Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF LATTICE INCLUSION OF CU AND FE IONS ON THERMAL DECOMPOSITION CHARACTERISTICS OF AMMONIUM PERCHLORATE
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: Experimental part is too simple to be a paper. The structure and melting point of the co-crystals should be demonstrated. Please see my comments for the details.
Validity of the findings: The data in the tables were not well explained.
Additional comments: Main logic:
How do you determine it is co-crystal? What are the types of interaction to form co-crystals? Does it affect the melting point or phase transition of the AP? Why the EDS show separation of different elements?
Manuscript preparation:
1.	Figure 1, the scale bar is too small. And most of the information under the bottom of the SEM Figures are not necessary, which should be cut off to make them clear.
2.	Figure 2, once again, the EDS images should have a scale bar or scale bars are too small to be seen. Also the labeling of elements should be clear and the color of same element from different samples should keep the same to avoid confusing. The original SEM images should be given beside these EDS results.
3.	Figure 3, standard AP pattern peaks should be given and marked. The peaks for the doped AP should be assigned.
4.	Figure 4, the markers should be hollow to avoid overlapping to each other. Such as, it is hard to distinguish the APFe-1 and APFe-2. DSC data is too overwhelming and the important peaks should be labeled.
5.	What method was used for the activation energy calculations? What is the different heating rate when do the calculations? The related information should be given in the main text.
6.	How do you know it “in turn improving the ballistic properties of the propellant”? (In abstract and conclusion). Any evidence?
7.	Table 1. The mole percentage of Cu or Fe should be given in addition to weight%.
8.	Table 1. How did you measure the bulk density and moisture and specific surface area?
9.	Please use µm instead of µ when describe the size.
10.	Table 2 and Figure 4. Important peaks should be labelled clearly in Figure 4.
11.	Line 160, “Copper present in the lattice is the initiation point for LTD and it get oxidized to CuO”, any evidence? Considering the oxidation temperature of Cu, it doesn’t make sense.
12.	The authors didn’t talk much about the numbers in their tables.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF LATTICE INCLUSION OF CU AND FE IONS ON THERMAL DECOMPOSITION CHARACTERISTICS OF AMMONIUM PERCHLORATE
Review round: 1
Reviewer: 2

Output:
Basic reporting: Less innovative
Experimental design: Less innovative
Validity of the findings: Less innovative
Additional comments: In this manuscript, the authors declared their successful synthesis of two co-crystal compounds incorporating ammonium perchlorate with copper perchlorate or Iron perchlorate. Then the authors studied the impact of the so-prepared materials on thermal decomposition characteristics of ammonium perchlorate. However, the characterization provided in the manuscript obviously cannot evidence the so-prepared materials as co-crystal compounds. Thus, I would reject the manuscript to be published:
(1) The authors provided the SEM images, Elemental mapping and XRD Patterns of Normal AP, ACuP-2 and AFeP-2, which cannot evidence these materials as co-crystal compounds especially the XRD Patterns. I think single-crystal structures and elemental analysis results of these materials are needed.
(2) In abstract, the authors mentioned “the Ammonium perchlorate (AP) is the universal oxidizer in use ... due to ... thermal decomposition without any solid residue”, which is inconsistent with the introduction of copper or Iron ions. Obviously, the solid residue created by copper or iron ions is not welcome in all the solid rocket propellant motors


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF LATTICE INCLUSION OF CU AND FE IONS ON THERMAL DECOMPOSITION CHARACTERISTICS OF AMMONIUM PERCHLORATE
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: 1. In co-crystallisation section, the detailed speed of agitator and cooling rate shoul be added.
2. For better understanding of the physical characteristics of AP, the particle size distribution of AP, AP doped with Cu and Fe should be given like D50 and distribution diagrams.
3. In Line 107, the authors said that the friability of the doped samples are higher compared to normal AP, but there is no evidence and how to prove this.
4. Figure.1 only shows the local SEM images at 10 μm, while single particle of AP, ACuP and AFeP should also be provided for better understanding of whole appearance and size.
5. As listed in Table 1, the weight percent of Cu or Fe is low, however, why is the crystal structure of AP doped with Cu/Fe quite different from normal AP?
6. It is hard to distinguish the line for each sample in Figure 4.
7. Throughout the paper there are many cases that words are either awkward or grammatically incorrect and some are listed below. These confuse the reader and the authors need to check he whole paper carefully.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF LATTICE INCLUSION OF CU AND FE IONS ON THERMAL DECOMPOSITION CHARACTERISTICS OF AMMONIUM PERCHLORATE
Review round: 2
Reviewer: 1

Output:
Basic reporting: fair
Experimental design: fair
Validity of the findings: fair
Additional comments: The authors made a lot of changes based on reviewers' comments. However, I found it is still not efficient to support their conclusion.

1. The current results could not support their co-crystal conclusion. The EDS results showed a separation of elements.

2. Why the authors didn't cite their previous paper:
https://pubs.acs.org/doi/10.1021/acsomega.9b03893

Such a small percentage of doped ions changed the XRD patterns dramatically, the authors didn't explain how the transition happens.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF LATTICE INCLUSION OF CU AND FE IONS ON THERMAL DECOMPOSITION CHARACTERISTICS OF AMMONIUM PERCHLORATE
Review round: 2
Reviewer: 2

Output:
Basic reporting: I noted that three reviewers have raised doubts about the formation of co-crystals in this work, but it is obvious that the author failed to provide sufficient evidence to support this in the improved version. Therefore, I insist on rejecting this submission.
Experimental design: I noted that three reviewers have raised doubts about the formation of co-crystals in this work, but it is obvious that the author failed to provide sufficient evidence to support this in the improved version. Therefore, I insist on rejecting this submission.
Validity of the findings: I noted that three reviewers have raised doubts about the formation of co-crystals in this work, but it is obvious that the author failed to provide sufficient evidence to support this in the improved version. Therefore, I insist on rejecting this submission.
Additional comments: I noted that three reviewers have raised doubts about the formation of co-crystals in this work, but it is obvious that the author failed to provide sufficient evidence to support this in the improved version. Therefore, I insist on rejecting this submission.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF LATTICE INCLUSION OF CU AND FE IONS ON THERMAL DECOMPOSITION CHARACTERISTICS OF AMMONIUM PERCHLORATE
Review round: 2
Reviewer: 3

Output:
Basic reporting: All of my conments have been well answered and this paper can be accepted now.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: All of my conments have been well answered and this paper can be accepted now.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IDENTIFICATION AND QUANTIFICATION OF FLAVONOIDS IN 207 CULTIVATED LOTUS (NELUMBO NUCIFERA) AND THEIR CONTRIBUTION TO DIFFERENT COLORS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript 71194v1, titled “Identification and quantification of flavonoids in 207 cultivated lotus (Nelumbo nucifera) and their contribution to different colors,” details the flowers of N. nucifera content in flavonoids and relates it with their color. It is attractive to other researchers and within the journal scope. A few points need to be clarified before acceptance.
1.	The authors can improve some English language. One example is the phrase in lines 15 to 17 that would be clearer if the authors used: This study performed a systematic qualitative and quantitative determination of five anthocyanins and 18 non-anthocyanin flavonoids from the petals of 207 lotus cultivars.
2.	Please clarify if the full name of the species is Nelumbo nucifera Gaertn.
Experimental design: The manuscript 71194v1, titled “Identification and quantification of flavonoids in 207 cultivated lotus (Nelumbo nucifera) and their contribution to different colors,” details the flowers of N. nucifera content in flavonoids and relates it with their color. It is attractive to other researchers and within the journal scope. A few points need to be clarified before acceptance.
3.	Explain the differences between your work presented and discussed in this manuscript and previous works cited between lines 39 to 50. In particular, the differences with the Deng et al. (2013) manuscript.
4.	The authors should probably join the Results and Discussion part.
Validity of the findings: The manuscript 71194v1, titled “Identification and quantification of flavonoids in 207 cultivated lotus (Nelumbo nucifera) and their contribution to different colors,” details the flowers of N. nucifera content in flavonoids and relates it with their color. It is attractive to other researchers and within the journal scope. A few points need to be clarified before acceptance.
5.	It would help the readers if table 1 included a column with references, and more ion fragments should be included. The accuracy used in the m/z values is probably less important than the indication of more ion fragments. For example, peak 8 identification is discussed in lines 183 to 187. It is indicated that the loss of m/z 146 and m/z 176 implies the linkage of the sugar unit to C-3. The reference supporting this information is Ablajan et al., 2006? The presence of m/z 447 in table 1 would help to understand.
6.	The authors did not detect the common flavonoid nucleus fragmentation?
7.	Table 2 is confusing. The abbreviations used for the compound’s names are indicated in Table 1, so there is no need to repeat them, and the footnote can be simplified. The footnote b should be for all results, not just for the first one. The table caption should probably indicate that each value is the mean ± standard deviation. Furthermore, it would be better if the notes and the letters used to identify differences differed. It will be less confusing for readers. Finally, are the differences within the column or the row?
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IDENTIFICATION AND QUANTIFICATION OF FLAVONOIDS IN 207 CULTIVATED LOTUS (NELUMBO NUCIFERA) AND THEIR CONTRIBUTION TO DIFFERENT COLORS
Review round: 1
Reviewer: 2

