Acid hydrolysis of saponins extracted in tincture

Jamie Love; Casey R. Simons

doi:10.1371/journal.pone.0244654

Peer Review History

Original SubmissionOctober 6, 2020
20 Nov 2020 Decision Letter - Pasquale Avino, Editor PONE-D-20-31354 Acid hydrolysis of saponins extracted in tincture PLOS ONE Dear Dr. Love, 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. Please submit your revised manuscript by Jan 03 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. 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Competing interests can arise in relationship to an organization or another person. Please follow this link to our website for more details on competing interests: http://journals.plos.org/plosone/s/competing-interests [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer #1: This manuscript describes attempts to improve sapogenin hydrolysis and purification from hydroalcoholic extracts of Smilax. The use of qNMR is innovative and provides a method that can be repeated easily in different While they have made progress to that end there are some issues that need to be addressed. 1) For any phytochemical study the plant identity needs to be confirmed. Usually this is done with a herbarium voucher that is identified by a taxonomist. At first mention, the botanical name is given with taxonomic authority and family in brackets Similar ornate Hook. (Smilacaceae). Thereafter it is referred to as S. ornata. 2) Major findings are mostly yields under different conditions (ex table 2,3 and figures 6and 7). These values need to be presented as a mean and standard error with number of replicates. Ideally a multiple comparison test would show which methods are statistically superior. For this reason also pie charts are rarely accepted in peer reviewed literature 3) The paper reads like a thesis rather than a focussed scientific paper with concise introduction, methods results and discussion. For example. a. The introduction is long and rambling. Avoid point form and subsections, quotations. b. Methods are presented in results c. Equations are not numbered d. Convert machine specific rpm to universal unit xg e. Ensure that appropriate units in brackets are placed above columns [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0244654.r001
Revision 1
4 Dec 2020 Author Response EDITORS RESPONSE TO REVIEWERS 1. Please ensure that your manuscript meets PLOS ONE's style requirements. RESPONSE: I believe we meet the requirements but if you believe we have not, please specify. 2. In your Data Availability statement, you have not specified where the minimal data set underlying the results described in your manuscript can be found. RESPONSE: All our data are found in the tables (Table 2 and Table 3) in the manuscript. This is not a data heavy report. 3. Thank you for stating the following in the Financial Disclosure section: 'The author(s) received no specific funding for this work.' We note that one or more of the authors are employed by a commercial company: Synapses Ltd, Mill of Fyall. a, b and c RESPONSE: Synapses is not a commercial company. A couple years ago I retired (from U of Edinburgh) but have not retired from research. I have a home lab (shed) in which I conduct all my work, including the work that contributed to this manuscript. As a “civilian” (no longer with the university), I had difficulty purchasing certain chemicals – chloroform, for example. Sigma, et al. will not ship to private individuals or residential addresses. Synapses, was the name I used as a consultant (independent of the university) since 1998, so I decided to incorporate it – and now I can order the chemicals I need. Synapses is my “hobby lab” where I tinker with projects that interest me. Synapses is non-profit and totally self-funded and I am currently unincorporating it. I have no competing or conflicting interest. I will not (cannot) profit from publishing “Acid hydrolysis of saponins extracted in tincture” in PLOS ONE. “This does not alter our adherence to PLOS ONE policies on sharing data and materials.” I would be happy to address any additional concerns. Reviewers' comments: Reviewer #1: This manuscript describes attempts to improve sapogenin hydrolysis and purification from hydroalcoholic extracts of Smilax. The use of qNMR is innovative and provides a method that can be repeated easily in different While they have made progress to that end there are some issues that need to be addressed. 1) – a - For any phytochemical study the plant identity needs to be confirmed. Usually this is done with a herbarium voucher that is identified by a taxonomist. RESPONSE: We are not trying to prove that sarsasapogenin is present in S. ornata. That was done in 1934 by Jacob and Simpson (and referenced). Nor are we trying to prove that smilagenin is produced from S. ornata. That was done in 1936 by Askew, Farmer and Kon. These observations have been confirmed since their first discovery by several investigators (including RE Marker, a luminary in steroidal chemistry). That said, I understand and appreciate the reviewer’s point. Proper identification is crucial and an herbal voucher is preferred. However, we do not have the resources to collect the specimens (in Central America) nor the skill to identify them. Instead, we must trust the supplier of the tincture to have used the correct plant. Our tincture was prepared