Sage Journals: Discover world-class research

Abstract

Sickle cell disease (SCD) remains a major public health burden, especially in sub-Saharan Africa, where access to conventional therapies is limited. Consequently, herbal medicines/medicinal plants are increasingly utilised for SCD management. However, standardisation and quality control of these medicinal plants are hindered by variability in phytochemical composition, underscoring the relevance of marker compounds. This systematic review applies the Herbal Marker Ranking System (Herb MaRS) to identify and prioritise appropriate marker compounds for quality control of the most frequently reported anti-sickling medicinal plants. Selected plants include Carica papaya, Terminalia catappa, Annona muricata, Ceiba pentandra, Ocimum gratissimum, Garcinia kola, and Rauvolfia vomitoria. Data on phytochemical composition, anti-sickling bioactivity, toxicity, compound concentration, and availability of analytical methods and standards were obtained through a systematic literature review. Marker compounds were ranked based on adapted Herb MaRS criteria prioritising anti-sickling activity, safety, and analytical feasibility. Overall, 35 compounds were identified, with rutin and ellagic acid achieving the highest scores (8/8), indicating their suitability as marker compounds. Quercetin (7/8), carpaine (6/8), gallic acid (6/8), campesterol (6/8), and stigmasterol (6/8) also ranked highly. Several compounds demonstrated toxicity concerns requiring monitoring, while others lacked sufficient data for full evaluation. This study demonstrates the utility of the Herb MaRS framework for systematic marker selection to support the quality control of herbal formulations for SCD management. It highlights promising chemical markers that can facilitate standardisation, advancing the safe and effective use of plant-based therapies for SCD.

Keywords

chemical markers sickle cell disease quality control medicinal plants herbal marker ranking system (Herb MaRS)

Introduction

Sickle cell disease (SCD) is a monogenic condition characterised by mutation in the beta-globin gene, leading to the formation of uncharacteristic haemoglobin S (HbS).¹ The abnormality results in deoxygenated HbS, which polymerises, causing erythrocytes of victims to assume a sickle shape.² The malformed cells contribute to vaso-occlusion, chronic haemolytic anaemia, organ damage, and a range of lethal complications.³ SCD is the most prevalent monogenic disorder globally, with an estimated 300 000 infants born annually with the condition; a figure projected to exceed 400 000 by 2050.⁴ This burden is unduly high in sub-Saharan Africa, which accounts for approximately 75% of global SCD births. Alarmingly, due to limited access to early diagnosis and adequate healthcare services, the majority of affected children in these regions do not survive beyond the age of five.^4–7 The management of SCD incurs substantial healthcare costs for individuals affected by the condition. In Sub-Saharan Africa, most individuals live below the poverty line and cannot afford conventional treatments such as hydroxyurea or blood transfusions.⁷ Consequently, indigenes in this region have increasingly resorted to medicinal plants to manage SCD and other diseases, leveraging traditional knowledge to access affordable and culturally accepted remedies.^8,9

Medicinal plants have been extensively used in Africa and other regions to manage SCD and its symptoms, offering affordable alternatives to conventional treatments. Ethnobotanical studies have reported these uses in countries including Nigeria^8,10,11, Cameroon⁹ and the Ivory Coast.¹² Some of these medicinal plants have been validated for SCD management, with Amponsah et al⁷ reporting a total of 117 plant species with anti-sickling activities. Out of the 117 reported species, seven (7), Carica papaya L. (Caricaceae), Terminalia catappa L. (Combretaceae), Annona muricata L. (Annonaceae), Ceiba pentandra (L.) Gaertn. (Malvaceae), Ocimum gratissimum L. (Lamiaceae), Garcinia kola Heckel (Clusiaceae), and Rauvolfia vomitoria Afzel. (Apocynaceae) were the most frequently reported. These plants contain various bioactive compounds which show promise as lead molecules for alternative product development for SCD management.

The efficacy and safety of herbal medicines depend on consistent quality, which is often affected by variability in phytochemical content due to environmental factors, harvesting practices, and processing methods.¹³ This underscores the importance of regular quality control measures in standardising the medicinal plants and their formulations. The usage of marker compounds has become a popular and dependable strategy among suggested quality control techniques.^14,15 Particularly in complex multi-herb formulations, marker compounds provide a dependable means of assessing batch-to-batch consistency, detecting product variability, and monitoring the concentrations of active constituents.¹⁶ Nevertheless, selecting appropriate markers for standardisation remains challenging due to the complex and diverse chemical composition of medicinal plants. This highlights the need for a structured approach, such as the Herbal Marker Ranking System (Herb MaRS).

The Herbal Marker Ranking System (Herb MaRS), created by Bensoussan et al¹⁷ and adapted by Addotey et al¹⁶, provides a systematic framework for selecting marker compounds. This selection focuses on biological activity, concentration of potential marker compounds in plant material, reports of analytical assays and standards, and safety profiles of potential markers. However, its application has not been explored in SCD management plants, leaving a critical gap in the standardisation of formulations incorporating such medicinal plants. Therefore, this study aims to identify and rank potential marker compounds for quality control of herbal medicinal products formulated from the most reported anti-sickling plant species reported by Amponsah et al⁷, using the Herb MaRS framework.

Method

Medicinal Plants Selection

This study builds upon the findings of a systematic review by Amponsah et al⁷, which identified medicinal plants most frequently reported for SCD management. The plants included Carica papaya, Terminalia catappa, Annona muricata, Ceiba pentandra, Ocimum gratissimum, Garcinia kola, and Rauvolfia vomitoria. The plants were chosen for Herb MaRS evaluation to rank and choose potential marker compounds for standardising medicinal plants and/or herbal medicines that incorporate them in formulations.

Data Collection Methods

A systematic, literature-based approach was employed to compile detailed information on the selected plants. A comprehensive desktop review was conducted to extract data on their anti-sickling properties, isolated compounds, safety profiles, and potential chemical markers. Data were retrieved from multiple scientific databases, such as Google Scholar, PubMed, ScienceDirect, Semantic Scholar and Web of Science. The terms included in the search were the botanical names of the plants in combination with keywords such as “anti-sickling activity,” “sickle cell disease management,” “phytochemicals,” and “bioactive compounds.” The date of publication of obtained articles was not constrained to any specific year, and only manuscripts that were published in the English language were considered.^16,18

Reports of Bioactivity and Toxicity of Medicinal Plants Selected

A database was compiled cataloguing the selected medicinal plants, with each entry detailing the plant's scientific name, family, and documented anti-sickling properties. An extensive review was conducted to gather evidence of biological activity related to SCD management, focusing on mechanisms such as inhibition of haemoglobin S polymerisation and erythrocyte membrane stabilisation. Safety profiles were assessed by reviewing toxicity data from sources such as the Pharmacopoeias (West African Herbal¹⁹ and the Ghana Herbal²⁰), peer-reviewed articles, and pharmacological databases. The compiled data comprised information on documented studies and, where available, clinical evidence on the activity and toxicity profile of selected plants.^16,18

Potential Chemical Markers Identification

Potential chemical markers for the selected plants were identified using a modified version of Herb MaRS¹⁷, adapted to prioritise compounds with anti-sickling activity.^16,18 The criteria for selection entailed reported biological (anti-sickling) activity, reported bioactivity pertinent to SCD management, reported concentration of compounds in respective plant materials, commercial reference standards’ availability, existence of validated analytical methods, and documented toxicity profiles.¹⁸

Bioactivity and Toxicity Profiles of Potential Markers

A thorough literature review was conducted in order to identify compounds that were obtained from the plants being studied and to compile a list of their documented anti-sickling properties.

The review focused on compounds with reported effects on haemoglobin polymerisation, oxidative stress, or erythrocyte stability, as these are critical mechanisms in SCD management. Toxicity profiles of the compounds were similarly systematically documented. Searches were conducted across Web of Science, PubMed, Google Scholar, and Semantic Scholar, using criteria combining plant names, compound names, and terms like “anti-sickling activity” and “sickle cell disease.” Only full-length peer-reviewed manuscripts written in the English language were considered. The date of publication was not a barrier to manuscript consideration.¹⁶

Review of Marker Compounds Concentration in Selected Plants and Analytical Methods Availability

Literature was reviewed to identify reports/studies quantifying marker compounds in the respective plant materials. Each marker compound's concentration in the respective plant was assessed, which is intended to facilitate reliable detection and standardisation. Additionally, reports on validated assay methods for assaying chemical markers were assessed to ensure their applicability in quality control. The analytical assay procedures were prioritised based on their sensitivity, reproducibility, and accessibility, particularly in resource-limited settings.^16,18

Review of Analytical Reference Standards Availability

The commercial accessibility of reference standards of potential chemical markers was reviewed and documented to support their use in quality control. This was done following the method by Amponsah et al,¹⁸ where the data was sourced from reputable suppliers. The unit prices and availability of standards were recorded to assess feasibility for routine analysis. Compounds lacking commercially available standards were noted as less suitable for immediate standardisation purposes¹⁸.

Prioritisation of Appropriate Markers for Standardisation

The catalogued compounds were prioritised using a modified scoring system adapted from Amponsah et al,¹⁸ with compounds scoring from 0 to 8. 0 represented the least ideal chemical marker, while 8 represented the most ideal chemical marker. Scoring was conducted as follows: evidence of anti-sickling activity (0 or 2 points); evidence of bioactivity related to SCD management (0 or 1 point); concentration of potential chemical marker in raw material (<5 µg/g: 1 point; 5-50 µg/g: 2 points; >50 µg/g: 3 points); availability of a commercial reference standard (1 point), and existence of a validated analytical method (1 point). Compounds with documented toxicity were flagged for cautious use and adherence to safety protocols. Consequently, they were assigned a score of 8 points. The highest-scoring compounds were prioritised as potential marker compounds for standardising herbal products formulated from these plants.

Results

Evidence of Bioactivity and Toxicity of Medicinal Plants Selected

There have been reports of the selected medicinal plants demonstrating anti-sickling activity, indicating their potential as therapeutic agents for managing SCD (Table 1). In addition to the anti-sickling activity, Garcinia kola also exhibits erythrocyte membrane stabilisation, a unique bioactivity that may enhance its efficacy in preventing sickling. Carica papaya, Terminalia catappa, Annona muricata, and Ceiba pentandra showed low acute toxicity with LD₅₀ values exceeding 2000-5000 mg/kg. Garcinia kola is markedly more toxic, with an LD₅₀ of 358 mg/kg, while doses above 300 mg/kg of Rauvolfia are associated with liver damage.

Table 1.

Evidence of Activity and Toxicity of Catalogued Plants.