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: Dear authors,
The manuscript intituled “Identification and quantification of flavonoids in 207 cultivated lotus (Nelumbo nucifera) and their contribution to different colors” is interesting and suitable to publish. However, it needs a few adjustments.
Introduction
Your introduction needs more detail. I suggest that you improve the description at lines 35- 38 using the lotus as an object of the studies. A description/explanation regarding the main cultivars (can be the ones that you described in your study) can be included in lines 30-33. Also, highlight the hypothesis to provide more justification for the research.
Materials & Methods
Lines 94-97: Please, include at the end of these lines which section the authors will describe the total content of non-anthocyanin flavonoids (TF) and the total content of anthocyanins (TA).
Line 148: Please, edit the information regarding the “quick RNA Isolation Kit po(, Beijing, China)”. Also, line 150.

Results and Discussion sections
For me, these sections are the most critical point of the study. Therefore, I suggest a rearrangement and an improvement in these sections. For instance, some descriptions could be included in the discussion section and are in the results section (regarding table 2, for example). Moreover, the results of Figure 6 were not described in the results section. Besides that, the discussion is shown in the section “Putative flavonoid biosynthesis pathway of lotus” could be improved. Also, a test of significance can be included in Figure 6.

Lines 385-387: Why could it have happened?
Line 405: Please, edit this sentence.
Table 2. Please, standardize the significant letters.
Figure 1: Why these data were obtained from the mixture of three cultivars: “Qiaoshou-I” (71), “Jinlingningcui” (151), and “Silian13-I” (178)?
Figure 6: Please, correct the spelling of the title on this figure.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IDENTIFICATION AND QUANTIFICATION OF FLAVONOIDS IN 207 CULTIVATED LOTUS (NELUMBO NUCIFERA) AND THEIR CONTRIBUTION TO DIFFERENT COLORS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Nothing to report.
Experimental design: Nothing to report.
Validity of the findings: Nothing to report.
Additional comments: The authors changed the manuscript accordingly to the previous comments and improved it significantly. So, the manuscript is suitable for publication.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IDENTIFICATION AND QUANTIFICATION OF FLAVONOIDS IN 207 CULTIVATED LOTUS (NELUMBO NUCIFERA) AND THEIR CONTRIBUTION TO DIFFERENT COLORS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The language was improved. They shared the raw data, but some columns are in Mandarin. I recommend that the authors translate everything into English.
Experimental design: As I described before in my last review, Figure 6 must be included a test of significance. I have included a comment in the PDF file regarding it.
Validity of the findings: The manuscript has new findings and can be accepted after revisions.
Additional comments: The manuscript is very interesting.
The authors accepted and made changes in this version but still need a broader discussion.
I recommend the authors consider describing the compounds and the findings more, so the manuscript can be more read and cited in the future.

My best regards,
The reviewer.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SPATIAL METABOLOMICS SHOWS CONTRASTING PHOSPHONOLIPID DISTRIBUTIONS IN TISSUES OF MARINE BIVALVES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript entitled “Spatial metabolomics reveals functional diversity of phosphonolipids in marine bivalves” explains the detection and localization of lipids in two species of marine mollusks. Overall, the manuscript has an interesting approach combining MS imaging, histology, and LC-MS/MS techniques.
However, the manuscript explains not so well the methodology, the ionization technique is mentioned as MALDI but the instrument used in AP-SMALDI10 (TransMIT GmbH). The authors did not go in detail in the imaging part, the quality of the figures should be improved in order to reflect much better what is in the title of the manuscript "spatial metabolomics". The results reflect moderately what they hypothesized, phosphonolipids as protective molecules present on the outside mollusk tissues but should be explained and discussed in more detail. There are references poorly written, incomplete, and even nonpresent in the bibliography list. The figures have poor quality and contain errors.
Experimental design: The question and experimental design in principle follow what is expected in this type of analysis including a complementary approach using LC-MS/MS and a histological analysis.
In the subsection: MALDI-MSI sample preparation, it is not clear nor explained why the authors used two different approaches to apply the matrix, two different matrices and two instruments e.g. gill and mantle sections 2,5-dimethoxy and 2-hydroxy-5-methoxybenzoic acid (Super-DHB, Sigma Aldrich) was applied by the SMALDIPrep sprayer (TransMIT GmbH). For M. edulis foot 2′,5′-Dihydroxyacetophenone (DHAP) was applied via a Sono-Tek SimCoat sprayer with ACCUMIST ultrasonic nebulizer (Sono-Tek Corporation). The authors did not explain the reason, not in the experimental design not in the results or discussion sections.
In the subsection: MALDI-MSI measurements, Datasets were acquired with an AP-SMALDI10 (TransMIT GmbH) ion source, the ionization method needs clarification: MALDI or Atmospheric Pressure MALDI. The authors must clarify the resolution, the data conversion, and how the data were analyzed (open software, commercial one). imzML data from the foot of Mytilus edulis is missing in Metaspace2020.
Validity of the findings: The results section, including the figures, does not represent remarkable results.  Figure 1, for example, is not a good figure and is poorly labeled (see Fig1a is written positive mode but in the caption, it is written negative mode), the figure does not contribute too much to be presented in the main text. Figure 2 uses too much space for the heat map and needs to be modified (see intensity bar). Figure 3 the intensity bar is illegible in all cases. In the caption "l" is erroneously added, but the last figure is marked as "k". More MS images of lipid distribution, comparison, and a discussion with possible protective function are expected in the results section.
Additional comments: The localization of molecules in tissues represents an extraordinary method to investigate and decipher the possible functions of molecules in organisms. I suggest that the authors re-evaluate the data already acquired, attention should be paid to the details, avoiding repetitive ideas. Improve the quality of the figures.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SPATIAL METABOLOMICS SHOWS CONTRASTING PHOSPHONOLIPID DISTRIBUTIONS IN TISSUES OF MARINE BIVALVES
Review round: 1
Reviewer: 2

Output:
Basic reporting: NO comment
Experimental design: No comment
Validity of the findings: The findings in this manuscript are novel and are of potential interest. The experiments are clearly presented, and the study question is certainly of great interest. The combined use of a metabolomic approach is a strength, much of the study is well done and the group is very capable of such laboratory analyses.
Additional comments: There are minor concerns that the authors need to address for the manuscript.
(1)	It is not clear to the readers why authors choose gill, mantle tissue, foot for sampling?
(2)	How do the authors rule out the possibility of the Phosphonolipid composition  could be due to different environmental conditions or due to differences in seasonal modifications?
(3)	Perhaps it was overlooked but could the authors please describe briefly about seasonal changes in phosphono lipid composition and also the interplay of environmental factors such as temperature and food quality.
(4)	The authors have to mention if their study obtained ethical approval


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SPATIAL METABOLOMICS SHOWS CONTRASTING PHOSPHONOLIPID DISTRIBUTIONS IN TISSUES OF MARINE BIVALVES
Review round: 1
Reviewer: 3

Output:
Basic reporting: It is difficult read some of the labels on figures (Figure 1 and Figure 2). Please render images at high resolution and increase the font size.
Experimental design: No comment
Validity of the findings: 1. It seems the authors have not thoroughly analyzed the relative abundance of Pne-Cer lipid subtypes in different organs or tissues of M. edulis and C.gigas. For eg: the localization of Pne-Cer 34:1 is not prominent in the mantle of C.gigas. Similarly, 34:2 is not abundant in the gill of C.gigas and 36:2 is not expressed in M.edulis. The authors need to evaluate these differential expressions within/in between the species and explain with respect to their phenotypes.

2.	The authors stated that phosphonolipids only exist in invertebrates (Line 115-116). However, phosphonolipids are known to exist in variety of other species such as Tetrahymena pyriformis and single celled organisms (protozoa) - Trypanosoma cruzi. The phosphonolipids appear less abundantly in various bovine tissues and even in human aorta. Can authors comment on chemical composition. nomenclature differences, or side chain dissimilarities of these phosphonolipids in comparison to the lipids in mollusks species?

