been established specifically for fresh edamame or immature soybean seed. As freeze drying does not affect nutritional composition in fresh samples and thus it was used as the reference in the study. Considering the impact of the shape of dried seed, as you mentioned above and discussed in the text, we also ground the seeds and used the powder to analyze. The results indicated that it was appropriate. Of course, it’s better to include analytical chemical data also. Unfortunately, we could not provide such data at this point because it would take a lot of addition time to achieve. Actually, it is an objective of our on-going projects, in which at least several hundreds of samples will be analyzed using both traditional chemical and NIR methods. We expect that the results will be available in few years. Once again, thank you very much for your time and advice.
Reviewer 3 Report
Line 28: All soybeans are “consumable”. Specify what type of consumption you are referring to.
Line 28: If the crop is “sold” then it is “commercial”, be specific about what you are talking about.
The text repeatedly says “The Author” did this and that. There are 3 listed authors. Does this mean that two did nothing? I think you should instead say “The authors” did this and that. It would be better to reword those sentences to the 3rd person.
Minor changes suggested in attached pdf file.
Recommendation: Accept with minor spelling and grammar changes
Comments for author File: 
Comments.pdf
Author Response
We sincerely appreciate your comments and suggestions.
Line 28: All soybeans are “consumable”. Specify what type of consumption you are referring to.
Response: A change has been made. Thanks.
Line 28: If the crop is “sold” then it is “commercial”, be specific about what you are talking about.
Response: Thank you. A change has been made.
The text repeatedly says “The Author” did this and that. There are 3 listed authors. Does this mean that two did nothing? I think you should instead say “The authors” did this and that. It would be better to reword those sentences to the 3rd person.
Response: Thank you for your advice. Yes, all the words “author” have been changed to “authors”.
Minor changes suggested in attached pdf file.
Recommendation: Accept with minor spelling and grammar changes
peer-review-26708749.v2.pdf
Response: Thank you very much for your comments and suggestions.
Minor points copied from the suggestions marked in manuscript:
Line 34, change have to has.
Response: Yes, changed.
Line 37, change values to value.
Yes, Changed.
Line 50, change composition in to component in soybean seeds.
Yes, changed.
Line 55, change labor-input to labor intensive.
Yes, changed as suggested.
Line 57, change breeding materials to crosses.
Yes, change it to breeding lines.
Line 70-72, Run on sentence?
Thanks for the comment. The sentence has been rewritten.
Line 78, change quality to protein quantitation.
Yes, changes.
Line 86-88, Run on sentence?
The sentence has been rewritten. Thank you.
Line 102, At what rate?
The information has been added as suggested. Thank you.
Line 193-195, Run on sentence? Very strange construction.
Thank you for your comment. We have rewritten the sentence.
Reviewer 4 Report
The research is very good, and it is useful for large-scale evaluation of amino acids in vegetable soybean seeds.
Some minor revisions are suggested as followed:
Line155：P should be italic
Line240：change“average”to“averaged”
The conclusions should be concise.
Author Response
The research is very good, and it is useful for large-scale evaluation of amino acids in vegetable soybean seeds.
Response: Thank you so much for your positive comments. We appreciate your time and suggestions.
Some minor revisions are suggested as followed:
Line155：P should be italic
Yes, changed.
Line240：change“average”to“averaged”
Yes, changed.
The conclusions should be concise.
Yes, modifications have been made to the conclusions as suggested. Once again, thank you.
