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    Article

    Anti-Inflammatory and Analgesic Effects of Bixa orellana Chloroform Extract

    Tapas Mishra1★, Sudhakar Kaushik1, Priyanka Bisht1, Bhawana Bhatt2★ Corresponding author

    1. 1Department of Pharmacology, School of Pharmaceutical Sciences, Shri Guru Ram Rai University, Dehradun, Uttarakhand, INDIA.
    2. 2Department of Pharmacognosy, School of Pharmaceutical Sciences, Shri Guru Ram Rai University, Dehradun, Uttarakhand, INDIA.

    CORRESPONDENCE

    Tapas Mishra

    Department of Pharmacology, School of Pharmaceutical Sciences, Shri Guru Ram Rai University, Dehradun-248001, Uttarakhand, INDIA.

    tapasmishra033@gmail.com

    Received: 03-09-2025; Revised: 29-10-2025; Accepted: 19-12-2025.

    Volume 18, Issue 2 · pp. 548–554 · PUBLISHED Apr-Jun 2026 · DOI: 10.5530/pres.20260084

    View on Pharmacogn. Res. original site ↗

    ABSTRACT

    Background Inflammation is a critical defence mechanism employed by the body in response to injury, infection, or harmful stimuli. While Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) are commonly used to manage pain and inflammation, their prolonged use is often associated with adverse effects, including gastrointestinal, cardiovascular, and renal complications. Consequently, there is a growing interest in exploring alternative therapeutic strategies. Among these, plant-derived natural compounds have garnered significant attention due to their rich content of bioactive constituents with potent analgesic and anti-inflammatory properties. Objectives This study aimed to evaluate the preliminary phytochemicals, anti-inflammatory, and analgesic activities of the Chloroform Extract of Bixa orellana leaves (CEBO). Materials and Methods Leaves were extracted using cold maceration with chloroform. Preliminary Phytochemical screening was performed to identify key bioactive compounds. Anti-inflammatory activity was assessed using the carrageenan-induced paw edema model. Analgesic activity was evaluated using Eddy’s hot plate and tail-flick methods. Statistical analysis was done using two-way ANOVA followed by Tukey’s multiple comparison test. Results Phytochemical analysis revealed the presence of flavonoids, alkaloids, terpenoids and phytosterols. Inflammation was significantly reduced in a dose-dependent manner in the carrageenan-induced paw edema model. The extract at 200 mg/kg showed comparable efficacy to indomethacin. CEBO also significantly increased pain threshold in both hot plate and tail-flick tests. Conclusion The chloroform extract of Bixa orellana leaves exhibited significant anti-inflammatory and analgesic activities. These findings support its traditional use and suggest the potential for developing Phyto-therapeutic agents. Further studies are recommended to isolate active constituents and explore their mechanisms of action.

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    FULL TEXT

    INTRODUCTION

    Inflammation is a fundamental biological process triggered by injury or infection, and it is often accompanied by pain as a key symptom. Inflammatory pain arises because of chemical mediators like prostaglandins, bradykinin, and cytokines, which sensitize nociceptors and intensify pain signaling. Nonsteroidal Anti-Inflammatory Drugs (NSAIDs) are widely used to manage inflammatory pain by inhibiting Cyclooxygenase (COX) enzymes, thereby reducing prostaglandin production. Despite their effectiveness, prolonged NSAID use is linked to gastrointestinal, renal, and cardiovascular complications, which emphasizes the need for safer therapeutic alternatives (Vane and Botting, 1998).

    Pain itself is a complex protective phenomenon, but it can become a clinical issue when it persists or becomes chronic. Various drug classes-including NSAIDs, opioids, anticonvulsants, and antidepressants-are utilized to address different pain mechanisms. However, concerns regarding side effects, tolerance, and dependence continue to push research toward safer and more targeted analgesic therapies (Scholz and Woolf, 2002).

