Journal of Biochemistry International

Vitamin and Mineral Constituents of Eupatorium odoratum (Obiar Ofu)

2026-01-08T04:56:48+00:00

Eupatorium odoratum is a tropical plant widely used in traditional medicine in Nigeria. While its therapeutic applications are well documented, information on its vitamin and mineral composition remains limited. This study investigated the vitamin and mineral content of Eupatorium odoratum leaves to evaluate its potential as a dietary supplement and functional food ingredient. Fresh leaves were collected from Enugu State University of Science and Technology, Agbani, Enugu State, Nigeria, identified and authenticated, and analyzed using UV spectrophotometry for vitamins and atomic absorption spectrophotometry for minerals. The vitamin analysis revealed that E. odoratum is a rich source of essential vitamins, with particularly high levels of vitamin C (68.059 mg/kg). Substantial amounts of fat-soluble vitamins were also detected, including vitamin E (21.850 mg/kg), vitamin D (10.476 mg/kg), and vitamin A (9.237 mg/kg). The plant further contained B-complex vitamins, such as vitamin B1 (0.044 mg/kg), B2 (0.126 mg/kg), B3 (0.476 mg/kg), B6 (0.238 mg/kg), and B12 (2.738 mg/kg), though in smaller quantities. Mineral analysis showed the presence of essential macro- and micro-elements, including sodium (7.676 mg/kg), calcium (5.098 mg/kg), potassium (4.267 mg/kg), magnesium (3.787 mg/kg), iron (1.422 mg/kg), zinc (0.564 mg/kg), and selenium (0.178 mg/kg), alongside trace amounts of other elements. Toxic metals such as arsenic and aluminum were not detected, while lead, cadmium, and mercury occurred at very low levels. the findings highlight Eupatorium odoratum as a vitamin- and mineral-rich plant with promising nutritional and health benefits, supporting its potential application in disease prevention, functional foods, and drug development.

Impact of Herbicides on Biochemical and Antioxidant Traits of Moth Bean (Vigna aconitifolia L.)

2026-01-13T12:46:31+00:00

Herbicides are widely used for effective weed control in legume crops, and their application may alter physiological and biochemical attributes. In the present investigation, the impact of pre and post emergence herbicides on major biochemical and antioxidant traits of Vigna aconitifolia L. was evaluated under field conditions during the Kharif seasons of 2024–25. The experiment was conducted in a Randomized Block Design with three replications using two genotypes viz., MH-65 and MH-45 and under the application of pendimethalin, imazethapyr, and quizalofop individually and in different combinations. The biochemical responses revealed that control plants maintained the highest levels of water soluble carbohydrates, protein, and leghemoglobin. In MH-65 and MH-45, it showed variation in the carbohydrate content (73.20 and 69.80 mg g^-1 FW), protein content (29.80 and 26.90 mg g^-1FW), and leghemoglobin (1.80 and 1.15 mg g^-1), respectively. Herbicide application, particularly in combination, resulted remarkable reductions in tested parameters whereas proline showed accumulation and superoxide dismutase activity increased significantly under herbicide exposure, with maximum values of 5.35 μmol g⁻¹ FW for proline and 30.20 U mg⁻¹ protein for SOD. Overall, genotype MH-65 exhibited greater metabolic stability, whereas MH-45 showed stronger stress-responsive biochemical activation.

Synergistic Antiulcerogenic Effects of Carica papaya and Chromolaena odorata Leaves in Varying Fractions in an Ethanol-Induced Rat for Gastric Ulcer

