Effects of Laser Irradiation at 488, 514, 532, 552, 660, and 785 nm on the Aqueous Extracts of Plantago lanceolata L.: A Comparison on Chemical Content, Antioxidant Activity and Caco-2 Viability

Round 1

Reviewer 1 Report

1. The manuscript does not indicate why these lasers were taken, and not any others. Why are these particular wavelengths and powers chosen?
2. Section 2.4 does not specify exactly how the lasers were used: at what distance are they installed? what was the mode of radiation generation (pulsed, continuous)? How long have the lasers been working? What was the telescopic system used for? For defocusing?
3. It is absolutely incorrect to compare lasers of different powers! If we evaluate the effectiveness of lasers of different wavelengths, then it is necessary to ensure the same irradiation (E, W/m²) of the working surface of the extracts. This is difficult, but feasible on the experimental setup given in the article.
4. In Figure 2, instead of position (9), there is position (10) in the caption.
5. The Discussion section, in fact, is not a discussion of the results, and the information provided in it would be appropriate in the Introduction section.

Author Response

First of all, thank you very much for your very careful review of our paper! Based on your comments and suggestions, the paper has been significantly improved. The answers are as follows:

1. The manuscript does not indicate why these lasers were taken, and not any others. Why are these particular wavelengths and powers chosen?                                                                                                           - Discussion section has been rebuild to meet the requirements at points 1, 3 and 5. Briefly, the consulted scientific data (before and after the effective laser experiments and project proposal)indicated a high interest for the selected wavelengths; - The laser wavelengths we have chosen cover a large spectral area, from 488nm to 785nm; - The lasers we used were affordable for us in terms of price. These lasers are manufactured by the same company Coherent Inc. (as OBIS family and platform) and offers the best performances on the market with respect to stability and beam quality. All the lasers belong to a platform that offers the power supply and the control for any configuration. The number of laser modules from these family could be increased in the future in order to cover an extended spectral domain and/or power levels. The outputs of every laser is delivered to the telescope by means of an optical fiber. Also, the beams could be combine to be a single optical fiber that collects the beams from every laser generator and lead it to the telescope; - Also, the lasers were used at maximum power to get significant results for every wavelength.     - Finally, the authors have analyzed and explained these wavelengths in terms of results on animal and plant matter, on living and non-living matter, and we believe that the Discussion section now brings a better understanding of the mode of use and selection of laser radiation in extractive biotechnology.
2. Section 2.4 does not specify exactly how the lasers were used: at what distance are they installed? what was the mode of radiation generation (pulsed, continuous)? How long have the lasers been working? What was the telescopic system used for? For defocusing? - The laser outputs (pigtailed) are directly driven by the optical fibers to the telescope so the distance from the lasers to the telescope is not relevant. We have used 2 m length optical fiber. All lasers benefit from the common platform to supply and control the lasers. - The lasers emit in continuous mode. The duration of irradiation is 1 hour (see 2.3 in our propose paper).The telescopic system collects the radiation from the lasers through an optical fiber connector and deliver an uniform, circular, 15 mm diameter laser beam almost similar for every wavelength.The telescope was placed at about 16 cm from the liquid surface. Since the beam is collimated, this distance does not influence the power density (W/m2). We have a large area of irradiation (approximately 176mm2) with perfect uniform and stable laser beams.
3. It is absolutely incorrect to compare lasers of different powers! If we evaluate the effectiveness of lasers of different wavelengths, then it is necessary to ensure the same irradiation (E, W/m2) of the working surface of the extracts. This is difficult, but feasible on the experimental setup given in the article. - We do not compare directly the efficiency of different lasers of different powers and different wavelengths,but we simply give the results we got at different regimes of irradiation for similar irradiated surface. An experimental study is envisaged, and the specific laser powers and wavelengths (photon energy) effects will be considered to be accurately compared.
4. In Figure 2, instead of position (9), there is position (10) in the caption. - REVISED;
5. The Discussion section, in fact, is not a discussion of the results, and the information provided in it would be appropriate in the Introduction section. - REVISED;

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments in attached file. 

Comments for author File: Comments.pdf

Author Response

First of all, thank you very much for your very careful review of our paper! Based on your comments and suggestions, the paper has been significantly improved. The answers are as follows: Line 13: antioxidant activity-missing - REVISED Line 17: It is enough to say control extract. Instead laser extract I suggest to use laser treated extract. - REVISED Line 53: pose less security - What do you mean be that?! - REVISED by exclusion (it refers to the safety of work in plant extraction laboratories, but it is not the object of the study, and therefore has been cut) Line 78-79: part of the text is in italic, why? - REVISED (is cited “...”) Line 166: there test samples were taken? - REVISED Line 241: ..total time is... Why did you use different extract for the analysis? For example for total phenolic, you used extract after 2h of extraction and for antioxidant activity after 1h of extract. 　　- The way in which the six series of extracts were obtained and analyzed, and pharmacological tested was imposed by the necessity of using 2 control samples: the control sample of each experiment(control samples at 1 hour) and the control sample at 2 hours (to measure the efficacy in time, and to assure identical total extraction time) together allowing vegetal extraction monitoring 1)in terms of optimal parameters and 2)real progress of laser irradiation. 　　-These explanations also were inserted in Section 3.2 Antioxidant activity results. Line 43-48: I understand what you wanted to say, however I think is should be rewritten to be more understandable. - REVISED Conclusion look like some up of the results which would better fit in discussion part. In conclusion try to say why one lesser treatment is better than the other ones. - REVISED Additional to the comment concerning results, manuscript is hard to read. . - REVISED.

Round 2

Reviewer 1 Report

The authors have significantly improved the article. At the same time, I believe that the wavelengths and laser powers were chosen without the necessary SCIENTIFIC justification. I recommend the authors to pay attention to this in their further research.