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Phytochemical and GC-MS analysis of Thevetia peruviana fruit methanol extract as an anti-rodenticide potential against balb C rats · 10.1016/j.heliyon.2024.e29012

Name: Verified Rigor Record — Phytochemical and GC-MS analysis of Thevetia peruviana fruit methanol extract as an anti-rodenticide potential against balb C rats · 10.1016/j.heliyon.2024.e29012
Item: Phytochemical and GC-MS analysis of Thevetia peruviana fruit methanol extract as an anti-rodenticide potential against balb C rats · 10.1016/j.heliyon.2024.e29012
Rating: 71
Author: Αlpha¹

Paper source

Αlpha¹ Editorial

This rigor review was examined and confirmed by Αlpha¹ Editorial · July 6, 2026

Αlpha¹ Rigor Review

Rigor Reviewed

WARN

Warnings to address

Eight dimensions — click a segment for details

Critical

High-priority

Model-reported confidence: HighReviewer agreement: highEngine: 7.5.0Source: full text

Not verified in this report

No reported statistical tests were found to recompute.
Declared data/code links were not checked for liveness or content.

This Αlpha¹ Rigor Review uses AI Rigor Reviewers trained on a curated corpus of high-fidelity and retracted papers, with expert supervision and curation. It can still make mistakes; verify each finding against the source before relying on it.

7 major claims checked against the paper's own evidence: 1 not fully backed by the presented evidence (unsupported or overstated).

7 integrity concerns flagged (1 high).

Rigor

Eight Dimensions1 pass · 7 warn

7 major claims1 overstated1 partial5 supported

The introduction cites multiple studies on T. peruviana toxicity, the presence of cardiac glycosides, and previous reports of antimicrobial, wound-healing, and anti-termite activities. The logical link from known toxicity to potential anti-rodenticide use is clear. The study aims to exploit toxicity that 'has not yet been fully exploited as an anti-rodenticide.' Limitations of prior synthetic rodenticides (environmental pollution, toxicity to non-targets) are described as motivation, but the paper does not explicitly discuss limitations of the prior T. peruviana studies themselves.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedINADEQUATE

Per reviewer (2) — the votes behind the verdict

PASSReviewer 1· 90% conf

The paper establishes a clear scientific premise for evaluating a natural anti-rodenticide from Thevetia peruviana, citing prior work on its toxicity and phytochemistry.

Evidence

direct quote[Introduction, paragraph 3]

“Knowledge about the toxicity of T. peruviana has been clear since ancient times, with fatal poisoning of insects, animals, and humans.”

direct quote[Introduction, last paragraph]

“The toxicity effect reported in T. peruviana leaf, root and fruit extract has not yet been fully exploited as an anti-rodenticide and this study aimed at exploiting the toxicity of T. peruviana fruit in developing an anti-rodent.”

direct quote[Introduction, paragraph 5]

“Various synthetic products used for rat control ... have heavy metals such as zinc and arsenic which contribute to environmental pollution and are highly toxic.”

PASSReviewer 2· 95% conf

The scientific premise is well-established: prior work on T. peruviana toxicity is cited, the rationale for testing its anti-rodenticidal potential is logical, and limitations of prior work are partially addressed.

Evidence

direct quote[Introduction, paragraphs 2-4]

“Toxicity of T. peruviana has been related to digitalis toxicity where pathophysiology involves direct inhibition of sodium ion (Na+) – potassium ion (K+) – ATpase pump of the heart”

direct quote[Introduction, paragraph 5]

absence[Introduction]

Limitations of prior T. peruviana studies are not discussed; only the drawbacks of synthetic rodenticides are addressed.

The Methods state that mice were 'randomly selected' but no randomization method or unit is given. Blinding is not mentioned at any stage. No power analysis is reported. The distinction between biological and technical replicates is not discussed (each mouse is a biological replicate; n=5 per group). Inclusion criteria are only 'body weights of 20–25 g' and '8–12 weeks old,' but no exclusion criteria. Outlier handling is not addressed. A control group (Cage 5 with no extract) is present, but no positive control (e.g., a known rodenticide) is described. Independent replication is not mentioned. The stepwise acute toxicity protocol is referenced from prior work, which partly mitigates the lack of detail.