3.	Although the authors precisely identify the distribution of PnE-Cer in the foot of M. edulis, they didn't correlate the functional significance. In order to do so, the authors must answer following questions:
       •	Why the phosphonolipid PnE-Cer (34:1) was absent in other foot cell types, such as musculature
            and gland cells.
       •	What is the relative distribution of phosphonolipid Pne-Cer (35:3) and others in the foot compared
            to gill and mantle of M.edulis?
Additional comments: 1.	The authors must differentiate the nomenclature of Pne-Cer phosphonolipids. For instance, Fig. 2 summarize the relative abundance of Pne-Cer lipids in mussels and mantle tissue. However, the sidechain nomenclature 34:2, 35:1, 36:2...etc are repeating throughout the plot.

2. Is there any chemical similarity between currently available therapeutics for shellfish hatcheries and phosphonolipids?

3.	It is interesting to see that the phosphonolipids protect mucous covered soft tissues against microbial pathogen invasions. Do the phosphonolipids also protect against harmful UV rays and other environmental pollutants?

4.	HMDB is wrongly abbreviated in the manuscript as HMBD. Please correct this minor thing.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SPATIAL METABOLOMICS SHOWS CONTRASTING PHOSPHONOLIPID DISTRIBUTIONS IN TISSUES OF MARINE BIVALVES
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors addressed the reviewers' questions and comments.
The authors clarified the experimental part, improved the figures and typing errors, and added important information in the introduction and discussion.
The manuscript improved considerably.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SPATIAL METABOLOMICS SHOWS CONTRASTING PHOSPHONOLIPID DISTRIBUTIONS IN TISSUES OF MARINE BIVALVES
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors have addressed all my concerns and thoroughly revised the manuscript. So, I don't have any additional comments.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DETERMINATION OF OPTICAL DENSITY (OD) OF OLIGODEOXYNUCLEOTIDE FROM HPLC PEAK AREA
Review round: 1
Reviewer: 1

Output:
Basic reporting: In this manuscript, they demonstrated a method for directly measuring the OD260 of ODNs using the HPLC peak area. They claimed that the method will save significant time and minimize the loss of oligonucleotide samples. However, they need to carefully check their accuracy and the affect of HPLC conditions on the standard curve. If they need to construct standard curve very often to get the accurate data, I did not see how this method will save significant time for researchers.
1. They should provide the original HPLC raw data. In addition, in Table 1 and Figure 1, they only performed once for each different sample. They should at least run triplicate samples to validate the repeatability.
2. For Figure 1, they should provide the standard curve equation. More importantly, they should expand their range of standard curves since they used the current standard curve to calculate the OD260 for ODN 1b. Obviously, the peak area of 1690.6 is out of the detection range.
3. To validate their method, they synthesize ODN 1b and 1c and compare the data obtained through the HPLC peak area and the data by measuring the absorbance at 260 nm on a UV-Vis spectrometer. While they only tested one sample for each ODN. They should at least test three samples for each by synthesizing at different scales. In addition, they should also calculate the accuracy to check whether they are in an acceptable range.
4. They claimed that by skipping the UV-Vis spectrometer measurement, they could be able to minimize the loss of oligonucleotide samples. While the volume of samples needed for Nanodrop is around 1-2 µL. Is this really significant to save this small amount of samples by risking the results may not be that accurate due to the poor quality of the standard curve that was caused by HPLC condition changes? They should explain more here.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DETERMINATION OF OPTICAL DENSITY (OD) OF OLIGODEOXYNUCLEOTIDE FROM HPLC PEAK AREA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Referencing is not appropriate and some original papers on DNA cleavage and purification needs to be cited. Currently, authors only cited their own their own previous works.
Experimental design: Experimental design is appropriate
Validity of the findings: Results seems to support their conclusions
Additional comments: In this paper, Chillar et al. reported a method to directly measure DNA optical density from their HPLC peak area.  This is an interesting method to measure DNA concentration and can be potentially useful. I believe this paper should be eventually accepted to be published by Peer J Analytical Chemistry journal after authors answer following questions.

Authors used water as eluent and therefore blank in their measurement. However, for many post synthesis modifications, purification and quantification is done with gradient flow of organic and aqueous solutions. How this method can be adopted to these cases? Authors need to show method applicability and accuracy under such conditions.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DETERMINATION OF OPTICAL DENSITY (OD) OF OLIGODEOXYNUCLEOTIDE FROM HPLC PEAK AREA
Review round: 1
Reviewer: 3

Output:
Basic reporting: Fang et al., developed a method to directly measure the concentration of oligonucleotides (ODNs) with the peak area obtained from the HPLC purification process. The authors established a linear correlation curve between the peak area and the absorbance of the same ODNs obtained from UV-Vis spectroscopy at 260 nm. With the correlation graph, the peak area of targeted ODNs could be directly converted to the absorbance at 260 nm. The authors tested three ODNs with different sequences and found the absorbances at 260 nm converted from the peak area aligned well with the ones from both UV-Vis spectrometer and Nanodrop. Overall, it is interesting to quantify the concentration of ODNs during the process of purification and save the step to separately measure it after drying the samples. However, there are several issues the authors need to address before publication:
1. The authors did not specify the mathematic approach they used to calculate the peak area and how accurate the calculation could be, which is quite important to the theory they claimed here.
2. The linear range of this method was not determined. Besides, the author claimed the concentration of the targeted ODNs is too high, the correlation between the peak area and the absorbance at 260 nm may not be linear and a simple solution to that was to collect fraction of the peak and add the datum to the graph. In this case, how accurate the graph can be used to determine the concentration of targeted ODNs with the peak area value in the nonlinear range?
3. There was only one data point for each injection volume and peak area included in Table 1 and Figure 1, and no error bars were included in Figure 1. More replicates need to be added to prove the reproducibility and consistence of the method.
4. The correlation coefficient (such as R2) of the two lines needs to be added in Figure 1.
5. The author mentioned the correlation graph between the peak area and the absorbance needed to be checked occasionally, especially when the column, eluents, or flow rate were changed. How long one correlation graph could be consistent to determine the absorbance of ODNs with the peak area? If all the parameters for the purification process were the same, yet the correlation graph could change, maybe due to day-to-day or sample-to-sample variations, would it be easier to measure the absorbance after drying the sample with UV-Vis spectroscopy?
6. It was indicated for ODN 1b and 1c, the absorbances at 260 nm obtained from the HPLC peak area and the UV spectrometer were quite different (1b is 12.16 and 11.20, while 1c is 16.92 and 15.40, respectively). The authors claimed it was possibly due to the loss between the transfer from HPLC eluent to UV measurements. What is the specific reason for the loss? Besides, does it mean the concentration of targeted ODNs obtained from the peak area would be different from the real final concentration since there was a loss in the process of solvent drying or centrifuge evaporation? How big the difference would be, and which result people should rely on?
7.Proper references need to be added to the introduction section.
8. The word “intuitively” from the last paragraph of the result section is not proper for a scientific article. Is there any specific reason or evidence that the fractions may not be the corresponding ones if without removal of the eluent collection tube?
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DETERMINATION OF OPTICAL DENSITY (OD) OF OLIGODEOXYNUCLEOTIDE FROM HPLC PEAK AREA
Review round: 2
Reviewer: 1

Output:
Basic reporting: After the revision, the manuscript can be accepted now.
Experimental design: After the revision, the manuscript can be accepted now.
Validity of the findings: After the revision, the manuscript can be accepted now.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DETERMINATION OF OPTICAL DENSITY (OD) OF OLIGODEOXYNUCLEOTIDE FROM HPLC PEAK AREA
Review round: 2
Reviewer: 2

Output:
Basic reporting: Clear and unambiguous, professional English used throughout
Experimental design: its properly designed and executed
Validity of the findings: Presented data supports their conclusions
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS OF PROTEIN-HEPARIN INTERACTIONS USING A PORTABLE SPR INSTRUMENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: This organization of the paper is good. The manuscript has been written very clearly in a very unambiguous and professional language. The figures have been appropriately described and labelled and relevant to the content of the article. Sufficient background information has been provided in the literature.
Experimental design: The methodology has been described in sufficient details with thorough investigation and well defined research question.
Validity of the findings: The results have been depicted convincingly with a well stated conclusion.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS OF PROTEIN-HEPARIN INTERACTIONS USING A PORTABLE SPR INSTRUMENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: English is clear
Literature sufficient
Articles structure generally OK, except that the authors go back & forth in citing Figures, particularly Figure 3-6
Experimental design: Original research, well described, relevant and meaningful
Validity of the findings: Supplemental/Raw data was not provided
Replicate information was not clear
Conclusions were clear
Additional comments: In this study, the authors have attempted to provide an optimized method/workflow for passivation and functionalization of a portable SPR instrument to be used in the field. Specifically, the authors use biotin-OEG::OEG in combination with streptavidin to generate the capture surface. The authors then provide examples of how it could be potentially applied in real-world scenarios.
Overall, the experimental design is simple, and the study appears convincing. The study could be published given the following minor comments are addressed:

Did the authors performed replicate injections and only the representative data were provided?