2026-01-23T12:23:43+00:00

Gastric ulcer, which is a type of peptic ulcer, is considered one of the major ailments affecting about 60% of adults and nearly 80% of the child population in tropical countries. Several plants have been traditionally used in the treatment of ulcers; however, this study utilized Carica papaya and Chromolaena odorata. This present study aims to evaluate the synergistic antiulcerogenic potentials of different fractions of these plants. The different fractions of the combination of Carica papaya and Chromolaena odorata were prepared using whatever method. The synergistic antiulcerogenic potential of the combination of plants was determined using sixty-five (65) rats weighing between 180g and 200g, which were randomized into thirteen (13) groups, with each group containing five (5) rats. Group 1 (normal control) was given normal rat chow and water with no treatment. Group 2 (negative control) was ulcer-induced without pretreatment. Group 3 (standard control) was ulcer induced and pretreated with 25mg/kg omeprazole. Group 4 was ulcer-induced and pre-treated with 100 mg/kg n-hexane fraction of C. papaya and C. odorata. Group 5 rats were induced with ulcers and given a pre-treatment of 200 mg/kg n-hexane fraction from C. papaya and C. odorata. Rats in group 6 were induced with ulcers and pretreated with 100 mg/kg of the chloroform fraction from C. papaya and C. odorata. Group 7 were induced with ulcers and pretreated with 200 mg/kg chloroform extracts of C. papaya and C. odorata. Group 8 were induced with ulcers and pretreated with 100 mg/kg of the ethyl-acetate fraction from C. papaya and C. odorata. Rats in group 9 were induced with ulcers and pretreated with 200 mg/kg of the ethyl-acetate fraction of C. papaya and C. odorata. Rats in group 10 were induced with ulcers and received a pretreatment of 100 mg/kg of butanol fraction from C. papaya and C. odorata. Group 11 were induced with ulcers and pre-treated using 200 mg/kg of butanol fraction from C. papaya and C. odorata. Group 12 were induced with ulcers and received a pretreatment of 100 mg/kg of ethanol extract from C. papaya and C. odorata. Group 13 rats were induced with ulcers and pretreated with 200 mg/kg of the ethanol fraction from C. papaya and C. odorata. The animals underwent a 24-hour fasting period prior to receiving the extract. Thirty (30) minutes after extract administration, 1ml of absolute ethanol was given to each animal. Two hours after the induction of ulcer with ethanol, the animals were sacrificed, and the stomach dissected to check the level of ulceration. The rats pretreated with different doses of each fraction of C. papaya and C. odorata leaves and Omeprazole significantly (p <0.05) reduced the gastric lesions as compared to the normal negative control with an exception to the group pretreated with 200 mg/kg of n-hexane fraction. The ethyl-acetate fraction (100 mg/kg and 200 mg/kg) produced the most potent inhibition of gastric lesions when compared to Omeprazole (20 mg/kg). These findings show that the different fractions of the combination of C. papaya and C. odorata leaves possess a synergistic antiulcerogenic potential; hence, justifies its traditional usage in the treatment of gastric ulcer.

Enzyme-Driven Phytate Degradation in Fermented Agro-Waste Enhances Protein Efficiency and Oxidative Stability for Broiler Production

2026-02-09T12:55:25+00:00

Background: The utilization of agro-waste in poultry nutrition is limited by anti-nutritional factors, particularly phytate, which reduces nutrient bioavailability. This study investigated the effects of enzyme-driven phytate degradation via yeast fermentation on protein utilization, in-vitro antioxidant potential of feeds, and organ integrity in broiler chickens.

Methods: Agro-waste feed ingredients were fermented using Saccharomyces cerevisiae to enhance endogenous phytase activity prior to feed formulation. Fifty (50) one-day-old Ross 308 broiler chicks were randomly allocated to three dietary groups: phytate-treated fermented feed, non-phytate-treated feed, and a commercial feed control, and reared for six weeks. Protein efficiency ratio (PER) was determined weekly. In-vitro antioxidant assays, including lipid peroxidation (TBARS), catalase, and superoxide dismutase activities, were performed on feed samples. Serum liver enzymes (ALT, AST, ALP, GGT), total protein, and histopathological changes in the liver, small intestine, and bursa of Fabricius were evaluated in birds.

Results: Broilers fed the phytate-treated fermented diet showed a progressive increase in PER from 0.06 ± 0.00 (week 1) to 0.82 ± 0.40 (week 6), which was significantly higher (p < 0.05) than values recorded for the non-phytate-treated diet (0.47 ± 0.11) and commercial feed (0.57 ± 0.05) at week 6. In-vitro lipid peroxidation values decreased with increasing feed concentration, with the phytate-treated feed showing significantly improved oxidative stability across concentrations. Catalase activity was highest in the phytate-treated feed at 40 mg/ml (4.22 ± 0.00 µmol/g, p < 0.05). Serum ALT (20.00–23.67 IU/L), AST (25.67–37.00 IU/L), ALP (22.33–26.33 IU/L), GGT (450.67–473.33 IU/L), and total protein (36.00–59.33 g/dL) remained within normal physiological ranges across all groups. Histopathological evaluation revealed better preservation of hepatic architecture, intestinal villi, and bursal structure in birds fed the phytate-treated fermented diet.

