A Targeted UHPLC-MS/MS Method Validated for the Quantification of Ergot Alkaloids in Cereal-Based Baby Food from the Belgian Market
Round 1
Reviewer 1 Report
The authors have described a new developed, optimized and validated UHPLC-MS/MS method for the simultaneous determination of six ergot alkaloids in 49 cereal-based baby food from the Belgian market.
In my opinion the manuscript is quite innovative, it is well written, presented and provides details of a method that may become useful in laboratories for monitoring and quality control of food.
This manuscript can be accepted after minor revision:
line 9: ,,-1” should be superscript
line 99: v/v should be in parentheses
Table1:
column 2: ,,R” should be subscript (tR)
line 126: should be 2.1.2.
line 156: ,,2.3. Method validation” should be from a new paragraph
line 186: there should be a dot at the end of the sentence
Table 2:
tR (min) should be in the second column
units (ng g-1) should be written above (first row) e.g. LOD (ng g-1)
Table 3:
column 2 and 3: ,,-1” should be superscript
column 5: ,,rep” should be subscript
Data shown in Table 3 (column 1 and 2) should be written with dots instead of commas
line 238: should be a pause (…...and the extraction….)
Data shown in Table 4 should be written with dots instead of commas
Sincerely yours,
Wioletta Parys
Author Response
User Menu
Home (/user/myprofile)
Manage Accounts (/user/manage_accounts)
Change Password (/user/chgpwd)
Edit Profile (/user/edit) Logout (/user/logout)
Submissions Menu
Submit Manuscript (/user/manuscripts/upload)
Display Submitted Manuscripts (/user/manuscripts/status)
Display Co-Authored Manuscripts (/user/manuscripts/co- authored)
English Editing
Journal Toxins (https://www.mdpi.com/journal/toxins) (ISSN 2072-6651) Manuscript ID toxins-1298379
Type Article Number of Pages 20
Title A targeted UHPLC-MS/MS method validated for the quantification of ergot alkaloids in cereal- based baby food from the Belgian market
Abstract Following pending new legislation in the European Union setting a maximum of 20 ng g-1 for the total sum of ergot alkaloids in dry cereal-based baby food, a new UHPLC-MS/MS method was developed. It is suitable for the quantification of six ergot alkaloids, ergocornine, ergocristine, er- gometrine, ergosine, ergotamine, α-ergocryptine, and their corresponding epimers. The method is able to reliably detect individual ergot alkaloids at a level as low as 0.5 ng g-1. The method uses a modified QuEChERS extraction approach before UHPLC-MS/MS analysis. The method showed good sensitivity, accuracy, and precision. It has been applied to 49 samples from the Belgian mar-ket. In 26 samples, not a single ergot alkaloid was detected, in 23 out of 49 samples at least one ergot alkaloid was detected with 2 samples containing 12 ergot alkaloids.
Ergometrine was the alkaloid most frequently detected i.e. 16 out of 49 samples. Only one sample, testing positive for all 12 ergot alkaloids, would be non-conform to the newly proposed MRL.
Author's Reply to the Review Report (Reviewer 1)
(/user/pre_english_article/status)
Discount Vouchers (/user/discount_voucher)
Invoices (/user/invoices)
LaTex Word Count (/user/get/latex_word_count)
* Author's Notes to
Reviewer
Please provide a point-by-point response to the reviewer’s comments and either enter it in the box below or upload it as a Word/PDF file. Please write down "Please see the attachment." in the box if you only upload an attachment. An example can be found here (/bundles/mdpisusy/attachments/Author/Example for author to respond reviewer - MDPI.docx? e9a1de4761e1496f).
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||
Reviewers Menu
Reviews (/user/reviewer/status)
Volunteer Preferences (/volunteer_reviewer_info/view)
Word / PDF No file chosen
Submit or Save as draft (submit later)
Review Report Form
Open Review (x) I would not like to sign my review report
( ) I would like to sign my review report
English language
and style
( ) Extensive editing of English language and style required
( ) Moderate English changes required
( ) English language and style are fine/minor spell check required
(x) I don't feel qualified to judge about the English language and style
|
Does the introduction provide sufficient
Can be improved
Must be improved
Not applicable
|
||||||||||||||||
(x) ( ) ( ) ( )
Comments and Suggestions for
Authors
The authors have described a new developed, optimized and validated UHPLC-MS/MS method for the simultaneous determination of six ergot alkaloids in 49 cereal-based baby food from the Belgian market.
