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. 2026 Jan 16;21(1):e0341070. doi: 10.1371/journal.pone.0341070

The quality and safety of Rhodiola rosea supplements on the U.S. market: An analysis of biomarkers, heavy metals, and pesticide residues

Steffen Porwollik 1, Mahtab Jafari 1,*
Editor: Mahmoud W Yaish2
PMCID: PMC12810810  PMID: 41544018

Abstract

Rhodiola rosea supplements have become popular in the U.S., with a $3.4 billion market and an annual growth of about 10% in sales in the past few years. While the health benefits of this plant have been evaluated in many scientific studies, the potential differences in quality of these botanical products on the U.S. market have not been studied in detail. Using reversed-phase ultra-performance liquid chromatography, we determined the concentrations of the biomarker molecules, rosavins and salidroside, in a small but representative sample of R. rosea dietary supplement products commercially available in the U.S. Concentrations of rosavins and salidroside ranged from 0.01% to 3.08% and 0.07% to 2.91%, respectively, including substantial aberrations from advertised biomarker amounts. One product showed an undisclosed likely addition of synthetic salidroside. We also assessed heavy metal contaminations via inductively coupled plasma mass spectrometry and pesticide contents by gas chromatography-mass spectrometry or liquid chromatography-mass spectrometry in these botanical products. While pesticide levels were below detection limits, all seven tested capsular products had trace amounts of arsenic, cobalt, and lead. Two of these contained notably elevated levels of cobalt and arsenic, where follow-up arsenic speciation would be required to verify whether they remain within minimal risk levels established by the U.S. Department of Health and Human Services. Overall, these results underscore the need for more stringent quality control in herbal supplements containing R. rosea available in the U.S.

Introduction

Rhodiola rosea (a plant in the Crassulaceae family) is also commonly known as golden root, rose root, or Arctic root, and is a widely used adaptogenic plant recognized for its potential health benefits. The plant has a long history of use in traditional medicine, particularly in Scandinavian countries, Russia, China, and other regions with cold climates. It has been utilized for centuries to enhance physical endurance, reduce fatigue, and promote resilience against environmental stressors [1,2].

Rhodiola rosea has been reported to improve exercise performance [3], to help with stress management [4,5], and to improve mental health [6]. It has also displayed anti-aging properties, increasing lifespan and improving healthspan, in various animal model systems, such as fruit flies, silkworms, and rats [710]. Due to these purported effects, R. rosea has been incorporated into a variety of increasingly popular dietary supplements. Global sales of R. rosea-containing supplements have experienced significant growth, and in the United States, these supplements have experienced a compound annual growth rate exceeding 10% between 2019 and 2024 [11].

Rhodiola rosea naturally thrives in high-altitude, dry, and cold regions, including the Arctic, the mountains of Central Asia, and parts of North America and Europe. It grows in rocky, nutrient-poor soils and exhibits remarkable adaptability to harsh environmental conditions. Due to increasing commercial demand, concerns about overharvesting and habitat degradation of wild R. rosea are voiced by environmental scientists [12,13]. Although R. rosea is currently not included in the International Union for Conservation of Nature’s Red List of Threatened Species, the plant has been added to the list of protected species by the Convention on International Trade in Endangered Species in late 2022 [13], making sure that its international trade is regulated to facilitate sustainability in its harvesting from the wild.

Rhodiola rosea contains a diverse array of phytochemicals, with approximately 140 different compounds identified, including glucosides, phenols, flavonoids, and monoterpenes [5,14]. Among the glucosides, the phenylpropanoid glycosides rosarin, rosin, and rosavin, collectively called rosavins, as well as salidroside, are recommended as key biomarker molecules and reported in HPLC-based analyses verifying the quality of this plant’s extracts [15,16]. However, the concentration of these compounds varies within different plant parts and between male and female specimens [17]. Additionally, environmental factors such as altitude, soil composition, and climate conditions significantly influence the phytochemical composition of R. rosea, further complicating standardization efforts [18]. Nevertheless, standardized pharmacopeial references for R. rosea exist in many countries, including Russia, Australia, and Northern European nations. The U.S. Pharmacopeia (USP Certificate for catalog number 1602580, Rhodiola rosea root and rhizome dry extract) includes a chromatographic profile specifying rosavins (rosin, rosarin, and rosavin) and salidroside as the main biomarkers of the plant, and the USP Herbal Medicines Compendium indicates lower limits of salidroside (0.08%), and rosavins (0.3%) in dried R. rosea roots and rhizomes [19]. Some of the adaptogenic effects of the plant, including its antioxidant properties, have been attributed to these biomarker molecules [20,21], but the beneficial effects cannot be consistently recapitulated with isolated rosavins or salidroside in laboratory settings. Other molecules, such as quercetin-glycosides, have also been identified as contributors [22]. Therefore, the adaptogenic properties are likely caused by the synergistic activity of at least some of the > 100 organic compounds of the plant.

While dietary supplements are not subjected to the same pre-market scrutiny and regulatory oversight as pharmaceuticals, their manufacturing regulations are governed by 21 CFR Part 111, a regulation set by the U.S. Food and Drug Administration (FDA), which covers specifications for identity, purity, strength, composition, and limits on contaminants. However, surveillance of these products, post-market, often simply relies on reports of adverse events and subsequent label modifications [23], and the quality of the R. rosea supplements in the U.S. has not been analyzed in detail. Previous studies have identified inconsistencies in R. rosea extract composition across the European market [2427], where widespread adulteration with other Rhodiola species or synthetic compounds has been reported, raising safety concerns and doubts about the products’ efficacy. Such fluctuations in quality, active ingredient concentrations, and labeling accuracy abound in the dietary supplement market, and have also been shown for CBD-containing melatonin gummies [28] and immune health dietary supplements [29] sold in the U.S.