Did the authors test percentage of biotin-OEG over >1%? If yes, could the data be shown? If not, why not?

The authors optimized/prepared surfaces using PBS as the running buffer but then switched to PBST for their FGF2 measurements. Is there a reason for it? If measuring FGF2 in PBST is important, I suggest that the surfaces be prepared in PBST as well.

Why the authors choose 50 nM GST-HS3ST3A1 for Figure 3B and not 100 nM as they did in Figure 4B? This is important for a proper comparison between streptavidin/OEG and heparin binding efficiencies.

In Figure 3C, a side-by-side comparison of SDS/HCl and NaCl/HCl will be useful for comparison.

What is the motivation for using 0.25% SDS in Figure 3C? The authors should explain this in the text as it appears as a surprise!

In Figure 3C, Ch_D dropped below baseline after HCl, why?


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS OF PROTEIN-HEPARIN INTERACTIONS USING A PORTABLE SPR INSTRUMENT
Review round: 1
Reviewer: 3

Output:
Basic reporting: This paper described the assembly of a surface of thiolated oleyl ethylene glycol/biotin oleyl ethylene glycol and its functionalisation with streptavidin and reducing end biotinylated heparin for a SPR instrument. Two examples of the analysis of heparin-binding proteins are presented. Overall the structure of the paper is clear, while a number of clarification and modifications are necessary.

The authors don't cite enough references to claim the necessary of using heparin of this research.

The flow rate of the running buffer is 500μL/min, while it is too fast for molecular reaction and cost a lot of reaction reagent in the SPR experiment The authors should explain the reason for choosing this flow rate which is always lower than 50 μL/min in other common SPR instrument.

Why two regeneration steps were conducted, and usually acid or alkali solution can achieve one-step regeneration. It should be more clearly discussed.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: no comment


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS OF PROTEIN-HEPARIN INTERACTIONS USING A PORTABLE SPR INSTRUMENT
Review round: 2
Reviewer: 1

Output:
Basic reporting: No negative remarks
Experimental design: No negative remarks
Validity of the findings: No negative remarks
Additional comments: No negative remarks


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DEVELOPMENT AND VALIDATION OF UV CHROMATOGRAPHIC METHOD FOR QUANTIFICATION OF COPPER AND COPPER NANOPARTICLES IN DIFFERENT MATRICES AND PHARMACEUTICAL PRODUCTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The work concerns the quantification of copper in biological matrices and pharmaceutical preparations. The RP-HPLC method with DAD detection was selected for analysis. The principle of detection is based on the formation of a complex of copper with citric acid and EDTA as known complexing agents. My doubts are raised by the lack of copper specifications, and although the article shows that the authors denote copper in a metallic form and in the form of nanoparticles, it is doubtful, because they use nitric acid for dissolution. No confirmation of the presence of nanoparticles in the tested matrices, e.g. using TEM or SEM. In my opinion, the authors actually mean Cu (II) and therefore in ionic form. The manuscript should be reworded appropriately and accurately name the form they denote. Validation meets the criteria and it is a pity to waste this effort. I don't really understand the choice of method. In fact, simple colorimetry would suffice in this case. What are the benefits of using HPLC which is a separation method as there is only one analyte? Authors should justify their choice.
There is also no comparison of the precision and accuracy of the method with the determinations described so far. Therefore, it is not known what benefits result from the methodology used.
It is difficult to recommend this article for publication in this form.
Fig. 2 - A peak that needs to be identified appears in the chromatogram.
Fig.3- There are actually two unresolved peaks in this chromatogram so the quantification is questionable.
No UV-VIs spectrum of the analyte determined.
Experimental design: Experimental design hould be improved.
Validity of the findings: It should be improved.
Additional comments: see above


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DEVELOPMENT AND VALIDATION OF UV CHROMATOGRAPHIC METHOD FOR QUANTIFICATION OF COPPER AND COPPER NANOPARTICLES IN DIFFERENT MATRICES AND PHARMACEUTICAL PRODUCTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: English language used is not professional and requires extensive editing with the help of a colleague proficient in English. Some errors are listed below:

Line 209: reference is not appropriate
Abstract:
•	This following sentence is not clear to understand:  “The use of citric acid as a green chelating agent and extraction time with pH adjustment afforded high Cu recovery in biological samples.”
•	Also correct for appropriate symbols in” The precision RSD% was ˂ 2.”

Title:
•	Line 2: Use “copper nanoparticles” instead of “nano copper”
Text:
•	et al., needs to be in italics in all references
•	line 54: ionizing agents and diagnostic imaging
•	line 57 and 60: inaccurate sentence structure
•	line 84: detectors
•	line 86: This study presents a method (this is not a review)
•	line 219: Cu1+
•	line 239: RP HPLC is used to quantify copper and Cu NPs via potent
•	line 211-213: inaccurate sentence structure
Experimental design: A reverse phase HPLC method to quantify copper and copper nanoparticles is reported in this study. However, this is not a novel method. Along with the references provided, SiELC website has a page with similar method using a different column is published on its website : https://www.sielc.com/Application-HPLC-UV-Analysis-of-Copper-Ions-in-Salt-Mixture.html. The authors have used an existing method and validated its use with different biopharmaceutical samples.
Validity of the findings: The validation of the method was executed and data presented accordingly.
Additional comments: Editing is required to accurately describe the results and scope of this study.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DEVELOPMENT AND VALIDATION OF UV CHROMATOGRAPHIC METHOD FOR QUANTIFICATION OF COPPER AND COPPER NANOPARTICLES IN DIFFERENT MATRICES AND PHARMACEUTICAL PRODUCTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The manuscript has been edited appropriately to address all the reviewer's comments. The manuscript is now more polished and ready for publication.
Some of the references have the journal name in italics and in other the journal name is not italicized.
Make all the references consistent in formatting.
Experimental design: The experimental design is clear to understand
Validity of the findings: The results are based on the data obtained and discussed accurately.
Additional comments: Publish after editing the references section


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: LIPID EXTRACTION FROM MICROALGAE USING PURE CAPROLACTAM-BASED IONIC LIQUIDS AND WITH ORGANIC CO-SOLVENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors here describe a protocol of extracting lipids from wet and dried Spirulina platensis microalgae using ionic liquids. This is indeed an interesting work towards a step to follow green methodology for finding resources of biofuels. The manuscript may be published in the PeerJ Analytical Chemistry after considering the points mentioned in the detailed report.
Experimental design: In some cases, the authors have mentioned the experimental procedure to be found in literatures, such as, 'Soxhlet extraction'. Authors may consider to provide a brief description of all such methods to help the readers to immediately have an idea while going through the paper.
Validity of the findings: The results are interesting which are validated by reproducible data set.
Additional comments: 1. The authors may clarify if the ionic liquids (ILs) used in the study are the room temperature ILs. Mentioning their melting point is important as a reader may find different uses of these molecules.

2. All the ILs have same cation but differing in their anions. The different effectiveness of extracting lipids seems to depend on the type of anions. Why so?