    Conventional treatment strategies, especially synthetic analgesics and anti-inflammatory agents, play an essential role in controlling pain and inflammation. Drugs like NSAIDs and opioids remain the mainstay due to their quick and significant relief. However, long-term use often brings notable adverse effects, including gastrointestinal irritation, renal impairment, cardiovascular risks, and the risk of addiction and tolerance associated with opioids (Lanas and Chan, 2017; Benyamin et al., 2008). These challenges have encouraged exploration of alternative therapies, especially those derived from medicinal plants.

    Phytochemicals-bioactive compounds present in plants-have shown promising anti-inflammatory and analgesic effects, often with fewer side effects. Compounds like flavonoids, alkaloids, terpenoids, and polyphenols are being extensively studied for their capacity to modulate inflammatory responses and influence pain perception, without the toxicity commonly observed in synthetic drugs (Calixto, 2000). As a result, there is a growing interest in integrating phytochemical-based medicine as complementary or alternative options for managing pain and inflammation.

    Bixa orellana, widely known as the lipstick plant, is traditionally used in several medicinal systems. Native to tropical regions of the Americas (Neotropics), it is now cultivated across tropical and subtropical zones worldwide, including India, Southeast Asia, the Caribbean, and parts of Africa (Prasad and Kashyap, 2013). This small evergreen tree or shrub typically grows 6-8 M tall, with simple, ovate leaves, entire margins, and fragrant bisexual flowers. The plant’s fruit is a spiny capsule containing many seeds covered with a vivid orange-red aril rich in the pigment bixin (Prasad and Kashyap, 2013; Vilar et al., 2010).

    Different parts of Bixa orellana, particularly the leaves and seeds, are abundant in phytochemicals such as carotenoids (notably bixin and norbixin), flavonoids, phenolic compounds, glycosides, tannins, saponins, steroids, and alkaloids (Rivera et al., 2017; Calixto, 2000). Bixin, the main pigment, is widely utilized as a natural food colorant (E160b) and also in cosmetics and textiles. Traditionally, the plant has been employed to address ailments including fever, sore throat, gastrointestinal disorders, jaundice, hypertension, diabetes, snakebite, and skin infections. In many cultures, it is valued for its anti-inflammatory, antimicrobial, antipyretic, analgesic, and anticonvulsant activities. Decoctions or infusions made from the leaves are often consumed orally or used for gargling, particularly in the treatment of respiratory and digestive conditions (Mbah et al., 2012). The present study was conducted to evaluate the anti-inflammatory and analgesic activities of Bixa orellana Chloroform leaves Extract (CEBO), administered orally, in animal models of inflammation and pain to validate ethnopharmacological claims.

    MATERIALS AND METHODS

    Drugs and chemicals

    All necessary chemicals, reagents, and laboratory facilities required for the experimental procedures were generously provided by Shri Guru Ram Rai University, Dehradun, India. The chemicals used in the study included Chloroform, Carrageenan, Indomethacin (standard anti-inflammatory drug), Ethanol, Vegetable oil, Carboxy Methyl Cellulose (CMC), Tween 80, and Distilled Water. All reagents and solvents were of analytical grade.

    Collection and extraction of plant material

    Bixa orenalla, leaves samples were obtained in the month of January 2025 from SGRRU Herbal garden Dehradun. S. K. Singh of BSI, Dehradun authenticate and granted a certificate under reference number BSI/ NRC/ Herb (Ident.)/ 2024-25/997.

    The coarsely powdered leaves of Bixa orellana was subjected to cold maceration using chloroform as the solvent. Approximately 100 g of dried leaves of Bixa orellana powder was immersed and macerated in 500 mL of chloroform in a stopped conical flask and kept at room temperature for 72 hr with intermittent stirring to facilitate maximum extraction of bioactive compounds. After maceration, the mixture was filtered through Whatman No. 1 filter paper. The obtained filtrate was then concentrated using a rotary evaporator under reduced pressure at 45 °C. The crude extract was stored in a well-closed container and protected from light (Bhatt et al., 2022; Morris et al., 2016; Syamkumar et al., 2023).