This study takes a snapshot of the quality of R. rosea supplements currently available on the U.S. market. Specifically, the study assesses the content of the biomarker molecules, rosavins and salidroside, of the plant as well as potential contaminations with heavy metals and pesticides of ten R. rosea products commercially available in the U.S. Heavy metal exposure can have dramatic detrimental effects on human health, particularly on kidney function [30] while pesticides may cause chronic health conditions, including asthma, cancer, or diabetes [31]. By analyzing these R. rosea products on the U.S. market, this study appraises their safety and variability, informing regulatory agencies, healthcare professionals, and, most importantly, consumers.

Materials & methods

Sample selection and preparation for analysis

The selection of ten Rhodiola rosea supplements from the U.S. market was based on brand diversity, the products’ cost (where we aimed to cover a broad range), and availability. In addition, products needed to be manufactured at separate sites. All capsules and tinctures were purchased from Amazon in 2024.

Capsular products were homogenized by creating a composite of capsule fill material, which was then sampled. Fill material from 3–10 capsules per product was used for the analysis. Prior to injection into chromatography systems, capsule and tincture samples were centrifuged and filtered using a 0.45-micrometer filter.

Chemical analysis

All chemical analyses were completed at the Eurofins Scientific testing facilities in Brea, CA, and Madison, WI, following all applicable procedure-specific industry standards, as required for accredited testing laboratories in the U.S. For the quantification of rosavins and salidroside, a standardized chromatography-based method was employed [19]. Samples were extracted in a mixture of phosphoric acid and methanol and analyzed via reversed-phase ultra-performance liquid chromatography (UPLC) equipped with a photodiode array detector on a Waters™ Acquity I-Class PLUS UPLC System, employing external standards for calibration.

For heavy metal ion analysis, methods 2015.01, 2011.19, and 993.14, as published by AOAC INTERNATIONAL, were followed, with minor modifications [32]. Briefly, samples were digested with nitric acid (HNO₃) using a microwave digestion system. Detection was conducted via inductively coupled plasma mass spectrometry (ICP-MS) on Agilent 7900 ICP-MS Systems, where digested samples were compared to calibration standards of known concentrations.

For pesticide determination, compound levels were extracted and processed essentially as described in the AOAC INTERNATIONAL method 2007.01 [32], following the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) strategy [33,34]. The protocol involves acetonitrile extraction, partitioning, and dispersive solid-phase extraction clean-up, followed by compound detection via gas chromatography-mass spectrometry (GC/MS) or liquid chromatography mass spectrometry (LC/MS) on Agilent 6400 Triple Quadrupole LC/MS, Agilent 7890 GC/MS, and Agilent 8890 GC/MS Systems.

Quality assurance

Certified reference materials and calibration standards were used in all chromatography assays to ensure data accuracy. Instruments were routinely subjected to quality control determinations to validate results. Method validation parameters, including limits of detection, quantification, and recovery rates, were incorporated where available to ensure analytical robustness.

For rosavins and salidroside quantification, the limits of quantitation (LOQs) were 50 μg/g (ppm), with a precision of approximately 5% relative standard deviation. For pesticide determination, LOQs were typically at or below 0.01 μg/g for most analytes, except for azinphos-methyl, endosulfans, and heptachlor endo-epoxide (0.02 μg/g) and pyrethrum (0.2 μg/g). For heavy metal ion analysis, LOQs varied depending on the matrix but were typically 5 ppb for cadmium, mercury, and lead, and 10 ppb for arsenic and cobalt.

Results

Biomarker molecules

The concentrations of key biomarker molecules in the ten commercial Rhodiola rosea products are illustrated in Fig 1 and Table 1 and reveal substantial variability.

Fig 1. Contents of rosavins and salidroside in 10 commercially available Rhodiola rosea products in the U.S. Where known, advertised concentrations of these biomarker molecules are also shown.

Fig 1

Table 1. Biomarker, heavy metal, and pesticide contents of ten Rhodiola rosea products available on the U.S. market. * = Biomarker content is significantly lower than advertised. N.D. = not detected, ppb = parts per billion.

Sample Name Rosavin [%] Rosarin [%] Rosin [%] Total Rosavins (advertised on product label) [%] Salidroside (advertised on product label) [%] Arsenic [ppb] Cadmium [ppb] Cobalt [ppb] Lead [ppb] Mercury [ppb] Pesticides
Capsule 1 1.010 0.370 0.211 1.590 0.661 22.8 34.7 46.4 75.1 < 5 N.D.
Capsule 2 0.754 0.507 0.246 1.510 (3.0)* 1.000

(1.0)
54.4 < 5 50.8 11.6 < 5 N.D.
Capsule 3 0.118 0.049 0.023 0.191 (1.0)* 0.217

(1.0) *
21 < 5 18 9 < 5 N.D.
Capsule 4 1.680 0.726 0.437 2.850 (3.0) 2.070

(1.0)
159 22 164 88 < 5 N.D.
Capsule 5 1.900 0.705 0.474 3.080 (3.0) 2.570

(1.0)
393 11 733 85 < 5 N.D.
Capsule 6 0.006 0.002 0.002 0.011 0.104 65.8 < 5 40.8 43.8 < 5 N.D.
Capsule 7 0.100 0.061 0.030 0.191

(0.25)
2.910

(3.0)
59.7 < 5 106 55.2 10.9 N.D.
Tincture 1 0.083 0.057 0.033 0.173 (0.183) 0.122

(0.131)
< 10 < 5 < 10 < 5 < 5 N.D.
Tincture 2 0.087 0.030 0.017 0.134 0.071 < 10 < 5 < 10 < 5 < 5 N.D.
Tincture 3 0.194 0.064 0.029 0.286 0.130 < 10 < 5 < 10 < 5 < 5 N.D.