Sub-criteria

controlsINADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitNOT REPORTED

inclusion exclusionINADEQUATE

randomization methodNOT REPORTED

replicate distinctionNOT REPORTED

independent replicationNOT REPORTED

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 75% conf

The study design has several inadequacies: lack of clear randomization method, no blinding, no power analysis, and no distinction between biological and technical replicates.

Evidence

direct quote[Section 2.7.2, paragraph 2]

“Balb C mice were randomly selected, marked to allow individual identification”

absence[Section 2.7]

No blinding reported.

absence[Section 2.7.2]

No power analysis reported.

WARNReviewer 2· 90% conf

The study design has several reporting gaps: randomization method and unit are not described, blinding is not mentioned, no a priori power analysis, inclusion/exclusion criteria are vague, outlier handling is absent, and independent replication is not reported. However, the acute toxicity test uses a stepwise method with a control group and some dose–response observations.

Evidence

direct quote[Section 2.7.2]

“Balb C mice were randomly selected, marked to allow individual identification”

absence[Methods]

Blinding is not mentioned.

direct quote[Table 5]

“Cage 5 (Control) 0.00 5 5 0%”

The paper states 'Adult males and females balb C rats' and later 'Female balb C mice used were nulliparous and non-pregnant', indicating both sexes were used. However, no scientific justification is given for using both sexes; this is not a 'single-sex' study, so sex_justified is not an applicable sub-criterion. Age (8–12 weeks) and weight (20–25 g) are reported. Species and strain are adequately stated (Balb C mice) but the source (SAFARI at JKUAT) is vague. Housing conditions are described briefly: 'well ventilated room with enough light and temperature (20–25 °C)' but no light cycle, enrichment, or welfare details. Demographics is not applicable for animal studies. Applicable criteria: sex_reported (adequate), age_weight_health (adequate), species_strain_source (adequate), housing_conditions (reported_but_inadequate). 3 of 4 applicable adequate = 75% → pass for scoring, but housing is inadequate, so overall status is warn due to the inadequate housing detail.

Sub-criteria

demographicsN/A

sex reportedADEQUATE

sex justifiedN/A

age weight healthADEQUATE

housing conditionsINADEQUATE

species strain sourceADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 80% conf

Sex, age, weight, and species/strain are reported, but sex is mixed without justification and housing conditions are underspecified.

Evidence

direct quote[Section 2.7.1]

“Adult males and females balb C rats were used for experiments.”

direct quote[Abstract]

“The animals 8–12 weeks' old were grouped into 5 cages (5 animals per cage) ... with body weights of 20–25 g.”

direct quote[Section 2.7.1]

“The mice were housed in cages (5 mice per cage) in a well ventilated room with enough light and temperature (20–25 °C).”

PASSReviewer 2· 90% conf

Sex (both males and females), age (8–12 weeks), body weight (20–25 g), species/strain (Balb C mice), and source (SAFARI at JKUAT) are reported. Housing conditions (ventilated room, light, temperature 20–25 °C, food and water) are described. Health status is not explicitly stated but can be inferred from acclimatization.

Evidence

direct quote[Section 2.7.1]

“Adult males and females balb C rats were used for experiments. A total of 25 male mice with body weights of 20–25 g were purchased from the Laboratory of Small Animal Facility for Research and Innovation (SAFARI) at Jomo Kenyatta University of Agriculture Technology, Kenya.”

direct quote[Section 2.7.2]

“Each mouse, at the commencement of its dosing, was between 8 and 12 weeks old”

The paper states: 'The animal management protocol and the experimental design was accepted by the Research Ethics Committee, Jomo Kenyatta University of Agriculture Technology, Kenya' but does not provide a protocol number or explicit statement of approval (e.g., 'approved'). It does not cite any regulatory framework (e.g., Guide for the Care and Use of Laboratory Animals) or institutional animal welfare guidelines. The study involves animal subjects (Balb C mice), so ethics criteria apply. The iacuc_statement (nameless body and no approval) is inadequate. Regulatory compliance is not reported.

Sub-criteria

iacuc statementINADEQUATE

informed consentN/A

irb ethics statementN/A

regulatory complianceNOT REPORTED

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 85% conf

An ethics committee is named (Research Ethics Committee, JKUAT) but no protocol number or approval statement is provided, and regulatory compliance is absent.

Evidence

direct quote[Section 2.7.1]

“The animal management protocol and the experimental design was accepted by the Research Ethics Committee, Jomo Kenyatta University of Agriculture Technology, Kenya”

absence[Section 2.7]

No regulatory compliance statement.