Medicinal plant	Evidence of activity associated with SCD management	Evidence of toxicity
Carica papaya L. (Caricaceae)	Anti-sickling activity, ie in vitro and in vivo^21,22	NOAEL ≥ 2 g/kg/b.w, LD₅₀ > 2000 mg/kg²³, 5000 mg/kg²⁴.
Terminalia catappa L. (Combretaceae)	Anti-sickling activity^25–27	LD₅₀ > 5000 mg/kg²⁸, Non-toxic²⁹.
Annona muricata L. (Annonaceae),	Anti-sickling activity^30,31	LD₅₀ > 5000 mg/kg³², Genotoxic³³
Ceiba pentandra (L.) Gaertn. (Malvaceae)	Anti-sickling activity^34,35	LD₅₀ > 5000 mg/kg³⁶
Ocimum gratissimum L. (Lamiaceae)	Anti-sickling activity^37,38	LD₅₀ > 5000 mg/kg³⁹
Garcinia kola Heckel (Clusiaceae)	Anti-sickling activity⁴⁰, Erythrocyte membrane stabilisation⁴¹	Very toxic with LD₅₀ of 358 mg/kg⁴²
Rauvolfia vomitoria Afzel. (Apocynaceae)	Anti-sickling activity⁴³	High doses exceeding 300 mg/kg could cause damage to liver, LC₅₀ = 1.205 mg/mL⁴⁴

Potential Chemical Markers, and Their Bioactivity, Toxicity and Analytical Assay Methods

Several catalogued compounds demonstrate various bioactivities implicated in the management of SCD. Compounds including rutin, quercetin, ellagic acid, and naringenin, primarily from Carica papaya, Terminalia catappa, and Annona muricata, demonstrate anti-sickling properties, positioning these plants as promising candidates for SCD management. Other compounds have been reported to exhibit bioactivities, including antihaemolytic, antiplatelet, antithrombotic, and erythrocyte membrane stabilisation effects, which play a supportive yet significant role in symptomatic management of SCD (Table 2).

Table 2.

Reported Bioactivities, Toxicity Profiles, and Analytical Methods of Compounds Isolated from Selected Plants.

Medicinal plant	Isolated compounds	Anti-sickling and other related bioactivities	Safety/toxicity profile	Reported analytical methods
Carica papaya L. (Caricaceae)	Carpaine	Docking reports of inhibiting the 2HbS protein⁴⁵	Non-cytotoxic⁴⁶, No adverse effects observed after 20 days of treatment⁴⁷	HPLC-UV⁴⁸, LC-MS⁴⁷
	Rutin	Anti-sickling⁴⁹, antihaemolytic⁵⁰, antioxidant⁵¹	Non-toxic in acute and chronic studies⁵², LD₅₀ > 1.51 g/kg⁵³, cytotoxic at 810 µM⁵⁴	HPLC, UV spectrometry⁵⁵
	Kaempferol	Antioxidant⁵⁶	No adverse effects in 28-day toxicity test⁵⁷, consumption of 50 mg daily is safe in humans⁵⁸	HPLC⁵⁹, LC-MS/MS⁶⁰
	Quercetin	Anti-sickling⁶¹, antioxidant⁶²	Safe in mice treated with 50 mg/kg for 14 weeks⁶³, evidence of carcinogenic activity in male rats at high doses⁶⁴	HPLC-DAD⁶⁵
	Isorhamnetin	Antiplatelet and antithrombotic effects⁶⁶, antioxidant	NA	HPLC-DAD⁶⁷
	Loliolide⁶⁸	Antioxidant⁶⁹	NA	GC-MS⁷⁰
	Myricetin⁷¹	Antioxidant⁷²	No evidence of genotoxic potential⁷³	UPLC-MS/MS⁷⁴, LC-MS/MS⁷⁵
	Luteolin⁷⁶	Antioxidant⁷⁷	NA	UHPLC-UV/PDA⁷⁷
Terminalia catappa L. (Combretaceae)	Punicalagin	Antioxidant⁷⁸	High doses are not toxic to rats⁷⁹, Doses > 25 mg/kg may induce liver toxicity⁷⁸	HPTLC⁸⁰, UPLC-MS⁸¹
	Punicalin	Antioxidant⁷⁸	Doses > 25 mg/kg could cause liver toxicity⁷⁸	NA
	Gallic acid	Antioxidant⁸²	LD₅₀ > 900 mg/kg⁸³	RP-HPLC⁸⁴
	Ellagic acid⁸⁵	Anti-sickling⁸⁶, antioxidant⁸⁷	Non-toxic^88,89	UFLC-DAD-MS⁹⁰, RP-HPLC⁹¹
	Quercetin	Anti-sickling⁶¹, antioxidant⁶²	Safe in mice treated with 50 mg/kg for 14 weeks⁶³, evidence of carcinogenic activity in male rats at high doses⁶⁴	HPLC-DAD⁶⁵
	Apigenin	Antioxidant⁹²	Doses > 200 mg/kg may induce hepatotoxicity⁹³	UHPLC - DAD⁹⁴
	Naringenin	Anti-sickling⁹⁵, erythrocyte membrane stabilisation⁹⁶, antioxidant⁹⁷	NA	UHPLC - DAD⁹⁴
	Isorhamnetin⁹⁸	Antiplatelet and antithrombotic effects⁶⁶, antioxidant⁹⁹	NA	HPLC-DAD⁶⁷
Annona muricata L. (Annonaceae)	Annonaine¹⁰⁰	Antioxidant with vasorelaxant properties^101,102	NA	HPLC-UV¹⁰³
	Quercetin	Anti-sickling⁶¹, antioxidant⁶²	Safe in mice treated with 50 mg/kg for 14 weeks⁶³, evidence of carcinogenic activity in male rats at high doses⁶⁴	HPLC-DAD⁶⁵
	Gallic acid	Antioxidant⁸²	LD₅₀ > 900 mg/kg⁸³	RP-HPLC⁸⁴
	Kaempferol	Antioxidant⁵⁶	No adverse effects in 28-day toxicity test⁵⁷, consumption of 50 mg daily is safe in humans⁵⁸	HPLC⁵⁹, LC-MS/MS⁶⁰
	Rutin¹⁰⁴	Anti-sickling⁴⁹, antihaemolytic⁵⁰, antioxidant⁵¹	Non-toxic in acute and chronic studies⁵², LD₅₀ > 1.51 g/kg⁵³, cytotoxic at 810 µM⁵⁴	HPLC, UV spectrometry⁵⁵
Ceiba pentandra (L.) Gaertn. (Malvaceae)	Vavain¹⁰⁵	Antioxidant¹⁰⁶	NA	NA
	Loliolide	Antioxidant⁶⁹	NA	GC-MS⁷⁰
	Campesterol	Antioxidant¹⁰⁷	NA	LC-MS¹⁰⁸
	Stigmasterol¹⁰⁹	Antioxidant¹⁰⁷	NA	LC-MS¹⁰⁸
Ocimum gratissimum L. (Lamiaceae)	Oleanolic acid	Antioxidant¹¹⁰	Antifertility¹¹¹, doses > 1350 mg/kg may cause cholestatic liver injury¹¹².	GC-FID¹¹³, HPLC¹¹⁴
	Stigmasterol	Antioxidant¹⁰⁷	NA	LC-MS¹⁰⁸
	β-sitosterol¹¹⁵	Antioxidant¹⁰⁷	Chronic administration of β-sitosterol in rats showed no clear toxicity¹¹⁶	HPLC¹¹⁷
	Eugenol	Antioxidant¹¹⁸	Doses of 20 & 30 µg/100 g body weight induced liver toxicity in rats¹¹⁹, highly toxic to human dental pulp fibroblasts even at very low concentrations¹²⁰, toxic at concentrations of 50-100 mg/L¹²¹.	RP-HPLC, GC-FID¹²²
	Thymol¹²³	Antioxidant¹²⁴	LD₅₀ for rats and mice were 2462.23¹²⁵ and 1350.9 mg/kg/bw¹²⁶ respectively, doses of 20 and 40 mg/kg can cause neurotoxicity, memory impairment, and blood-brain barrier breakdown in mice¹²⁷.	HPTLC¹²⁸
	Luteolin¹²⁹	Antioxidant⁷⁷	NA	UHPLC-UV/PDA⁷⁷
Garcinia kola Heckel (Clusiaceae)	Thymol	Antioxidant¹²⁴	LD₅₀ for rats and mice were 2462.23¹²⁶ and 1350.9 mg/kg/bw¹²⁵ respectively, doses of 20 and 40 mg/kg can cause neurotoxicity, memory impairment, and blood-brain barrier breakdown in mice¹²⁷.	HPTLC¹²⁸
	Squalene¹³⁰	Antioxidant¹³¹	NA	HPLC-DAD-MS¹³², GC-FID¹³³
	Kolaflavonone	Antioxidant¹³⁴	NA	LC-DAD¹³⁵
Rauvolfia vomitoria Afzel. (Apocynaceae)	Reserpine	Erythrocyte membrane stabilisation¹³⁶	Hypothermia-related testicular toxicity¹³⁷, CNS toxicity¹³⁸	HPTLC¹³⁹
Rauvolfia vomitoria Afzel. (Apocynaceae)	Yohimbine¹⁴⁰	Antioxidant¹⁴¹	Neurotoxicity¹⁴², hyperadrenergic effects¹⁴³	HPLC-UV-API/MS¹⁴⁴

Compounds, such as carpaine, kaempferol, rutin, ellagic acid and β-sitosterol, are reported as non-toxic or safe at specific doses, with some showing no adverse effects in acute, chronic, or human studies. However, compounds like punicalagin and punicalin, eugenol, thymol, reserpine, and yohimbine pose significant toxicity risks, including liver, neurological, reproductive, and carcinogenic effects at higher doses (Table 2).

Analytical methods for identifying and quantifying the compounds are well-developed for most, with high-performance liquid chromatography (HPLC) and its variants (eg, HPLC-UV, HPLC-DAD, RP-HPLC) being the most prevalent due to their precision and versatility (Table 2).

Concentration of Potential Marker Compounds in Plant Materials and the Availability, Cost, and Purity of Their Analytical Standards

The concentrations of the potential markers in plant materials vary widely, with some compounds present in trace amounts and others in higher quantities. For example, the concentration of campesterol is 99 200 µg/g in seed oil, and loliolide is 55 800 µg/g in leaves, indicating high natural abundance that could facilitate extraction. Conversely, kaempferol and apigenin occur at low concentrations, posing challenges for cost-effective isolation. Compounds including quercetin, isorhamnetin, myricetin, and luteolin, which are present across multiple plants, lack reported concentration data, suggesting limited quantitative studies on their natural occurrence (Table 3).

Table 3.

Summary of the Availability, Cost, Purity, and Concentration of Analytical Standards for Selected Marker Compounds.