Conclusion: Yeast-mediated phytate degradation improves feed oxidative quality, enhances protein utilization, and supports organ integrity in broiler chickens, highlighting fermented agro-waste as a sustainable poultry feed alternative.

Anti-Diabetic Property of Ethanol Extracts of Mimosa pudica, Cissampelos pareira and Their Combinations in Alloxan-induced Diabetic Rats

2026-02-10T11:55:22+00:00

Mimosa pudica and Cissampelos pareira are perennial subshrubs and creeping herbs respectively which have been reported to have potency in curing various diseases in traditional medicine, including diabetes mellitus. This study aims to evaluate the anti-diabetic property of Mimosa pudica, Cissampelos pareira and their combinations in alloxan-induced diabetic rats. The rats were fasted overnight for 16 hours and diabetes was induced by a single intraperitoneal injection of 150mg/kg alloxan monohydrate. Acute toxicity test (LD₅₀) was determined in two phases using Lorke’s method. Diabetes was checked at three days intervals using One Touch Glucometer and test strips. Lipid profiling was done using standard diagnostic methods. Low doses of M. pudica, did not show any sign of toxicity in the animals, high dosage showed signs of toxicity. Low doses of C. pareira, did not show any sign of toxicity in the animals while high dosage showed sign of toxicity and death was observed after twelve (12) hours. The lower doses of the combination, did not show any sign of toxicity in the animals, higher doses showed signs of toxicity. The control group and the groups that were administered Mimosa pudica and Cissampelos pareira and their combinations showed a significant increase (p<0.05) in their body weight after 21 days of treatment compared to diabetic untreated. Throughout the period of study, the diabetic untreated control rats showed a significant increase (p<0.05) in fasting blood glucose level. The percentage decrease in the fasting blood glucose concentration of the treated groups when compared to the initial fasting blood glucose (FBG) after 21 days, Group F (100mg/kg C. pareira) recorded 65.83% which accounts for the highest percentage reduction. The animals treated with M. pudica and C. pareira and their combinations showed a significant reduction in the levels of total cholesterol, triglycerides, low-density lipoprotein (LDL) and very-low-density lipoprotein (VLDL) when compared with that of diabetic untreated control. In conclusion, M. pudica and C. pareira and their combinations showed significant antidiabetic activities in alloxan-induced diabetic rats. The results of the fasting blood glucose levels indicate improvement in glucose metabolism, reduced weight loss, and managing blood glucose levels in diabetic models.

Synergistic in-vitro Anti-inflammatory and Anti-platelet Activities of Combined Ethanol Leaf Extract of Ficus capensis and Mentha spicata with Phytochemical Characterization

2026-03-11T09:57:09+00:00

Inflammation is a key factor in the development of many chronic diseases, and the search for safer, plant-based anti-inflammatory agents remains important. This study investigated the synergistic in-vitro anti-inflammatory and antiplatelet potential of the combined ethanol leaf extract of Ficuscapensis and Mentha spicata, alongside its phytochemical compositions. Phytochemical screening showed that the combined extract is rich in phenolics and flavonoids, with moderate levels of tannins, saponins, alkaloids, terpenoids, and steroids, and trace amounts of glycosides. The anti-inflammatory activity was assessed using phospholipase A₂ (PLA₂) induced haemolysis, hypotonic solution induced human red blood cell (HRBC) membrane stabilization, and platelet aggregation assays. The extract significantly (P <.001) reduced haemoglobin release caused by PLA₂, demonstrating a concentration-dependent effect with a maximum inhibition of 89.4% at 1.0 µg/ml. It also protected red blood cells against hypotonic-induced lysis, showing inhibition values comparable to diclofenac. In addition, the extract significantly (P <.001) decreased platelet aggregation, with the highest inhibition (80.3%) observed at the highest concentration tested. These findings imply that the combined extract exerts its protective effects through membrane stabilization, inhibition of inflammatory enzymes, and antioxidant activity linked to its rich phytochemical constituents. This study addresses an important research gap by evaluating the synergistic activity of the combined extracts rather than assessing each plant individually. By addressing the limited scientific evidence on their synergistic action, this research supports the traditional use of plant combinations and highlights their potential as natural alternatives for managing inflammation. Further in-vivo and structural elucidation studies are recommended to confirm these promising results.

