PONE-D-21-40356Non-Invasive Skin Sampling Detects Systemically Administered Drugs in HumansPLOS ONE

Dear Dr. Tsunoda,

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Panitchpakdi et al. report detection of diphenhydramine and its metabolites on skin. They take advantage of a well-established swab sampling method and untargeted mass spectrometric detection. The protocol seems to be detailed sufficiently. The report may be of interest to some other scientists focused on drug metabolism and pharmacokinetics.

Comments:

- How do we know if the amounts of skin excretions collected by skin swabbing are always the same? Moreover, sweating rates may vary among the subjects and over time. Does this variability affect the results?

- The mean age of participants (26.3) is rather low. How does it affect the results?

- Can the peak areas in Figure 1 be converted to concentrations of these analytes in sweat?

- The report does not mention some relevant previous reports and reviews, e.g. J. Mass Spectrom. 2015, 50, 1321; JALM 2020, 5, 877; Trends Endocrinol. Metab. 2021, 32, 66.

- Reference 4 is a review on hydrogel-based devices for biomedical applications. Based on its content, it is not justified to cite it after the words “and hydrogel micro patches”. Only reference 5 (or other relevant papers) should be cited in this place.

- The authors point out the limitations of the other skin sampling methods (“but these are limited to testing topical, hydrophilic substances secreted with sweat”). There is no evidence that the sampling method used in this study performs better because there is no systematic comparison.

- “co-eluting ion of m/z 418.222”: Is this species in ionic form already in the column? Or, does it become ion in the ion source, i.e. after being eluted from the column?

- Some of the text/numbers in Figure 2 are too small to read.

- Some lines in Figure 2 are so thin that they are barely visible.

Reviewer #2: This article describes the use of liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) for the analysis of swabbed skin samples to detect orally administered diphenhydramine and its metabolites. The article is brief, but well written and includes a nicely designed study combining human plasma measurements as well as non-invasive skin sampling. I only have minor comments and questions that I hope will be of use:

Specific Comments:

1. What is the clinical relevance/usage of diphenhydramine?

2. References to studies examining exogenous compounds from latent fingerprints are relevant to the current work and should be cited, including:

a. Hinners, O’Neil, Lee, “Revealing Individual Lifestyles through Mass Spectrometry Imaging of Chemical Compounds in Fingerprints,” Scientific Reports, 2018, 8, 5149.

b. Bailey, et al. “Rapid detection of cocaine, benzoylecgonine and methylecgonine in fingerprints using surface mass spectrometry,” Analyst, 2015, 140, 6254-6259.

c. Guinan, Vedova, Kobus, Voelcker, “Mass spectrometry imaging of fingerprint sweat on nanostructured silicon,” Chemical Communications, 2015, 51, 6088–6091.

d. Lauzon, Dufresne, Chauhan, Chaurand, “Development of laser desorption imaging mass spectrometry methods to investigate the molecular composition of latent fingermarks,” Journal of the American Society of Mass Spectrometry, 2015, 26, 878–886.

e. Groeneveld, de Puit, Bleay, Bradshaw, Francese, “Detection and mapping of illicit drugs and their metabolites in fingermarks by MALDI MS and compatibility with forensic techniques,” Scientific Reports, 2015, 5, 11716.

f. Kaplan-Sandquist, LeBeau, Miller, “Chemical analysis of pharmaceuticals and explosives in fingermarks using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry,” Forensic Science International, 2014, 235, 68–77.

3. The m/z values at which the mass resolving powers (i.e., 35,000 and 17,500) are measured should be reported.

4. The figures are a bit blurry and the data difficult to read.

5. “Further, spectral evidence (exact mass) indicated…” should read “Further, spectral evidence (accurate mass) indicated…” (i.e., exact mass is theoretical, accurate mass refers to a measured value).

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Reviewer #1: No

Reviewer #2: No

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Non-Invasive Skin Sampling Detects Systemically Administered Drugs in Humans

PONE-D-21-40356R1

Dear Dr. Tsunoda,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Timothy J Garrett, PhD

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: (No Response)

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

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6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: Most of my previous comments have been addressed. The only piece of information to add to the manuscript is the m/z value at which the mass resolving powers (i.e., 35,000 and 17,500) are measured. Resolving power is defined as m/Δm, so the m (i.e., m/z) of the resolving power is an important figure to report (along with how Δm is measured), especially in instruments for which the resolving power varies over the mass range. On Thermo instrument, the resolving power reported in the software is usually defined at m/z 200 and Δm is measured as full-width at half maximum (FWHM).

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: No

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