Medicinal plant	Compounds	Commercial sources	Commercial unit available	Cost (Euros -€)	Purity	Concentration of compound in plant material
Carica papaya L. (Caricaceae)	Carpaine	NA	NA	NA	NA	0.93 g/kg in the leaf⁴⁸
	Rutin	SA	50 mg	247.00	PRS	100-160 µg/g in fruit¹⁴⁵
	Kaempferol	SA	20 mg	438.00	PRS	30 µg/g in leaf¹⁴⁶.
	Kaempferol	CC	25 mg	NA	≥98%	30 µg/g in leaf¹⁴⁶.
	Quercetin	SA	20 mg	270.00	PRS	40 µg/g in leaf¹⁴⁶.
	Isorhamnetin	SA	20 mg	184.00	PRS	NA
	Isorhamnetin	CC	1 mg	NA	≥98%	NA
	Loliolide	SA	1 mg	402.00	≥90%	55 800 µg/g in the leaf¹⁴⁷
	Loliolide	CC	1 mg	NA	≥98%	55 800 µg/g in the leaf¹⁴⁷
	Myricetin	SA	20 mg	351.00	PRS	NA
	Myricetin	CC	10 mg	NA	≥98%	NA
	Luteolin	SA	20 mg	392.00	PRS	NA
	Luteolin	CC	10 mg	NA	≥98%	NA
Terminalia catappa L. (Combretaceae)	Punicalagin	SA	1 mg	107.00	≥98% (HPLC)	NA
Terminalia catappa L. (Combretaceae)	Punicalagin	CC	1 mg	NA	≥95%	NA
	Punicalin	SA	10 mg	526.00	PRS	NA
	Punicalin	CC	1 mg	NA	≥95%	NA
	Gallic acid	SA	100 mg	368.00	PRS	510 μg/g in stem¹⁴⁸
	Gallic acid	CC	10 g	NA	≥98%	510 μg/g in stem¹⁴⁸
	Ellagic acid	SA	50 mg	281.00	PRS	1285 ug/g in stem¹⁴⁸
	Ellagic acid	CC	100 mg	NA	≥95%	1285 ug/g in stem¹⁴⁸
	Quercetin	SA	20 mg	270.00	PRS	NA
	Apigenin	SA	25 mg	351.00	PRS	6.5 ug/g in stem¹⁴⁸
	Apigenin	CC	25 mg	NA	≥98%	6.5 ug/g in stem¹⁴⁸
	Naringenin	SA	25 mg	292.00	PRS	NA
	Naringenin	CC	1 g	NA	≥98%	NA
	Isorhamnetin	SA,	20 mg	184.00	PRS	NA
	Isorhamnetin	CC	1 mg	NA	≥98%	NA
Annona muricata L. (Annonaceae)	Annonaine	NA	NA	NA	NA	5760 µg/g in the leaf¹⁴⁹
	Quercetin	SA	20 mg	270.00	PRS	NA
	Gallic acid	SA	100 mg	368.00	PRS	1600 μg/g in the fruit¹⁵⁰
	Gallic acid	CC	10 g	NA	≥98%	1600 μg/g in the fruit¹⁵⁰
	Kaempferol	SA	20 mg	438.00	PRS	29.86 mg/mL in fruit juice¹⁵¹
	Kaempferol	CC	25 mg	NA	≥98%	29.86 mg/mL in fruit juice¹⁵¹
	Rutin	SA	50 mg	247.00	PRS	1780 ug/g in leaf¹⁵²
Ceiba pentandra (L.) Gaertn. (Malvaceae)	Vavain	NA	NA	NA	NA	NA
	Loliolide	SA	1 mg	402.00	≥90%	NA
	Loliolide	CC	1 mg	NA	≥98%	NA
	Campesterol	SA	10 mg	548.00	PRS	99 200 µg/g in seed oil¹⁵³
	Campesterol	CC	1 mg	NA	≥98%	99 200 µg/g in seed oil¹⁵³
	Stigmasterol	SA	100 mg	247.00	PRS	20 000 µg/g in the seed oil¹⁵³
	Stigmasterol	CC	1 g	NA	≥90%	20 000 µg/g in the seed oil¹⁵³
Ocimum gratissimum L. (Lamiaceae)	Oleanolic acid	SA	50 mg	317.00	PRS	NA
Ocimum gratissimum L. (Lamiaceae)	Oleanolic acid	CC	100 mg	NA	≥95%	NA
	Stigmasterol	SA	100 mg	247.00	PRS	NA
	Stigmasterol	CC	1 g	NA	≥90%	NA
	β-sitosterol	SA	10 mg	383.00	Synthetic, ≥95%	NA
	Eugenol	SA	250 mg	50.90	Analytical standard	NA
	Eugenol	CC	1 mg	NA	≥95%	NA
	Thymol	SA	1 g	145.00	Analytical standard	NA
	Thymol	CC	100 mg	NA	≥95%	NA
	Luteolin	SA	20 mg	392.00	PRS	NA
	Luteolin	CC	10 mg	NA	≥98%	NA
Garcinia kola Heckel (Clusiaceae)	Thymol	SA	1 g	145.00	Analytical standard	NA
Garcinia kola Heckel (Clusiaceae)	Thymol	CC	100 mg	NA	≥95%	NA
	Squalene	SA	1 g	405.00	Analytical standard	NA
	Squalene	CC	50 g	NA	≥95%	NA
	Kolaflavonone	NA	NA	NA	NA	NA
Rauvolfia vomitoria Afzel. (Apocynaceae)	Reserpine	SA	50 mg	232.00	PRS	689.5 ug/g in the root¹⁵⁴
Rauvolfia vomitoria Afzel. (Apocynaceae)	Reserpine	CC	1 g	NA	≥98%	689.5 ug/g in the root¹⁵⁴
	Yohimbine	SA	100 mg	276.00	PRS	NA
	Yohimbine	CC	500 mg	NA	≥98%	NA

SA, Sigma-Aldrich; CC, Cayman Chemical; NA, Not available; PRS, Phyproof® reference substance.

Several compounds are commercially available, primarily through reputable suppliers like Sigma-Aldrich and Cayman Chemical, with unit sizes ranging from 1 mg to 50 g and costs spanning €50.90 to €548.00. Quercetin, kaempferol, rutin, and luteolin, which are present in multiple plants, are commercialised with high purity (≥95% or phyproof^® Reference Standard) but at higher prices (ranging between €247.00 and €438.00). Gallic acid and ellagic acid are similarly available at high purity (≥95%) but moderate costs (€281.00-€368.00). Carpaine, annonaine, vavain and kolaflavonone lack commercial sources, highlighting their limited accessibility for research or industrial use (Table 3).

Prioritisation of Appropriate Markers Using the Herb MaRS Framework

Most of the potential markers scored 1/1 for exhibiting bioactivity relevant to SCD management (A = 1). Carpaine, rutin, quercetin, ellagic acid, and naringenin scored 2/2 due to reported anti-sickling activities (B = 2). The variations in the concentrations of compounds in their respective plant materials led to different scores (C = 1, 2 or 3). About 19 compounds lack reported data on concentration in plant materials, and thus, scored 0/3 (C = 0). Availability of reference standards was recorded for 33 compounds, scoring 1/1 (D = 1), while carpaine, annonaine, and vavain scored 0/1 (D = 0). Additionally, analytical methods have been reported for 34 compounds (E = 1), except for punicalin and vavain (E = 0).

Toxicity (F = 8) is reported for 16 compounds, including rutin, quercetin, punicalagin, gallic acid, eugenol, thymol, and reserpine, while 21 compounds, such as carpaine, kaempferol, ellagic acid, and naringenin, show no evidence of toxicity (F = 0).

The Herb MaRS scores range from 1 to 8, with rutin and ellagic acid achieving the maximum score of 8/8. Quercetin scores 7/8, while carpaine, gallic acid, kaempferol, loliolide, campesterol, stigmasterol, and reserpine score 6/8. Punicalin and vavain had low scores of 2/8 and 1/8, respectively (Table 4). Overall, the chemical markers prioritised for standardising the selected medicinal plants include carpaine, rutin, kaempferol, quercetin, loliolide, gallic acid, ellagic acid, apigenin, naringenin, campesterol, stigmasterol, thymol, reserpine, yohimbine and annonaine (Figure 1).

Figure 1.

Chemical Structures of Selected Marker Compounds Isolated from Medicinal Plants.

Table 4.

Scoring and Ranking of Potential Markers for Medicinal Plants Selected for the Study.

Medicinal plant	Isolated compound	Evidence of bioactivity related to SCD management (A)	Evidence of anti-sickling activity (B)	Concentration of compound in plant material (C)	Availability of reference standard (D)	Availability of analytical method (E)	Herb MaRS score / 8	Evidence of toxicity (F)
Medicinal plant	Isolated compound	YES – 1NO - 0	YES - 2NO - 0	NA-0<5 µg/g- 15 - 50 µg/g- 2> 50 µg/g -3	YES – 1NO - 0	YES – 1NO - 0	A + B + C + D + E	Evidence of toxicity (F)
Carica papaya L. (Caricaceae)	Carpaine	0	2	3	0	1	6	0
	Rutin	1	2	3	1	1	8	8
	Kaempferol	1	0	2	1	1	5	0
	Quercetin	1	2	2	1	1	7	8
	Isorhamnetin	1	0	0	1	1	3	0
	Loliolide	1	0	3	1	1	6	0
	Myricetin	1	0	0	1	1	3	0
	Luteolin	1	0	0	1	1	3	0
Terminalia catappa L. (Combretaceae)	Punicalagin	1	0	0	1	1	3	8
	Punicalin	1	0	0	1	0	2	8
	Gallic acid	1	0	3	1	1	6	8
	Ellagic acid	1	2	3	1	1	8	0
	Quercetin	1	2	0	1	1	5	8
	Apigenin	1	0	2	1	1	5	8
	Naringenin	1	2	0	1	1	5	0
	Isorhamnetin	1	0	0	1	1	3	0
Annona muricata L. (Annonaceae)	Annonaine	1	0	3	0	1	5	0
	Quercetin	1	2	0	1	1	5	8
	Gallic acid	1	0	3	1	1	6	8
	Kaempferol	1	0	3	1	1	6	0
	Rutin	1	2	3	1	1	8	8
Ceiba pentandra (L.) Gaertn. (Malvaceae)	Vavain	1	0	0	0	0	1	0
	Loliolide	1	0	0	1	1	3	0
	Campesterol	1	0	3	1	1	6	0
	Stigmasterol	1	0	3	1	1	6	0
Ocimum gratissimum L. (Lamiaceae)	Oleanolic acid	1	0	0	1	1	3	8
	Stigmasterol	1	0	0	1	1	3	0
	β-sitosterol	1	0	0	1	1	3	0
	Eugenol	1	0	0	1	1	3	8
	Thymol	1	0	0	1	1	3	8
	Luteolin	1	0	0	1	1	3	0
Garcinia kola Heckel (Clusiaceae)	Thymol	1	0	0	1	1	3	8
	Squalene	1	0	0	1	1	3	0
	Kolaflavonone	1	0	0	1	1	3	0
Rauvolfia vomitoria Afzel. (Apocynaceae)	Reserpine	1	0	3	1	1	6	8
Rauvolfia vomitoria Afzel. (Apocynaceae)	Yohimbine	1	0	0	1	1	3	8

Discussion

Carica papaya L. (Caricaceae) has been widely studied for its potential in managing sickle cell disease (SCD), particularly its leaves, seeds, and fruit. Ethnobotanical studies, including Amponsah et al⁷ document Carica papaya as a key component in polyherbal formulations for SCD in Ghana and Nigeria, often used as leaf decoctions or seed extracts. Studies have confirmed the anti-sickling, membrane-stabilising and sickling reversal effects of papaya leaf extracts.^21,22 Additionally, Singh et al¹⁵⁵ reported that papaya leaf extract significantly decreased sickle cell crises by enhancing antioxidant defences, attributed to its high phenolic content. Carica papaya is generally considered safe at therapeutic doses, with minimal toxicity reported.^23,24 However, excessive consumption of unripe papaya or high doses of seed extracts may cause gastrointestinal irritation due to benzyl isothiocyanate, a compound requiring monitoring in formulations.¹⁵⁶ Carica papaya contains bioactive compounds such as flavonoids (quercetin, kaempferol, myricetin, luteolin), terpenoids (loliolide), and alkaloids (carpaine), which contribute to its therapeutic potential. Muhammad et al⁶¹ has demonstrated that quercetin inhibits sickle cell haemoglobin polymerisation by stabilising the oxygenated state of haemoglobin, thus reducing sickling. Similarly, rutin has shown anti-sickling effects in vitro by modulating red blood cell membrane stability.⁴⁹ These compounds align with Herb MaRS criteria, scoring high (quercetin at 7/8 and rutin at 8/8) due to their anti-sickling activity, adequate plant concentrations (>50 µg/g), and availability of analytical standards for quality control (Table 4). It should be noted that both compounds scored 8/8 for toxicity, warranting caution in their use. Carpaine, a dimeric piperidine alkaloid, showed the highest inhibitory activity in a molecular docking study against the 2HBS protein, which drives sickling and associated complications.⁴⁵ It scored 6/8 in the analysis owing to limited commercial standards and insufficient concentration data. Similarly, loliolide, a monoterpenoid lactone, scored 6/8 but lacks reported data on anti-sickling activities. It, however, exhibits antioxidant and anti-inflammatory properties, which may indirectly benefit SCD management by reducing oxidative stress and inflammation, key drivers of sickle cell crises.⁶⁹ Likewise, kaempferol, lacking reported anti-sickling data, scored 5/8, while isorhamnetin, myricetin and luteolin each scored 3/8 (Table 4). The application of Herb MaRS to Carica papaya highlights quercetin and rutin as ideal chemical markers.