WARNReviewer 2· 85% conf

An ethics approval statement is present but does not name the specific committee and lacks a protocol number, making it reported_but_inadequate. No animal welfare regulatory compliance is stated. Informed consent and IRB are not applicable (animal study). IACUC statement is the applicable criterion but is not adequately reported.

Evidence

direct quote[Section 2.7.1]

“The animal management protocol and the experimental design was accepted by the Research Ethics Committee, Jomo Kenyatta University of Agriculture Technology, Kenya [].”

absence[Methods]

Regulatory compliance not reported.

The study uses a methanol extract of T. peruviana fruit as the investigational product. The source is described (JKUAT botanical garden, voucher specimen AIN–JKUATBH/001/2021), but no batch/lot number, purity, or storage conditions are provided for the extract. The FTIR (Shimadzu FTS-8000) and GC-MS (Agilent 5975) instruments are named, but no software versions or RRIDs are given. No antibodies, cell lines, or organisms other than the study animals were used. Mycoplasma testing is not applicable. Reagents (methanol, Folin-Ciocalteu, etc.) are listed but without catalog numbers or suppliers. Software (e.g., for probit analysis, ANOVA) is not identified.

Sub-criteria

mycoplasma testingN/A

reagents identifiedINADEQUATE

organisms identifiedINADEQUATE

antibodies identifiedN/A

cell line authenticationN/A

software tools identifiedNOT REPORTED

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 85% conf

The investigation product (T. peruviana fruit extract) is identified, but no details on preparation batch, lot, or purity are given; analytical instruments are named but software versions are missing.

Evidence

direct quote[Section 2.2]

“T. peruviana fruits were collected from Jomo Kenyatta University of Agriculture and Technology Botanical Garden ... voucher specimens were deposited ... AIN–JKUATBH/001/2021”

direct quote[Section 2.6.1]

“The Shimadzu Fourier Transform Infrared spectrophotometer (FT-IR) (FTS- 8000, Japan) was used to analyze functional groups”

direct quote[Section 2.1]

“Chemicals: 99.8 % Methanol, Gallic Acid, 5 % sodium nitrite, 1 N Folin-Ciocalteu reagent”

WARNReviewer 2· 80% conf

The study uses chemicals, plant material, extraction solvents, and a GC-MS instrument but does not provide catalogue numbers or RRIDs for reagents, nor software versions. The extract is the investigational product; its preparation is described. Software (e.g., NIST 17 database) is mentioned but version and vendor details are sparse.

Evidence

direct quote[Section 2.1]

“Chemicals: 99.8 % Methanol, Gallic Acid, 5 % sodium nitrite, 1 N Folin-Ciocalteu reagent, 0.1 mM DPPH solution, 10 % aluminium chloride, tannic acid, Mayer reagent, 5 % sodium carbonate, Lead acetate, Ferric chloride, 4 % sodium hydroxide, Alkaline reagent, Iodine solution, sodium hydroxide, Wagner's reagent, and Molisch's reagent.”

direct quote[Section 2.6.2]

“GC–MS analysis of crude T. peruviana was evaluated using a shimadzu GC–MS”

direct quote[Table 4]

“NIST 17 spectral data base”

Section 2.9: 'The data was subjected to probit analysis and ANOVA.' No details on which software, which ANOVA model (one-way?), post-hoc tests, or normality checks. Table 5 presents mortality counts per cage (n=5 each), which is adequate for an acute toxicity study (no individual data points needed per se). The paper does not provide p-values, effect sizes, or confidence intervals for any comparison. Statistical software is not identified. The mathematical plausibility of the mortality counts is fine: all numbers are whole, percentages sum correctly. However, the statement 'as the concentration increases, death rate also reduces' contradicts the table (Cage 4 at 20% extract shows 80% mortality, lower than Cage 2 and 3 at 100%, so some reduction, but this is not tested statistically).

Sub-criteria

tests namedINADEQUATE

exact p valuesNOT REPORTED

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedNOT REPORTED

assumptions verifiedNOT REPORTED

mathematical plausibilityPASS

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 80% conf

Statistical methods are vaguely described ('probit analysis and ANOVA'), assumptions not verified, exact p-values not reported, and data presentation lacks individual points.

Evidence

direct quote[Section 2.9]

“The data was subjected to probit analysis and ANOVA.”

absence[Section 3]

No exact p-values reported.

absence[Section 3.5]

No effect sizes or confidence intervals.

