Review Reports - Valorisation of the Halophyte Cakile maritima as a Food Resource for Human Consumption

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript presents a comprehensive characterisation of the nutritional profile of the facultative halophyte Cakile maritima across several plant organs (stems, leaves, immature fruits, roots and flowers). The study includes proximate composition, mineral analysis (ICP-OES, with sodium determined by flame photometry), vitamin C, total antioxidant capacity (DPPH) and total polyphenol content. The comparison of two leaf morphotypes is also interesting, and the analytical part is usefully complemented by an online consumer survey (n = 147) and a sensory evaluation of 11 dishes (n = 31). Overall, the topic is relevant and the combination of compositional, physiological and consumer-related data is one of the strengths of the manuscript.

Before the manuscript can be accepted, however, several numerical inconsistencies and a few errors in the presentation of the results should be clarified. Most of these issues appear to be readily correctable, but some concern the interpretation or consistency of the reported data. My comments below refer only to points that can be checked directly against the submitted text, tables and figures.

A. Data and internal consistency

1. Phosphorus content in roots (Table 1; line 292)
The phosphorus value reported for roots (4710.47 mg/100 g dw, corresponding to approximately 4.71% dw) does not appear to be compatible with the reported ash content of 7.4% dw. When expressed as P₂O₅, 4.71% P corresponds to roughly 10.8% dw, which would already exceed the total ash fraction. Since phosphorus should remain in the mineral residue after ashing, these two values cannot both be correct as currently reported. The authors should therefore re-check the phosphorus result, including the dilution factor, units and calibration, as well as the ash value. It would also be worth checking the entire phosphorus column, since the reported concentrations appear rather high for plant tissues.

2. Sodium comparisons in the text do not correspond to Table 1 (lines 299–301)
The manuscript states that sodium in leaves (680.81 mg/100 g dw) is approximately three times higher than in flowers and four times higher than in roots. According to the values in Table 1, however, the ratios are approximately 5.0 for leaves/flowers (680.81/136.69) and 3.0 for leaves/roots (680.81/229.9). The comparisons with stems (about 1.8-fold) and fruits (about 3.8-fold) are consistent with the table. It therefore seems that the comparisons for flowers and roots have simply been interchanged.

3. Mercury significance (Table 1; line 305)
The text indicates that Mn, Mo, Se and Hg did not differ significantly among plant organs. This is not consistent with Table 1, where the Hg values are assigned different significance letters: stems are labelled “a” (0.04), whereas the other organs are labelled “b” (0.02–0.03). The statement in the text or the lettering in the table should be checked and corrected accordingly.

4. Figure reference for protein data (lines 327–330)
The paragraph discussing total protein refers to “Figure 3D”. Protein data are shown in Figure 2D, whereas Figure 3 contains TAC, vitamin C and polyphenol results. The figure references in this paragraph should therefore be Figure 2D–F.

5. Interpretation of Table 4
The meaning of the three rows in Table 4 is not sufficiently clear. For the bitter attribute, the table gives Detected = 158, Negative = 152 and Positive = 103. The Positive and Negative values correspond to the totals obtained from Figure 5 (103 and 152), but their sum is 255 rather than 158. This suggests that “Detected” is calculated using a different approach, possibly as the number of detections rather than an intensity-weighted score. If so, this needs to be defined explicitly in the Methods or in the table legend. At present, the three categories cannot be interpreted unambiguously.

6. Bitterness of dish 10 (lines 476–478 and 488–489; Figure 5J)
There is a contradiction in the description of bitterness. The manuscript first states that bitterness was detected in all dishes except dishes 4 and 10. Later, however, bitterness in dish 10 is described as predominantly positive (15 vs 1), which agrees with Figure 5J, where bitterness is clearly reported for this dish. The first statement should therefore be revised.

7. Hedonic scale (line 268 vs line 513)
The scale is described differently in the Materials and Methods and Results sections. Line 268 states that responses were converted to numerical values from 0 to 7, whereas line 513 refers to a hedonic scale ranging from 1 to 7. The correct range should be stated consistently throughout the manuscript.

8. Cadmium and arsenic
The list of analytical wavelengths in lines 182–185 includes P, Hg, Se, Zn, Fe, B, Mn, Mo, Mg, Ca, Cu, Pb and K, with Na measured separately by flame photometry. Cd and As do not appear to have been determined. Considering that the manuscript discusses C. maritima as a potential edible plant, the absence of data for these two elements should at least be mentioned as a limitation of the study. If samples and analytical capacity are still available, their determination would further strengthen the safety assessment.