Terminalia catappa L. (Combretaceae), commonly known as Indian almond or tropical almond, is a medicinal plant widely reported in ethnobotanical studies for its use in SCD management.^7,25,26 It exhibits significant anti-sickling activity, primarily attributed to its ability to inhibit haemoglobin S (HbS) polymerisation. Studies have demonstrated its efficacy in both in vitro and in vivo models, where extracts from its leaves and seeds reduce red blood cell sickling and improve erythrocyte stability.^25–27 The plant has a favourable safety profile, with an LD₅₀ exceeding 5000 mg/kg in animal models^28,29, indicating low acute toxicity. Key bioactive compounds include quercetin, apigenin, ellagic acid, naringenin, isorhamnetin, gallic acid, punicalagin, and punicalin, which contribute to its anti-sickling and antioxidant properties (Table 2). Punicalagin and punicalin, respectively, scored 3/8 and 2/8, owing to limited data on anti-sickling activity and concentrations in plant materials (Table 4). Gallic acid, although lacking direct anti-sickling activity, protects against oxidative stress-induced osmotic fragility in erythrocytes, supporting symptomatic relief in SCD.⁸² It scored 6/8 in the analysis, signifying a valuable marker for standardisation. Moreover, gallic acid has a favourable safety profile, with an LD₅₀ exceeding 900 mg/kg in animal models, indicating low acute toxicity. This was further confirmed in sub-acute toxicity studies in albino mice.⁸³ Nevertheless, caution should be applied in its use, especially at high doses. Ellagic acid, a polyphenolic compound, exhibits significant anti-sickling activity by directly inhibiting haemoglobin S (HbS) polymerisation.⁸⁶ Studies, including those using Berkeley mice, show that ellagic acid reduces pathophysiological manifestations of SCD, such as vaso-occlusion, by stabilising erythrocyte membranes and mitigating oxidative stress.^86,87 Its antioxidant properties further protect erythrocytes from oxidative damage, complementing its anti-sickling effects.⁸⁷ In the absence of reported toxicity^88,89, ellagic acid serves as the ideal chemical marker, scoring 8/8 in the Herb MaRS analysis (Table 4). Apigenin and naringenin, both flavonoids, each received a score of 5/8 in the analysis (Table 4). While there have not been any reports on the anti-sickling activity of apigenin, naringenin exhibits significant anti-sickling activity in vitro on human sickle red blood cells, where it inhibits haemoglobin S (HbS) polymerisation.⁹⁵ Additionally, contrary to apigenin, which has studies indicating potential hepatotoxicity at doses exceeding 200 mg/kg⁹³, limited toxicity data are available for naringenin, with no comprehensive studies reporting acute or chronic toxicity in animal or human models (Table 3). From our study, gallic acid, ellagic acid, apigenin, naringenin and quercetin emerge as ideal markers for quality control purposes. However, owing to toxicity concerns, ellagic acid and naringenin are recommended for standardisation.

Annona muricata L. (Annonaceae), commonly known as soursop, is a medicinal plant widely used in sub-Saharan Africa for SCD management, particularly in countries like Nigeria and Cameroon.^7,9,30 It exhibits significant anti-sickling activity, as demonstrated in vitro and in vivo, with its extracts inhibiting haemoglobin S (HbS) polymerisation.^30,31 The plant has a relatively safe profile, with an LD₅₀ exceeding 5000 mg/kg in animal models.³² However, some studies report genotoxic potential, particularly with prolonged use or high doses, necessitating careful monitoring.³³ Key bioactive compounds, including quercetin, rutin, gallic acid, and kaempferol, contribute to its anti-sickling and antioxidant properties (Table 2). Annonaine, an isoquinoline alkaloid, exhibits antioxidant and vasorelaxant properties, which are relevant to SCD management by reducing oxidative stress and improving vascular function.^101,102 Limited data exist on annonaine's toxicity profile, with no comprehensive studies reporting acute or chronic toxicity in animal or human models, resulting in 0/8 in the toxicity scoring. It, however, scored 5/8, indicating potential as a chemical marker. Quercetin, likewise, scored 5/8, kaempferol and gallic acid each scored 6/8, and rutin scored 8/8 (Table 4), signifying their ideal selection as markers for Annona muricata in SCD management formulations.

Ceiba pentandra L. (Malvaceae), commonly known as kapok or silk-cotton tree, is a tropical tree widely used in traditional medicine in sub-Saharan Africa, particularly in the Democratic Republic of Congo, for SCD management.⁷ It exhibits significant anti-sickling activity, with aqueous extracts from its trunk bark and branches demonstrating the ability to inhibit HbS polymerisation.³⁴ The plant also shows antithrombin effects by activating heparin cofactor II and modulating fibrinolytic activity. This reduces the secretion of tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA), and plasminogen activator inhibitor type-1 (PAI-1) without affecting their synthesis.³⁵ These properties help mitigate thrombotic crises, a major complication in SCD. The LD₅₀ exceeds 5000 mg/kg in animal models, indicating low acute toxicity.³⁶ Bioactive compounds such as campesterol, stigmasterol, vavain, and loliolide contribute to its anti-sickling, antioxidant, and membrane-stabilising effects (Table 2). Vavain exhibits antioxidant properties, which are relevant for SCD management by reducing oxidative stress, a major contributor to erythrocyte damage and vaso-occlusive crises.¹⁰⁶ Additionally, its ability to inhibit cyclooxygenase-1 and -2 (COX-1 and COX-2) catalysed prostaglandin biosynthesis suggests anti-inflammatory effects that may support symptomatic relief in SCD.¹⁰⁵ That notwithstanding, the limited data on vavain led to the scoring of 1/8 in the analysis (Table 4). Campesterol and stigmasterol each scored 6/8 (Table 4), indicating their suitability as chemical markers for C. pendandra and its formulations. Although direct evidence of anti-sickling activity is limited, they can stabilise erythrocyte membranes, supporting the prevention of haemolysis, a common complication in SCD.¹⁰⁷ Additionally, the suitability is enhanced by the absence of reported toxicity data.

Ocimum gratissimum L. (Lamiaceae), commonly known as African basil, exhibits significant anti-sickling activity, with methanolic leaf extracts inhibiting HbS polymerisation in vitro.^37,38 Furthermore, the antioxidant properties of Ocimum are attributed to compounds such as stigmasterol, β-sitosterol, eugenol, thymol, and luteolin (Table 2), which mitigate oxidative stress, reducing erythrocyte damage and vaso-occlusive events.^{77,107,118,124} Eugenol and thymol also demonstrate membrane-stabilising effects, further supporting SCD management by preventing haemolysis.^118,124 With an LD₅₀ exceeding 5000 mg/kg in animal models, Ocimum has a generally safe toxicity profile.³⁹ However, specific compounds present risks: oleanolic acid may cause cholestatic liver injury at doses above 1350 mg/kg and has antifertility effects^111,112, while eugenol¹¹⁸ and thymol¹²⁴ exhibit liver and neurotoxicity at high doses. β-sitosterol shows no clear toxicity in chronic studies, but careful dose monitoring is needed for safe therapeutic use.¹¹⁶ All the compounds scored 3/8 in the Herb MaRS analysis, indicating low scores for potential chemical markers. However, consideration was given to their reported toxicity profiles, which are crucial in selecting safe doses for therapy. Thus, luteolin, stigmasterol and β-sitosterol were prioritised to serve as potential markers owing to their safety profiles.

Garcinia kola Heckel (Clusiaceae), commonly known as bitter kola, is a tree indigenous to West and Central Africa, widely used in traditional medicine for managing SCD.^7,10 Kola extracts exhibit anti-sickling activity, with methanolic leaf extracts and aqueous fractions demonstrating inhibition of HbS polymerisation in vitro at concentrations of 0.1-10 mg/mL, comparable to p-hydroxybenzoic acid.⁴⁰ The erythrocyte membrane stabilisation properties of the aqueous extracts of the seeds have also been reported.⁴¹ The methanol extract of the stem bark has been reported as very toxic, with an LD₅₀ of 358 mg/kg⁴², highlighting the need for cautious dosing in therapeutic applications. Bioactive compounds isolated from G. kola, including thymol, squalene, and kolaflavonone, all demonstrate antioxidant activity (Table 2). However, thymol exhibits significant toxicity, with LD₅₀ values of 2462.23 mg/kg in rats¹²⁵ and 1350.9 mg/kg in mice¹²⁶, and doses of 20-40 mg/kg may induce neurotoxicity, memory impairment, and blood-brain barrier breakdown in mice.¹²⁷ This toxicity necessitates careful monitoring in the selection of doses for SCD management. Per the Herb MaRS criteria, thymol scored 3/8, which is below the acceptable score for a marker for standardisation. However, its 8/8 score for toxicity could be used to select it as a toxicological marker for the plant. Thus, formulations incorporating bitter kola could be assayed for toxicity by monitoring the levels of thymol. Squalene and kolaflavonone lack comprehensive toxicity data, and kolaflavonone is not commercially available as an analytical standard, limiting its immediate use for standardisation (Table 3).

Rauvolfia vomitoria exhibits anti-sickling activity, with root and leaf extracts inhibiting HbS polymerisation in vitro, reducing sickling of erythrocytes under hypoxic conditions.⁴³ The plant has an LC₅₀ of 1.205 mg/mL, and doses >300 mg/kg may cause liver damage.⁴⁴ Its biological activities have been attributed to indole alkaloids, including reserpine and yohimbine, which mitigate oxidative stress (Table 2). Reserpine, in particular, has erythrocyte membrane stabilisation effects¹³⁶, which indirectly contribute to the symptomatic management of SCD. Its use should, however, be monitored owing to reports of hypothermia-related testicular toxicity¹³⁷ and CNS toxicity.¹³⁸ Reserpine scored 6/8 (Table 4) in the analysis, indicating a suitable marker for standardising R. vomitoria and formulations that incorporate it. Yohimbine, on the other hand, scored 3/8, mainly due to a lack of reports on its concentration in the plant material. It, however, scored 8/8 for reported toxicity (Table 2). Both reserpine and yohimbine are therefore prioritised as markers, serving as a toxicological marker for the plant.