In my opinion the manuscript is quite innovative, it is well written, presented and provides details of a method that may become useful in laboratories for monitoring and quality control of food.
This manuscript can be accepted after minor revision: line 9: ,,-1” should be superscript
Adapted
line 99: v/v should be in parentheses
Adapted
Table1:
column 2: ,,R” should be subscript (tR)
In tR the R is already in subscript but as t is in uppercase and R is in lower case they appear identical in word.
In the beginning of the previous phrase it also shows that at least to the naked eye there both normal case.
I have set “t” in letter size 10 and “R” in letter size 8.
line 126: should be 2.1.2.
Adapted
line 156: ,,2.3. Method validation” should be from a new paragraph line 186: there should be a dot at the end of the sentence
I have added “and”.
Table 2:
tR (min) should be in the second column
I have added tR and again R in a smaller letter size.
units (ng g-1) should be written above (first row) e.g. LOD (ng g-1)
Adapted
Table 3:
column 2 and 3: ,,-1” should be superscript column 5: ,,rep” should be subscript
Data shown in Table 3 (column 1 and 2) should be written with dots instead of commas
Table 3 has been adapted on all 4 points
line 238: should be a pause (…...and the extraction….)
Adapted as: “The conditions chosen for both the preparation of standards and the extraction , i.e. alkaline conditions combined with acetonitrile, should minimize epimerization.”
Data shown in Table 4 should be written with dots instead of commas
All commas have been replaced by points.
Sincerely yours, Wioletta Parys
Submission Date 29 June 2021
Date of this review 05 Jul 2021 09:47:50
© 1996-2021 MDPI (Basel, Switzerland) unless otherwise stated Disclaimer Terms and Conditions (https://www.mdpi.com/about/terms-and- conditions) Privacy Policy (https://www.mdpi.com/about/privacy)
Author Response File: 
Author Response.pdf
Reviewer 2 Report
This manuscript summarizes a well designed study that provides detailed methods for a sensitive, validated assay that addresses difficult to detect toxin derived from fungi. I see no issues with the work presented, the study design, the methods or the results. The article is concise, well-written, and thoughtful. The assay is robust and more sensitive than the guidelines outlined by the governing agencies. Minor edits are needed for formatting and word choice, otherwise, I see no reason not to accept the manuscript.
Author Response
Journals (https://www.mdpi.com/about/journals/)
Information (https://www.mdpi.com/guidelines) Initiatives
About (https://www.mdpi.com/about/)
User Menu
Home (/user/myprofile)
Manage Accounts (/user/manage_accounts)
Change Password (/user/chgpwd)
Edit Profile (/user/edit) Logout (/user/logout)
Submissions Menu
Submit Manuscript (/user/manuscripts/upload)
Display Submitted Manuscripts (/user/manuscripts/status)
Display Co-Authored Manuscripts (/user/manuscripts/co- authored)
English Editing
Journal Toxins (https://www.mdpi.com/journal/toxins) (ISSN 2072-6651) Manuscript ID toxins-1298379
Type Article Number of Pages 20
Title A targeted UHPLC-MS/MS method validated for the quantification of ergot alkaloids in cereal- based baby food from the Belgian market
Abstract Following pending new legislation in the European Union setting a maximum of 20 ng g-1 for the total sum of ergot alkaloids in dry cereal-based baby food, a new UHPLC-MS/MS method was developed. It is suitable for the quantification of six ergot alkaloids, ergocornine, ergocristine, er- gometrine, ergosine, ergotamine, α-ergocryptine, and their corresponding epimers. The method is able to reliably detect individual ergot alkaloids at a level as low as 0.5 ng g-1. The method uses a modified QuEChERS extraction approach before UHPLC-MS/MS analysis. The method showed good sensitivity, accuracy, and precision. It has been applied to 49 samples from the Belgian mar-ket. In 26 samples, not a single ergot alkaloid was detected, in 23 out of 49 samples at least one ergot alkaloid was detected with 2 samples containing 12 ergot alkaloids.
Ergometrine was the alkaloid most frequently detected i.e. 16 out of 49 samples. Only one sample, testing positive for all 12 ergot alkaloids, would be non-conform to the newly proposed MRL.