3. The authors primarily have described their extracted lipids to be used for biofuel. Are they suitable for preparing model membrane, such as, vesicles? A discussion may be added to extend the uses of the lipids.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: LIPID EXTRACTION FROM MICROALGAE USING PURE CAPROLACTAM-BASED IONIC LIQUIDS AND WITH ORGANIC CO-SOLVENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: Species names should be properly mentioned (following the guidelines and put in italic, etc).
Furthermore, I suggest the authors not to overgeneralize the statement from literature and I invite the authors to perform critical reflections on the results.
Experimental design: The water content was unfortunatley not reported, thus the different ionic liquids are not directly comparable.
I especially disagree with the method to determine the yield, which I address several times in the annotated PDF. This could result in misleading interpretation, serious claims and wrong conclusion.
Validity of the findings: Due to the improper method (described above), the authors obtained peculiar data, which seems to be 'too good to be true'. A critical reflection on the obtained results is not observed from the report.
Replicates or standard deviation in some sections were not reported.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: LIPID EXTRACTION FROM MICROALGAE USING PURE CAPROLACTAM-BASED IONIC LIQUIDS AND WITH ORGANIC CO-SOLVENT
Review round: 2
Reviewer: 1

Output:
Basic reporting: Done in first review
Experimental design: Good
Validity of the findings: During the first review, there were multiple comments and suggested correction. The revised manuscript, the authors has taken care of all of them. Now the manuscript is ready to publish.
Additional comments: During the first review, there were multiple comments and suggested correction. The revised manuscript, the authors has taken care of all of them. Now the manuscript is ready to publish.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EXTENDED CHARACTERIZATION OF PETROLEUM AROMATICS USING OFF-LINE LC-GC-MS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is an interesting paper describing a new selective and efficient method for high-resolution fractionation of complex mixtures of petroleum aromatics which are contained in crude oils in a wide range of concentrations. The method is based on semi-preparative liquid chromatography and subsequent GC-MS analysis of collected fractions. The method enables not only an efficient isolation of the more abundant aromatics, but also the separation, isolation and preconcentration of trace aromatic components. This significantly increases the number of aromatic species which can be identified in crude oil samples compared to traditional methods like SPE-GC-MS or comprehensive GCxGC.

The manuscript is written in professional, unambiguous language. The article includes a sufficient introduction and the relevant prior literature is appropriately referenced. I consider the paper suitable for publication after some corrections in order to improve its quality. Specific remarks are listed below.
Experimental design: The subject is well within the scope of Peer J An. Chem and the manuscript structure conforms to PeerJ standards.

The method and all experiments are clearly described and the method is properly validated. The advantages and disadvantages (e.g. relatively long fractionation time) of the method are pointed out and well documented. The presented results confirm that the method can serve as a complementary tool to traditional sample preparation techniques or comprehensive GCxGC. HPLC fractionation of 14 crude oils with subsequent GC-MS analysis demonstrated the method suitability for identification of trace aromatics including the tentative identification of several compounds not observed by comparable routinely used methods.
Validity of the findings: All relevant data, on which the conclusions are based, are provided in tabulated (three tables) or graphical form (ten figures). The conclusions are safe and well supported by experimental results.
Additional comments: Specific remarks:

1.	Experimental results are presented in three tables but two of them (Tables 1 and 2) are not mentioned anywhere in the text relevant for Table 1 (lines 216 to 230, and for Table 2 (lines 239 to 242).

2.	There is a discrepancy between the figure numbers (Figures 5–9) listed in related manuscript paragraphs and the numbers and content of attached figures:
line 244: Figure 4 should be corrected to Figure 5
line 245: Figure 5 should be corrected to Figure 6
line 274: Figure 6 should be corrected to Figure 7
lines 301 and 328: Figure 7 should be corrected to Figure 8
line 312: Figure 8 should be corrected to Figure 9
Consequently, figure number should be corrected also in the Supplemental file:
Figure_5_Full_chromatograms to Figure_6_Full_chromatograms

3.	In Table 3 title GCxGC should be added. The first sentence should read: Comparison of the number of tentatively identified species in SPE-GC and GCxGC versus LC-GC.

4.	Figure 1: SPE enables separation of saturates and aromatics (not saturates and saturates).

5.	Line 72: corect (M. Pampanin & O. Sydnes, 2017) to (Pampanin & Sydnes, 2017)

6.	Lines 145 and 146: correct µM to µm

7.	Line 236: correct BTEX-type monoaromatics (benzene, toluene, xylene) to BTEX-type monoaromatics (benzene, toluene, ethylbenzene, xylenes)

8.	Line 244: delete the word secondary

9.	Line 270: correct (Countryman, Kelly & Garriques) to (Countryman, Kelly & Garriques, 2005)

10.	Lines314 and 316: abbreviation SPE/LC-GC is confusing; I would suggest writing SPE-GC and LC-GC instead.

11.	References:
Line 408: correct Countryman S, Kelly K, Garriques M. to Countryman S, Kelly K, Garriques M. 2005.

12.	Line 410: correct Davis JM, Giddings JCalvin. to Davis JM, Giddings JC.

13.	Line 442: correct M. Pampanin D, O. Sydnes M to Pampanin DM, Sydnes MO.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EXTENDED CHARACTERIZATION OF PETROLEUM AROMATICS USING OFF-LINE LC-GC-MS
Review round: 1
Reviewer: 2

Output:
Basic reporting: Tables 1 and 2 are not referenced in the body of the text

Figure 9 is not referenced in the text

The supplemental figure 6 has a file name denoting figure 5

Line 181 - there is an erroneous "29,30"

Is the Python script you used for the data processing available?

The figure legends need more information. As they stand, the figures require far too much reader intepretation to guess what they are showing.
Experimental design: An interesting premise, a DB-5 column was used for the GC portion of the experiment. Why was this preferred to the more routinely used DB-1 column? Or even a PAH column as this separates all isomers? Would using a different column have an impact on the identification on isolated chemicals?

You used one type of SPE sorbent for the SPE experimentation. Were other SPE sorbents tested and did these have any affect on the extraction?
Validity of the findings: no comment
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EXTENDED CHARACTERIZATION OF PETROLEUM AROMATICS USING OFF-LINE LC-GC-MS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors have successfully revised the original manuscript by taking into account all the reviewers' suggestions and comments. In the accompanying letter they clearly explained all the changes made in the manuscript text, tables, and figures. They have also addressed and satisfactorily commented some issues raised by the reviewers such as the choice of the GC column and SPE sorbent. In my opinion this manuscript can be accepted for publication in the present form.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: No comment.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DETECTION-CONFIRMATION-STANDARDIZATION-QUANTIFICATION: A NOVEL METHOD FOR THE QUANTITATIVE ANALYSIS OF ACTIVE COMPONENTS IN TRADITIONAL CHINESE MEDICINES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The English language should be improved to ensure that an international audience can clearly understand your text. Some examples where the language could be improved include lines 292, 322 – the word ‘reliability’ makes comprehension difficult, suggest replacing with’dependency’.

Lines 91- Is it necessary the word ‘directly’ because the preparative HPLC is different from the above-mentioned quantitative HPLC.

Please check:
Lines 169- The signals for the H-15 of cryptotanshinone (4.884 ppm, t, 1H), 1H or 2H？

Lines 218- times of 17.2, 19.0, and 26.1 min, It doesn't match the coordinates in Fig. 2 (A2 and B2);

Lines 304- the doublets in 2b and 2f (left side) belonged to 305 tanshinone I, the doublet in 2f (right side) belongs to tanshinone IIA, there is a contradiction with the annotation of Figure 7

Lines 309- in 1c and 2f (right side) (Fig. 7)  1c or 1b?

Lines 315- tanshinone IIA calculated from the NMR spectra of the MCRS with five different concentrations, five or three?
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: DETECTION-CONFIRMATION-STANDARDIZATION-QUANTIFICATION: A NOVEL METHOD FOR THE QUANTITATIVE ANALYSIS OF ACTIVE COMPONENTS IN TRADITIONAL CHINESE MEDICINES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper reported a novel strategy for the quantitative analysis of active components in traditional Chinese medicines, without individual reference standard. The proposed strategy was used for the quantitative analysis of cryptotanshinone, tanshinone I, and tanshinone IIA in Danshen Samples. I think it is worthy research work and has potentials in traditional Chinese medicines. However, there are still some details and issues as follows that need further clarification and discussion. Thus, I recommend the acceptance of this work after major revisions.

1.Accurate quantitative analysis of target analytes in complex matrices is very important, so, quantitative results of cryptotanshinone, tanshinone I, and tanshinone IIA in Danshen samples should be provided in the text.
2.The NMR spectra of complex mixtures often exhibit severe peak overlap, which can affect the accuracy of the quantity of analyte. The separation degree of the peaks in 1b, 2f (Figure 7) and 4a should be calculated.
3.The experiment process seems a little complicated, whether this will cause the loss of the target analyte?
4.The results of the proposed method should be verified by using individual reference standards and evaluated by using statistical methods.
Experimental design: sss
Validity of the findings: sss
Additional comments: The paper reported a novel strategy for the quantitative analysis of active components in traditional Chinese medicines, without individual reference standard. The proposed strategy was used for the quantitative analysis of cryptotanshinone, tanshinone I, and tanshinone IIA in Danshen Samples. I think it is worthy research work and has potentials in traditional Chinese medicines. However, there are still some details and issues as follows that need further clarification and discussion. Thus, I recommend the acceptance of this work after major revisions.