The application of Herb MaRS for the identification and prioritisation of chemical markers for the quality control of medicinal plants used in the management of SCD represents a significant advancement in the standardisation of such herbal formulations. A major contribution of this study is the systematic adaptation of the Herb MaRS framework to prioritise markers specifically associated with anti-sickling activity, thereby addressing a critical gap in existing literature. This targeted approach improves the precision and relevance of marker selection for complex herbal products intended for SCD management. However, certain limitations were identified, including the dependence on secondary, literature-derived data, which may overlook unpublished findings or region-specific studies. Additionally, the absence of concentration data for 19 compounds restricted their full evaluation within the Herb MaRS framework. Future studies should incorporate primary phytochemical investigations to address these limitations and undertake clinical validation of the proposed markers.

This study has certain limitations that warrant acknowledgement. As a systematic review and analytical application of the Herb MaRS framework, the findings depend largely on published literature, and as such, minor but pharmacologically potent compounds may be underrepresented where their activities have not yet been reported. Furthermore, the analysis may not fully capture geographical and environmental influences, including soil type, climate, harvest season, and genetic variation, which can significantly affect phytochemical composition. The use of the Herb MaRS framework also introduces a reductionist dimension, as it focuses on individual chemical markers for standardisation and quality control. While this approach does not encompass the holistic principles and synergistic interactions fundamental to traditional herbal medicine, it serves as a complementary scientific tool to support the consistency, safety, and reliability of herbal formulations. Future studies should therefore incorporate region-specific phytochemical analyses, experimental validation, and integrative approaches that bridge traditional therapeutic concepts with modern quality control systems.

Conclusion

The application of the Herbal Marker Ranking System (Herb MaRS) has enabled the systematic identification and prioritisation of chemical markers for quality control of medicinal plants used in SCD management. The study highlights several compounds as ideal markers due to their demonstrated anti-sickling activity, high concentrations in respective plant materials and analytical accessibility. Compounds including thymol, reserpine and yohimbine are also prioritised as toxicological markers for their respective plant materials. Despite promising outcomes, data gaps persist regarding the concentration and toxicity of several compounds, underscoring the need for further phytochemical and clinical investigations. Overall, this approach enhances standardisation efforts, ensuring the quality, safety, and efficacy of herbal medicines used for managing SCD.

Footnotes

Acknowledgements

The authors wish to acknowledge the Departments of Pharmacognosy and Herbal Medicine, Faculty of Pharmacy and Pharmaceutical Sciences, College of Health Sciences, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana.

ORCID iDs

Desmond Nkrumah

Reinhard Isaac Nketia

Michael Kwesi Baah

Linda Mensah Sarpong

George Adjei-Hinneh

Bernard Kofi Turkson

Isaac Kingsley Amponsah

Ethical Approval

Ethical Approval is not applicable for this article.

Statement of Human and Animal Rights

This article does not contain any studies with human or animal subjects.

Statement of Informed Consent

There are no human subjects in this article and informed consent is not applicable.

Author Contributions

Desmond Nkrumah: Conceptualisation, Methodology, Data curation, Writing – original draft. Reinhard Isaac Nketia: Writing – review & editing, Formal analysis. Michael Kwesi Baah: Writing – review & editing, Methodology. Linda Mensah Sarpong: Writing – review & editing, Data curation. George Adjei-Hinneh: Data curation, Formal analysis. Bernard Kofi Turkson: Supervision, Data curation, Formal analysis. Isaac Kingsley Amponsah: Conceptualisation, Writing – review & editing, Supervision.

Funding

The authors received no financial support for the research, authorship, and/or publication of this article.

Declaration of Conflicting Interests

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Data Availability

All datasets used and/or analysed during the current study are available in the manuscript and could also be requested from the corresponding author.

References

Cordovil

. Sickle cell disease: a genetic disorder of beta-globin. In: Thalassemia and Other Hemolytic Anemias. InTech; 2018.; 98-113

Eaton

Bunn

. Treating sickle cell disease by targeting HbS polymerization. Blood. 2017;129(20):2719–2726. doi:10.1182/blood-2017-02-765891

Kato

Piel

Reid

, et al. Sickle cell disease. Nat Rev Dis Prim. 2018;4(1):1–22. doi:10.1038/nrdp.2018.10

Mettananda

. Sickle cell disease. In: A Guide to Paediatric Red Blood Cell Disorder. Vol. 376. N Engl J Med; 2020:61–70. doi:10.47363/jmhc/2021(3)162

Modell

Darlison

. Global epidemiology of haemoglobin disorders and derived service indicators. Bull World Health Organ. 2008;2008(6):480–487. doi:10.2471/BLT.06.036673

Colombatti

Hegemann

Medici

Birkegård

. Correction to: systematic literature review shows gaps in data on global prevalence and birth prevalence of sickle cell disease and sickle cell trait: call for action to scale up and harmonize data collection. J Clin Med. 2024;13(10):2893. doi:10.3390/jcm13102893

Amponsah

Opoku-Kwabi

Armah

Addotey

Turkson

Kontoh

. A systematic review of medicinal plants and their compounds validated as agents for the management of sickle cell disease. Nat Prod Commun. 2024;19(9). doi:10.1177/1934578X241265889

Gbadamosi

. An inventory of ethnobotanicals used in the management of sickle cell disease in Oyo State, Nigeria. Bot Res Int. 2015;8(4):65–72.

Kotue

Pieme

Fokou

. Ethnobotanicals usages in the management of sickle cell disease (SCD) in some localities of Cameroon. Pharmacophore. 2016;7(4):192–200. Accessed May 7, 2025. http://www.pharmacophorejournal.com/.

10.

Amujoyegbe

Idu

Agbedahunsi

Erhabor

. Ethnomedicinal survey of medicinal plants used in the management of sickle cell disorder in Southern Nigeria. J Ethnopharmacol. 2016;185:347–360. doi:10.1016/j.jep.2016.03.042

11.

Famojuro

Moody

. Survey of medicinal plants used in the management of sickle cell disease by traditional medical practitioners of Gbonyin Local Government Area of Ekiti State, Nigeria. Niger J Nat Prod Med. 2015;19:78–84. doi:10.4314/njnpm.v19i0.8

12.

Akakpo-Akue

Kplé

Kiyinlma

, et al. Ethnobotanical study of medicinal plants used against sickle cell anaemia in the eastern part of the Côte d’Ivoire. J Anim Plant Sci. 2020;45(1):7839–7852. doi:10.35759/janmplsci.v45-1.7

13.

Pandey

. Harvesting and post-harvest processing of medicinal plants: problems and prospects standardization of non-destructive harvesting practices of medicinal plants view project project on Andrographis paniculata view project harvesting and post-harvest process. Pharma Innov J. 2017;6(12):229–235. Accessed May 7, 2025. https://www.researchgate.net/publication/322076894

14.

Sahoo

Manchikanti

Dey

. Herbal drugs: standards and regulation. Fitoterapia. 2010;81(6):462–471. doi:10.1016/j.fitote.2010.02.001

15.

WHO. WHO guidelines for selecting marker substances of herbal. WHO Tech Rep Ser No 1003. Published online 2017:71–86. http://apps.who.int/medicinedocs/en/m/abstract/Js23240en/

16.

Addotey

Boadu

Kontoh

, et al. Unlocking chemical markers for the standardization of antimalarial medicinal plants and products: application of the herbal marker ranking system (Herb MaRS). Sci Afr. 2025;27(January):e02538. doi:10.1016/j.sciaf.2025.e02538

17.

Bensoussan

Lee

Murray

, et al. Choosing chemical markers for quality assurance of complex herbal medicines: development and application of the herb MaRS criteria. Clin Pharmacol Ther. 2015;97(6):628–640. doi:10.1002/cpt.100

18.

Amponsah

Owusu

Sarkodie

, et al. Application of the herbal marker ranking system (Herb MaRS) affords chemical markers for the standardization of medicinal plants used for male vitality herbal products. Phytomedicine Plus. 2025;5(5):100864. doi:10.1016/J.PHYPLU.2025.100864

19.

WAHO. West African Herbal Pharmacopoeia. KS Printcraft Gh. Ltd; 2013.

20.

Ghana Herbal Pharmacopoeia. Science and Technology Policy Research Institute (STEPRI), Council for Scientific and Industrial Research (CSIR); 2007. Accessed March 27, 2025. https://books.google.com/books/about/Ghana_Herbal_Pharmacopoeia.html?id=mpXhzQEACAAJ

21.

Naiho

Okonkwor

Okoukwu

. Anti-sickling and membrane stabilizing effects of carica papaya leaf extract. Br J Med Med Res. 2015;6(5):484–492. doi:10.9734/bjmmr/2015/14608

22.

Adetayo Modupe

Shokunbi

Oyelese

Adetayo

. In-vitro antisickling and sickling-reversal activities of Carica papaya fruit at different stages of ripening. Babcock Univ Med J. 2020;3(2):10–18. doi:10.38029/bumj.v3i2.38

23.

Halim

Abdullah

Afzan

Abdul Rashid

Jantan

Ismail

. Study of acute toxicity of Carica papaya leaf extract in Sprague Dawley rats. J Med Plants Res. 2011;5(10):1867–1872.

24.

Dinda Nadiyah

Kharisma

Yuniarti

. Penentuan Derajat Toksisitas Akut Ekstrak Air Buah Pepaya (Carica papaya L.) Muda Pada Mencit Menggunakan Purposed New Recommended Method. J Jamu Indones. 2016;1(2):15–19. doi:10.29244/jjidn.v1i2.30608

25.

Samuel

Olaniyi

Olakunle

Michele

Okogun

. Phytochemical and anti-sickling activities of Terminalia catappa Linn. J Phytomedicine Ther. 2009;14:31–36. doi:10.4314/jopat.v14i1.69407

26.

Mgbemene

Ohiri

. Anti-sickling potential of Terminalia catappa leaf extract. Pharm Biol. 1999;37(2):152–154. doi:10.1076/phbi.37.2.152.6090

27.

Moody

Segun

Aderounmu

Omotade

. Antisickling activity of Terminalia catappa leaves harvested at different stages of growth. Niger J Nat Prod Med. 2004;7(1). doi:10.4314/njnpm.v7i1.11701

28.

Mshelia Halilu

Ugwah-Oguejiofor

Oduncuoğlu

Gamde Matthias

. Physicochemical, toxicity and antioxidant activity of terminalia catappa kernel oil in mice. Pharmacognosy Res. 2022;15(1):119–127. doi:10.5530/097484900304

29.

Iheagwam

Okeke

De Campos

Adegboye

Ogunlana

Chinedu

. Toxicopathological, proinflammatory and stress response evaluation of Terminalia catappa extract in male Wistar rats. Toxicol Reports. 2021;8:1769–1776. doi:10.1016/j.toxrep.2021.10.005

30.

Onyegeme-Okerenta

Essien

. In vitro antisickling potentials of ethanol extract of Annona muricata, Delonix regia and Senna alata. 2019;2(1):7–18. doi:10.5281/zenodo.3458736

31.

Agu

Ayevbuomwan

Imade

, et al. Biochemical investigation of the upstream anti-sickling mechanisms of soursop (Annona muricata): 15-acetyl guanacone as an inhibitor of deoxyhaemoglobin polymerisation. J Biomol Struct Dyn. 2022;40(4):1503–1520. doi:10.1080/07391102.2020.1828171

32.

Arthur

Larbie

Woode

. Evaluation of acute and subchronic toxicity of Annona Muricata (Linn.) aqueous extract in animals Toxicology View project Cure Epilepsy Project View project. Artic Eur J Exp Biol. 2011;1(4):115–124. Accessed May 20, 2025. https://www.researchgate.net/publication/225280339_Evaluation_of_acute_and_subchronic_toxicity_of_Annona_Muricata_Linn_aqueous_extract_in_animals%0Awww.pelagiaresearchlibrary.com

33.

