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Reviewer Recommendation and Comments for Manuscript Number CARBPOL-D-13-02147

Monosaccharide composition of glycans based on Q-HSQC NMR

Original Submission
Arildo Jos Braz de Oliveira, Ph.D (Reviewer 1)

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Recommendation: Minor Revisions for acceptance Overall Manuscript Rating (1-100): 90

Manuscript Question(s): Scale Rating

Please rate on a scale of 1-3 whether the Highlights are a meaningful and accurate
representation of the article. 1 = Meaningful; 2 = Not Meaningful; 3 = Not Provided. [1-3] 1
For more information, see www.elsevier.com/highlights.

Reviewer Blind Comments to Author:

1. The paper describe an original solution to the problems encountered in terms of resolution when trying to do
the experiment with a mixture of monosaccharides by direct analysis by NMR. The paper is of good quality and
suitable subject to the scope of this journal and in general mode of presentation is adequate, but then does some
questions to the authors that must be answered.

2. Line 58 change "1. INTRODUCTION" to 1. Introduction and all situations always don't use capitals letter.

3. Line 99 change "2. EXPERIMENTAL" to 2. Materials and methods;

Line 102 describes how pH was adjusted.

4. Results and discussion: Line 185 to define what experiments had a good correlation; Line 187 Which
concentration of standards here (1 mg/ml or 5 mM?); Lines 252-253 "For all tested glycans, the correlation
between the NMR and the GC-MS-Filisetti-Cozzi protocol showed very similar results, giving robustness for
quantification by Q-HSQC" Analyze in a more consistent form, not only use the term very similar, but is based on
some parameter statistician (eg correlation coefficient, t-test, anova); Line 258 Please highlight the signs of
aldobiouronic acids in Figure 6a and remove the citatao for Figure 8a because it does not exist; Lines 253-255
Why TFA was used for hydrolysis of the samples and no H2SO4 or D2SO4 which as reported by Souza et al, 2013
(hydrolysis conditions of Seaman) are most effective for hydrolysis of very strong linkages in polysaccharides
containing acid groups as EPS and pineapple gum and at the same time it is mild toward the monomers?

5. Line 262 change "2. CONCLUSION" to 4. Conclusion; Line 275 Quantification is absolute or relative, because
it was not clear in the experimental part and the results are being analyzed for the relative concentrations
between monosaccharides or the concentration of each individual?

6. Figure Captions: Figure 1. In the item of materials and methods used in the temperature experiment was
described was 27 degrees Celsius and not 23 as written in the legend, by the standards of the journal would not
have to be in Kelvin? As the legend to be self-explanatory desirable have information about the solvent used and
the pH value in the analysis.

Reviewer Confidential Comments to Editor:

A note formatting of topics is not in accordance with the standards of the journal, but nothing that compromises
the paper.

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