Comparison of Different Methods for Determining 5-hydroxymethylfurfural in Assessing the Quality of Honey from Greater Poland

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Review on manuscript:

Comparative analysis of methods for determining HMF in honey from Greater Poland in order to create more sustainable consumption and production patterns

 

by Joanna Zembrzuska, , Łukasz Pakulski, Bożena Karbowska, Jarosław Bartoszewicz, Edyta Janeba-Bartoszewicz, Jarosław Selech and Przemysław Kurczewski

 

submitted to Sustainability

 

In the manuscript submitted for review, the authors provide the results of the determination of HMF content in seven honey samples performed using three methods. The purpose of the research undertaken by the authors is unclear. If the authors wanted to compare three different determination methods, they should use standard statistical tests that can be used to demonstrate, for example, that the results obtained (mean, SD) are different or not different. On the other hand, if the aim of the study was to monitor the occurrence of HMF in honey from a given area, seven honeys definitely do not constitute a representative sample.

 

Additional recommendations:

The title is unclear, what is the correlation between HMF determination methods and production and consumption patterns?

Abstract – should contain results in the form of numerical data,

Introduction – should be shortened,

Figure 1 – this structure is well known so it is not needed,

lines 155-164 – the purpose of the research should be clarified, the mere determination of HMF in honey samples cannot be a scientific goal,

lines 167-178 – this part of the work should be significantly shortened,

line 182 – seven honeys is not a representative sample for such an area,

lines 187-188 – what does it mean that the authors know the production process?

lines 210-224 – such a detailed description of the preparation of reagents is not needed,

line 295 – what is the purpose of the Ishikawa diagram shown,

line 318 – for the remaining two methods, the authors should provide LOD and LOQ,

line 327 – the qualitative analysis section does not contribute anything important, the same results from table 5,

line 395 – why suddenly four methods?

line 416-424 – these are not conclusions that result from the authors' research.

Author Response

Reviewer# 1

First of all, the authors would like to thank reviewer for his/her apt comments on their manuscript. A detailed, point-by-point response to each comment is follows:

1. In the manuscript submitted for review, the authors provide the results of the determination of HMF content in seven honey samples performed using three methods. The purpose of the research undertaken by the authors is unclear. If the authors wanted to compare three different determination methods, they should use standard statistical tests that can be used to demonstrate, for example, that the results obtained (mean, SD) are different or not different. On the other hand, if the aim of the study was to monitor the occurrence of HMF in honey from a given area, seven honeys definitely do not constitute a representative sample.

The aim of the research was to compare three research methods. All methods are intended to determine the HMF content in honey. Each method has different sensitivity of HMF detection, which is an indicator of honey quality in the article. The results depend on the sensitivity of the method, its accuracy or repeatability, the presence of HMF derivatives or the presence of compounds that facilitate the ionization of HMF in the ion source. Individual measurement methods should be selected for different types of honey. This is the main cognitive effect.

2. The title is unclear, what is the correlation between HMF determination methods and production and consumption patterns?

The title of the article is formulated in general terms and partly covers a fragment of the area of sustainable honey processing.

3. Abstract – should contain results in the form of numerical data,
4. The determined HMF values in all honey samples ranged from 0 to 28.5 mg HMF/kg honey.
5. Introduction – should be shortened,

The introduction has been shortened as suggested by most reviewers

5. Figure 1 – this structure is well known so it is not needed

We agree with the reviewer. We have removed Figure 1.

6. lines 155-164 – the purpose of the research should be clarified, the mere determination of HMF in honey samples cannot be a scientific goal

Sentences added:

The presented research aims to improve the state of knowledge in this area by comparing research methods and indicating the scope of their use.

7. lines 167-178 – this part of the work should be significantly shortened.

Thank you for your attention. The fragment has been shortened.

8. line 182 – seven honeys is not a representative sample for such an area

In the case of the tested samples, the authors were only interested in their diverse characteristics and not in a full description of the honeys available in the region.

9. lines 187-188 – what does it mean that the authors know the production process?

The fragment was removed as irrelevant to the article.

10. lines 210-224 – such a detailed description of the preparation of reagents is not needed,

The authors accept the suggestion, but found it appropriate to leave it in view of the possibility of repeating the experiment in other conditions.

11. line 295 – what is the purpose of the Ishikawa diagram shown,

The Ishikawa diagram assesses the error with which we determine the HMF content in honey using the LC-MS/MS technique

12. line 318 – for the remaining two methods, the authors should provide LOD and LOQ,

White metod: LOQ = 4 µg/g, Winkler method LOQ = 2 µg/g

13. line 327 – the qualitative analysis section does not contribute anything important, the same results from table 5,

Subsections 3.2 and 3.3 have been combined. Their division brings nothing except the conviction of two cases whose results were omitted.

14. line 395 – why suddenly four methods?

Thank you for your attention. We improved.

15. line 416-424 – these are not conclusions that result from the authors' research.

That's true. This is a general summary and will be deleted.