B. Editorial, referencing and typographical issues

9. Figure 4 legend (lines 463–464)
The abbreviation “P” is used for both “price” and “pests”. Different abbreviations should be used to avoid confusion.

10. Chef’s name
The name is written as “Paco Brocal Rubio” at line 246 and “Paco Borcal Rubio” in the Acknowledgments (line 736). The correct spelling should be used consistently.

11. Reference 25
The author “Santos, Fernanda F.” appears four times consecutively in the author list. The reference should be checked against the original publication.

12. Language and typographical corrections
There are several minor language errors that should be corrected. Examples include “cricifers” → “crucifers” (reference 15), “significatively” → “significantly” (lines 378, 389 and 390), “stablished” → “established” (line 586), and the phrase “obtained for to stems” (lines 342–343). A careful language revision of the full manuscript would be advisable.

Author Response

Reviewer 1

Data and internal consistency

Phosphorus content in roots (Table 1; line 292)
The phosphorus value reported for roots (4710.47 mg/100 g dw, corresponding to approximately 4.71% dw) does not appear to be compatible with the reported ash content of 7.4% dw. When expressed as P₂O₅, 4.71% P corresponds to roughly 10.8% dw, which would already exceed the total ash fraction. Since phosphorus should remain in the mineral residue after ashing, these two values cannot both be correct as currently reported. The authors should therefore re-check the phosphorus result, including the dilution factor, units and calibration, as well as the ash value. It would also be worth checking the entire phosphorus column, since the reported concentrations appear rather high for plant tissues.

Thank you very much for this important comment. During the revision of the manuscript, we carefully rechecked the raw data and calculations and identified an error in the arrangement of the mineral data presented in the original table. With the exception of sodium, the values corresponding to flowers and roots had been inadvertently interchanged. The same error was identified in Supplementary Table 1 for the data expressed on a fresh-weight basis, and Supplementary Table 2 for the data corresponding to different leaf morphotypes. We have corrected the three tables, and we highlighted these changes for clarification in the NO track changes version of the MS. This issue, however, does not solve the incompatibility of phosphorous concentration and ash content that you noticed.

We checked raw data, dilutions and calculations, and everything is ok, therefore it is highly likely that spectral overlap and interferences are occurring at the phosphorus measurement wavelength. Consequently, phosphorus measurements for all samples have been repeated using spectrophotometric techniques, and this methodology is indicated in the M&M section of the corrected MS (lines 198-201). The values have been adjusted based on the new methodology. Nevertheless, phosphorus levels remain noticeable higher than previous reports in other species, including halophytes, but lower than previously determined by ICP. We have therefore included a paragraph in the discussion section of the revised manuscript (lines 658-670).

Sodium comparisons in the text do not correspond to Table 1 (lines 299–301)
The manuscript states that sodium in leaves (680.81 mg/100 g dw) is approximately three times higher than in flowers and four times higher than in roots. According to the values in Table 1, however, the ratios are approximately 5.0 for leaves/flowers (680.81/136.69) and 3.0 for leaves/roots (680.81/229.9). The comparisons with stems (about 1.8-fold) and fruits (about 3.8-fold) are consistent with the table. It therefore seems that the comparisons for flowers and roots have simply been interchanged.

Thanks for this observation. We made corresponding correction in the revised MS (lines 333-335).

Mercury significance (Table 1; line 305)
The text indicates that Mn, Mo, Se and Hg did not differ significantly among plant organs. This is not consistent with Table 1, where the Hg values are assigned different significance letters: stems are labelled “a” (0.04), whereas the other organs are labelled “b” (0.02–0.03). The statement in the text or the lettering in the table should be checked and corrected accordingly.

Thanks for this observation. We made corresponding correction in the revised MS (lines 340-342).

Figure reference for protein data (lines 327–330)
The paragraph discussing total protein refers to “Figure 3D”. Protein data are shown in Figure 2D, whereas Figure 3 contains TAC, vitamin C and polyphenol results. The figure references in this paragraph should therefore be Figure 2D–F.

Thanks for this observation. We made corresponding correction in the revised MS (line 369).

Interpretation of Table 4
The meaning of the three rows in Table 4 is not sufficiently clear. For the bitter attribute, the table gives Detected = 158, Negative = 152 and Positive = 103. The Positive and Negative values correspond to the totals obtained from Figure 5 (103 and 152), but their sum is 255 rather than 158. This suggests that “Detected” is calculated using a different approach, possibly as the number of detections rather than an intensity-weighted score. If so, this needs to be defined explicitly in the Methods or in the table legend. At present, the three categories cannot be interpreted unambiguously.