For rosavins, measured concentrations ranged from 0.01% to 3.08%, with rosavin contributing between half and two-thirds, rosarin between one-fifth and one-third, and rosin accounting for the remainder. The three tinctures exhibited similar rosavins content, ranging from 0.13% to 0.28%. Only one of these, tincture 1, declared target concentrations of these biomarkers on their label, which closely matched our measurements. Among the seven capsule products, rosavins concentrations varied greatly. Three capsules advertised a target concentration of 3% rosavins, where two closely matched that target while capsule 2 fell about two-fold short. Furthermore, capsule 3, labeled as containing 1% rosavins, had measured concentrations nearly five times lower than declared. One product, capsule 6, contained only 0.01% rosavins, suggesting an unexpectedly low R. rosea extract content.

For salidroside, a similarly wide concentration range was observed. The three tinctures contained salidroside levels between 0.07% and 0.13%, whereas capsule concentrations varied from 0.10% to 2.9%. Four capsule products advertised a 1% salidroside content. Three of these (capsules 2, 4, and 5) either matched or exceeded that target, while capsule 3 missed it by almost five-fold. Capsule 7, labeled as containing 3% salidroside, had a measured concentration of 2.9%, closely matching its claimed value.

Heavy metals and pesticides

We quantified ions from five heavy metals—arsenic, cadmium, cobalt, lead, and mercury—in all ten products. Because most heavy metal ions do not readily extricate from plant material into liquid extracts [35], the tinctures were not expected to contain problematic amounts of cadmium, cobalt, lead, or mercury ions. As expected, none of the three tinctures had detectable levels of these metal ions, nor of arsenic ions. However, the seven capsule products exhibited considerable variation in contamination levels (Table 1, Fig 2). Mercury ions were only detected in capsule 7 (10.9 ppb), while cadmium ions were present in three capsules (11.0–34.7 ppb). Ions from the remaining three metals were detected in all capsules: arsenic (21–393 ppb), cobalt (18–733 ppb), and lead (9–88 ppb). While capsule 4 had elevated amounts of arsenic and cobalt ions, capsule 5 exhibited the greatest levels of ions of these two heavy metals, with concentrations over 6.5-fold and over 14-fold higher, respectively, than the median values detected across all capsules.

Fig 2. Heavy metal content in seven commercial capsule preparations of Rhodiola rosea.

Fig 2

Furthermore, we explored whether the products contained any of the 70 pesticides listed in the U.S. Pharmacopeia (USP) chapter <561 > . All quantified pesticide compounds—including DDT, chlorpyrifos, heptachlor, and hexachlorobenzene—remained below detection limits. However, USP < 561 > focuses on contaminants historically relevant to herbal products and botanical pharmaceuticals, and does not include glyphosate or neonicotinoids, two pesticide classes of growing concern in agricultural and environmental contexts.

Discussion

The ten Rhodiola rosea supplements analyzed in this study exhibited substantial variability in biomarker molecule content. While this is the first study exploring R. rosea products available in the U.S., our observations align with prior studies of dry extracts commercially available in Europe [26], where the lowest salidroside concentrations were approximately nine times higher than the lowest concentrations detected in our study among capsular R. rosea products. While rosavins levels in some European samples exceeded 5%, we observed low amounts of this biomarker in several samples. A low rosavins content has been linked to adulteration with other Rhodiola species that lack this biomarker compound, such as Rhodiola crenulata [25]. Our findings identified three capsular products in which rosavins concentrations were lower than their corresponding salidroside amounts, raising concerns of potential adulteration. In one case, capsule 7, the low rosavins content coincided with a 15-fold higher salidroside concentration, suggesting adulteration, blending with other Rhodiola species, or manipulation of biosynthetic pathways in Rhodiola cell culture suspensions [36]. Although the product label accurately reported the salidroside concentration, it did not disclose the addition of synthetic salidroside or the presence of other Rhodiola species. Notably, one tested capsular product advertised a rosavins concentration of 1%, yet measured levels were approximately five times lower. In contrast, two other products claiming a 3% rosavins content contained levels very close to the target concentration, while one other product exhibited rosavins at 1.5%, a two-fold lower concentration than the stated content.

Each milliliter of the tincture products tested in this study is reported to be made from 300–500 mg of dried root of R. rosea. These tinctures did not contain pesticides and heavy metals, but displayed lower concentrations of rosavins and salidroside than most other products surveyed here and elsewhere [19,25]. However, the biological activity and adaptogenic properties of R. rosea may not be solely dependent on its biomarker molecule content. Over the past two decades, we have tested the biological activity of R. rosea products in fruit flies and mice [8,9,37,38]. Our recent lifespan assays in Drosophila melanogaster demonstrated comparable life-extending effects between R. rosea extracts with lower concentrations of rosavins and an R. rosea extract with a higher concentration of rosavins (S1 File). Whether this equivalence extends to other purported benefits of the plant, such as enhanced energy, mental health support, stress reduction, and exercise performance, remains to be determined. Although salidroside and rosavins are frequently cited as bioactive compounds, their physiological bioavailability, synergy with other plant compounds, or the presence of other molecules in commercially developed R. rosea extracts may affect their functional profile in vivo. In fact, total R. rosea extracts are known to perform better than isolated biomarkers [18].