WARNReviewer 2· 85% conf

Statistical methods are named (probit analysis, ANOVA) but not detailed. Assumptions are not verified. Effect sizes and confidence intervals are not reported. Exact p-values are not given. Software is not identified. Data presentation (Table 5) shows group mortality counts but no individual data points or dispersion. The GRIM test cannot be applied to these data because the outcome is mortality (counts) and no means of continuous variables are used for inferential tests. Most applicable criteria are reported_but_inadequate or not_reported.

Evidence

direct quote[Section 2.9]

“The data was subjected to probit analysis and ANOVA.”

absence[Results]

No p-values or effect sizes reported.

direct quote[Table 5]

“Cage 1 (1.25% extract) 0.25 5 5 4 (80%)”

The statement says data are not in a repository but are included in the article/supplementary. The article itself contains the mortality table and figures; no raw data (e.g., individual animal weights, histopathology scores) are provided as supplementary. The statement does not explain why data are not in a repository or where they could be obtained. For a preclinical animal study with small n, inclusion in the article is typical but a more explicit statement (e.g., 'available from corresponding author upon reasonable request') would be adequate. Criteria: data_availability_statement is reported_but_inadequate; repository_deposit and accession_numbers are not_applicable (no sequencing or large datasets); code_sharing is not_applicable (no custom code).

Sub-criteria

code sharingN/A

accession numbersN/A

repository depositN/A

data availability statementINADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 90% conf

A data availability statement is present but vague; no data or code in a public repository.

Evidence

direct quote[Data availability statement]

“Data associated with this study has not been publicly availed in any repository. The data has been included in the Article/Supplementary Material/Referenced in article.”

absence[Data availability statement]

No repository deposit.

WARNReviewer 2· 95% conf

A data availability statement is present but vague: 'Data associated with this study has not been publicly availed in any repository. The data has been included in the Article/Supplementary Material/Referenced in article.' This is reported_but_inadequate because it does not specify a concrete route or repository. No code was shared; no accession numbers. Repository deposit and code sharing are not applicable because data are included in the article.

Evidence

direct quote[Data availability statement]

“Data associated with this study has not been publicly availed in any repository. The data has been included in the Article/Supplementary Material/Referenced in article.”

The methods section describes extraction, phytochemical analysis, and animal procedures, but lacks sufficient detail for replication (e.g., exact dosing regimen, random sequence generation). No trial registration is applicable (animal study). No reporting guideline (e.g., ARRIVE) is mentioned. All outcomes are reported (LD50, histopathology, phytochemical content), but negative results (e.g., non-significant findings) are not addressed. Limitations are not explicitly discussed in a dedicated section. Conclusions are largely proportional, stating that T. peruviana extract 'can be utilized as an anti-rodenticide' based on the observed toxicity. Funding sources and conflict of interest are declared.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessINADEQUATE

all outcomes reportedINADEQUATE

limitations discussedNOT REPORTED

conclusions proportionalADEQUATE

Per reviewer (2) — the votes behind the verdict

WARNReviewer 1· 85% conf

Methods are partially complete, but no trial registration or reporting guideline is followed; limitations are briefly discussed and conclusions are proportional.

Evidence

direct quote[Section 2.7.2]

“Balb C mice were randomly selected, marked to allow individual identification”

absence[Section 2.7]

No reporting guideline mentioned.

direct quote[Declaration of competing interest]

“The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.”

WARNReviewer 2· 85% conf

Methods are described in fair detail for a basic acute toxicity study, but some steps are ambiguous. The study is not a clinical trial, so trial registration is not applicable. No reporting guideline (ARRIVE) is referenced. All outcomes mentioned (mortality, histopathology, behavior) are reported qualitatively. Limitations are briefly implied but not explicitly discussed. Conclusions are proportional to the data: the extract caused toxicity and can be used as anti-rodenticide. Funding/COI statement is present.

Evidence

paraphrase[Section 2.3]

“The extraction yielded a solid extract but the percentage yield is not reported.”

absence[Methods]

No reporting guideline referenced.

absence[Discussion]

Limitations not discussed.

20 copyedit issues flagged (3 major): mostly grammar, typo, consistency.

Checked 38 references: 12 verified — 26 unresolved.

Full text (39,784 chars)Analyzed: Jul 6, 2026

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