Thank you for your comment. The section 2.6 of material and methods was improved in order to clarify how we obtained these data (lines 292-296). Additionally, we incorporated a sentence to explain this fact in lines 523-526 of results section of the corrected MS.

Bitterness of dish 10 (lines 476–478 and 488–489; Figure 5J)
There is a contradiction in the description of bitterness. The manuscript first states that bitterness was detected in all dishes except dishes 4 and 10. Later, however, bitterness in dish 10 is described as predominantly positive (15 vs 1), which agrees with Figure 5J, where bitterness is clearly reported for this dish. The first statement should therefore be revised.

Thank you for your observation. We changed this statemen according to our results (lines 512-516).

Hedonic scale (line 268 vs line 513)
The scale is described differently in the Materials and Methods and Results sections. Line 268 states that responses were converted to numerical values from 0 to 7, whereas line 513 refers to a hedonic scale ranging from 1 to 7. The correct range should be stated consistently throughout the manuscript.

Thank you for your observation. Hedonic scale ranged from 0 to 7. We changed the text (line 553) accordingly.

Cadmium and arsenic
The list of analytical wavelengths in lines 182–185 includes P, Hg, Se, Zn, Fe, B, Mn, Mo, Mg, Ca, Cu, Pb and K, with Na measured separately by flame photometry. Cd and As do not appear to have been determined. Considering that the manuscript discusses C. maritima as a potential edible plant, the absence of data for these two elements should at least be mentioned as a limitation of the study. If samples and analytical capacity are still available, their determination would further strengthen the safety assessment.

We measured Cd levels, although these were under the limit of detection for all samples tested. In the corrected MS, wave length for Cd detection was incorporated in line 194 of the M&M section, Cd results were included in lines 343-345 of results section. We did not measure, however, As levels. We discussed in lines 672-678 the importance of measuring toxic elements such as Cd and As.

Editorial, referencing and typographical issues

Figure 4 legend (lines 463–464)
The abbreviation “P” is used for both “price” and “pests”. Different abbreviations should be used to avoid confusion.

Pr and Pe were used for price and pests, respectively. We modified the legend of figure 4 and the labels within the panels 4 D and F of figure 4 (lines 515-516).

Chef’s name
The name is written as “Paco Brocal Rubio” at line 246 and “Paco Borcal Rubio” in the Acknowledgments (line 736). The correct spelling should be used consistently.

The correct surname “Brocal” was used. Thank you very much for noticing this.

Reference 25
The author “Santos, Fernanda F.” appears four times consecutively in the author list. The reference should be checked against the original publication.

This reference is corrected in the new version of the MS (ref 26; line 929-931).

Language and typographical corrections
There are several minor language errors that should be corrected. Examples include “cricifers” → “crucifers” (reference 15), “significatively” → “significantly” (lines 378, 389 and 390), “stablished” → “established” (line 586), and the phrase “obtained for to stems” (lines 342–343). A careful language revision of the full manuscript would be advisable.

Thanks for your observation. Language errors were corrected.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript presents an interesting and timely evaluation of Cakile maritima as a potential food resource for saline agriculture. The combination of nutritional characterization, antioxidant analyses, consumer survey, and sensory evaluation adds value to the study and broadens its relevance.

The manuscript is generally well structured and the results support the conclusions. I only have a few minor suggestions that could further improve the paper:

Consider including one or two key quantitative findings in the Abstract to better highlight the significance of the results.

A brief discussion of the limitations of the consumer survey (snowball sampling and respondent representativeness) would strengthen the interpretation of the findings.

The sensory evaluation results are interesting; however, a short comment on the relatively small size of the consumer panel would be helpful.

Please check the manuscript for consistency in terminology (e.g., "proximate analysis" rather than "proximal analysis").

A light language revision is recommended, as a few grammatical and stylistic issues remain throughout the text.

Overall, this is a valuable contribution.

Recommendation: Minor Revision

Comments on the Quality of English Language

The manuscript is generally clear and understandable. Only minor language polishing is recommended before publication.

Examples include:

replacing "proximal analysis" with "proximate analysis";
replacing "significatively" with "significantly";
replacing "consisting on" with "consisting of";
improving a few long sentences for readability.

Overall, the English is acceptable, and only minor editorial corrections are required.

Author Response

Reviewer 2

The manuscript is generally well structured and the results support the conclusions. I only have a few minor suggestions that could further improve the paper:

Consider including one or two key quantitative findings in the Abstract to better highlight the significance of the results.

A couple of sentences were added in the abstract summarizing quantitative findings (lines 24-26 and 29-33).