Protective Effect of Aqueous Extract of Fermented Prosopis africana Seeds (Okpeye) in Wistar Rats their Topical Anti-Inflammatory and Antibacterial Activities

2026-03-12T10:41:49+00:00

Background: Anti-inflammatory medications are classified as either steroidal or non-steroidal. Non-steroidal anti-inflammatory drugs (NSAIDs) relieve pain, lower fever, inhibit blood clot formation, and, at higher doses, reduce inflammation. Their adverse effects vary by drug but commonly include an increased risk of gastrointestinal ulcers and bleeding, as well as potential cardiovascular and kidney complications. Oxidative stress has been recognized as a key pathogenic factor in ulcer development, directly disrupting cellular functions and causing damage to organelles such as mitochondria, lysosomes, and the nucleus.

Aims: The aim of this study was to evaluate the anti-inflammatory and antiulcer effects of aqueous extract of fermented seeds of Prosopis africana (okpeye) in Wistar rats.

Study Design: The study design monitors the therapeutic potential of aqueous extract of fermented seeds of Prosopis africana by assessing its anti-inflammatory and antiulcer effects on experimentally induced rats.

Place and Duration of Study: Department of Biochemistry, University of Nigeria, Nsukka. Between December 2023 and September 2024.

Methodology: The induction of gastric ulcer was carried out using the method of Urushidani. Twenty (20) adult rats of both sexes were used, randomly divided into 5 groups of 4 rats each. They were treated orally with normal saline and varying doses of the aqueous extract of fermented seeds of Prosopis africana. Cimetidine was used as the standard anti-ulcer drug. The In vivo anti-inflammatory study was done using fifteen (15) male Wistar rats. They were divided into five (5) groups of three (3) rats each. Diclofenac was used as the standard anti-inflammatory drug.

Results: The result showed a significant (P < 0.05) decrease of ulcer formation in group 2 animals treated with 100 mg/kg cimetidine (before induction) with respect to the control group 1. The result also showed a significant (P < 0.05) decrease of ulcer formation in groups 4 and 5 animals treated with 100 mg/kg and 200 mg/kg of extract respectively, when compared with the animals in the control group 1. In the first and second phase of anti-inflammatory study, the result shows that the paw oedema of the rats treated with the extract, decreased significantly (P < 0.05) when compared with the animals in the control group 1.

Conclusion: This result shows that aqueous extract of fermented seeds of Prosopis africana (okpeye) aided in the healing of the induced inflammation and chronic ulcer. Hence, the fermented seeds of Prosopis africana (okpeye) has both antiulcer and anti-inflammatory activity.

Exploring Phytochemical Composition and Antioxidant Efficacy of a Polyherbal Formulation

2026-04-07T08:11:00+00:00

This study investigated the phytochemical composition and antioxidant potential of a polyherbal extract from the leaves of Morindageminata, Kalanchoe pinnata, and Solanum americanum using phytochemical screening and multiple in vitro antioxidant assays. Phytochemical analysis revealed high levels of phenolics, flavonoids, tannins, and terpenoids, while saponins, alkaloids, glycosides, and steroids were present in moderate amounts, indicating diverse phytochemical constituents with strong therapeutic potential. The polyherbal extract exhibited strong antioxidant activity across all assays. In the DPPH radical scavenging activity, it showed a concentration dependent increase (62.1%–84.8%) with a low EC₅₀ value (1.285) and a strong correlation coefficient (R² = 0.949), comparable to ascorbic acid, the standard drug used EC₅₀ value (1.174) and correlation coefficient (R² = 0.980). Total antioxidant capacity (TAC) ranged from 0.546 ± 0.01 to 1.720 ± 0.00 expressed as ascorbic acid equivalent (AAE), indicating strong overall reducing power. In the Ferric reducing antioxidant power (FRAP) assay, a significant (P < 0.001) and concentration dependent increase in ferric reducing ability, confirmed the extract efficient electron donating ability through efficient conversion of Ferric (Fe³⁺) to Ferrous (Fe²⁺) ion. The results indicate that the polyherbal extract possesses strong and broad spectrum antioxidant capacity across multiple models assayed. This is likely attributable to its rich phenolic compounds, terpenoids and flavonoid content, and possible synergistic interactions among the bioactive constituent of the medicinal plants. The consistent dose dependent responses across all assays further confirm the reliability and potency of its antioxidant effects. These findings support the potential use of this polyherbal formulation as a natural source of antioxidant agents and provide a scientific basis for its traditional medicinal applications. Further studies are recommended to isolate the active compounds and evaluate in vivo efficacy and safety.