Heavy metal contamination is a longstanding concern in herbal supplements. A global survey found that approximately 30% of 1,700 tested samples of 86 herbal medicines contained at least one heavy metal above regulatory limits [39], and over a quarter of 2,300 herbal products sold in 37 countries showed signs of adulteration [40], a practice that occurs in even higher proportions in popular supplements such as ginkgo and black cohosh [41], and in 29% of herbal products sold in the US [42]. In our study, all seven tested capsular products contained trace amounts of arsenic, cobalt, and lead ions. While most levels were low, capsule 5 contained approximately 200 ng of arsenic and 360 ng of cobalt per serving (500 mg), while capsule 4 had about 40 ng of arsenic and cobalt per serving (250 mg). The U.S. Department of Health and Human Services established a minimal risk level (MRL) for oral cobalt ingestion at 30 ng/kg/day [43]. Notable arsenic compounds include inorganic arsenic, monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA). For inorganic arsenic, MRLs are 5 ng/kg/day for acute-duration ingestion (14 days or less) and 0.3 ng/kg/day for chronic-duration ingestion (> 1 year) [44]. For MMA, MRLs are higher (100 ng/kg/day and 10 ng/kg/day for acute-duration and chronic-duration ingestion, respectively), and the established MRL for chronic ingestion of DMA is 20 ng/kg/day [44]. Our detection procedure does not include arsenic speciation, and we are therefore unable to verify whether capsules 4 and 5 exceed established MRLs or not.

None of the ten tested products contained pesticide residues above our detection limits. However, given the small sample size, we cannot exclude the presence of such contaminants in other R. rosea products available on the market. Regular, comprehensive safety assessments would be instrumental in addressing public concerns regarding the quality and safety of these supplements. Enhanced regulatory oversight and standardized quality control measures are essential to ensure consumer safety and product efficacy. Enforcing Good Manufacturing Practices, ensuring transparent supply chains, and implementing third-party verification could further safeguard consumers while supporting the conservation of this valuable medicinal plant in its natural habitat.

In summary, this study represents the first investigation of biomarker molecule variability and contamination levels in commercially available R. rosea products in the U.S. While biomarker compound concentrations varied considerably, the impact of this variability on product efficacy remains unclear. More critically, we detected (occasionally considerable) heavy metal contamination in all capsular products. Consumers should therefore be vigilant about product sourcing, manufacturer transparency, ingredient traceability, and independent third-party validations. The results of our survey urge more frequent and widespread testing and improved quality control strategies for U.S. R. rosea products, to ensure both product quality and consumer safety.

Supporting information

S1 File. Lifespan extension of Drosophila melanogaster with Rhodiola rosea extracts containing different biomarker concentrations.

(XLSX)

pone.0341070.s001.xlsx (283.9KB, xlsx)

Acknowledgments

We are grateful to Eurofins Scientific, USA, for their technical and scientific support and critical review of the manuscript. This study was funded by an unrestricted gift from the Kay Family Foundation at Irvine, CA.

Data Availability

All relevant data are within the manuscript and its Supporting Information files.

Funding Statement

The author(s) received no specific funding for this work.

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Decision Letter 0

Mahmoud Yaish

30 Jul 2025

Dear Dr. Jafari,

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.

The manuscript requires some revisions before it can be resubmitted for peer review.

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Academic Editor

PLOS ONE

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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?

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: N/A

Reviewer #2: N/A

**********

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

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: Yes

**********

Reviewer #1: Adulteration of botanical ingredients and their products is a big concern for the botanical dietary supplement and related industries. Therefore information about the quality and authenticity of commercial products is important to gather. For this manuscript, the authors have used standard analytical test methods to evaluate the levels of phenolic marker compounds, some heavy metals, and pesticides in 10 commercial samples labeled to contain Rhodiola rosea root/rhizome.

The text is easy to read and follow, and the experimental part is well done. There are a few minor suggested edits, and suggestions for additional publications that may be helpful to include. But overall, the manuscript can be accepted after a few modifications.

Line 40: Please add the botanical family

Line 45: “is considered an emerging botanical”… I am not familiar with this term. Presumably the authors mean to write that rhodiola is a botanical with growing consumer interest, but this would be an odd way to state this.

Line 50: During which time period did the AGR exceed 10%?

Lines 56 and 57: According to one source (Bejar, 2017), “in some geographical areas, the two most frequently used species, R. crenulata and R. rosea, are becoming vulnerable or at-risk (one source uses the terms “threatened” and “critically endangered” when referring to specific areas).” Hence, I don’t think the sentence that R. rosea is not classified as endanegers is accurate. It depends on the location.

Line 67: What do you mean by USP Catalog No: 1602580? Please indicate which USP monograph you refer to.

Line 75: “surveillance of these products, post-market, is sporadic at best,…How do you know that? Please cite some references to support that post-market surveillance is sporadic. I am not aware of such data, but there is a lot of prejudice about what is going on in the industry.

Line 89: How did you determine “brand-diversity”. Please specify what criteria you used.

Line 90: Well-regarded distributors…like whom? Please specify which distributors you refer to. And what are your criteria for well-regarded?