    Phytochemical Screening

    Phytochemical screening of the chloroform extract of Bixa orellana leaves (CEBO) was performed as per Shaikh and Patil (2020) to identify different phytoconstituents like flavonoids, alkaloids, phenols, carbohydrate, tannins, phytosterols, and terpenoids (Shaikh et al., 2020).

    Animal and Ethical Aspects

    Wistar albino rats (150-180 g) were used as experimental models, with six rats per group. They were given a 7-day acclimatization period to the laboratory environment and were provided with food and water ad libitum. The work protocol was approved by the Institutional Animal Ethics Committee (IAEC No. 264/ CPCSEA/IAEC/2025/08), and the study was conducted in the School of Pharmaceutical Sciences, Shri Guru Ram Rai University, Dehradun, in 2025 (Nyiramugisha and Odoma, 2021; Hassan et al., 2024; Yong et al., 2013).

    Anti-Inflammatory Activity

    Carrageenan Induced paw edema

    The anti-inflammatory activity of the chloroform extract of Bixa orellana leaves was investigated using the carrageenan-induced paw edema model in rats. The animals were randomly assigned into six groups (n=6 per group): Group A, the normal control, received 0.9% saline orally at a dose of 10 mL/kg; Group B served as the carrageenan control without any treatment; Group C was administered indomethacin at 10 mg/kg orally as the reference anti-inflammatory agent; and Groups D, E, and F received the Bixa orellana chloroform extract orally at doses of 50, 100, and 200 mg/kg respectively. Inflammation was induced by injecting 0.1 mL of carrageenan (prepared as a 1% suspension in sterile normal saline) into the rats' left hind paws to cause edema. All treatments were given orally 1 hr prior to the carrageenan injection. Paw volume was measured with a plethysmometer at intervals of 0, 2, 4, 6, and 24 hr after injection to evaluate edema and the extract’s anti-inflammatory effects (Morris, 2003).

    Analgesic Activity

    Eddy’s hot plate

    Rat of either sex with an initial weight of 150-180 g are used for each dose. The hot plate, which is commercially available, consists of an electrically heated surface. The temperature was maintained at 55±0.2ºC. The animals are placed on the hot plate and the time until either licking or jumping occurs is recorded by a stopwatch. The latency time is recorded before and after 0, 30, 60, 90 and 120 min (Mian et al., 2025; Savla et al., 2025).

    Tail flick test

    Radiant heat was applied to a single spot on the proximal third of the tail using an analgesiometer. The time taken for the animal to withdraw (flick) its tail was recorded as the reaction time. The standard drug or test substance was administered after measuring the baseline reaction times at intervals of 0, 30, 60, 90 and 120 min (Kaushik et al., 2025; Cecchi et al., 2008).

    Statistical Analysis

    The results were analysed using the statistical program GraphPad, with two-way ANOVA employed for group comparisons. A p-value of less than 0.05 (p<0.05) was considered statistically significant. Data are presented as Mean±S.E.M. (Standard Error of the Mean).

    RESULTS

    Phytochemical Screening

    In extracts from chloroform extract of Bixa orellana leaves, a variety of metabolites were discovered (Table 1). Preliminary phytochemical study of the chloroform extract of Bixa orellana leaves, showed the presence of alkaloids, flavonoids, phytosterols and terpenoids.

    Table 1: Preliminary Phytochemical Screening of chloroform extract of Bixa orellana leaves.
    TestObservation
    AlkaloidsPresent
    FlavonoidsPresent
    Phenolic CompoundsAbsent
    CarbohydrateAbsent
    TanninsAbsent
    PhytosterolsPresent
    TerpenoidsPresent

    Anti-Inflammatory Activity- Carrageenan Induced Paw edema

    The anti-inflammatory activity of the Chloroform Extract of Bixa orellana leaves (CEBO), evaluated in rats using carrageenan-induced paw edema, and compared with the standard anti-inflammatory drug indomethacin (10 mg/kg) summarizes in Table 2 and Figure 1. The changes in paw thickness observed at various time points demonstrated that the Chloroform Extract of Bixa orellana leaves (CEBO) produced a dose-dependent anti-inflammatory effect. Notably, the 200 mg/kg dose exhibited significantly greater efficacy compared to the lower doses. These findings suggest that CEBO possesses promising anti-inflammatory properties, potentially supporting its use in the development of plant-based therapeutic agents.