1.Accurate quantitative analysis of target analytes in complex matrices is very important, so, quantitative results of cryptotanshinone, tanshinone I, and tanshinone IIA in Danshen samples should be provided in the text.
2.The NMR spectra of complex mixtures often exhibit severe peak overlap, which can affect the accuracy of the quantity of analyte. The separation degree of the peaks in 1b, 2f (Figure 7) and 4a should be calculated.
3.The experiment process seems a little complicated, whether this will cause the loss of the target analyte?
4.The results of the proposed method should be verified by using individual reference standards and evaluated by using statistical methods.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NON-TARGET SCREENING OF ORGANIC COMPOUNDS IN OFFSHORE PRODUCED WATER BY GC×GC-MS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The basic reporting is good and the manuscript is structured well. The authors should double check for typos (although they are only minor) - for example, line 25 "furtthermore" should be corrected to "furthermore."
Experimental design: While this is an interesting manuscript and good concept, I think the experimental design is flawed. From my understanding of the manuscript, you are using the NIST library to identify compounds. However, you only use TMS derivatisation to derivatise your samples for running on the GCxGC-MS. This means you are missing many samples or will be unable to identify them using the NIST library. The prime example is fatty acids. Those greater than C12 in length, in particular. While they can and are routinely TMS derivatised, this is not the gold standard for derivatisation and, therefore, much of this data is missing/lacking from the NIST library. These compounds are rountinely derivatised to make methyl esters and it is the MS data from these that are found in the NIST library. For such as scoping study, it would be better to use LC-MS as derivatisation isn't (always) needed.
Validity of the findings: Unfortunately, not all supplemental data was available to me. There was only jpeg images of 2D scans with no figure legends. Therefore, it was difficult to assess many of the points in the manuscript where it said to look at supplementary data.
Additional comments: This is a very interesting study and does show that there is more compounds than we think in produced water.

Line 69 - reference requires a date
Line 76 - "(LLE)" needs to be placed after the word "extraction"
Line 130 - why was 25 ppm chosen? This is not made clear in the manuscript
Line 130 - this information is not in the supplemental data
Line 132 - it says the synthetic water was extracted 4 times, but it doesn't say how. More detailed is required, even if it is just to say "using the above method."
Line 158 - the script isn't in the supplementary information provided
Line 186 - there is no need to say "1.5 mL of n-hexane" because this is actually a false number, because in the actual protocol you are summarising 1 mL is transferred to be derivatised

Figure legends need to be improved to make it easier for the reader to understand what is happening in the figure and they should be able to stand alone of the text.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NON-TARGET SCREENING OF ORGANIC COMPOUNDS IN OFFSHORE PRODUCED WATER BY GC×GC-MS
Review round: 1
Reviewer: 2

Output:
Basic reporting: See comments grouped below.
Experimental design: See comments grouped below.
Validity of the findings: See comments grouped below.
Additional comments: The author study the important problem of produced water, released into the environment after treatment, using GCxGC-MS tools.

- lines 80-91: the authors just mentioned GCxGC, now they write about GC and LC. The authors should clarify if this text (lines 80-91) is about unidimensional or multidimensional GC and LC.
- lines 107-114: were the samples more or less regularly spaced (in time) during this period? Were there some replicate samples?
- line 112: might the plastic from the bottles have affected the results (partitioning of hydrophobic molecules to the plastic?)
- line 116: "extracted in three experimental replicates". Maybe clarify if three "analysis replicates" or "aliquot replicates" are meant. The use of the word "experimental" feels a bit unclear it seems. (Or was there always three replicate samples collected on the platform at each time point?) (If what is described here is the same as at lines 180-181, the text at lines 180-181 is more clear.)
- lines 107-114: it would help to specify at what stage in the process the samples were taken (upon entering the test separator? upon leaving it (effluent)?). What were the different steps of processing before the sample was taken, and are there further steps that are performed on the platform before release into the sea? (It is basically somehow unclear if the sample corresponds to raw water coming out of the well, partly-treated water, or treated water ready for release/disposal). It is also unclear what is meant with "test separator".
additionally: it would help to specify the sample volume.
- line 177 (Figure 1): could the authors specify in the caption to that figure (or in the text) which ones of the bottles on Figure 1 were rejected? Also, could the authors clarify if the nine samples mentioned at line 107 were the number of samples that passed this test, or if (how many?) of these nine samples were excluded because of a visible oil layer.
- line 177 (Figure 1): the caption to Figure 1, it is said that these are the samples "as received". Are these really plastic bottles (line 112)? (I may have a wrong feeling, it looks like glass bottle with a plastic cap...)
- line 179 (Figure 2): the caption to Figure 2 is unclear. What are the two panels?
- lines 183-184: what about small aromatics like benzene, toluene, etc.? Are these not expected to be present? Are these lost (volatilized) during the produced water treatment on the platform?
- lines 195-197: is retention time shifting also a problem (peak deformation, leading to shifting of the peak apex, which is sometime taken as peak position). Or are retention times irrelevant in the authors's analysis? (Line 257 mentions Kovats retention indices, and I think these must be calculated from retention times (?)).
- lines 204-205: what is "synthetic produced water"? clarify the water used..
- line 207: "peak volumes": the authors may like to clarify how peak volumes were determined. This may be relatively software-independent for such well-separated peaks, but in some cases peak volumes may be very dependent on the approach use (e.g. software, baseline approach, etc., see Samanipour et al., 2015).
- line 210: I believe that the authors mean "relative standard", not "relative-standard".
- line 211: "the model compounds": do the author mean "peak volumes" of these compounds(?)
- line 247 (and throughout): "experimental replicates": as written above, is "experimental" really the best choice of word?
- line 253: this is shown in Figure 4, not Figure 3. (By the way, for figure 3, up to the authors, but it might help to annotate the chromatogram to show the positions of a few compounds known to the authors--it seems they describe some in the text. I am personally more familiar with a different set of columns and with crude oil chromatograms, and I'd love to be provided such information on that figure to guide my understanding... (this is a mere suggestion, maybe resulting from my ignorance...))
- around line 294: it would help to provide additional explanation of the two contrasting studies. Was the study by Sorensen et al. for produced water containing droplets? Was the sample at a different stage of processing of the produced water? Was there any difference in the analysis method that can explain that?
- line 318: relative concentrations: it might be appropriate for the authors to provide a quantitative statement? ("high" is very unspecific...)
- line 328: "from the same well and only three days apart,": there is a lack of detail about the sampling times in the methods, which limit the ability to appreciate this statement...
- line 129: "sampling and test separator status" is unclear.
- supplementary files: the chromatograms are given file names that probably pertain to sample ID, but this info is (it seems) not available to the author. Maybe add a table providing understanding for the readers.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NON-TARGET SCREENING OF ORGANIC COMPOUNDS IN OFFSHORE PRODUCED WATER BY GC×GC-MS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: I think that the reasoning for the experimental design has been addressed adequately.
Validity of the findings: No comment
Additional comments: A very interesting paper. Thank you for clarifying the experimental design and making the suggested changes.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF HETEROZYGOUS HEMOGLOBIN E ON SIX COMMERCIAL METHODS FOR HEMOGLOBIN A1C MEASUREMENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: General comments:  The authors examine interference from HbAE using six commercial HbA1c methods.  They compared results with  an LC-MS/MS.  There were no statistically difference results for HbAE for 4 of these methods.  However, for two methods (VII Turbo 2.0 and Cobas b101) there were statistically significant differences despite manufacturer’s claim s to the contrary.  This is an important paper that adds to the information we have on variant interference with HbA1c methods. This is especially important since there are discordant data in the literature for the VII T2.0 on this interference.