A brief discussion of the limitations of the consumer survey (snowball sampling and respondent representativeness) would strengthen the interpretation of the findings. The sensory evaluation results are interesting; however, a short comment on the relatively small size of the consumer panel would be helpful.

A paragraph discussing these limitations was added in the discussion section (lines 799-813).

Please check the manuscript for consistency in terminology (e.g., "proximate analysis" rather than "proximal analysis").

Thank you for this observation. The term “proximate” was used along the text.

A light language revision is recommended, as a few grammatical and stylistic issues remain throughout the text.

Overall, this is a valuable contribution.

Recommendation: Minor Revision

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The authors conducted an interesting study, but it is not without a number of correctable shortcomings.

Comments for author File: Comments.pdf

Author Response

Reviewer 3

Review on article «Valorisation of the halophyte Cakile maritima as a food resource for human consumption».

Authors: Ricardo Mir, María Dolores García-Martínez,, Monica Boscaiu,, Oscar Vicente, Jaime Prohens, María Dolores, Raigón-Jiménez

The authors conducted an interesting study, but it is not without a number of correctable shortcomings.

Comment: With respect to Line 29 and in Introduction: the phrase «food resource for saline agriculture» raises some concerns. In our opinion, it is more appropriate to use the phrase «plant resource» since the word «food» has multiple meanings, including those related to technologically processed products.

Response: In order to avoid confusion, we replace “food resource” for “species” (line 34).

Comment: In addition in our view, it would be more appropriate to acknowledge that salinization can be either natural or anthropogenic.

Response: Natural and anthropogenic salinization concepts are incorporated in lines 42 and 44.

Comment: In Line 52 - The statement that halophytes neutralize reactive oxygen species (ROS) is not unique to these plants, as this ability is common to plants in general. It appears that a crucial premise or sentence has been omitted here. We would like to point out that living organisms, and plants in particular, are capable of eliminating ROS through both polyphenolic compounds and enzymatic activity. The authors are therefore advised to revise this part of the introduction accordingly.

Response: A sentence in line 58-59 was added to indicate the efficient ROS detoxification system of halophytes, although non-exclusive of such as species.

Comment: Then in Line 56 - The text makes an abrupt transition to ascorbic acid. However, this compound is only one of many antioxidant substances found in plant tissues. Moreover, different plant species rely on different ratios of these antioxidants, and their composition can vary considerably. We would recommend reworking and expanding this fragment to provide a more balanced and comprehensive overview.

Response: We thank the reviewer for this observation. We agree that the antioxidant system of plants comprises a broad range of enzymatic and non-enzymatic components, including glutathione, carotenoids and tocopherols in addition to polyphenols and ascorbate. We have therefore revised the Introduction to describe the diversity of plant antioxidant systems, with specific emphasis on polyphenols and vitamin C as representative non-enzymatic antioxidant compounds analysed in our work (lines 61-67).

Comment: In Line 60 - It is not entirely justified to directly equate high antioxidant activity – or high concentrations of antioxidant compounds with a beneficial effect on human health. While such an effect is possible, it is by no means guaranteed and should not be stated as a foregone conclusion. This claim requires either more cautious wording or supporting evidence.

Response: “may be” is used instead of “are” (line 70-71)

Comment: in Lines 77-79 should be understood as indicating that this species behaves as an aggressive invasive plant in some areas. We suggest that the authors explicitly point out this fact and note that using this plant as food may help partially regulate its population. For reference, we provide an example of a relevant article addressing the possible invasiveness of this species: «Reassessment of the invasion history of two species of Cakile (Brassicaceae) in Australia»// Cunninghamia. 2013. V.13. P. 275-290… DOI: 10.7751/cunninghamia.2013.005. The authors may also find otherexamples on this matter.

Response: We incorporated this information to the new version of the MS (lines 86-89), as well as the suggested reference (ref 17).

Comment: In lines 207 and 216, it is unclear why the authors used fresh weight. Is this due to the fact that the plant is edible? If so, this should be clearly noted in the text. Otherwise, this may confuse readers, as calculations based on fresh weight are highly dependent on the moisture content of the sample.

Response: We thank the reviewer for this observation. Fresh weight (FW) was used to express antioxidant compound concentrations because C. maritima is being evaluated as a potential food resource, and FW-based values provide a more direct estimation of the amount of antioxidant compounds present in the fresh plant material as consumed. We acknowledge that FW-based concentrations are influenced by tissue moisture content and have therefore clarified this rationale in the revised manuscript (line 232-234).