    Figure 1: Graphical representation of effect of chloroform extract (doses 50, 100 and 200 mg/kg) of Bixa orellana leaves carrageenan induced hind paw edema. Indomethacin (10 mg/kg) was used as a standard drug. Values are Mean±S.E.M, * Significance Indicators: a*=vs. Group A (Control), b*=vs. Group B (Positive Control), c*=vs. Group C (Standard), d*=vs. Group D (CEBO 50 mg/kg), e*=vs. Group E (CEBO 100 mg/kg), *p<0.05, **p<0.01, ***p<0.001.
    Table 2: Effect of chloroform extract of Bixa orellana leaves carrageenan-induced paw edema.
    GroupTreatmentDosePaw volume(mL) at different time interval
    0 Hr2 Hr4 Hr6 Hr24 Hr
    Group A (Control)0.9% Saline10 mL/kg0.961±0.0050.960±0.0110.968±0.0070.966±0.0080.960±0.008
    Group B (Positive Control)Carrageenan (1% w/v)-0.970±0.0101.976±0.004 a***1.993±0.005 a***1.966±0.005 a***1.952±0.004 a***
    Group C (Standard)Indomethacin10 mg/kg0.968±0.0071.396±0.008 a***,b***1.416±0.007 a***,b***1.297±0.011 a***,b***1.059±0.009 a***,b***
    Group D (CEBO)Bixa orellana Extract50 mg/kg0.958±0.0111.760±0.014 a***,b***, c***1.769±0.013 a***,b***, c***1.589±0.003 a***,b***, c***1.209±0.005 a***,b***, c***
    Group E (CEBO)Bixa orellana Extract100 mg/kg0.956±0.0041.662±0.021 a***,b***, c*,d*1.679±0.013 a***,b***, c***,d*1.459±0.012 a***,b***, c***,d***1.161±0.013 a***,b***, c**
    Group F (CEBO)Bixa orellana Extract200 mg/kg0.966±0.0181.436±0.007 a***,b***, c*,d***, e***1.453±0.006 a***,b***, c*,d***,e***1.320±0.006 a***,b***, d***,e***1.113±0.004 a***,b***, c*,d***

    Analgesic Activity

    Eddy’s hot plate

    The Chloroform Extract of Bixa orellana leaves (CEBO) produced a significant, dose-dependent increase in reaction time, indicative of central analgesic activity, as presented in Table 3 and Figure 2. Among the extract-treated groups, CEBO at 50 mg/kg exhibited a moderate analgesic effect, while higher doses (100 mg/kg and 200 mg/kg) produced more pronounced responses. Notably, the 200 mg/kg dose showed analgesic efficacy comparable to the standard drug, Indomethacin.

    Figure 2: Graphical Representation of Effect of chloroform extract of Bixa orellana leaves (doses 50, 100 and 200mg/kg) using Hot Plate. Indomethacin (10 mg/kg) used as a standard. All values are expressed as Mean±SEM. Significance Indicators: a*=vs. Group A (Control), b*=vs. Group B (standard), c*=vs. Group C (CEBO 50mg/kg), d*=vs. Group D (CEBO 100 mg/kg), *p<0.05, **p<0.01, ***p<0.001.
    Table 3: Effect of chloroform extract of Bixa orellana leaves by hot plate test, all the values are in second.
    Sl. No.Treatment and DoseReaction Time(sec)
    0 Min30 Min60 Min90 Min120 Min
    1Control3.542 ±0.0693.8±0.0423.818±0.1124.044±0.0363.93±0.025
    2Standard3.4±0.0437.624±0.123 a***10.11±0.285 a***12.963±0.282 a***11.311±0.172 a***
    3CEBO 50mg/kg3.235 ±0.0675.932±0.066 a***,b***6.837±0.148 a***, b***8.653±0.159 a***,b***7.209±0.277 a***,b***
    4CEBO 100mg/kg3.337 ±0.1136.912±0.093 a***,b**,c***8.181±0.294 a***, b**, c*9.989±0.235 a***,b***,c**8.579±0.353 a***,b**
    5CEBO 200mg/kg3.555 ±0.1407.770±0.074 a***,c***,d***9.593±0.331 a***, c***12.437±0.318 a***,c***,d**10.650±0.227 a***,c***,d**