Specific comments:

1.	The authors should clarify up front whether or not the LC-MS/MS method they use as a reference method is part of the IFCC network.  If not, they need to show some data to indicate comparability with the IFCC network, which is monitored on a regular basis.
2.	Line 126-127: 36% seems a bit high.  I calculate ~31% of participants.
3.	Table3: it would be helpful to highlight those over 6%.
4.	Figure 1: label x-axes
5.	Figure 1: would it be possible to include the data points so that the distribution of points can be seen.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF HETEROZYGOUS HEMOGLOBIN E ON SIX COMMERCIAL METHODS FOR HEMOGLOBIN A1C MEASUREMENT
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors wrote a clear manuscript with acceptable English and well-organized structure. However, the title of x-axis is required in fugure 1.
Experimental design: If possible, the average of blood sugar levels from both EA and A2A groups should provide in the manuscript. The authors should specify the type of patients such as diabetic patients for replacing patients in the study subjects.
Validity of the findings: No comment
Additional comments: The authors of the manuscript report the impact of Hb E heterozygote on 6 commercial methods for Hb A1c measurement and IDMS was used as reference laboratory method. EA and A2A  Hb types with different levels of Hb A1c were used to determined Hb A1c levels by 6 selected commercial methods. The manuscript is well written. However, there are some points shoud be addressed before publication.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF HETEROZYGOUS HEMOGLOBIN E ON SIX COMMERCIAL METHODS FOR HEMOGLOBIN A1C MEASUREMENT
Review round: 2
Reviewer: 1

Output:
Basic reporting: acceptable
Experimental design: acceptable
Validity of the findings: acceptable
Additional comments: the authors have addressed all of my concerns except that I still think that the X-axes in figure 1 should be labeled within the figure itself.  I will leave this up to the editors.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IMPACT OF HETEROZYGOUS HEMOGLOBIN E ON SIX COMMERCIAL METHODS FOR HEMOGLOBIN A1C MEASUREMENT
Review round: 2
Reviewer: 2

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: According to the objective of this study, this study examined the impact of heterozygous HbE on HbA1c measurements using six commonly used commercial methods and a modified isotope dilution mass spectrometry (IDMS) reference method, the authors need to revise the conclusions for linking to original research question.
Additional comments: There is minor revision of the manuscript to be considered before further publication.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SCREENING OF 49 ANTIBIOTIC RESIDUES IN AQUATIC PRODUCTS USING MODIFIED QUECHERS SAMPLE PREPARATION AND UPLC-QTOFMS ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. The English language should be improved, such as line 49, lines 120-121, lines 129-130；
2.  The format of references should meet the requirements of the Journal；
Experimental design: No comment
Validity of the findings: 1. Line 20, RSDs were between 1.6% and 14.0% in different samples were inconsistent with the datas given in Table 4, should check it；
2. Figure 1a-1c, the title of the Y-axis were unclear；
3, form Figure 3, the effects of C18 and PSA-C18 nearly the same, even in grass Carp and Scylla serrata higher average recoveries were achieved using C18, why select PSA-C18 as the sorbents？
4, In Table 1, the structure of target compounds was unclear.
Additional comments: The reviewed manuscript describes the development and application of a new analytical method for determining antibiotic residues in aquatic products by an optimised QuEChERS method and UPLC-QToFMS determination. The information in systematically well described and organised. The design of experiments for the QuEChERS and cleanup steps is well organized. The interest of the manuscript is high and it could be published in PeerJ after minor modifications.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SCREENING OF 49 ANTIBIOTIC RESIDUES IN AQUATIC PRODUCTS USING MODIFIED QUECHERS SAMPLE PREPARATION AND UPLC-QTOFMS ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The figure quality should be improved, the reference format should be consistent with the journal.
Experimental design: 1. May be modified as “Screening of 49 antibiotic residues in aquatic products using modified QuEChERS sample preparation procedure and UPLC-QToFMS analysis
2. Abstract, Line 19 to 20, The recoveries of target antibiotics at the different spiked levels ranged from 60.6% to 117.9% …, 60.6% should be changed to 60.2% from table 4.
3. Introduction, a small description of regulatory limits of pesticide residues in China/EU/Codex/USA may be given to justify the LOQ and LOD requirement of the method.
4. Materials and methods, a separate section may be given for a brief description on different treatments used with respect to each optimization parameters may be given for the clarity of readers.
Validity of the findings: 1. The spiked concentration selected is 10 μg/L, 50 μg/L, and 100 μg/L, however, as the LOQ of the most of the pesticides are 1.0 μg/L, first level recovery should be of 1.0 μg/L. Further ensure that, the highest calibration level should be 200 μg/L or more.
2. In a multi-residue method, the crux of the method is the optimization of sample preparation, especially in a matrix like aquatic products. Hence, more emphasis should be given for sample preparation optimization, matrix effect minimization, clean-up optimization, sample size optimization, sample preprocessing optimization and further validation.
Additional comments: The authors presented a good piece of work on analysis of multi-residue antibiotics in aquatic products. The manuscripts used sound analytical technology (UPLC-QToFMS) for antibiotic residue analysis. However, some of the gaps detailed below should be addressed carefully before publishing the paper in PeerJ.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A SIMPLE SPECTROPHOTOMETRIC EVALUATION METHOD FOR THE HYDROPHOBIC ANTICANCER DRUG PACLITAXEL
Review round: 1
Reviewer: 1

Output:
Basic reporting: I find a report about the spectrophotometric determination of PTX (International Journal of Advances in Pharmaceutical Sciences 2(1):29-32, 2011). please explain about the advantage of your work than this report.
Experimental design: It is good.
Validity of the findings: Some analytical parameters of the proposed method such as limit of detection and quantification are not calculated.
Additional comments: The manuscript well designed. The below references about the spectrophotomeric and HPLC determinations can be added to the introduction.
1)	Heydari, R., Hosseini, M., Alimoradi, M., Zarabi, S. A simple method for simultaneous spectrophotometric determination of brilliant blue FCF and sunset yellow FCF in food samples after cloud point extraction. Journal of the Chemical Society of Pakistan, 38, 2016, 438-445
2)	Heydari, R., Hosseini, M., Zarabi, S. A simple method for determination of carmine in food samples based on cloud point extraction and spectrophotometric detection. Spectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy, 150, 2015, 786-791
3)	R. Heydaria, F. Bastami, M. Hosseini, M. Alimoradi, Simultaneous determination of Tropaeolin O and brilliant blue in food samples after cloud point extraction. Iranian Chemical Communication, 5, 2017, 242-251
4)	Heydari, R., Shamsipur, M., Naleini, N. Simultaneous determination of EDTA, sorbic acid, and diclofenac sodium in pharmaceutical preparations using high-performance liquid chromatography. AAPS PharmSciTech, 14, 2013, 764-769


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A SIMPLE SPECTROPHOTOMETRIC EVALUATION METHOD FOR THE HYDROPHOBIC ANTICANCER DRUG PACLITAXEL
Review round: 1
Reviewer: 2

Output:
Basic reporting: In some parts of the article English must be improve and some sentences should be rewritten to be more understandable.
I consider some more references must be added.
Experimental design: Some comments and suggestions:

Introduction
In my opinion, the introduction should be more complete. For example, it can be completed taking into account these considerations:
- Authors can cite some more new references. In line 56 you can cite some more current article about the determination of paclitaxel by HPLC.
- I your article you demonstrate that PTX can be quantified spectrophotometrically. However this method will be only useful when one analyte is present in the sample, for example in drugs For the determination of PTX in plasma you will need HPLC. In my opinion in the introduction you should clarify this point.
- There are other articles where spectrophotometric methods have been proposed to quantify PTX. You should mention them in the introduction and underline the advantages of your method over these.
Figure 1 has a lot of test to be a Figure. The test should be remove from the Figure and placed in the test.

Materials and methods
- In line 114 you mention, “The concentration of PTX in each test sample was subsequently determined using the same conditions as those used to generate the standard calibration curve”, however you have not mentioned before what the test samples are. I think you should explain before what you will use as test samples.
- Line 156 and 157 describe some of the results found: “there was no significant difference between the spectrometry and HPLC values”, in my opinion these comment must be placed in Results.

Results
- In Figure 2 the title of the X Axis should be PTX concentration (µg/mL) and clarify that 40 % , 50 %, 60 % is the % of methanol. Moreover, you can use different colours for a better differentiation between the calibrations curves. Moreover, it would be useful if you add the slope and intercept value in this figure or at least in the table related to this figure.

- In “Calibration curve” you should clarify if you have used methanol and the different % of methanol (in each case) as blank or not. Because the absorbance of the different dissolvent should be taken into account. This analyte can have different absorbance in the different solvents but also the solvents themselves may have different absorbance. I consider this point should be clarify in the article.
. I also miss a figure with the spectrum of PTX in different solvents. Could it be added in a new figure?
- In line 177 and 178 as it is written it is not clear if you have used several samples of PTX-saturated saline solution with different % of methanol, if you have used the different calibration curves prepared in these solvents, or both things to be in the same solvent the sample and the calibration curve. I think this sentence should be rewritten.
- In Figure 3, the title of the X Axis should be MeOH concentration (%)
- Line 193: the regression curve should be express with the slope and intercept deviation.
- Figure 4A: PTX (+) or PTX (-) must be replace by dilution with or without PTX in order to be more understandable.