Ferreira

Quaresma

ACS

Brandão

DLDN

, et al. Evaluation of genotoxicity and toxicity of Annona muricata L. seeds and in silico studies. Molecules. 2023;28(1):1–16. doi:10.3390/molecules28010231

34.

José

Marie

Christian

Mizu

Josué

. Phytochemical profile of an anti-sickling fraction from the trunk bark and branches of Ceiba pentandra. Phytomedicine Plus. 2022;2(4):100352. doi:10.1016/j.phyplu.2022.100352

35.

Miezi Nsimba

. In vitro reversal of deformity and inhibition of aggregation of sickle red blood cells by two Congolese herbal medicines. J Med Plants Res. 2012;6(31). doi:10.5897/jmpr11.376

36.

Ibrahim

Abubakar

. Toxicological studies of Ceiba pentandra Linn. Afr J Biochem Res. 2009;3(7):279–281. Accessed May 20, 2025. http://www.academicjournals.org/AJBR

37.

Tshilanda

Onyamboko

Babady-Bila

, et al. Anti-sickling activity of ursolic acid isolated from the leaves of Ocimum gratissimum L. (Lamiaceae). Nat Products Bioprospect. 2015;5(4):215–221. doi:10.1007/s13659-015-0070-6

38.

Tshilanda

Mutwale

Onyamboko

DVN

, et al. Chemical fingerprint and anti-sickling activity of rosmarinic acid and methanolic extracts from three species of Ocimum from DR Congo. J Biosci Med. 2016;04(01):59–68. doi:10.4236/jbm.2016.41008

39.

Assih

Essotolom

Jocelyn

, et al. In-vitro and in-vivo toxicological studies of hydroethanolic leaf extract of Ocimum gratissimum Linn. (Lamiaceae) in Wistar rats. Adv Med Plant Res. 2022;10(2):30–38. doi:10.30918/ampr.102.22.012

40.

Adejumo

Ayoola

Kolapo

Orimoyegun

Olatunji

. Antisickling activities of extracts of leaf, seed and seed pod of Garcinia kola Heckel. African J Pharm Pharmacol. 2011;5(1):48–52. doi:10.5897/ajpp10.052

41.

Elekwa

Monanu

Anosike

. Effects of aqueous extracts of Garcinia kola seeds on membrane stability of HbAA, HbAS and HbSS human erythrocytes. Glob J Med Sci. 2004;2(2). doi:10.4314/gjms.v2i2.10116

42.

Kagbo

Ejebe

. Phytochemistry and preliminary toxicity studies of the methanol extract of the stem bark of Garcinia kola (Heckel). Internet J Toxicol. 2010;7(2). doi:10.5580/2540

43.

Abere

Ojogwu

Agoreyo

Eze

. Antisickling and toxicological evaluation of the leaves of Rauwolfia vomitoria Afzel (Apocynaceae). J Sci Pract Pharm. 2014;1(1):11–15.

44.

Adomefa

Essotolom

Diallo

, et al. Toxicological study of the aqueous extract of the stem bark of Rauvolfia vomitoria (Apocynaceae). Asian J Biol Sci. 2024;17(2):169–178. doi:10.3923/ajbs.2024.169.178

45.

Das

Kumar

Dash

. Molecular docking and its application in search of antisickling agent from Carica papaya. J Appl Biol Biotechnol. 2020;8(1):105–116. doi:10.7324/JABB.2020.80117

46.

Teng

Chan

Suwanarusk

, et al. In vitro antimalarial evaluations and cytotoxicity investigations of carica papaya leaves and carpaine. Nat Prod Commun. 2019;14(1):33–36. doi:10.1177/1934578X1901400110

47.

Zunjar

Dash

Jivrajani

Trivedi

Nivsarkar

. Antithrombocytopenic activity of carpaine and alkaloidal extract of Carica papaya Linn. leaves in busulfan induced thrombocytopenic Wistar rats. J Ethnopharmacol. 2016;181:20–25. doi:10.1016/j.jep.2016.01.035

48.

Wang

Chen

Wang

. Isolation and identification carpaine in carica papaya L. Leaf by HPLC-UV method. Int J Food Prop. 2015;18(7):1505–1512. doi:10.1080/10942912.2014.900785

49.

Muhammad

Waziri

Forcados

, et al. Sickling-preventive effects of rutin is associated with modulation of deoxygenated haemoglobin, 2,3-bisphosphoglycerate mutase, redox status and alteration of functional chemistry in sickle erythrocytes. Heliyon. 2019;5(6):e01905. doi:10.1016/j.heliyon.2019.e01905

50.

Ramani

Basak

Puneeth

Mahato

Malathi

. Anti hemolytic activity of rutin incase of phenyl hydrazine induced hemolysis. Curr Trends Biotechnol Pharm. 2021;15(5):467–470. doi:10.5530/ctbp.2021.3s.41

51.

Salam

Arif

Sharma

Mahmood

. Protective effect of rutin against thiram-induced cytotoxicity and oxidative damage in human erythrocytes. Pestic Biochem Physiol. 2023;189:105294. doi:10.1016/j.pestbp.2022.105294

52.

Wilson

Mortarotti

Doxtader

. Toxicity studies on rutin. Proc Soc Exp Biol Med. 1947;64(3):324–327. doi:10.3181/00379727-64-15781

53.

Hamid

Sahib

. The acute toxicity of rutin in mice. Iraqi J Pharm Sci. 2021;30(2):231–240. doi:10.31351/vol30iss2pp231-240

54.

Cristina Marcarini

Ferreira Tsuboy

Cabral Luiz

Regina Ribeiro

Beatriz Hoffmann-Campo

Ségio Mantovani

. Investigation of cytotoxic, apoptosis-inducing, genotoxic and protective effects of the flavonoid rutin in HTC hepatic cells. Exp Toxicol Pathol. 2011;63(5):459–465. doi:10.1016/j.etp.2010.03.005

55.

Moghaddasian Behnaz Eghdami Anoosh . Determination of rutin content in Caper (Capparis spinosa) by three analytical methods. Sch Res Libr Ann Biol Res. 2012;2012(9):4303–4306. Accessed June 19, 2025. http://scholarsresearchlibrary.com/archive.html

56.

Liao

Chen

Jiao

Wang

. Protective effects of kaempferol against reactive oxygen species-induced hemolysis and its antiproliferative activity on human cancer cells. Eur J Med Chem. 2016;114:24–32. doi:10.1016/j.ejmech.2016.02.045

57.

Yangzom

Amruthanand

Sharma

, et al. Subacute 28 days oral toxicity study of kaempferol and biochanin-A in the mouse model. J Biochem Mol Toxicol. 2022;36(8). doi:10.1002/jbt.23090

58.

Akiyama

Mizokami

Ito

Ikeda

. A randomized, placebo-controlled trial evaluating the safety of excessive administration of kaempferol aglycone. Food Sci Nutr. 2023;11(9):5427–5437. doi:10.1002/fsn3.3499

59.

Zhang

. Sensitive determination of kaempferol in rat plasma by high-performance liquid chromatography with chemiluminescence detection and application to a pharmacokinetic study. J Chromatogr B Anal Technol Biomed Life Sci. 2009;877(29):3595–3600. doi:10.1016/j.jchromb.2009.08.046

60.

Zhang

Wang

Zhou

, et al. Determination of free and glucuronidated kaempferol in rat plasma by LC-MS/MS: application to pharmacokinetic study. J Chromatogr B Anal Technol Biomed Life Sci. 2010;878(23):2137–2140. doi:10.1016/j.jchromb.2010.06.002

61.

Muhammad

Waziri

Forcados

, et al. Antisickling effects of quercetin may be associated with modulation of deoxyhaemoglobin, 2, 3-bisphosphoglycerate mutase, redox homeostasis and alteration of functional chemistry in human sickle erythrocytes. Ann Sci Technol. 2019;4(1):38–47. doi:10.2478/ast-2019-0005

62.

Sánchez-Gallego

López-Revuelta

Hernández-Hernández

, et al. Comparative antioxidant capacities of quercetin and butylated hydroxyanisole in cholesterol-modified erythrocytes damaged by tert-butylhydroperoxide. Food Chem Toxicol. 2011;49(9):2212–2221. doi:10.1016/j.fct.2011.06.014

63.

Cunningham

Patton

VanderVeen

, et al. Sub-chronic oral toxicity screening of quercetin in mice. BMC Complement Med Ther. 2022;22(1):1–12. doi:10.1186/s12906-022-03758-z

64.

National Toxicology Program. Toxicology and carcinogenesis studies of quercetin (CAS No. 117-39-5) in F344 rats (feed studies). Natl Toxicol Program Tech Rep Ser. 1992;409:1–171. Accessed June 19, 2025. http://www.ncbi.nlm.nih.gov/pubmed/12621521

65.

Carvalho

Jesus

Pinho

Oliveira

Moreira

Oliveira

. Validation of an HPLC-DAD method for quercetin quantification in nanoparticles. Pharmaceuticals. 2023;16(12):1–14. doi:10.3390/ph16121736

66.

Rodríguez

Badimon

Méndez

, et al. Antiplatelet activity of isorhamnetin via mitochondrial regulation. Antioxidants. 2021;10(5):1–12. doi:10.3390/antiox10050666

67.

Subhi Sammani

Clavijo

González

Cerdà

. High-performance liquid chromatographic method for the simultaneous determination of four flavonols in food supplements and pharmaceutical formulations. Anal Lett. 2019;52(8):1298–1314. doi:10.1080/00032719.2018.1536138

68.

Hamed

ANE

Abouelela

El Zowalaty

Badr

Abdelkader

MSA

. Chemical constituents from Carica papaya Linn. leaves as potential cytotoxic, EGFRwt and aromatase (CYP19A) inhibitors; a study supported by molecular docking. RSC Adv. 2022;12(15):9154–9162. doi:10.1039/d1ra07000b

69.

Yang

Kang

Lee

Kang

Lee

Jeon

. Antioxidant activity and cell protective effect of loliolide isolated from Sargassum ringgoldianum subsp. coreanum. ALGAE. 2011;26(2):201–208. doi:10.4490/algae.2011.26.2.201

70.

Percot

Yalçin

Aysel

Erduǧan

Dural

Güven

. Loliolide in marine algae. Nat Prod Res. 2009;23(5):460–465. doi:10.1080/14786410802076069

71.

Nugroho

Heryani

Choi

Park

. Identification and quantification of flavonoids in Carica papaya leaf and peroxynitrite-scavenging activity. Asian Pac J Trop Biomed. 2017;7(3):208–213. doi:10.1016/j.apjtb.2016.12.009

72.

Mahmoud

. Protective effect of myricetin on proteins and lipids of erythrocytes membranes. Asian J Biol Sci. 2012;6(1):76–83. doi:10.3923/ajbs.2013.76.83

73.

Hobbs

Swartz

Maronpot

, et al. Genotoxicity evaluation of the flavonoid, myricitrin, and its aglycone, myricetin. Food Chem Toxicol. 2015;83:283–292. doi:10.1016/j.fct.2015.06.016

74.