Line 197: Two additional papers on the frequency of adulteration of botanicals:

- Ichim MC. The DNA-based authentication of commercial herbal products reveals their globally widespread adulteration. Front Pharmacol. 2019;10:1227. 10.3389/fphar.2019.01227

- Orhan N, Gafner S, Blumenthal M. Estimating the extent of adulteration of the popular herbs black cohosh, echinacea, elder berry, ginkgo, and turmeric – its challenges and limitations. Nat Prod Rep. 2024;41(10):1604-1621. https://doi.org/10.1039/d4np00014e

Other publications worth considering:

- Bejar E, Upton R, Cardellina II JH. Adulteration of rhodiola (Rhodiola rosea) rhizome and root extract. Botanical Adulterants Prevention Bulletin. Austin TX, ABC-AHP-NCNPR Botanical Adulterants Prevention Program. 2017;1-8. http://doi.org/10.59520/bapp.bapb/PenB1664

- Gemma S, Multari G, Turco L, Gallo FR. A densitometric HPTLC method for the simultaneous quali-quantitative determination of rosavin and salidroside in Rhodiola rosea L. based commercial products: A quick screening by HPTLC-based fingerprint and antioxidant evaluation. J Liq Chromatogr Rel Techn. 2025. 10.1080/10826076.2025.2462263

Figures 1 and 2 need to be submitted as high-resolution graphs.

Reviewer #2: Introduction:

1. add full botanical name of Rodiola rosea

2. any information on the mechanism of action is missing

3. it will be beneficial to add information about the negative effect of the potential contaminations.

Materials and methods:

1. It will be beneficial to add if the product sample was a convinience sample or you used any other criteria. When were the supplements purchased?

2. Why did you choose 7 capsule products and 3 tinctures?

Discussion:

1. it will be very beneficial to add a comment on the biological activity of the supplements based on the different biomarker molecules content.

**********

what does this mean? ). If published, this will include your full peer review and any attached files.

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Reviewer #1: Yes:  Stefan Gafner

Reviewer #2: No

**********

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PLoS One. 2026 Jan 16;21(1):e0341070. doi: 10.1371/journal.pone.0341070.r002

Author response to Decision Letter 1


22 Aug 2025

August 22, 2025

Ref: Submission PONE-D-25-18554; The quality of Rhodiola rosea supplements on the U.S. market: An analysis of biomarker molecule, heavy metal, and pesticide contents

Dear Dr. Yaish,

We sincerely thank both reviewers for their careful assessment of our manuscript and the many useful suggestions that they made. Their efforts have improved our manuscript substantially. We have carefully considered all comments and have made appropriate revisions to the manuscript. For each reviewer comment, we provide a detailed response and indicate any corresponding changes made to the manuscript.

As requested, we have modified the manuscript format to adhere to the PLoS One guidelines. Both authors agree to full publication of all underlying data related to this manuscript. Regarding our previous use of “data not shown”, we have now prepared a supplementary file that contains the underlying data in support of the relevant statement.

After further review and considering the reviewers’ comments, we have retained anonymity of all products and removed any identifying reference (e.g., specific sources such as Alaskan Rhodiola rosea) to keep the focus on the analytical findings.

We have carefully considered all comments and have made appropriate revisions to the manuscript. Please see our point-by-point response below. Reviewers’ comments are copied and listed in black, and our responses are listed in blue. For each reviewer comment, we provide a detailed response and indicate any corresponding changes made to the manuscript.

We hope these revisions address the reviewers’ comments and demonstrate our commitment to strengthening the scientific contribution of this study.

Sincerely,

Mahtab Jafari, PharmD

Professor of Pharmaceutical Sciences

Director, The UC Irvine Center for Healthspan Sciences

University of California, Irvine - School of Pharmacy and Pharmaceutical Sciences

Reviewer #1

Adulteration of botanical ingredients and their products is a big concern for the botanical dietary supplement and related industries. Therefore information about the quality and authenticity of commercial products is important to gather. For this manuscript, the authors have used standard analytical test methods to evaluate the levels of phenolic marker compounds, some heavy metals, and pesticides in 10 commercial samples labeled to contain Rhodiola rosea root/rhizome.

The text is easy to read and follow, and the experimental part is well done. There are a few minor suggested edits, and suggestions for additional publications that may be helpful to include. But overall, the manuscript can be accepted after a few modifications.

REPLY: Thank you so much for this positive feedback.

Line 40: Please add the botanical family

REPLY: The botanical family Crassulaceae was added, as suggested.

Line 45: “is considered an emerging botanical”… I am not familiar with this term. Presumably the authors mean to write that rhodiola is a botanical with growing consumer interest, but this would be an odd way to state this.

REPLY: Thank you for pointing this out. We have removed this phrase.

Line 50: During which time period did the AGR exceed 10%?

REPLY: We have now clarified the CAGR by specifying that “…in the United States, these supplements have experienced a compound annual growth rate exceeding 10% between 2019 and 2024 [11].”

Lines 56 and 57: According to one source (Bejar, 2017), “in some geographical areas, the two most frequently used species, R. crenulata and R. rosea, are becoming vulnerable or at-risk (one source uses the terms “threatened” and “critically endangered” when referring to specific areas).” Hence, I don’t think the sentence that R. rosea is not classified as endanegers is accurate. It depends on the location.

REPLY: Thank you so much for this thoughtful comment. We agree with your comment and have addressed this ambiguity by stating that “…R. rosea is currently not included in the International Union for Conservation of Nature’s Red List of Threatened Species.”

Line 67: What do you mean by USP Catalog No: 1602580? Please indicate which USP monograph you refer to.

REPLY: We apologize for this ambiguity. We referred to the US Pharmacopeial Convention’s Certificate for catalog number 1602580, Rhodiola rosea root and rhizome dry extract. We have now added the full name to the text.