Comment: Regarding line 208, why did the authors choose the Folin-Ciocalteu method when other alternative methods are available? The literature describes that in some cases this method may not be selective solely with respect to the phenolic components of the raw material, so we recommend that the authors justify their choice or discuss this limitation.

Response: We thank the reviewer for this valuable comment. We selected the Folin–Ciocalteu method because it is one of the most widely used and well-established methods for estimating total phenolic content, and it is routinely employed in our laboratory, allowing comparison with our previous analyses and with a substantial body of published literature. We agree, however, that this method is not fully specific to phenolic compounds, as other reducing substances present in plant matrices may also contribute to the measured response. We have therefore clarified this limitation in the revised manuscript.

Comment: in section «2.5 Online survey», the authors indicate that they used online forms for the survey. However, several important points remain unclear.

Were there any questions about the participants' age and place of residence, such as urban or rural? We assume that the social status and professional position of the respondents, even if anonymous, are also unknown, which is relevant because the proposed dishes may be consumed and perceived differently depending on a person's social status and level of affluence.

Response: Regarding personal information, in the online questionnaire we only asked for age range and gender, and this information is included in lines (lines 447-451). Regarding social status and professional position, we agree these could influence the fact of consuming wild species (either positively or negatively). This in fact would be an interesting point of further research. However, with the limited number of responses, and the lack of this information, it is not possible to analyse this at this moment.

Comment: Additionally, were the questions in the online form presented linearly in a fixed order or randomized? This distinction matters because there is a risk of accidental or careless responses, particularly if two people were taking the survey at the same time on the same device.

Response: Questions were presented in a specific order, although there is not control about the real order of answering, being possible that participants answer in a distinct order. This is, however, unlikely. Moreover, participants answer the questionnaire mostly using personal self-phones or computers. Thus, two people taking the questionnaire at the same time using the same device is unlikely.

Comment: Finally, we strongly recommend that the authors provide a margin of error for their sociological research, which can be estimated based on the approximate size of the population residing in the territory where the survey was conducted.

Response: We are aware about the limitation of the citizen science approach realized in our work, both the online questionnaire and the tasting event. For this reason, we incorporated a paragraph in the discussion section (lines 799-813) with this regard.

Comment: In lines 245-255, it is worth specifying the approximate proportion of Cakile maritima in the recipes, for example, by weight per dry or wet weight. The point is that in some dishes, the proportion may be so small that the consumer will perceive this plant simply as a decoration or an embellishment for the dish, rather than as an actual ingredient.

Response: Providing an approximate proportion of C. maritima is not feasible for the dishes included in the sensory evaluation, as the species was incorporated using different organs and culinary preparations, and its contribution to each dish varied accordingly. Moreover, the sensory impact of C. maritima was strongly dependent on the preparation method and on its combination with other ingredients. Some preparations, such as pickling, may modify or attenuate the characteristic flavour of the plant, while combinations with other ingredients may mask its bitterness, requiring a greater proportion of C. maritima for its flavour to be perceived. Conversely, preparations using raw leaves may require only a small amount of plant material for its bitter flavour to become evident. Therefore, the amount of C. maritima used cannot be directly equated with its sensory contribution across the different dishes. On the other hand, in culinary practice, visual appearance and presentation are important components of food perception, and the incorporation of flowers and other aerial organs may enhance the attractiveness and perceived novelty of a dish, beyond their contribution to flavour and aroma. Thus, we consider visual presence of C. maritima to be part of its gastronomic valorisation rather than merely a decorative use.

AComment: dditionally, it is stated that 31 participants took part in the tasting, but in Figure 5, the total number of negative and positive ratings for each taste characteristic exceeds 31 in some cases. How can the authors explain this discrepancy?

Response: The values presented in figure 5 correspond to scores, calculated as the number of times that a specific flavour was identified, multiply by 1 (light) or 2 (intense). For this reason, scores can be greater than 31. This is clarified in lines 291-296 of the corrected MS.

Comment: It is also unclear why, in Figure 6, some parts of the graphs show 31 data points, others show 32, and in some instances the number is less than 30. What accounts for these variations? In some cases, the total number of points is indeed less than 31, which may suggest that not all tasters wanted to try a particular dish. The entire figure requires a more detailed discussion in the text to clarify these inconsistencies.

Response: Thank you very much for this observation. The final number of participants is 32, not 31 as indicated in the original version of the MS. We changed it accordingly in the new version of the MS (lines 28, 129, 274). On the other hand, the fact that in some cases the plots display 31 data points (F1, F4, F5, F7, F8, F9, T2, T4, T5, T7, T8, T9), or less (F11, T10 and T11), is due to some of the participants not scoring flavour or texture for a specific dish. Thus, this data is not available.