    Tail-flick test method

    As summarized in Table 4 and Figure 3, the Chloroform Extract of Bixa orellana leaves (CEBO) showed significant, dose-dependent analgesic activity in rats. These results demonstrate that CEBO produces a significant, dose-dependent increase in pain threshold, indicating strong central analgesic activity comparable to the standard drug at higher doses.

    Figure 3: Graphical Representation of Effect of chloroform extract of Bixa orellana leaves (doses 50, 100 and 200mg/kg) using Tail Flick Method. Indomethacin (10 mg/kg) used as a standard. Values are Mean±S.E.M.Significance Indicators: a*=vs. Group A (Control), b*=vs. Group B (standard), c*=vs. Group C (CEBO 50mg/kg), d*=vs. Group D (CEBO 100 mg/kg), *p<0.05, **p<0.01, ***p<0.001.
    Table 4: Effect of chloroform extract of Bixa orellana leaves by tail-flick test, all the values are in second.
    Sl. No.Treatment and DoseReaction Time (sec)
    0 Min30 Min60 Min90 Min120 Min
    1Control2.216±0.1302.34±0.3252.42±0.2122.36±0.0952.36±0.198
    2Standard2.3±0.1415.68±0.185 a***7.5±0.178 a***10.24±0.151 a***8.927±0.164 a***
    3CEBO 50mg/kg2.249±0.1173.77±0.060 a*,b***4.917±0.054 a***,b***5.72±0.304 a***,b***5.522±0.099 a***,b***
    4CEBO 100mg/kg2.356±0.1524.606±0.091 a**,b**,c***6.018±0.070 a***,b**,c***7.83±0.150 a***,b***,c**6.9±0.135 a***,b***,c***
    5CEBO 200mg/kg2.361±0.1285.303±0.139 a***,c***,d*7.309±0.144 a***,c***d***9.761±0.229 a***,c***d***8.060±0.032 a***,b*,c***,d**

    DISCUSSION

    The present study was utilize to determine the preliminary phytochemical analysis, anti-inflammatory and analgesic activities of the chloroform extract of Bixa orellana leaves (CEBO). The qualitative phytochemical analysis revealed the presence of alkaloids, flavonoids, phytosterols and terpenoids compounds-each known for distinct pharmacological actions, particularly in relation to inflammation and pain modulation. The presence of key bioactive compounds are well documented for their medicinal properties (Pietta, 2000; Russo, 2008; Gertsch et al., 2008).

    CEBO exhibited dose-dependent anti-inflammatory effects in the carrageenan-induced paw edema model. At 100 mg/kg and 200 mg/kg, CEBO significantly inhibited paw swelling with efficacy comparable to the reference drug indomethacin. This anti-inflammatory activity may be attributed to inhibition of pro-inflammatory mediators such as prostaglandins and leukotrienes, membrane stabilization by phytosterols, and antioxidative effects of flavonoids mitigating reactive oxygen species (Pietta, 2000; Bouic and Lamprecht, 1999; Middleton et al., 2000).

    Analgesic activity was demonstrated by increased latency in Eddy’s hot plate and tail-flick tests, indicating central and peripheral analgesic mechanisms. The highest dose (200 mg/kg) showed analgesic effects approaching those of indomethacin. Terpenoids in CEBO may exert analgesic effects through CB2 receptor activation, while flavonoids likely modulate TRPV1 channels involved in pain transmission (Gertsch et al., 2008; Vriens et al., 2008; Wagner and Elmadfa, 2003; Paduch et al., 2007; Gupta, 1980).

