PONE-D-23-26541Drugs in blood and urine samples from suspected spiked drink victims: a prospective observational study from Oslo, NorwayPLOS ONE

Dear Dr. Vallersnes,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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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: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: Partly

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

Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

Reviewer #4: N/A

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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

Reviewer #3: Yes

Reviewer #4: No

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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

Reviewer #3: Yes

Reviewer #4: No

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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: Dear Authors,

It was my pleasure to review the article titled Drugs in blood and urine samples from suspected spiked drink victims: a prospective

observational study from Oslo, Norway, a solid and well researched article centered around drug-facilitated sexual assault (or less commonly other crimes targeting an uncconscious ibìncapacitated victim). The study encompasses a 100-patient sample and focuses on detection of unreported drugs.

The article has noteworthy strengths: it is thorough in terms of pursuing its stated objective; it is relevant and overall a worthy contribution to toxicology research; it has an element of novelty in its design, and relies on sound methodology as far as I was able to determine.

It is my belief that the article, particularly in its discussion, could benefit from a slightly higher degree of contextualization and broader scope when it comes to screening and detection techniques. It would certainly make the article more comprehensive and well-rounded to draw comparisons and outline distinctive features of various techniques and their analytical value in terms of tackling the spread of illegal/misused substances, both for recreational use and as a tool to victimize others. Broader remarks on the legal and law enforcement potential of detection efforts, at least within the European context, could also be advisable. The following sources should be drawn upon and cited:

Brunetti P, Lo Faro AF, Di Trana A, Montana A, Basile G, Carlier J, Busardò FP. β'-Phenylfentanyl Metabolism in Primary Human Hepatocyte Incubations: Identification of Potential Biomarkers of Exposure in Clinical and Forensic Toxicology. J Anal Toxicol. 2023 Jan 24;46(9):e207-e217. doi: 10.1093/jat/bkac065.

Di Trana A, Brunetti P, Giorgetti R, Marinelli E, Zaami S, Busardò FP, Carlier J. In silico prediction, LC-HRMS/MS analysis, and targeted/untargeted data-mining workflow for the profiling of phenylfentanyl in vitro metabolites. Talanta. 2021 Dec 1;235:122740. doi: 10.1016/j.talanta.2021.122740.

Lo Faro AF, Venanzi B, Pilli G, Ripani U, Basile G, Pichini S, Busardò FP. Ultra-high-performance liquid chromatography-tandem mass spectrometry assay for quantifying THC, CBD and their metabolites in hair. Application to patients treated with medical cannabis. J Pharm Biomed Anal. 2022 Aug 5;217:114841. doi: 10.1016/j.jpba.2022.114841.

Busardò FP, Zaami S, Baglio G, Indorato F, Montana A, Giarratana N, Kyriakou C, Marinelli E, Romano G. Assessment of the stability of exogenous gamma hydroxybutyric acid (GHB) in stored blood and urine specimens. Eur Rev Med Pharmacol Sci. 2015 Nov;19(21):4187-94.

The tables and figures are qite well crafted and effective at conveying key points and relevant findings.

Although the article is well-written overall, I recommend further proofreading by a native speaker of English.

Reviewer #2: The article is competently assembled and straightforward enough overall. Its aim is clearly delineated and the methodology appears to be sound. tables and figures contribute to the conveyance of key elements providing substance and clarity to the article' s fundamental reasoning and conclusions.

The Discussion I believe should be further developed to better highlight the article's relevance in toxicology research. More meaningful sources ought to be used in order to better elaborate on detection techniques and their value in tackling substance abuse and in upholding public health. The following should be looked at and cited in that regard: DOI: 10.23750/abm.v92i6.12696; DOI 10.1016/j.jpba.2020.113335. Furthermore, I suggest the authors brefly address legislative aspects as well against the backdrop of the major threat posed by novel psychoactive substances DOI: 10.1002/hup.2727; DOI: 10.1007/164_2018_160; DOI: 10.26355/eurrev_201911_19529). Such additions will provide an extra degree of elaboration which will highlight the importance and value of the article's conclusions and findings.

Sincerely.

Reviewer #3: Comments:

1. In abstract (line no 47), Check the sentence consistency.

2. Remove drink spiking word from the keyword.

3. Literature survey regarding analysis of drugs in case of drug facilitated crimes (DFC) is insufficient. The author needs to discuss the recently published analytical methods. The following papers may be cited and discussed in literature survey:

• Exploiting the potential of fabric phase sorptive extraction for forensic food safety: analysis of food samples in cases of drug facilitated crimes. Food chemistry, 432, 137191.

• Cellulose Paper Sorptive Extraction (CPSE) Combined with Gas Chromatography–Mass Spectrometry (GC–MS) for Facile Determination of Lorazepam Residues in Food Samples Involved in Drug Facilitated Crimes. Separations, 10(5), 281.

4. Line no 186; the author should mention the temperature at which the urine and blood samples were stored.

5. The author mustprovide the instrumental specifications of UHPLC-MS/MS and GC-MS used in determining drugs in blood and urine samples within the main manuscript.