The authors would like to thank you very much for all your comments.

Reviewer 2 Report

Comments and Suggestions for Authors

I am very grateful you for the invitation to review the manuscript sustainability-2977723 by Teixeira and coauthors "Comparative analysis of methods for determining HMF in honey from Greater Poland in order to create more sustainable consumption and production patterns”. The aim of the research was to determine the HMF content in Greater Poland honey available on the Polish market using three different methods and to compare the obtained results. The work is interesting but needs several adjustments to increase the quality of the material.

 

Comments:

- Abstract: Please indicate a better step-by-step about the work.

- Introduction: Authors must include appropriate references in the introduction. There are many paragraphs without any citation.  

- Line 58-59: better specify contamination aspects.

- Introduction: Include information about the honey market in the country.

- Line 78: Change “of honey.. 5-hy-” to “of honey. 5-hy-’.

- Line 180: Review the title of Figure 2.

- Standardize the use of units throughout the text (ml / mL, etc).

- Line 281: Review the writing throughout the text. There are many cut sentences, like “Determination of the linearity range”.

- Line 369 and throughout the text: Observe the correct form of citation.

- Title, especially “in order to create more sustainable 3

consumption and production patterns” have no relation to the study.

- Results and discussion: It should be reviewed, since there is no discussion regarding the results of the different types of honey analyzed and the impacts on quality.

- Introduction: It must be revised and condensed, as there is excess information. Part of the information should be used in discussing the results.

- Discussion: Discussion should be better presented to avoid appearing merely as an experimental script."

- Title: must be reviewed and directed more precisely to what was done in the work.

- Conclusion: Direct only to what was proposed as an objective. The sentence contained in lines 394-399 can be removed.

 

- Lines 416-424: Rewrite appropriately, as commonly presented in conclusion items.

Author Response

Reviewer# 2

First of all, the authors would like to thank reviewer for his/her apt comments on their manuscript. A detailed, point-by-point response to each comment is follows:

1. Abstract: Please indicate a better step-by-step about the work.

he abstract has been supplemented with the tasks performed.

2. Introduction: Authors must include appropriate references in the introduction. There are many paragraphs without any citation.

As suggested by other reviewers, the abstract was shortened to include redundant elements and those without detailed citations.

3. Line 58-59: better specify contamination aspects.

At the request of another reviewer, this fragment was removed from the article, perhaps to its detriment. The absence of this fragment does not result in very significant consequences for the article, although it may be worth mentioning the types of contaminants often to make them more permanent.

4. Introduction: Include information about the honey market in the country.

We have added maxims in chapter 2.2:

According to data from 2022 published by The Food and Agriculture Organization (FAO), honey production in Poland in 2021 amounted to 18.4 thousand. tons and was higher by approx. 5.8 thousand tons than in 2020. In Poland, the most expensive and rarest are heather and coniferous honeydew honeys, and the cheapest and most popular are multifloral and rapeseed honeys. Greater Poland is the second out of sixteen honey producers in Poland with an amount of 1.97 thousand. tons in 2021.

5. Line 78: Change “of honey.. 5-hy-” to “of honey. 5-hy-’.

Thank you for your attention. We have made changes

6. Line 180: Review the title of Figure 2.

Thank you very much, the title is now correct.

7. Standardize the use of units throughout the text (ml / mL, etc).

Thank you for your observation, the symbols have been reviewed and corrected. However, the White and Winkler methods refer to grams or kilograms because they are mass methods.

8. Line 281: Review the writing throughout the text. There are many cut sentences, like “Determination of the linearity range”.

Thank you for your attention. We have made changes

9. Line 369 and throughout the text: Observe the correct form of citation.

Thank you for your attention. We have made changes

10. Title, especially “in order to create more sustainable consumption and production patterns” have no relation to the study.

The title has been changed.

11. Results and discussion: It should be reviewed, since there is no discussion regarding the results of the different types of honey analyzed and the impacts on quality.

According to the European Council standard, the obtained results of HMF content range up to 40 mg/kg. This means that they meet the normative requirements, but not all methods gave consistent results.

12. Introduction: It must be revised and condensed, as there is excess information. Part of the information should be used in discussing the results.

The introduction has been shortened.

Discussion: Discussion should be better presented to avoid appearing merely as an experimental script."

13. Title: must be reviewed and directed more precisely to what was done in the work.

The title has been changed.

14. Conclusion: Direct only to what was proposed as an objective. The sentence contained in lines 394-399 can be removed.

In the case of this paragraph, the reviewers' opinions are divided, so after a minor correction we left it in the article.

15. Lines 416-424: Rewrite appropriately, as commonly presented in conclusion items.

At the request of another reviewer, this fragment was removed.

The authors would like to thank you very much for all your comments.