Dang

Lin

Xie

, et al. Quantitative determination of myricetin in rat plasma by ultra performance liquid chromatography tandem mass spectrometry and its absolute bioavailability. Drug Res (Stuttg). 2013;64(10):516–522. doi:10.1055/s-0033-1363220

75.

Latiff

Suan

Sarmidi

Ware

Rashid

SNAA

Yahayu

. Liquid chromatography tandem mass spectrometry for the detection and validation of quercetin-3-O-rutinoside and myricetin from fractionated Labisia pumila var. Alata. Malaysian J Anal Sci. 2018;22(5):817–827. doi:10.17576/mjas-2018-2205-09

76.

Soib

Ismail

Husin

Abu Bakar

Yaakob

Sarmidi

. Bioassay-guided different extraction techniques of Carica papaya (Linn.) leaves on in vitro wound-healing activities. Molecules. 2020;25(3):517. doi:10.3390/molecules25030517

77.

Zain

MSC

Osman

Lee

Shaari

. UHPLC-UV/PDA method validation for simultaneous quantification of luteolin and apigenin derivatives from elaeis guineensis leaf extracts: an application for antioxidant herbal preparation. Molecules. 2021;26(4):1084. doi:10.3390/molecules26041084

78.

Lin

Hsu

Lin

Hsu

. Antioxidant and hepatoprotective effects of punicalagin and punicalin on acetaminophen-induced liver damage in rats. Phyther Res. 2001;15(3):206–212. doi:10.1002/ptr.816

79.

Cerdá

Cerón

Tomás-Barberán

Espín

. Repeated oral administration of high doses of the pomegranate ellagitannin punicalagin to rats for 37 days is not toxic. J Agric Food Chem. 2003;51(11):3493–3501. doi:10.1021/jf020842c

80.

Gadkari

Daharwal

. Quantification of punicalagin in pomegranate peels from high-performance thin-layer chromatography. Biomed Biotechnol Res J. 2022;6(4):586–590. doi:10.4103/bbrj.bbrj_312_22

81.

Solakyildirim

. Fast punicalagin content analysis of various brands of pomegranate (Punica granatum L.) juices by UPLC-MS. Hacettepe J Biol Chem. 2019;47(3):267–275. doi:10.15671/hjbc.626949

82.

Ramkumar

Vijayakumar

Vanitha

, et al. Protective effect of gallic acid on alloxan-induced oxidative stress and osmotic fragility in rats. Hum Exp Toxicol. 2014;33(6):638–649. doi:10.1177/0960327113504792

83.

Variya

Bakrania

Madan

Patel

. Acute and 28-days repeated dose sub-acute toxicity study of gallic acid in albino mice. Regul Toxicol Pharmacol. 2019;101:71–78. doi:10.1016/j.yrtph.2018.11.010

84.

Kardani

Gurav

Solanki

Patel

. RP-HPLC method development and validation of gallic acid in polyherbal tablet formulation. J Appl Pharm Sci. 2013;3(5):37–42. doi:10.7324/JAPS.2013.3508

85.

Terças

Monteiro

ADS

Moffa

, et al. Phytochemical characterization of Terminalia catappa Linn. extracts and their antifungal activities against Candida spp. Front Microbiol. 2017;8(APR):595. doi:10.3389/fmicb.2017.00595

86.

Gour

Kour

Pandian

, et al. Ellagic acid exerts dual action to curb the pathophysiological manifestations of sickle cell disease and attenuate the hydroxyurea-induced myelosuppression in Berkeley mice. ACS Pharmacol Transl Sci. 2023;6(6):868–877. doi:10.1021/acsptsci.3c00026

87.

Deepika Maurya

. Protective effect of ellagic acid on erythrocytes subjected to oxidative stress during human ageing. Indian J Nat Prod Resour. 2023;14(1):107–111. doi:10.56042/ijnpr.v14i1.1129

88.

Da Graça Sampaio

Gontijo

AVL

Araujo

Koga-Ito

. In vivo efficacy of ellagic acid against Candida albicans in a Drosophila melanogaster infection model. Antimicrob Agents Chemother. 2018;62(12). doi:10.1128/AAC.01716-18

89.

Loo

WTY

Jin

Cheung

MNB

Chow

LWC

. Evaluation of Ellagic acid on the activities of oral bacteria with the use of adenosine triphosphate (ATP) bioluminescence assay. African J Biotechnol. 2010;9(25):3938–3943. . doi:10.5897/AJB2010.000-3270

90.

Girme

Saste

Pawar

, et al. Development and validation of a sensitive method for analysis of ellagic acid in dietary supplements from Punica granatum. Curr Top Nutraceutical Res. 2020;19(1):90–105. doi:10.37290/ctnr2641-452x.19:90-105

91.

Mane

Aloorkar

Ghorpade

Shinde

Kolekar

Gore

. Development and validation of a bioanalytical method for estimating ellagic acid in Wistar rat plasma by RP-HPLC using a gradient elution technique. BIO Integr. 2024;5(1):975. doi:10.15212/bioi-2024-0029

92.

Cao

Wang

. Attenuation of oxidative stress of erythrocytes by the plant-derived flavonoids vitexin and apigenin. Pharmazie. 2015;70(11):724–732. doi:10.1691/ph.2015.5665

93.

Singh

Mishra

Noel

Sharma

Rath

. Acute exposure of apigenin induces hepatotoxicity in Swiss mice. PLoS One. 2012;7(2):e31964. doi:10.1371/journal.pone.0031964

94.

Willer

Jöhrer

Greil

Zidorn

Çiçek

. Cytotoxic properties of Damiana (Turnera diffusa) extracts and constituents and a validated quantitative UHPLC-DAD assay. Molecules. 2019;24(5):855. doi:10.3390/molecules24050855

95.

Lone

Sharma

Kour

, et al. In-vitro anti-sickling potential of baicalin and naringenin isolated from Oroxylum indicum and Citrus aurantium on human sickle red blood cells. Nat Prod Res. 2023;37(22):3902–3908. doi:10.1080/14786419.2022.2158330

96.

Ajdžanovic

Jakovljevic

Milenkovic

, et al. Positive effects of naringenin on near-surface membrane fluidity in human erythrocytes. Acta Physiol Hung. 2015;102(2):131–136. doi:10.1556/036.102.2015.2.3

97.

Zaidun

Thent

Latiff

. Combating oxidative stress disorders with citrus flavonoid: naringenin. Life Sci. 2018;208:111–122. doi:10.1016/j.lfs.2018.07.017

98.

Yakubu

Lee

Shaari

. Chemical profiles of Terminalia catappa LINN nut and Terminalia subspathulata KING fruit. Pertanika J Trop Agric Sci. 2021;44(4):795–823. doi:10.47836/PJTAS.44.4.06

99.

Jiayi

Tianyi

Dan

Tingguo

Qingfeng

Qianqian

. Isorhamnetin protects endothelial cells model CRL1730 from oxidative injury by hydrogen peroxide. Pak J Pharm Sci. 2019;32(1):131–136.

100.

Hasrat

De Bruyne

De Backer

Vauquelin

Vlietinck

. Isoquinoline derivatives isolated from the fruit of Annona muricata as 5-HTergic 5-HT(1A) receptor agonists in rats: unexploited antidepressive (lad) products. J Pharm Pharmacol. 1997;49(11):1145–1149. doi:10.1111/j.2042-7158.1997.tb06058.x

101.

Costa

Da Cruz

PEO

De Lourenço

De Souza Moraes

De Lima Nogueira

Salvador

. Antioxidant and antimicrobial activities of aporphinoids and other alkaloids from the bark of Annona salzmannii A. DC. (Annonaceae). Nat Prod Res. 2013;27(11):1002–1006. doi:10.1080/14786419.2012.688044

102.

Chen

Liu

Chen

. The pharmacological activities of (-)-anonaine. Molecules. 2013;18(7):8257–8263. doi:10.3390/molecules18078257

103.

Ethiraj

Sridar

. Phytochemical screening, antioxidant activity and extraction of active compound (Anonaine) from fruit peel extract of annona reticulata l. Asian J Pharm Clin Res. 2018;11(11):372–377. doi:10.22159/ajpcr.2018.v11i11.27838

104.

Nawwar

Ayoub

Hussein

, et al. A flavonol triglycoside and investigation of the antioxidant and cell stimulating activities of Annona muricata Linn. Arch Pharm Res. 2012;35(5):761–767. doi:10.1007/s12272-012-0501-4

105.

Noreen

Serrano

Perera

Bohlin

. Flavan-3-ols isolated from some medicinal plants inhibiting COX-1 and COX-2 catalysed prostaglandin biosynthesis. Planta Med. 1998;64(6):520–524. doi:10.1055/s-2006-957506

106.

Fitria

Afrizal

. Isolation and characterization of antioxidative constituent from stem bark extract of Ceiba pentandra L. J Chem Pharm Res. 2010;7(10):257–260. Accessed June 20, 2025. https://www.jocpr.com/abstract/isolation-and-characterization-of-antioxidative-constituent-from-stem-bark-extract-of-ceiba-pentandra-l-6393.html

107.

Yoshida

Niki

. Antioxidant effects of phytosterol and its components. J Nutr Sci Vitaminol (Tokyo). 2003;49(4):277–280. doi:10.3177/jnsv.49.277

108.

Bedner

Schantz

Sander

Sharpless

. Development of liquid chromatographic methods for the determination of phytosterols in standard reference materials containing saw palmetto. J Chromatogr A. 2008;1192(1):74–80. doi:10.1016/j.chroma.2008.03.020

109.

Menaga

Geetha

. Bioactive compounds in Ceiba pentandra: gC-MS analysis and molecular docking insights into potential anticancer properties. Next Res. 2025;2(3):100500. Published online June 11, 2025. doi:10.1016/J.NEXRES.2025.100500.

110.

Senthil

Sridevi

Pugalendi

. Cardioprotective effect of oleanolic acid on isoproterenol-induced myocardial ischemia in rats. Toxicol Pathol. 2007;35(3):418–423. doi:10.1080/01926230701230312

111.

Rajasekaran

Bapna

Lakshmanan

Ramachandran Nair

Veliath

Panchanadam

. Antifertility effect in male rats of oleanolic acid, a triterpene from Eugenia jambolana flowers. J Ethnopharmacol. 1988;24(1):115–121. doi:10.1016/0378-8741(88)90142-0

112.

Wan

Liu

. Repeated oral administration of oleanolic acid produces cholestatic liver injury in mice. Molecules. 2013;18(3):3060–3071. doi:10.3390/molecules18033060

113.

Rada

Castellano

Perona

Guinda

. GC-FID determination and pharmacokinetic studies of oleanolic acid in human serum. Biomed Chromatogr. 2015;29(11):1687–1692. doi:10.1002/bmc.3480

114.

Noolvi

Singh

Yadav

. Quantification of oleanolic acid in the flower of Gentiana olivieri Griseb. by HPLC. J Basic Clin Pharm. 2012;3(2):241. doi:10.4103/0976-0105.103814

115.

Nganteng

DND

Melong

Mbiekop

, et al. Chemical constituents and cytotoxic activity of Ocimum gratissimum L. South African J Bot. 2022;150:330–333. doi:10.1016/j.sajb.2022.07.029

116.

Malini

Vanithakumari

. Rat toxicity studies with β-sitosterol. J Ethnopharmacol. 1990;28(2):221–234. doi:10.1016/0378-8741(90)90032-O

117.