Comment: In lines 288-299, the authors discuss the content of micro- and macroelements, but there is no accompanying discussion regarding the possibility of using specific parts of the plant as food in relation to the content of particular nutrients. For what type of diet is the plant suitable? For example, is it appropriate for people who abstain from salt consumption, or for those with insufficient calcium intake? How suitable is it in each of these cases? This part of the reasoning needs to be substantially expanded in the text.

Response: In the revised version of the manuscript, we have included data on the potential contribution of specific edible organs, particularly leaves and flowers, to the Nutrient Reference Values (NRVs) established by the EU (lines 329-332), including the corresponding reference (number 36). In addition, we have expanded the Discussion section (lines 644–656) with a specific paragraph addressing the potential suitability and limitations of C. maritima for specific dietary requirements in relation to its mineral composition. Importantly, however, the potential dietary contribution of this species should be interpreted in the context of its expected culinary use, as C. maritima is not intended to be consumed in large quantities, but rather as a complementary ingredient in specific dishes. Furthermore, as the bioavailability of the different mineral elements was not assessed in the present study, our results should be interpreted in terms of their potential contribution to dietary mineral intake rather than as evidence of their actual nutritional availability. This fact is discussed specially for phosphorous (line 666-669).

Comment: Figure 2 is difficult to read due to the similarity of the chosen colors and the thick border used for the graphs. We recommend changing the color scheme of the figure to improve clarity and distinguishability of the data.

Response: Thank you for your advice. Accordingly, we change colors and bar border line thickness of figures 2 and 3.

Comment: Table 1 and Supplementary Table 2 have the same legend but contain similar values, which may mislead the reader. It is necessary to provide a more detailed explanation of why both tables are needed, as their presence currently creates a sense of redundancy in the materials and a lack of sufficient description on the part of the authors. The same concern applies to Figure 3, which also requires additional clarification regarding its purpose and relation to other presented data.

Response: Table 1 correspond to element quantification of C. maritima with no distinction of the morphotype used (n = 6 for stems, leaves and fruits), whereas data for element quantification in Sup Table 2 is separated in both morphotypes (n=3 for stems, leaves and fruits). Data are obtained from the same experimental procedure, for this reason values are similar (average of pinnatifid and entire values in Sup Table 2 correspond to values on Table 1). However, the meaning of both tables is different (general element composition of C. maritima, and composition of each leaf morphotype). This comment applies for Figure 3 and table 3. To clarify the experimental sampling and procedure table legends were improved, and the way of sampling and sample analysis was clarified in materials section (lines 154-158).

Comment: Regarding section 3.5, we have several questions. Why did the authors not choose a sequential test with repetitions for their research, such as a multi-stage like A/B- testing procedure? This approach might have provided more robust statistical control.

Response: Thank you for this comment. In this work we aim to valorise C. maritima as an ingredient. This species is quiet unknown for general public and flavour is special (strong bitter, not too common in current cuisine), so we focused on showing different elaborations or ideas with this species, rather than an A/B testing procedure. In our opinion, A/B testing would be more appropriated to compare flavour of different morphotypes, or plants grown under different growing conditions. This, however, was not the aim of this pioneer tasting event.

Comment: Furthermore, did the authors provide the tasters with the names of the dishes or were they encrypted? This is important because for some people, the name and ingredients may influence their expectations and consequently affect the resulting scores in advance.

Response: Different dishes were encrypted. Only ingredient related to general intolerances were mentioned before the tasting event for safety reasons. An explanation of the names, elaboration procedure and specific ingredients of the different dishes was given by the professional chef once the tasting event was concluded. This information is included in the corrected MS (lines 279-284).

Comment: Did the order of serving dishes change between tasters, and could the tasters see each other's reactions while trying the dishes? Both factors could introduce bias into the sensory evaluation.

Response: The order of the serving dishes was the same for all testers, since this is in fact part of the degustation design (notice that it implies entrees, main dishes, desert and tea). Tasting of each dish was performed simultaneously in two groups of 14 and 18 testers. The separation of participants in two groups was made for logistical reasons. This is included in the new version of the MS (lines 279-284).

Comment: Was a lie scale also included in the survey to determine the approximate level of truthfulness of the respondents? Such a scale would help assess the reliability of the self-reported data.

Response: No lie scale was included in the questionnaire. The survey was anonymous, and participants were instructed that there were no correct or incorrect answers and that they should report their own sensory perceptions and preferences. As the evaluation was based on immediate responses to coded food samples, we considered a lie scale unnecessary. Importantly, the tasting included a control dish prepared without C. maritima (dish number 4). This control allowed us to contextualize participants’ sensory and hedonic responses to the Cakile-containing dishes under the same tasting conditions. We have clarified these aspects in the revised manuscript.