6. What is “IQR”? The author must furnish information regarding this term for better understanding of readers.

7. The author must include the chromatograms in the main manuscript for the better understanding of the readers.

Reviewer #4: The manuscript entitled “Drugs in blood and urine samples from suspected spiked drink victims: a prospective observational study from Oslo, Norway” describes an important subject.

However, the manuscript should be corrected to be written in more standard English. There are many typos and unclear definitions (unreported drugs, spiked drinks, voluntary taken etc.), and the manuscript is generally unfocused on the aim and clear conclusions are lacking. It is unclear whether the purpose is to identify which substances are used in these cases or whether the purpose is to make a comparison of identified substances in blood and urine. The authors are encouraged to include more relevant drug classes such as sedative antihistamines, antidepressants etc. to increase the value of the study.

Words/terms which advantageously could be rephrased/changed in the manuscript:

• The authors use the term “patients” throughout the manuscript. I see the point using the term “patients” when the individuals are hospitalized or involved with the emergency medicine services. However, in some sentences it would be more correct to use individuals or victims of drink spiking. As being the case in the objective “patients regularly contact”

• Rephrase the objectives where the sentence say “patients with suspected spiked drinks”. Maybe write suspected victims of drink spiking instead.

• The authors should be consistent with the term drink spiking/spiked drink/drink-spiking throughout the manuscript. They could consider to use drink spiking and victims of dink spiking throughout the manuscript instead of spiked drink victims. Also consider to correct the title.

• Consider to rephrase “unreported drugs” as this in many sentences is confusing. Consider to use “additional drugs which was not self-reported”

• Consider to change the key-words as some are also included in the title. Use only drink spiking or spiked drinks (Depending on which definition is used throughout the manuscript). Consider to include LC-MS/MS in keywords. The keywords deviates from manuscript to the front template.

Selected comments

Abstract:

• In objectives please, clarify the aim of the study.

• The conclusion in the abstract should be changed as this does not really include all important conclusion from the paper. Could the additional drugs measured, which have not been self-reported, potentially contribute to the drink spiking cases?

Introduction:

• The introduction should be shortened and focusing more on the aim of the study.

Materials and methods:

• It is unclear whether the substances are only screened or whether they are also verified.

• Include a more detailed description on the analytical methods used.

Result:

• Line 234 rephrase: we obtained biological samples in blood.

• Table 2: Misleading numbers in the table: is total number of cases in the reported and unreported 100 in each? while the total number is also 100.

• Table 3: Why mention the symptoms and clinical features if they are not used in the classification of "drink spiking".

Discussion:

• The authors write on page 20 line 413 that antidepressants and antihistamines and other drugs not uncommonly found in DFSA cases have not been included in the study. According to SOFT and the drug-facilitated crimes (DFC) committee both antihistamines and antidepressants are among the common DFC drugs in urine samples. Maybe there would be more than 15% of the cases with additional drugs than the self-reported if these compounds have been included. Please elaborate more on that in the manuscript.

• What can this study be used for? What do the reader achieve by reading the paper?

• The study indicates that ethanol could potential be used in drink spiking, and this conclusion is based upon no additional drug findings in the samples and more alcohol in the samples with no additional drug findings. However, this cannot be proven with methods and therefore not give any answers to the victims. What about the samples with other drug findings, could they potentially contribute to the drink spiking cases? More about the actual findings are missing. And could drugs that are commonly taken (antihistamines and antidepressants?) not be used in drink spiking cases? Please elaborate on this also.

• Part conclusions contain too many non-scientific claims such as "The patient may have smokey what they thought was nicotine" line 351 and "Ethanol might have been used to spike alcoholic drinks" line 425 and section 358–371.

• Elaborate more on the ethanol results according to sampling time interval. Many of the samples were collected late, which could be the reason for lack of ethanol detection.

Conclusion:

• What about the actual drug findings, these are not really elaborated?

• Elaborate on the aim.

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Reviewer #1: No

Reviewer #2: No

Reviewer #3: No

Reviewer #4: No

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Drugs in blood and urine samples from victims of suspected exposure to drink spiking: a prospective observational study from Oslo, Norway

PONE-D-23-26541R1

Dear Dr. Vallersnes,

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.

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Academic Editor

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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 #1: All comments have been addressed

Reviewer #3: All comments have been addressed

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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 #1: Yes

Reviewer #3: Yes

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

Reviewer #1: Yes

Reviewer #3: 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 #1: Yes

Reviewer #3: 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 #1: Yes

Reviewer #3: 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 #1: I have read your article and the responses to the inquiries from other reviewers with particular attention and interest. These inquiries are quite pertinent and have undoubtedly enriched the scientific content of your work. Personally, I find the impact of this study to be interesting and commend you for the scholarly presentation. In conclusion, the study certainly has interesting points as it addresses a topic of great interest. The objective is clear and adhered to. The main question addressed by the research is clear and entirely agreeable. I believe the information provided is sufficient and represents useful elements to encourage the development of new scientific work.

Reviewer #3: Manuscript Title: Drugs in blood and urine samples from victims of suspected exposure to drink spiking: a prospective observational study from Oslo, Norway

The authors have addressed my comments and now the manuscript can be accepted for publication.

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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 #1: Yes: Giuseppe Basile

Reviewer #3: No

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