Finally, I consider that if you are proposing a new analytical method, you should provide some analytical parameters such as the detection and quantification limit.
Validity of the findings: Authors must clarify the novelty of this methods and compare it with other spectrophotometric methods previously published.
Additional comments: This paper describe a simple study with basic experiments for the determination of paclitaxel. Some important parameters have been appropriately optimized and the study was well lead. However, some important issues must be clarify and improve in order to make the article more completed and understandable.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A SIMPLE SPECTROPHOTOMETRIC EVALUATION METHOD FOR THE HYDROPHOBIC ANTICANCER DRUG PACLITAXEL
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The article is important and reports interesting results for accurately quantification of hydrophobic anticancer paclitaxel (PTX) concentrations. No scientific changes required.
Some minor recommendations:
1.	Line 56-57: Add more recent references to methods for quantification od PTX (maybe something from 2010-2019)
2.	The sensitivity of the method depends on the amount of organic solvent  (Fig. 2) please add to the text in the chapter „Calibration curve” the values of the sensitivity factor
3.	Table 1: Add quantitative concentration from spectrophotometry measurement


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A SIMPLE SPECTROPHOTOMETRIC EVALUATION METHOD FOR THE HYDROPHOBIC ANTICANCER DRUG PACLITAXEL
Review round: 2
Reviewer: 1

Output:
Basic reporting: ok
Experimental design: ok
Validity of the findings: ok
Additional comments: I consider this article can be accepted only considering these minor aspects.
- You have added the value of the limit of detection and quantification, however you should add which criterium you have used to calculate it (IUPAC, Clayton, Long and Winerfordner, etc)

- On the other hand, regression curve continue without be expressed with the corresponding deviation


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ESTABLISHING AN OPTIMIZED METHOD FOR THE SEPARATION OF LOW AND HIGH ABUNDANCE BLOOD PLASMA PROTEINS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Professional article structure, figures, tables. Raw data shared.
Experimental design: Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.
Validity of the findings: All underlying data have been provided; they are robust, statistically sound, & controlled.
Additional comments: The manuscript titled “Establishing an optimized method for the preparation of low and high abundance blood plasma proteins” from Henian Yang et al tested the efficiency and reproducibility of a method for optimal separation of low and high abundant proteins in blood plasma. The authors are attempting to describe an appropriate conditions to separate low and high abundance plasma proteins from three different aspects. And then an optimized method is presented.
In my opinion, there are some points should be addressed as following.
(1) Although this paper provided the detailed protocol for three methods to concentrate the eluted plasma fraction and the bound proteins, no analysis of their differences and similarities among them were mentioned. The characteristics among them should be systematically clarified, which made it inconvenient for readers to understand these methods. And in the part of “Results and Discussion”, the reason of why samples treated with TCA precipitation did not generate good quality 2-D gels should be properly explained.
(2) The result of protein quantification in this study is only analyzed by the method of BCA protein assay. If there is another way to proof the final result, it will be more convinced for the quality of this manuscript.
(3) In the use of the Seppro spin column and the ProteoExtract Albumin/IgG removal kit to clarify the reproducibility of abundant protein depletion, the parallel control should be designed among the same spin columns.
(4) The figures in this article need to be more organized and accessible. For example, if the lanes in figure1A are divided clearly, it will be easier to understand.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ESTABLISHING AN OPTIMIZED METHOD FOR THE SEPARATION OF LOW AND HIGH ABUNDANCE BLOOD PLASMA PROTEINS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The language is clear enough. Can be improved slightly in terms of grammar and flow. Intro and Background are clearly written. There are appropriate references in the literature. Figures are good quality. We failed to detect any issues with the images.
Experimental design: The research work is original to the best of my knowledge. Research question is meaningful and well defined but is not relevant owing to omission of LC-MS. The researchers identify a knowledge gap but failed to fill it completely. The investigation is of good technical and ethical standard. The methods have been sufficiently described.
Validity of the findings: All underlying data provided. No speculative statements encountered. Conclusions are well drawn and in line with experimental findings.
Additional comments: The authors are commended for a very useful article. They have provided detailed coverage of various commercial protein separation strategies. These even include repeated use performance of the same. We think this manuscript will definitely add to the field. However, some crucial concerns remain, as listed below.
After reading the manuscript, we suggest the title be appropriately modified. This reviewer feels preparation be replaced with separation in the title. The Abstract is quite confusing. This reviewer suggests adding 1-2 lines stating the best method/combination of methods for optimized preparation of low and high abundance plasma proteins.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ESTABLISHING AN OPTIMIZED METHOD FOR THE SEPARATION OF LOW AND HIGH ABUNDANCE BLOOD PLASMA PROTEINS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment.
Experimental design: No comment.
Validity of the findings: No comment.
Additional comments: In the revised manuscript, the authors addressed most of the major questions raised by Reviewers improving both the main structure and quality of the present paper. I have no further additional comments.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ESTABLISHING AN OPTIMIZED METHOD FOR THE SEPARATION OF LOW AND HIGH ABUNDANCE BLOOD PLASMA PROTEINS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The language is clear enough. Intro and Background are clearly written. There are appropriate references in the literature. Figures are good quality. We failed to detect any issues with the images.
Experimental design: The research work is original to the best of my knowledge. Research question is meaningful and well defined but is not relevant owing to omission of LC-MS. The researchers identify a knowledge gap but failed to fill it completely. The investigation is of good technical and ethical standard. The methods have been sufficiently described.
Validity of the findings: All underlying data provided. No speculative statements encountered. Conclusions are well drawn and in line with experimental findings.
Additional comments: The authors are commended for a very useful article. They have provided detailed coverage of various commercial protein separation strategies. These even include repeated use performance of the same. We think this manuscript will definitely add to the field. I am satisfied with the author responses and recommending for the acceptance.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: FINDING A RELATIONSHIP BETWEEN PHYSICOCHEMICAL CHARACTERISTICS AND IONIC COMPOSITION OF RIVER NWORIE, IMO STATE, NIGERIA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper is difficult to read and several aspects has to be reconsidered, from the experimental plan to the data interpretation.

References can  be improved by introducing more recent papers.

Experimental data are poor. No data regarding the methods and data quality.

Figures with low resolution and not very conclusive for the study.

English language has to be well improved.
Experimental design: It is a classical monitoring study of water quality.

The manuscript has a poor experimental plan. Less samples - only five - were considered for the study. No relevant conclusion can be draw.

Methods have been poor described.
Validity of the findings: There are no novelty in this study.

The data do not support a comprehensive study for establishing a relation between the physicochemical parameters and the ionic composition of water.
Additional comments: The title of manuscript in not really consistent with the text. In my opinion the paper is only a simple monitoring study of water quality in the River Nworie, Nigeria. It is a classical study analyzing specific quality indicators of water quality by standard methods. What is the novelty of the work?
The paper has many gaps and I cannot recommended for publication in its present form. The manuscript is difficult to read.
- The abstract has to be reconsidered, it has to highlight the main findings regarding the relationship between physicochemical characteristics and ionic composition of the Nworie River waters, as is stated in the title.
- The study was performed during the dry season period only on 5 samples.  Why only 5 samples were considered? Normally, a more comprehensive study has to be done to draw real conclusions. Therefore, the experimental plan has to be reconsidered.
- A detailed description of the anthropogenic activities along the river has to be performed.
- Data on materials and methods used is very poor.
- Figures has low resolution.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: FINDING A RELATIONSHIP BETWEEN PHYSICOCHEMICAL CHARACTERISTICS AND IONIC COMPOSITION OF RIVER NWORIE, IMO STATE, NIGERIA
Review round: 1
Reviewer: 2

Output:
Basic reporting: I strongly recommend the authors to edit the language errors in the current version of the manuscript. This will help the readers to follow and understand the observations and discussions presented in this article.
Please double-check the figure numbers in the main body, figure captions as well as in the actual figure are consistent and are accurate. For instance, in line#146 it is indicated that the “percentage of ions are presented in Figure 2” which is not correct.
Experimental design: Although the authors had emphasized the importance of studying the relationship between physicochemical properties and chemical properties, the focus of the current study is not stated explicitly in the introduction of the manuscript.
Validity of the findings: In the manuscript, I don’t find any data for UV-Vis absorbance as well as other measurements based on which the relationship analysis is done. It is very important to show such experimental results that allow the readers to understand the measurement and estimation procedures followed by the authors. Due to the lack of any such experimental data in the manuscript, I find it difficult to follow the discussion and verify the conclusions derived through this study.
Additional comments: In tables 2 and 3, I would suggest using “WHO permissible limit (2007)” instead of just “WHO (2007)”


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: FINDING A RELATIONSHIP BETWEEN PHYSICOCHEMICAL CHARACTERISTICS AND IONIC COMPOSITION OF RIVER NWORIE, IMO STATE, NIGERIA
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: No additional comments