High Performance Liquid Chromatography (HPLC) with Fluorescence Detection for Quantification of Steroids in Clinical, Pharmaceutical, and Environmental Samples: A Review

Round 1

Reviewer 1 Report

Overall this review paper is written well and has lots of useful information, and a minor revision is needed before it is decisioned for acceptance. 

• - L1 - HPLC should be expanded form, although it is widely known. 
  - L12 synthesized form
  - L13-14 - should be combined, please rewrite.
  - L17 - please write, and check the grammar mistakes.
  - L17- FLD should be expanded.
  - The abstract contains limited details about HPLC, especially information given in L15, please elaborate if possible. 
  - L41 - List out the diseases.
  - L42 - list out the techniques either in brackets or without them.
  - L59 - expand the MS and abbreviation.
  - L61 - quantitative in nature seems repetitively used. please avoid using the same phrases often.
  - Introduction is lack of why steroid needs to be quantified, what if steroid concentration increase or decrease could cause to? please report it.
  - Also include more details that relate to Fig 1 in terms of the derivatizing agent in the introduction part.
  - L98 - the author should provide a schematic diagram for direct and indirect methods.
  - L109 - will be? or "should be"?
  - L150 - check reference format
  - L152-154 - related to the discussion in L150? please provide a reference or rewrite to connect the previous sentence.
  - Fig 3 is not consistent with other figure presentations, please provide names for every chemical in the chemical reactions.
  - Author should provide detailed information along with several results support for sections 4 and 5 in a table.

Author Response

Please see the attachment

Author Response File: Author Response.docx

Reviewer 2 Report

Comments:

The manuscript with reference number molecules-1619371 entitled “HPLC with Fluorescence Detection for Quantification of Steroids in Clinical, Pharmaceutical, and Environmental Samples” summarizes HPLC-FLD methods reported in the literature for the quantification of steroids in clinical, pharmaceutical, environmental and food applications. The authors focus on the introduction of fluorophores into targeted steroids using various derivatization methods and discuss aspects related to analytical methodology, including sample preparation, derivatization reagents, and chromatographic conditions. There are still some conspicuous problems in the format. Therefore, this manuscript was recommended for publication after solving the following problems.

1. The Keywords “high performance liquid chromatography” “HPLC” are repetition, you should remove one.
2. Perhaps you should check the page numbers of the manuscript.
3. The beginning of a paragraph should be consistent
4. The format of references marked "77" at line 465 of the manuscript should be consistent with that of other sections.
5. The header “Table 1:……” “Table 2:……” and their contents should be rearrange so that they fit together rather than leaving a lot of white space.
6. Sentence at line 114 of the manuscript should be checked for completeness.

Author Response

Please see the attachment

Author Response File: Author Response.docx

Reviewer 3 Report

A very interesting article, where is evidenced the importance of HPLC analysis. After some minor revisions, the article should be considered to be published.

Minor revisions:

1. The authors should try to format table 1 and the respective legend in the same page. Instead of a blank page only with the legend of table 1.

 

2. In each figure, since figure 4, the name of the resultant product of chemical reaction must be indicated.

 

3. Please verify the letter and paragraph format into lines 431-433, it seems a little bit different.

 

4. The authors should try to format table 2 and the respective legend in the same page. Instead of a blank page only with the legend of table 2.

Author Response

Please see the attachment.

Author Response File: Author Response.docx