Author's Reply to the Review Report (Reviewer 2)
(/user/pre_english_article/status)
Discount Vouchers (/user/discount_voucher)
Invoices (/user/invoices)
LaTex Word Count (/user/get/latex_word_count)
* Author's Notes to
Reviewer
Please provide a point-by-point response to the reviewer’s comments and either enter it in the box below or upload it as a Word/PDF file. Please write down "Please see the attachment." in the box if you only upload an attachment. An example can be found here (/bundles/mdpisusy/attachments/Author/Example for author to respond reviewer - MDPI.docx? e9a1de4761e1496f).
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||
Reviewers Menu
Reviews (/user/reviewer/status)
Volunteer Preferences (/volunteer_reviewer_info/view)
Word / PDF No file chosen
Submit or Save as draft (submit later)
Review Report Form
Open Review (x) I would not like to sign my review report
( ) I would like to sign my review report
English language
and style
( ) Extensive editing of English language and style required
( ) Moderate English changes required
(x) English language and style are fine/minor spell check required
( ) I don't feel qualified to judge about the English language and style
|
Does the introduction provide sufficient
Can be improved
Must be improved
Not applicable
|
||||||||||||||||
(x) ( ) ( ) ( )
Comments and Suggestions for
Authors
This manuscript summarizes a well designed study that provides detailed methods for a sensitive, validated assay that addresses difficult to detect toxin derived from fungi. I see no issues with the work presented, the study design, the methods or the results. The article is concise, well-written, and thoughtful. The assay is robust and more sensitive than the guidelines outlined by the governing agencies. Minor edits are needed for formatting and word choice, otherwise, I see no reason not to accept the manuscript.
Submission Date 29 June 2021
Date of this review 23 Jul 2021 21:00:17
© 1996-2021 MDPI (Basel, Switzerland) unless otherwise stated Disclaimer Terms and Conditions (https://www.mdpi.com/about/terms-and- conditions)
Author Response File: 
Author Response.docx
Reviewer 3 Report
The authors of the manuscript titled “A targeted UHPLC-MS/MS method validated for the quantifi-2 cation of ergot alkaloids in cereal-based baby food from the 3 Belgian market” have presented a very interesting work about ergot alkaloids in cereal-based baby food. The manuscript is well written and presented.
However, there are some minor revision to be clarified (reported below), which should be revised before the publication of the manuscript.
Line 7: Please, use colon after six ergot alkaloids before the list.
Line 15: Please, specify the full name before the acronym “MRL”.
Line 40: References should be added always in the same way in the manuscript (at the end of the sentence, before the comma).
Line 47: “They have a double bond between either C-8 and C-9 or C-9 and C-10”. Is this correct? Please, verify.
Line 94: “A QuEChERS based method was first applied to ergot alkaloids by Malachova UHPLC-MS/MS”. Do you mean by Malachova and co-workers? Please, remove UHPLC-MS/MS.
Line 99 and 115: Please, write “v/v” in italic form, and correct in the whole manuscript.
Line 151: The authors have reported the modified QuEChERS approach used to purify the sample; however, no information about the protocol were detailed described. Please, could the authors briefly describe the protocol?
Line 156: Please, move “2.3. Method validation” in the line below.
Table 2: Please, write in the first column “Tr (min)” instead only min, as retention times were reported below.
Line 293: maybe the conclusion should be reported at the end of the manuscript, after materials and methods section (unless the journal guidelines suggested to put here).
Author Response
Journals (https://www.mdpi.com/about/journals/)
Information (https://www.mdpi.com/guidelines) Initiatives
About (https://www.mdpi.com/about/)
User Menu
Home (/user/myprofile)
Manage Accounts (/user/manage_accounts)
Change Password (/user/chgpwd)
Edit Profile (/user/edit) Logout (/user/logout)
Submissions Menu
Submit Manuscript (/user/manuscripts/upload)
Display Submitted Manuscripts (/user/manuscripts/status)
Display Co-Authored Manuscripts (/user/manuscripts/co- authored)
English Editing
Journal Toxins (https://www.mdpi.com/journal/toxins) (ISSN 2072-6651) Manuscript ID toxins-1298379
Type Article Number of Pages 20
Title A targeted UHPLC-MS/MS method validated for the quantification of ergot alkaloids in cereal- based baby food from the Belgian market
Abstract Following pending new legislation in the European Union setting a maximum of 20 ng g-1 for the total sum of ergot alkaloids in dry cereal-based baby food, a new UHPLC-MS/MS method was developed. It is suitable for the quantification of six ergot alkaloids, ergocornine, ergocristine, er- gometrine, ergosine, ergotamine, α-ergocryptine, and their corresponding epimers. The method is able to reliably detect individual ergot alkaloids at a level as low as 0.5 ng g-1. The method uses a modified QuEChERS extraction approach before UHPLC-MS/MS analysis. The method showed good sensitivity, accuracy, and precision. It has been applied to 49 samples from the Belgian mar-ket. In 26 samples, not a single ergot alkaloid was detected, in 23 out of 49 samples at least one ergot alkaloid was detected with 2 samples containing 12 ergot alkaloids.