Line 75: “surveillance of these products, post-market, is sporadic at best,…How do you know that? Please cite some references to support that post-market surveillance is sporadic. I am not aware of such data, but there is a lot of prejudice about what is going on in the industry.

REPLY: Thank you for your question and suggestion. We were indeed imprecise. We have now modified the statement and say that “…surveillance of these products, post-market, often simply relies on reports of adverse events and subsequent label modifications [23].” The supporting reference is Li W, Wertheimer A. 2023. Innov Pharm. PMCID: PMC10686678.

Line 89: How did you determine “brand-diversity”. Please specify what criteria you used.

REPLY: We assembled a panel of products, where each product was from a different brand and produced at a different manufacturing site. We now add this information to our selection criteria.

Line 90: Well-regarded distributors…like whom? Please specify which distributors you refer to. And what are your criteria for well-regarded?

REPLY: Thank you for pointing this out. This characterization was not based on published criteria and has therefore been removed.

Line 197: Two additional papers on the frequency of adulteration of botanicals:

- Ichim MC. The DNA-based authentication of commercial herbal products reveals their globally widespread adulteration. Front Pharmacol. 2019;10:1227. 10.3389/fphar.2019.01227

- Orhan N, Gafner S, Blumenthal M. Estimating the extent of adulteration of the popular herbs black cohosh, echinacea, elder berry, ginkgo, and turmeric – its challenges and limitations. Nat Prod Rep. 2024;41(10):1604-1621. https://doi.org/10.1039/d4np00014e

REPLY: Thank you so much, we have now added these references to the manuscript.

Other publications worth considering:

- Bejar E, Upton R, Cardellina II JH. Adulteration of rhodiola (Rhodiola rosea) rhizome and root extract. Botanical Adulterants Prevention Bulletin. Austin TX, ABC-AHP-NCNPR Botanical Adulterants Prevention Program. 2017;1-8. http://doi.org/10.59520/bapp.bapb/PenB1664

- Gemma S, Multari G, Turco L, Gallo FR. A densitometric HPTLC method for the simultaneous quali-quantitative determination of rosavin and salidroside in Rhodiola rosea L. based commercial products: A quick screening by HPTLC-based fingerprint and antioxidant evaluation. J Liq Chromatogr Rel Techn. 2025. 10.1080/10826076.2025.2462263

REPLY: We sincerely thank the reviewer for bringing these references to our attention. We now include the highly relevant Bejar publication in our revised manuscript, as suggested.

Figures 1 and 2 need to be submitted as high-resolution graphs.

REPLY: Thank you for pointing this out. We investigated the resolution of our figures.

The uploaded versions of Figures 1 and 2 have high-resolution. It appears that their resolution changes once embedded in the PDF file. We communicated this issue with the PLOS One editorial staff, and they suggested the following:

“Since reviewers and the Academic Editor will have access to your File Inventory, they can download the files in their original resolution as necessary while evaluating the manuscript. If the manuscript is accepted, the figures or images in question will be linked in their full resolution. “

Reviewer #2

Introduction:

1. add full botanical name of Rodiola rosea

REPLY: Thank you. We have added the botanical family of Rhodiola rosea (Crassulaceae) to the manuscript text.

2. any information on the mechanism of action is missing

REPLY: Thank you for pointing this out. We had refrained from such a statement because the exact mechanism of action of R. rosea is still under investigation. Most previous studies centered on possible effects of the biomarker molecules salidroside and rosavins, which are thought to contribute to the adaptogenic effects of the plant. However, not all of R. rosea’s biological activities can be attributed to these biomarker molecules, and the benefits are likely imparted by the synergistic contribution of several of the over 140 organic compounds that have been identified in the plant. We very much look forward to future studies characterizing the impact of the various compounds on the plant’s adaptogenic properties. For now, we have added the following text to the manuscript describing the current state of knowledge on the mechanism of action of the plant. “Some of the adaptogenic effects of the plant, including its antioxidant properties, have been attributed to these biomarker molecules [20, 21], but the beneficial effects cannot be consistently recapitulated with isolated rosavins or salidroside in laboratory settings. Other molecules, such as quercetin-glycosides, have also been identified as contributors [22]. Therefore, the adaptogenic properties are likely caused by the synergistic activity of at least some of the > 100 organic compounds of the plant.”

3. it will be beneficial to add information about the negative effect of the potential contaminations.

REPLY: Thank you so much for this comment. We have now expressed the dangers of heavy metal and pesticide contamination with the following section: “Heavy metal exposure can have dramatic detrimental effects on human health, particularly on kidney function [30], while pesticides may cause chronic health conditions, including asthma, cancer, or diabetes [31].” In addition, we also point out that contamination with other Rhodiola species may raise doubts regarding the product’s efficacy.

Materials and methods:

1. It will be beneficial to add if the product sample was a convinience sample or you used any other criteria. When were the supplements purchased?

REPLY: Thank you for pointing out our oversight in characterizing the selection in more detail. All capsules and tinctures were purchased in 2024, and we have now added this information to the text. The samples were bestselling R. rosea products on Amazon.com, and included products in different price ranges. We added this description to the text.

2. Why did you choose 7 capsule products and 3 tinctures?

REPLY: The inclusion of tinctures provided better coverage of the different product types on the U.S. market. We selected more capsules than tinctures because more R. rosea capsule products are sold on the market, compared to tinctures. We estimated that this small sample of products, which included the current online bestsellers in different price ranges, would provide a good snapshot of the quality of the R. rosea products on the U.S. market.