by and purchased from a trusted, experienced and long-established local herbalist who knows how to source the marc (plant material) and judge its quality – important skills that we lack. BTW – sarsasapogenin has been reported to be in the roots of Smilax lanceolata and S. rotundifolia, Dioscorea colletii (a yam), nineteen species of yucca (Yucca spp.), Anemarrhena asphodeloide (used in traditional Chinese medicine) and several species of asparagus (Asparagus spp) including the domesticated species, as well as the seeds and leaves of Trigonella foenum-graecum. We are collecting powder made from all these plants (for a follow up paper) but, again, must rely upon the suppliers for identification of most of them. 1) -b -At first mention, the botanical name is given with taxonomic authority and family in brackets Hook. (Smilacaceae). Thereafter it is referred to as S. ornata. RESPONSE: My mistake. I’ve added the authority and family at first mention and corrected the spelling errors (ornata to ornate). Thank you. 2) -a -Major findings are mostly yields under different conditions (ex table 2,3 and figures 6and 7). These values need to be presented as a mean and standard error with number of replicates. RESPONSE: Table 2 shows the raw data, mean and standard deviation from four batches of tincture. Our intention in this table is not to find the average (mean) of each batch but the average across four batches. Table 3 shows the raw data with (only) a single data point for each of the 16 experiments. We did not run replicates because the volume of tincture needed to run (say four) replicates of each point would have exceeded the amount of tincture in a single batch. We wanted to make comparisons across all four conditions (combinations of temperature and acidity) at four different timepoints using the same (single) batch of tincture. The raison d'être for Table 3 is not accuracy (which would require replicates) but in comparisons between the conditions. We believe that others who replicate our Table 3 in their own labs using a single batch of S. ornata tincture, will come to similar conclusions about the relationship of between the conditions, but unlikely to get the same values. 2) - b - Ideally a multiple comparison test would show which methods are statistically superior. RESPONSE: We are not trying to find the statistically superior method. We are using the data in Table 3 to display trends. Importantly, we are not getting across our point and apologize for the confusion. Therefore, we have included a new table (Table 4. Diagram illustrating changes in maximum yields at 80 oC or 70 oC and 6N or 2N HCl) to highlight the conclusions that others may use to guide them in finding conditions that work with the saponin of their interest. 2) – c - For this reason also pie charts are rarely accepted in peer reviewed literature RESPONSE: The pie charts have been removed. 3) The paper reads like a thesis rather than a focussed scientific paper with concise introduction, methods results and discussion. For example. RESPONSE: It felt like a thesis too! We are working across several specialties, so try to be complete but has been trimmed and specific concerns are addressed below. a. The introduction is long and rambling. Avoid point form and subsections, quotations. RESPONSE: Edited to be succinct and removed some parts. Points and quotes are removed and subsections limited. I hope it reads better. b. Methods are presented in results RESPONSE: I assume the reviewer is referring to the demonstration of the oil in different concentrations of alcohol. I’ve moved the method, but not the result, into the Methods section. c. Equations are not numbered RESPONSE: The equation for qNMR is now numbered [eq 1] and Imassume that is what the reviewer had in mind. (Elsewhere, I show my maths to make calculations, but I believe they do not need to be numbered (and would look awkward if I did). I hope that is satisfactory. d. Convert machine specific rpm to universal unit xg RESPONSE: Added. e. Ensure that appropriate units in brackets are placed above columns RESPONSE: I don’t see an error. Can you be specific? ALSO: I felt that the data and discussion about background was poorly explained and upon reflection, decided it added nothing of value to the paper – so I removed the data from Table 3 (the last three rows) about the background crude and its discussion later on. I found and corrected a few typos. Thank you. Jamie Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0244654.r002
15 Dec 2020 Decision Letter - Pasquale Avino, Editor Acid hydrolysis of saponins extracted in tincture PONE-D-20-31354R1 Dear Dr. Love, 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. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Pasquale Avino, Ph.D. Academic Editor PLOS ONE Reviewers' comments: Reviewer #1: The authors have responded positively to the review with revisions and explanations. The manuscript is acceptable for publication. https://doi.org/10.1371/journal.pone.0244654.r003
Formally Accepted
18 Dec 2020 Acceptance Letter - Pasquale Avino, Editor PONE-D-20-31354R1 Acid hydrolysis of saponins extracted in tincture Dear Dr. Love: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Professor Pasquale Avino Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0244654.r004

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