Comment: Regarding Figure 6, we suggest that the authors consider constructing a clustering of dishes in the form of a dendrogram based on the data they have collected. This would allow for a visual representation of similarities and differences between the dishes and could add valuable analytical depth to the discussion.

Response: Thank you very much for this suggestion. A supplementary figure including a dendrogram clustering together with hierarchical heatmap, from data collected in the tasting event is included in the MS. Accordingly, corresponding text in results section (lines 560-570) was added.

Comment: In line 553, the authors refer to their own work. However, we were unable to find any significant discussions regarding the shelf life of this plant. The statement «does not appear to compromise their shelf» should be corrected, as this topic has essentially not been studied in the present work. The same applies to lines 564-566. Since the issue has not been investigated, this thesis should be softened or presented not as an imperative conclusion, but rather as a mild assumption or tentative suggestion on the part of the authors.

Response: We agree with reviewer comment. We softened both sentences in the new version of the MS, the one regarding to the self-stability (lines 594-596) and the one related to nutritional features of root-derived products (lines 607-609).

Comment: In lines 619-620, authors refer to work number 58, comparing the antioxidant activity you obtained with data obtained by other authors for the same species. However, it would be better to include other types of plants used in cooking for comparison, for

example, plants that were used in the dishes you prepared that did not contain Cakile maritima. There is no need to be afraid of such a comparison. It is necessary to openly compare these plant resources with each other. Ultimately, you could simply compare your results with leafy crops such as spinach or onion leaves, using data available from the literature.

Response: A new section in discussion was added in the new version of the MS (4.3. Antioxidant potential of Cakile maritima, lines 687- 721).

Comment: In lines 689-690, it would be worth adding some discussion about the possibility of masking the bitter taste of this plant in dishes. For example, what types of flavor combinations can smooth out the bitter taste of the plant based on your data? This can be done by assessing the relationship between positive responses and the predominant taste in a particular dish. This point is especially important for scaling the product on the market and attracting new consumers, as bitterness is often a barrier to acceptance of novel food ingredients.

Response: Thanks for this comment. A paragraph in the new version of the MS was added to the revised MS in this regard (lines 790-797).

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Authors,

The revised manuscript is considerably clearer, and most of the points raised in the previous round have been addressed. I have only a few remaining comments that should be resolved before the manuscript is accepted.

The timing of the ethical approval needs clarification. The online survey was conducted between 1 December 2024 and 13 February 2025, while the Institutional Review Board Statement gives 30 June 2025 as the date of approval for the study involving human participants. Could the authors clarify whether an earlier approval covered the survey, whether this date refers to a later amendment, or whether the date reported here is incorrect?
Please check the number of participants in the sensory evaluation. The Abstract and the Methods state that 312 participants took part, but the Methods later specify 14 participants in the first tasting session and 18 in the second, which gives a total of 32. The correct number should be used consistently throughout the manuscript.
There is still a contradiction regarding bitterness in dish 10. The text first states that bitter flavour was not detected in dishes 4 and 10, but the following sentence includes dish 10 among the dishes in which bitterness was detected, although it was not the dominant taste. Later, the bitter attribute for dish 10 is reported as predominantly positive (15 vs 1). This part should be checked against the original sensory data and corrected accordingly.
I would also check the statement in the Abstract that bitterness was perceived as pleasant by “fewer than 50% of participants.” In the Results, the corresponding information is presented as weighted positive and negative scores (103 and 152), rather than as the percentage of individual participants reporting a pleasant perception. If the percentage was calculated separately from the participant-level data, this should be made clear; otherwise, the wording in the Abstract should be adjusted.
The method used for phosphorus determination is still not entirely clear. Phosphorus is included among the elements analysed by ICP-OES, with a wavelength of 177.434 nm, but later in the same section the authors state that phosphorus was determined spectrophotometrically at 430 nm. Please indicate which procedure generated the phosphorus values reported in the tables, or explain the purpose of the two measurements if both were used.
The name of the chef should be checked once more. It appears as “Paco Brocal Rubio” in the Methods and “Paco Borocal Rubio” in the Acknowledgments. The correct spelling should be used in both places.
Finally, the first page still contains the unrelated heading “Fruit- and Vegetable-Derived Exosome-like Nanovesicles: A Review” above the actual manuscript title. This appears to be a leftover template/title and should be removed from the final version.