Protective Antioxidant Effects of Azadirachta indica Extract in Retina Tissue of a Diabetic Rat Model

2026-04-07T10:18:34+00:00

Aims: The present study aimed to evaluate the protective antioxidant effects of ethanol leaf extract of Azadirachta indica (A.I) leaves in the retina of streptozotocin (STZ)-induced diabetic rats.

Study Design: This research was an experimental, randomized, controlled in vivo study.

Place and Duration of Study: Department of Biochemistry, Lead City university, Ibadan, Nigeria, between September 2024 and July 2025.

Methodology: Diabetes was induced in male Wistar rats by a single intraperitoneal injection of STZ (55 mg/kg). Diabetic rats were treated daily with A.I leaf extract at doses of 200 and 400 mg/kg for 21 days. Metformin (70 mg/kg) served as a positive control, while untreated diabetic rats served as negative control, and non-diabetic rats served as normal control groups. Retinal homogenates were assayed for malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) activity, catalase (CAT) activity, and total protein using standard colorimetric/kit methods. Fasting blood glucose (FBG) was monitored. Data were analyzed by one-way ANOVA followed by Tukey's post hoc test (P < 0.05).

Results: Streptozotocin-induced diabetes (n = 5 per group) significantly increased retinal malondialdehyde (MDA) from 2.115 ± 0.03175 to 2.292 ± 0.00354 μM and reduced total protein from 29.30 ± 1.68 to 17.95 ± 1.68 mg/g tissue (P < 0.05). Antioxidant markers also declined, including glutathione (GSH) (1.594 ± 0.4791 to 0.6349 ± 0.1350 mM), superoxide dismutase (SOD) (0.02875 ± 0.00984 to 0.002622 ± 0.00016 U/mg), and catalase (CAT) (6.179 ± 0.505 to 1.615 ± 0.684 U/mg) (P < 0.05). Treatment with A.I extract (200 mg/kg) reduced MDA to 2.069 ± 0.01343 μM and restored SOD (0.02957 ± 0.01220 U/mg) and CAT (4.083 ± 0.812 U/mg) toward control levels (P < 0.05 vs. diabetic), with partial recovery of GSH (0.6979 ± 0.02581 mM) and total protein (25.22 ± 2.21 mg/g tissue). The 400 mg/kg dose showed moderate effects, while metformin produced comparable improvements across all parameters. Although all treatments significantly improved oxidative stress markers compared with the untreated diabetic group (P < 0.05), no statistically significant differences were observed between the extract-treated groups and metformin (P > 0.05), indicating comparable efficacy.

Conclusion: Ethanol extract of A.I leaves exerts significant antioxidant protection in the diabetic retina by attenuating lipid peroxidation, restoring enzymatic and non-enzymatic antioxidant defenses, and preserving retinal protein integrity. These findings support its potential as an affordable natural therapeutic agent for mitigating oxidative stress in early diabetic retinopathy, warranting further mechanistic, dose-optimization, and translational studies.

Combined Ethanol Extract of Piper guineense Leaf and Zingiber officinale Rhizome Attenuates Aluminium Chloride Induced Dementia in Wistar Rats by Inhibiting NADPH Oxidase Driven Oxidative Stress and Neuroinflammation

2026-05-13T12:35:20+00:00

Background: Alzheimer's disease (AD) is a progressive neurodegenerative disorder with limited treatment options. Neuroinflammation and oxidative stress, driven by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation, play central roles in AD pathogenesis. This study evaluated the safety profile and neuroprotective efficacy of a combined ethanol extract of Piper guineense leaf and Zingiber officinale rhizome in an aluminium chloride (AlCl₃)-induced dementia model in Wistar rats. Methods: Acute toxicity was assessed using a modified Lorke's method. Thirty male Wistar rats were divided into six groups (n=5): normal control, AlCl₃ control (120 mg/kg), extract control (800 mg/kg), low-dose extract (400 mg/kg + AlCl₃), high-dose extract (800 mg/kg + AlCl₃), and donepezil (5 mg/kg + AlCl₃). The extract doses (400 and 800 mg/kg) were selected based on neuroprotective doses reported in previous studies on the individual plants, and the LD₅₀ was determined to confirm safety. After 21 days, brain homogenates were analysed for NADPH oxidase, nitric oxide metabolites (NOx), nitrate, malondialdehyde (MDA), antioxidant enzymes (SOD, CAT, GPx), and cytokines (TNF-α, IL-6, IL-1β).