Discussion:

1. it will be very beneficial to add a comment on the biological activity of the supplements based on the different biomarker molecules content.

REPLY: This is a great idea, but it is difficult to provide such an estimate. As outlined above, and mentioned in our discussion, the biological activity of the Rhodiola rosea supplements cannot be reliably based on biomarker molecule content alone, and is, instead, a result of complex (and hitherto incompletely characterized) interactions of the many organic compounds of the plant.

Therefore, we largely refrain from speculating on the biomarkers’ biological activity. However, we included in our discussion that “Our recent lifespan assays in Drosophila melanogaster demonstrated comparable life-extending effects between R. rosea extracts with lower concentrations of rosavins and an R. rosea extract with a higher concentration of rosavins (S1 File). Whether this equivalence extends to other purported benefits of the plant, such as enhanced energy, mental health support, stress reduction, and exercise performance, remains to be determined.”

Rhodiola rosea products are standardized according to their rosavins and salidroside content, which is why we refer to them as biomarker molecules. Our goal was to verify the composition of the products on the market in the U.S., and to estimate their safety by checking for contamination with some of the most important health hazards. A survey of the biological activity of the products or the biomarker molecules was beyond the scope of this study. We wholeheartedly agree that future studies focusing on the biological activity of these products and their biomarkers would be highly informative. These future studies may include assays measuring the biological activities of a standardized amount of each product in cell cultures, as well as physical performance and stress evaluations in laboratory animals.

Attachment

Submitted filename: 250822 Rhodiola Quality PLOS ONE Response to Reviewers .docx

pone.0341070.s003.docx (34.8KB, docx)

Decision Letter 1

Mahmoud Yaish

20 Nov 2025

Dear Dr. Jafari,

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.

The manuscript is important and suitable for publication, but it still needs significant improvements in clarity, methodological detail, formatting, and data presentation before it can be accepted.

Please submit your revised manuscript by Jan 04 2026 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.

  • A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.

  • A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.

  • An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols .

We look forward to receiving your revised manuscript.

Kind regards,

Mahmoud W. Yaish, Ph.D.

Academic Editor

PLOS ONE

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If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. 

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Additional Editor Comments:

The manuscript is important and suitable for publication, but it still needs significant improvements in clarity, methodological detail, formatting, and data presentation before it can be accepted.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

Reviewer #3: (No Response)

**********

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

Reviewer #3: (No Response)

**********

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

Reviewer #3: (No Response)

**********

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

The PLOS Data policy

Reviewer #3: (No Response)

**********

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

Reviewer #3: (No Response)

**********

Reviewer #3: The manuscript with the title: The quality of Rhodiola rosea supplements on the U.S. market: An analysis of biomarker molecule, heavy metal, and pesticide contents”, the study is valuable, timely, and relevant to public health, pharmacognosy, and dietary supplement quality control. The work is interesting and the work is suitable for the scope of PLOS one. Here are some comments for improving the manuscript.

1- The title could be changed to Quality and safety of Rhodiola rosea supplements on the U.S. market: variability in biomarkers, heavy metals and pesticide residues

2- In the abstract: Add the analytical methods used in the study (e.g., UPLC, ICP-MS, QuEChERS) to improve clarity in the abstract.

3- in the abstract, the detected concentration range is missing, please adda in the abstract.

4- Keywords is missing, authors should add the keywords in the revised version.

5- All abbreviation should be mentioned in the revised manuscript before the introduction section.

6- Line 58, add suitable reference.

7- in materials and methods section: Please specify how many capsules were pooled per product for analysis?

8- The models of instruments such as ICP-MS, should be mentioned in M& M.

9- Most of the references of the methods are missing in the materials and methods.

10- There is no mentioned about the number of replication in the experiments (duplicate or triplicate).

11- The samples which do not meet the USP minimium limits should be highlighted in the table.

12- Figures 1 & 2: If replicates were performed, add error bars.

13- heavy metals should be in the form heavy metals ions.

14- Figures need high resolution.

15- References, some references the journals are abbreviated , other journals in full name, authors should follow the journal instruction.

**********

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 #3: Yes:  Sedky Hassan

**********

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PLoS One. 2026 Jan 16;21(1):e0341070. doi: 10.1371/journal.pone.0341070.r004

Author response to Decision Letter 2


30 Dec 2025

Reviewer #3

The manuscript with the title: The quality of Rhodiola rosea supplements on the U.S. market: An analysis of biomarker molecule, heavy metal, and pesticide contents”, the study is valuable, timely, and relevant to public health, pharmacognosy, and dietary supplement quality control. The work is interesting and the work is suitable for the scope of PLOS one. Here are some comments for improving the manuscript.

Thank you for the kind evaluation of our work.

1- The title could be changed to Quality and safety of Rhodiola rosea supplements on the U.S. market: variability in biomarkers, heavy metals and pesticide residues

Thank you for this suggestion. We have modified the title and incorporated most of the requested changes. Since there was no variability in pesticide residues, we changed the title to “The quality and safety of Rhodiola rosea supplements on the U.S. market: An analysis of biomarkers, heavy metals, and pesticide residues.”

2- In the abstract: Add the analytical methods used in the study (e.g., UPLC, ICP-MS, QuEChERS) to improve clarity in the abstract.

Thank you for this excellent idea. We have now modified the abstract to include these technical details.

3- in the abstract, the detected concentration range is missing, please adda in the abstract.

Thank you for this suggestion. We have added the concentration ranges to the abstract, as suggested.

4- Keywords is missing, authors should add the keywords in the revised version.