Reviewer 3 Report

Comments and Suggestions for Authors

The authors compared the measuring results of HMF in the honey produced in Great Poland by two spectrophotometric method and LC-MS/MS technique. This work is useful and supplies some guidelines for the human health. However, it is failed to clear explain the novelty or contribution to the technology improvement. I suggest a major revision and give some comments below:

1.       Introduction section should be re-written, the words should be reduced to pay more attentions on the measuring importance for HMF, as well as the related measuring methods and techniques; more research papers should be reviewed and compared to illustrate the contribution of this work;

2.       In section 2 materials and methods section: the research region should be removed in my opinion;

3.       In addition to the tables, the measuring or experimental results should be shown by some curves, which will help the readers to overview the results easier;

4.       Whether the different storage conditions, storage times, etc. will affect the concentration of HMF in honey needs to be discussed and analyzed.

Author Response

Reviewer# 3

First of all, the authors would like to thank reviewer for his/her apt comments on their manuscript. A detailed, point-by-point response to each comment is follows:

1. Introduction section should be re-written, the words should be reduced to pay more attentions on the measuring importance for HMF, as well as the related measuring methods and techniques; more research papers should be reviewed and compared to illustrate the contribution of this work;

The introduction has been edited in line with the suggestions of all four reviewers.

2. In section 2 materials and methods section: the research region should be removed in my opinion;

Section 2 has been shortened as suggested by two reviewers.

3. In addition to the tables, the measuring or experimental results should be shown by some curves, which will help the readers to overview the results easier;

As suggested, the most important data from Table 5 are included in an additional chart.

4. Whether the different storage conditions, storage times, etc. will affect the concentration of HMF in honey needs to be discussed and analyzed.

It is common knowledge that storage time and storage conditions influence the concentration of HMF in honey. This is already well documented in the literature, e.g.

Tichonow, A.I.; Bondarenko, L.A.; Jarnych, T.G.; Szpyczak, O.S.; Kowal, W.M.; Skrypnik-Tichonow, R.I. Natural honey in medicine and pharmacy. Origin, properties, application, medicinal preparations. Sadecki Bartnik. 2017, ISBN: 978-83-61904-13-7;

Gokmen, V.; Acar, O.C.; Serpne, A.; Morales, A.F.J. Effect of leaving agents and sugars on the formation of hydroxymethylfurfural in cookies during baking. European Food Research and Technology. 2008, 226, 1031. https://doi.org/10.1007/s00217-007-0628-6

Turhan, K. Effects of thermal treatment and storage on hydroxymethylfurfural (hmf) content and diastase activity of honeys collected from middle Anatolia on Turkey. Innovations in Chemical Biology. 2009, 23, 233. https://doi.org/10.1007/978-1-4020-6955-0_23

Mironescu, M.; Fratila, L.; Hupert, A.; Mironescu, I.D. Obtaining and Characterization of Starch-Based Edible Films Incorporating Honey, Propolis and Bee Bread. Acta Universitatis Cibiniensis. 2019, 23, 193. https://sciendo.com/article/10.2478/aucft-2019-0023

Thank you for this attention. Determining the effect of storage conditions was not the purpose of this study. We shortly described this problem in the introduction.

The authors would like to thank you very much for all your comments.

Reviewer 4 Report

Comments and Suggestions for Authors

Major revision

1.     In samples collection, authors have to describe what kind of honey samples were collected in detail because different honey is difference in producing HMF. It not clear that the authors described honeydew honey from forest areas.

2.     The authors also need to describe the procedure of LC-MS/MS such as designing the measures.

Author Response

Reviewer# 4

First of all, the authors would like to thank reviewer for his/her apt comments on their manuscript. A detailed, point-by-point response to each comment is follows:

1. In samples collection, authors have to describe what kind of honey samples were collected in detail because different honey is difference in producing HMF. It not clear that the authors described honeydew honey from forest areas.

The honey samples collected for analysis concerned seven different types of honey obtained in Greater Poland. In Greater Poland, apiaries in forests without access to meadows, arable fields and orchards are very rare. Therefore, saffron and multi-fruit honeys predominate. In the case of honeydew honey, food companies do not have clean substrates from coniferous, deciduous and other forests. The producer meets national requirements regarding the classification of these honeys as honeydew honey. However, scientific honesty leads us to claim that the tested honeydew honey is mixed honeydew honey.

2. The authors also need to describe the procedure of LC-MS/MS such as designing the measures.

LC-MS/MS procedure is described in the section "Apparatus and reagents" - what equipment was used and in the section "The LC-MS/MS method" - conditions of the separation and determination process.

The authors would like to thank you very much for all your comments.

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

In my opinion, the authors responded appropriately to my comments and the manuscript was improved, but it is a pity that the authors did not use more advanced statistical tests to compare the analytical methods used.

Reviewer 2 Report

Comments and Suggestions for Authors

I confirm that the authors have made all the necessary corrections according to the reviewers' instructions and that this version of the paper has been improved in relation to the previous one.

Reviewer 3 Report

Comments and Suggestions for Authors

The authors have replied the comments and improved the manuscript. I recommend its acceptance now.

Reviewer 4 Report

Comments and Suggestions for Authors

No further comments are given.