Results: The LD₅₀ was 7745.97 mg/kg, indicating a wide safety margin. AlCl₃ administration significantly upregulated NADPH oxidase (73.66±1.17 vs. 47.96±0.99 pg/mg in control), elevated total NOx, depleted nitrate levels, reduced SOD (from 26.84±0.90 to 12.58±0.26 u/g), CAT (from 11.67±0.29 to 5.71±0.12 u/g), and GPx (from 38.96±0.97 to 20.17±0.75 u/g), and increased MDA three-fold (0.60±0.03 vs. 0.20±0.01 mmol/g). Pro-inflammatory cytokines were markedly elevated. High-dose extract (800 mg/kg) significantly inhibited NADPH oxidase (50.00±0.26 pg/mg), restored nitrate levels (3.48±0.14 µmol/g), normalized antioxidant enzymes (SOD: 24.48±0.40 u/g; CAT: 10.24±0.24 u/g; GPx: 34.31±0.66 u/g), reduced MDA (0.28±0.01 mmol/g), and suppressed cytokine levels, with efficacy comparable to donepezil.

Conclusion: The combined extract demonstrates a favourable safety profile and exerts neuroprotective effects by inhibiting NADPH oxidase, restoring redox homeostasis, and suppressing neuroinflammation, supporting its potential as a multi-target therapeutic agent for neurodegenerative conditions.

Antioxidant Properties of Thespesia garckeana (Gorontula) Pulp and Seed

2026-05-27T07:50:45+00:00

The accumulation of free radicals causes oxidative damage to biomolecules, contributing to various metabolic and degenerative diseases. The need for effective antioxidants has led to the exploration of natural sources, particularly medicinal plants. Thespesia garckeana, a plant with ethnomedicinal value, has been traditionally used for various ailments. This study aimed to evaluate the antioxidant properties of T. garckeana pulp and seed. The plant materials were sourced from Gombe, Nigeria, and authenticated by a taxonomist. Various antioxidant assays, including DPPH, ABTS, superoxide, hydrogen peroxide, and hydroxyl radical scavenging activities, as well as the FRAP Assay, were carried out using standard procedures. The results revealed that T. garckeana pulp and seed extracts possess significant antioxidant properties, with the seed extract exhibiting higher DPPH scavenging activity and the pulp showing stronger ABTS, Superoxide scavenging activities, Hydroxyl radical and FRAP assay while both pulp and seed show the extracts' abilities to neutralize hydrogen peroxide at different concentrations when compared to the standard revealing among the extracts, the pulp showing higher antioxidant activity than the seeds. The extracts demonstrated considerable scavenging abilities against various reactive oxygen species (ROS), indicating their potential to mitigate oxidative stress. The findings support the traditional use of T. garckeana in managing oxidative stress-related diseases and suggest its potential as a natural antioxidant source. In conclusion, T. garckeana pulp and seed extracts are promising natural antioxidants, meriting further investigation into their therapeutic potential.

Biochemical and Antioxidant Responses of Dunaliella salina to Thermal and Nutrient Stress

2026-06-02T12:13:12+00:00

Dunaliella salina is a halophilic green microalga belonging to the genus Dunaliella of the phylum Chlorophyta, widely recognized as a premier natural source of high-value metabolites, particularly carotenoids and lipids, due to its remarkable adaptability to extreme environments. In this study, the microalgal strain D. salina CCAP 19/18 was utilized to investigate the accumulation of carotenoids, lipids, phenolics, as well as estimation of antioxidant capacity, under the induction of various thermal regimes and nutrient (NPK) stress. The results demonstrated that D. salina microalgae enhanced the synthesis of carotenoids, lipids, phenolic compounds and alongside increased antioxidant capacity under the stimulation of thermal and nutrient (NPK) stress. These metabolite contents reached peak levels at temperatures ranging from 10 to 25 ^oC across all nutrient (NPK) stress. Conversely, high temperature stress at 45^oC proved unsuitable for both growth and metabolite accumulation in D. salina microalgae. Consequently, thermal and nutrient (NPK) stress were identified as key factors in inducing the synthesis of bioactive compounds in D. salina microalgae.