While keywords are not requested or shown in PLoS One’s author guidelines, we now provide five of them.

5- All abbreviation should be mentioned in the revised manuscript before the introduction section.

PLoS One’s instructions for authors at https://journals.plos.org/plosone/s/submission-guidelines.com recommend to “[d]efine abbreviations upon first appearance in the text.” We have chosen to follow this guideline in our submission. We have also removed some abbreviations that only occurred once in the manuscript.

6- Line 58, add suitable reference.

We have expanded the text in the manuscript to clarify the way Rhodiola rosea trade regulations were instituted by the Convention on International Trade in Endangered Species, and have added this reference, as suggested by the reviewer.

7- in materials and methods section: Please specify how many capsules were pooled per product for analysis?

Thank you for this suggestion. We have added the following sentence to the Sample selection and preparation section in Materials & methods: “Fill material from 3 – 10 capsules per product was used for the analysis.”

8- The models of instruments such as ICP-MS, should be mentioned in M& M.

We have added the instruments used for the ICP-MS (Agilent 7900), the UPLC (Waters Acquity I-class), the GC/MS (Agilent 7880 and 8890), and the LC/MS protocols (Agilent 6400), as requested.

9- Most of the references of the methods are missing in the materials and methods.

Thank you for pointing out this omission. We have now included additional specific references from the AOAC compendium used in industrial settings for heavy metal and pesticide detection protocols. Please note that these procedures are routine and standardized industrial protocols used in U.S.-accredited and certified laboratories such as Eurofins.

10- There is no mentioned about the number of replication in the experiments (duplicate or triplicate).

Measurements were only performed once, a strategy also applied in similar high-profile publications evaluating biological compounds in dietary products, such as Cohen et al., 2023 (Quantity of melatonin and CBD in melatonin gummies sold in the US. JAMA. 2023;329(16):1401–1402. doi:10.1001/jama.2023.2296) or Crawford et al., 2022 (Analysis of select dietary supplement products marketed to support or boost the immune system. JAMA Netw Open. 2022;5(8):e2226040. doi:10.1001/jamanetworkopen.2022.26040).

The analyses reported in our manuscript were performed by Eurofins Scientific. The technical error in all applied methods was precisely controlled by reference standards sourced from qualified vendors, and measurements were automatically repeated when system suitability criteria were not met. Furthermore, all SOPs and associated documentation were tightly followed in each method, minimizing potential measurement errors.

11- The samples which do not meet the USP minimium limits should be highlighted in the table.

Thank you for this comment. Please note that the USP minimum limits are set for dried plant roots and rhizomes, as indicated in the Introduction. Those limits do not apply to dietary products containing Rhodiola rosea. Therefore, we refrained from implementing the suggested highlights. However, we have now added the advertised biomarker content on the products’ labels (where known) to Table 1, and marked with an asterisk the two capsular products where measured biomarker amounts were significantly lower than advertised.

12- Figures 1 & 2: If replicates were performed, add error bars.

As explained in our response to comment #10, replicate analyses were not performed due to the controlled precision of the measurements. Therefore, no error bars are shown.

13- heavy metals should be in the form heavy metals ions.

Thank you for this comment. We have rephrased the corresponding section in our manuscript (Heavy metals and pesticides in the Results chapter) and altered the Y-axis title in Figure 2 to reflect that the ions of heavy metals were detected in our analyses.

14- Figures need high resolution.

We agree and hope that our original images fulfill these criteria. We tested our graphs on Newgen Art Analysis, as suggested in PLoS One’s Instructions for Authors, and they passed. We are uncertain why the images appear in such low resolution after upload onto the PLoS One server, and we will work with the editor if this problem persists.

15- References, some references the journals are abbreviated , other journals in full name, authors should follow the journal instruction.

Thank you for pointing this out. We have now corrected this oversight.

Attachment

Submitted filename: 251226 PLoS One Rhodiola Response to reviewer.docx

pone.0341070.s004.docx (24.6KB, docx)

Decision Letter 2

Mahmoud Yaish

2 Jan 2026

The quality and safety of Rhodiola rosea supplements on the U.S. market: An analysis of biomarkers, heavy metals and pesticide residues

PONE-D-25-18554R2

Dear Dr. Jafari,

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,

Mahmoud W. Yaish, Ph.D.

Academic Editor

PLOS One

Additional Editor Comments (optional):

The authors have provided satisfactory responses to all reviewer comments and have adequately addressed the identified concerns. The manuscript is now substantially improved, and its scientific merits support acceptance for publication.

Reviewers' comments:

Acceptance letter

Mahmoud Yaish

PONE-D-25-18554R2

PLOS One

Dear Dr. Jafari,

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS One. Congratulations! Your manuscript is now being handed over to our production team.

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on behalf of

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Academic Editor

PLOS One

Associated Data

    This section collects any data citations, data availability statements, or supplementary materials included in this article.

    Supplementary Materials

    S1 File. Lifespan extension of Drosophila melanogaster with Rhodiola rosea extracts containing different biomarker concentrations.

    (XLSX)

    pone.0341070.s001.xlsx (283.9KB, xlsx)
    Attachment

    Submitted filename: 250822 Rhodiola Quality PLOS ONE Response to Reviewers .docx

    pone.0341070.s003.docx (34.8KB, docx)
    Attachment

    Submitted filename: 251226 PLoS One Rhodiola Response to reviewer.docx

    pone.0341070.s004.docx (24.6KB, docx)

    Data Availability Statement

    All relevant data are within the manuscript and its Supporting Information files.


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