Author Response

Comment: The timing of the ethical approval needs clarification. The online survey was conducted between 1 December 2024 and 13 February 2025, while the Institutional Review Board Statement gives 30 June 2025 as the date of approval for the study involving human participants. Could the authors clarify whether an earlier approval covered the survey, whether this date refers to a later amendment, or whether the date reported here is incorrect?

Response: Thank you for pointing this out. We apologise for this error in the manuscript. The dates originally reported referred to the period during which the questionnaire was designed and developed (1 December 2024 to 13 February 2025), rather than to the period of data collection. We believe this information is not relevant.

The questionnaire was not made available for public participation until September 2025, following approval by the Ethics Committee. Data collection subsequently took place between September and December 2025. We have corrected this information in the revised version of the manuscript (lines 246–247), which now states:

“Data collection was conducted between September and December 2025, and all responses were securely stored”

Comment: Please check the number of participants in the sensory evaluation. The Abstract and the Methods state that 312 participants took part, but the Methods later specify 14 participants in the first tasting session and 18 in the second, which gives a total of 32. The correct number should be used consistently throughout the manuscript.

Response: We would like to clarify that the total number of participants in the sensory evaluation was 32, with two separate tasting sessions comprising 14 and 18 participants, respectively.

The discrepancy may have arisen from the consultation of a version of the manuscript with track changes enabled, in which previous versions of the text may still have been visible. In the final version of the manuscript, as well as in the version uploaded for this second round of revision, the total number of participants is consistently reported as 32, both in the Abstract (line 28) and in the Materials and Methods section (line 276).

We hope this clarification resolves the discrepancy.

Comment: There is still a contradiction regarding bitterness in dish 10. The text first states that bitter flavour was not detected in dishes 4 and 10, but the following sentence includes dish 10 among the dishes in which bitterness was detected, although it was not the dominant taste. Later, the bitter attribute for dish 10 is reported as predominantly positive (15 vs 1). This part should be checked against the original sensory data and corrected accordingly.

Response: Similarly, this discrepancy might be due to checking errors. In the MS we uploaded in the first round of revision, and in the recently uploaded it states in lines 511-515:

“Per dish, bitter flavour was detected in all dishes, with the exception of dish 4 (Figure 5 D, J), which corresponded to the control dish which did not include C. maritima as an ingredient. For dishes 2, 3, 5 and 6, bitter flavour was by far the most identified one (Figure 5 B, C, E, F), whereas for dishes 1, 7, 9, 10 and 11, although detected, it was not the main taste attribute (Figure 5)”

Accordingly, bitterness in dish 10 is detected, and positive perception is generally registered (15 positive versus 1 negative; Figure 5J).

Comment; I would also check the statement in the Abstract that bitterness was perceived as pleasant by “fewer than 50% of participants.” In the Results, the corresponding information is presented as weighted positive and negative scores (103 and 152), rather than as the percentage of individual participants reporting a pleasant perception. If the percentage was calculated separately from the participant-level data, this should be made clear; otherwise, the wording in the Abstract should be adjusted.

Response: Thank you very much for this comment. We agree the explanation in the abstract is not accurate enough. We rephrase is as follows:

“Sensory evaluation revealed a prominent bitter flavour amongst dishes containing C. maritima, with weighted scores for negative perceptions slightly exceeding those for positive ones, suggesting that its sensory profile may limit its acceptance by the general public while offering potential for specific consumer segments”.

Comment: The method used for phosphorus determination is still not entirely clear. Phosphorus is included among the elements analysed by ICP-OES, with a wavelength of 177.434 nm, but later in the same section the authors state that phosphorus was determined spectrophotometrically at 430 nm. Please indicate which procedure generated the phosphorus values reported in the tables, or explain the purpose of the two measurements if both were used.

Response: Thanks for this comment. Data for P quantification shown in this MS is collected only with spectrophotometric approach. Thus, we eliminated “177.434 nm for P” from line 195 of the uploaded MS.

Comment: The name of the chef should be checked once more. It appears as “Paco Brocal Rubio” in the Methods and “Paco Borocal Rubio” in the Acknowledgments. The correct spelling should be used in both places.

Response: Correct spelling “Brocal” is incorporated in both places (line 265 and 912) in the previous version and in the newly uploaded.

Comment: Finally, the first page still contains the unrelated heading “Fruit- and Vegetable-Derived Exosome-like Nanovesicles: A Review” above the actual manuscript title. This appears to be a leftover template/title and should be removed from the final version.

Response: This comment should be an error, since we do not observe such an unrelated heading. The MS was prepared on a recently downloaded template.