Ergometrine was the alkaloid most frequently detected i.e. 16 out of 49 samples. Only one sample, testing positive for all 12 ergot alkaloids, would be non-conform to the newly proposed MRL.
Author's Reply to the Review Report (Reviewer 3)
(/user/pre_english_article/status)
Discount Vouchers (/user/discount_voucher)
Invoices (/user/invoices)
LaTex Word Count (/user/get/latex_word_count)
* Author's Notes to
Reviewer
Please provide a point-by-point response to the reviewer’s comments and either enter it in the box below or upload it as a Word/PDF file. Please write down "Please see the attachment." in the box if you only upload an attachment. An example can be found here (/bundles/mdpisusy/attachments/Author/Example for author to respond reviewer - MDPI.docx? e9a1de4761e1496f).
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||
Reviewers Menu
Reviews (/user/reviewer/status)
Volunteer Preferences (/volunteer_reviewer_info/view)
Word / PDF No file chosen
Submit or Save as draft (submit later)
Review Report Form
Open Review (x) I would not like to sign my review report
( ) I would like to sign my review report
English language
and style
( ) Extensive editing of English language and style required
( ) Moderate English changes required
( ) English language and style are fine/minor spell check required
(x) I don't feel qualified to judge about the English language and style
|
Does the introduction provide sufficient
Can be improved
Must be improved
Not applicable
|
|||||||||||||||||||||
(x) ( ) ( ) ( )
Comments and Suggestions for
Authors
The authors of the manuscript titled “A targeted UHPLC-MS/MS method validated for the quantifi-2 cation of ergot alkaloids in cereal-based baby food from the 3 Belgian market” have presented a very interesting work about ergot alkaloids in cereal-based baby food. The manuscript is well written and presented.
However, there are some minor revision to be clarified (reported below), which should be revised before the publication of the manuscript.
Line 7: Please, use colon after six ergot alkaloids before the list.
This has been adapted
Line 15: Please, specify the full name before the acronym “MRL”.
This has been adapted
Line 40: References should be added always in the same way in the manuscript (at the end of the sentence, before the comma).
This has been adapted
Line 47: “They have a double bond between either C-8 and C-9 or C-9 and C-10”. Is this correct? Please, verify.
I’m pretty sure this is at least so for the ergot alkaloids discussed here.
Line 94: “A QuEChERS based method was first applied to ergot alkaloids by Malachova UHPLC- MS/MS”. Do you mean by Malachova and co-workers? Please, remove UHPLC-MS/MS.
UHPLC-MS/MS has been removed
Line 99 and 115: Please, write “v/v” in italic form, and correct in the whole manuscript.
It has been adapted at 5 places: lines 99, 118, 159, 349 and 414.
Line 151: The authorshave reported the modified QuEChERS approach used to purify the sample; however, no information about the protocol were detailed described. Please, could the authors briefly describe the protocol?
The following sentence was added : In this modified QuEChERS approach 4g of sample is extrcated for at least 1 hour with a 30 mLsolution of 84% acetonitril and 16% (both in volume, not mass) water containing 200mg L-1.
Line 156: Please, move “2.3. Method validation” in the line below.
This has been adapted
Table 2: Please, write in the first column “Tr (min)” instead only min, as retention times were reported below.
In tR the R is already in subscript but as t is in uppercase and R is in lower case they appear identical in word.
In the beginning of the previous phrase it also shows that at least to the naked eye there both normal case.
I have set “t” in letter size 10 and “R” in letter size 8.
Line 293: maybe the conclusion should be reported at the end of the manuscript, after materials and methods section (unless the journal guidelines suggested to put here).
We did indeed do this following the example-template given on the website
Submission Date 29 June 2021
Date of this review 22 Jul 2021 19:55:15
© 1996-2021 MDPI (Basel, Switzerland) unless otherwise stated Disclaimer Terms and Conditions (https://www.mdpi.com/about/terms-and- conditions)
Author Response File: Author Response.docx