Khonsa

Setyaningrum

Saputro

Amelia

Ibrahim

Damayanti

. Analysis of β-sitosterol in supplement using high-performance liquid chromatography: development and validation. Rasayan J Chem. 2022;15(3):1997–2003. doi:10.31788/RJC.2022.1536752

118.

Kiyoshi

. Inhibitory effect of eugenol on Cu2+-catalyzed lipid peroxidation in human erythrocyte membranes. Int J Biochem. 1989;21(7):745–749. doi:10.1016/0020-711X(89)90205-X

119.

Soundran

Namagiri

Manonayaki

Vanithakumari

. Hepatotoxicity of eugenol. Anc Sci Life. 1994;13(3-4):213–217. http://www.ncbi.nlm.nih.gov/pubmed/22556649%0Ahttp://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=PMC3336525

120.

Escobar-García

Rodríguez-Contreras

Ruiz-Rodríguez

Pierdant-Pérez

Cerda-Cristerna

Pozos-Guillén

. Eugenol toxicity in human dental pulp fibroblasts of primary teeth. J Clin Pediatr Dent. 2016;40(4):312–318. doi:10.17796/1053-4628-40.4.312

121.

Gueretz

Somensi

Martins

Souza

APD

. Avaliação da toxicidade do eugenol em bioensaios com organismos-teste. Cienc Rural. 2017;47(12). doi:10.1590/0103-8478cr20170194

122.

YN MG, KC C, Setty SR. A validated comparative study of RP-HPLC, GC-FID and UV spectrophotometric methods for the quantification of eugenol isolated from Syzygium Aromaticum L. J Pharmacogn Phytochem. 2024;13(5):544–550. doi:10.22271/PHYTO.2024.V13.I5H.15123

123.

Paula-Freire

LIG

Molska

Andersen

Carlini

ELDA

. Ocimum gratissimum essential oil and its isolated compounds (Eugenol and Myrcene) reduce neuropathic pain in mice. Planta Med. 2015;82(3):211–216. doi:10.1055/s-0035-1558165

124.

Mahmud

Mauro

Föller

Lang

. Inhibitory effect of thymol on suicidal erythrocyte death. Cell Physiol Biochem. 2009;24(5-6):407–414. doi:10.1159/000257433

125.

Gad

MMES

Mohammad

TGM

. Acute and repeated-doses (28 days) toxicity of thymol formulation in male albino rats. Aust J Basic Appl Sci. 2013;7(10):915–922.

126.

André

WPP

Cavalcante

Ribeiro

WLC

, et al. Anthelmintic effect of thymol and thymol acetate on sheep gastrointestinal nematodes and their toxicity in mice. Rev Bras Parasitol Vet. 2017;26(3):323–330. doi:10.1590/S1984-29612017056

127.

Baldissera

Souza

De Matos

AFIM

, et al. Blood-brain barrier breakdown, memory impairment and neurotoxicity caused in mice submitted to orally treatment with thymol. Environ Toxicol Pharmacol. 2018;62:114–119. doi:10.1016/j.etap.2018.06.012

128.

Foudah

Shakeel

Alqarni

, et al. Determination of thymol in commercial formulation, essential oils, traditional, and ultrasound-based extracts of thymus vulgaris and Origanum vulgare using a greener HPTLC approach. Molecules. 2022;27:1164.

129.

Lin

Shi

Wang

Shen

. Luteolin, a flavonoid with potential for cancer prevention and therapy. Curr Cancer Drug Targets. 2008;8(7):634–646. doi:10.2174/156800908786241050

130.

Christian Emeka

Chikodi

Chinwendu Mirian

. The medicinal qualities of the bioactive compounds in Garcinia kola. Int J Adv Eng Manag. 2022;4(3):21. doi:10.35629/5252-04032123

131.

Farvin

KHS

Anandan

Sankar

Nair

PGV

. Protective effect of squalene against isoproterenol-induced myocardial infarction in rats. J Clin Biochem Nutr. 2005;37(2):55–60. doi:10.3164/jcbn.37.55

132.

Jiang

Chen

. Determination of squalene using high-performance liquid chromatography with diode array detection. Chromatographia. 2004;59(5-6):367–371. doi:10.1365/s10337-003-0176-6

133.

Budge

Barry

. Determination of squalene in edible oils by transmethylation and GC analysis. MethodsX. 2019;6:15–21. doi:10.1016/j.mex.2018.12.001

134.

Adaramoye

Farombi

Adeyemi

Emerole

. Comparative study on the antioxidant properties of flavonoids of Garcinia kola seeds. Pakistan J Med Sci. 2005;21(3):331–339.

135.

Tshibangu

Kapepula

Kapinga

MJK

, et al. Fingerprinting and validation of a LC-DAD method for the analysis of biflavanones in Garcinia kola-based antimalarial improved traditional medicines. J Pharm Biomed Anal. 2016;128:382–390. doi:10.1016/j.jpba.2016.04.042

136.

Seeman

Weinstein

. Erythrocyte membrane stabilization by tranquilizers and antihistamines. Biochem Pharmacol. 1966;15(11):1737–1752. doi:10.1016/0006-2952(66)90081-5

137.

Sato

Kitaura

Minami

Matsumoto

Fukuda

. Hypothermia-related testicular toxicity of reserpine in mice. Exp Toxicol Pathol. 2007;59(3-4):187–195. doi:10.1016/j.etp.2007.05.006

138.

Pfeifer

Greenblatt

Koch Weser

. Clinical toxicity of reserpine in hospitalized patients: a report from the Boston collaborative drug surveillance program. Am J Med Sci. 1976;271(3):269–276. doi:10.1097/00000441-197605000-00002

139.

Negi

Bisht

Bhandari

Bisht

Singh

. Quantification of reserpine content and antibacterial activity of Rauvolfia serpentina (L.) Benth. ex Kurz. African J Microbiol Res. 2014;8(2):162–166. doi:10.5897/ajmr2013.5847

140.

Xiaojiang

. Indole alkaloids from Rauwolfia vomitoria. Nat Prod Res Dev. 2007;19(3):235–239.

141.

Neha Ansari

Khan

. Yohimbine hydrochloride ameliorates collagen type-II-induced arthritis targeting oxidative stress and inflammatory cytokines in Wistar rats. Environ Toxicol. 2017;32(2):619–629. doi:10.1002/tox.22264

142.

Giampreti

Lonati

Locatelli

Rocchi

Campailla

. Acute neurotoxicity after yohimbine ingestion by a body builder. Clin Toxicol. 2009;47(8):827–829. doi:10.1080/15563650903081601

143.

Cimolai

. Yohimbine use for physical enhancement and its potential toxicity. J Diet Suppl. 2011;8(4):346–354. doi:10.3109/19390211.2011.615806

144.

Zanolari

Ndjoko

Ioset

Marston

Hostettmann

. Qualitative and quantitative determination of yohimbine in authentic yohimbe bark and in commercial aphrodisiacs by HPLC-UV-API/MS methods. Phytochem Anal. 2003;14(4):193–201. doi:10.1002/pca.699

145.

Rivera-Pastrana

Yahia

González-Aguilar

. Phenolic and carotenoid profiles of papaya fruit (Carica papaya L.) and their contents under low temperature storage. J Sci Food Agric. 2010;90(14):2358–2365. doi:10.1002/jsfa.4092

146.

Canini

Alesiani

Arcangelo

Tagliatesta

. ARTICLE IN PRESS gas chromatography – mass spectrometry analysis of phenolic compounds from Carica papaya L. Leaf. J Food Compos Anal. 2007;20:584–590. doi:10.1016/j.jfca.2007.03.009

147.

Maungchanburi

Chaithada

Rattanaburi

, et al. Antiproliferative activity and GC−MS analysis from the leaves extract of different cultivars Carica papaya. ASEAN J Sci Technol Rep. 2024;27(6):e254526. doi:10.55164/ajstr.v27i6.254526

148.

Singh

Bajpai

Kumar

. Comparative profiling of phenolic compounds from different plant parts of six Terminalia species by liquid chromatography – tandem mass spectrometry with chemometric analysis. Ind Crop Prod. 2016;87:236–246. doi:10.1016/j.indcrop.2016.04.048

149.

Mohitha

Shanmugam

. Chemometric analysis and quantification of major anti-cancer activity compounds from leaves of soursop (Annona muricata Linn.). Chem Pap. 2023;77(10):6069–6081. doi:10.1007/s11696-023-02922-0

150.

Misra

Srivastava

Rawat

AKS

. A validated reversed-phase over-pressured layer chromatography-ultraviolet method for the quantification and optimum recovery of gallic acid in annona muricata L. J Planar Chromatogr - Mod TLC. 2016;29(2):127–131. doi:10.1556/1006.2016.29.2.6

151.

Ezerioha

Ibegbulem

Onuoha

Akudinobi

Dike

Morah

. Bioactive phytochemicals in Annona muricata fruit juice ethanol extract. GSC Biol Pharm Sci. 2024;26(2):017–023. doi:10.30574/gscbps.2024.26.2.0034

152.

Cercato

Araújo

JMD

Oliveira

, et al. Reduced cutaneous inflammation associated with antioxidant action after topical application of the aqueous extract of Annona muricata leaves. Inflammopharmacology. 2021;29(1):307–315. doi:10.1007/s10787-020-00735-1

153.

Anwar

Rashid

Shahid

Nadeem

. Physicochemical and antioxidant characteristics of kapok (Ceiba pentandra Gaertn.) seed oil. JAOCS, J Am Oil Chem Soc. 2014;91(6):1047–1054. doi:10.1007/s11746-014-2445-y

154.

Bindu

Rameshkumar

Kumar

Singh

Anilkumar

. Distribution of reserpine in rauvolfia species from India - HPTLC and LC-MS studies. Ind Crops Prod. 2014;62:430–436. doi:10.1016/j.indcrop.2014.09.018

155.

Singh

Kumar

Mathan

, et al. Therapeutic application of Carica papaya leaf extract in the management of human diseases. DARU, J Pharm Sci. 2020;28(2):735–744. doi:10.1007/s40199-020-00348-7

156.

Nakamura

Yoshimoto

Murata

, et al. Papaya seed represents a rich source of biologically active isothiocyanate. J Agric Food Chem. 2007;55(11):4407–4413. doi:10.1021/jf070159w

Chemical Markers for Quality Control of Medicinal Plants Used in Sickle Cell Disease Management: Application of the Herbal Marker Ranking System (Herb MaRS)

Abstract

Keywords

Introduction

Method

Medicinal Plants Selection

Data Collection Methods

Reports of Bioactivity and Toxicity of Medicinal Plants Selected

Potential Chemical Markers Identification

Bioactivity and Toxicity Profiles of Potential Markers

Review of Marker Compounds Concentration in Selected Plants and Analytical Methods Availability

Review of Analytical Reference Standards Availability

Prioritisation of Appropriate Markers for Standardisation

Results

Evidence of Bioactivity and Toxicity of Medicinal Plants Selected

Potential Chemical Markers, and Their Bioactivity, Toxicity and Analytical Assay Methods

Concentration of Potential Marker Compounds in Plant Materials and the Availability, Cost, and Purity of Their Analytical Standards

Prioritisation of Appropriate Markers Using the Herb MaRS Framework

Discussion

Conclusion

Footnotes

Acknowledgements

ORCID iDs

Ethical Approval

Statement of Human and Animal Rights

Statement of Informed Consent

Author Contributions

Funding

Declaration of Conflicting Interests

Data Availability

References