    These findings corroborate traditional claims regarding Bixa orellana and highlight its potential as a promising source of natural anti-inflammatory and analgesic agents. Further studies are warranted to isolate the active constituents and elucidate the underlying mechanisms of action.

    CONCLUSION

    The chloroform extract of Bixa orellana leaves contains a diverse array of phytochemicals that have demonstrated notable anti-inflammatory and analgesic activities. However, further pharmacological investigations are essential to elucidate the precise mechanisms of action and explore the full spectrum of biological activities associated with this plant. The present study reinforces the potential of plant-based therapeutics and underscores the importance of integrating traditional knowledge with modern scientific approaches to develop safe and effective medicines.

    REFERENCES

    As published

    Showing references and in-text citations exactly as published.

    1. 1.Benyamin, R.; Trescot, A. M.; Datta, S.; Buenaventura, R.; Adlaka, R.; Sehgal, N. et al. Opioid complications and side effects. Pain Physician. 2008;11(2):S105–S120. https://doi.org/10.36076/ppj.2008/11/s105DOIGOOGLE SCHOLAR
    2. 2.Bhatt, B.; Chaurasia, H.; Singh, R.; Kaushik, S.. Phytochemical profile and in vitro sun-protective activity of Polyalthia longifolia (Sonn.) Thwaites bark extracts. Tropical Journal of Natural Product Research. 2022;6(8):1174–1177. https://doi.org/10.26538/tjnpr/v6i8.1DOIGOOGLE SCHOLAR
    3. 3.Bouic, P. J. D.; Lamprecht, J. H.. Plant sterols and sterolins: A review of their immune-modulating properties. Alternative Medicine Review. 1999;4(3):170–177.GOOGLE SCHOLAR
    4. 4.Calixto, J. B.. Efficacy, safety, quality control, marketing and regulatory guidelines for herbal medicines (phytotherapeutic agents). Brazilian Journal of Medical and Biological Research. 2000;33(2):179–189. https://doi.org/10.1590/S0100-879X2000000200004DOIGOOGLE SCHOLAR
    5. 5.Cecchi, M.; Capriles, N.; Watson, S. J.; Akil, H.. Differential responses to morphine-induced analgesia in the tail-flick test. Behavioural Brain Research. 2008;194(2):146–151. https://doi.org/10.1016/j.bbr.2008.06.034DOIGOOGLE SCHOLAR
    6. 6.Gertsch, J.; Leonti, M.; Raduner, S.; Rácz, I.; Chen, J.-Z.; Xie, X.-Q. et al. β-caryophyllene is a dietary cannabinoid. Proceedings of the National Academy of Sciences of the United States of America. 2008;105(26):9099–9104. https://doi.org/10.1073/pnas.0803601105DOIGOOGLE SCHOLAR
    7. 7.Gupta, M. B.; Nath, R.; Srivastava, N.; Shanker, K.; Kishor, K.; Bhargava, K. P.. Anti-inflammatory and antipyretic activities of β-sitosterol. Planta Medica. 1980;39(2):157–163. https://doi.org/10.1055/s-2008-1074919DOIGOOGLE SCHOLAR
    8. 8.Hassan, M.; Zahra, N.; Shafi, A.; Shahzadi, S.; Moustafa, A.; Kloczkowski, A.. Investigation of anti-inflammatory, antipyretic and analgesic activities of Citrullus colocynthis in albino rats through in vivo and pharmacoinformatics studies. Recent Advances in Anti-infective Drug Discovery. 2024;19(2):119–136. https://doi.org/10.2174/2772434418666230412105317DOIGOOGLE SCHOLAR
    9. 9.Kaushik, S.; Bhandari, P.; Bhandari, A.; Bhatt, B.; Mittal, H.; Babbar, A.. Analgesic and anti-inflammatory properties of Oxalis dehradunensis Raizada in rats. Pharmacognosy Research. 2025;17(1):185–191. https://doi.org/10.5530/pres.20251985DOIGOOGLE SCHOLAR
    10. 10.Lanas, A.; Chan, F. K. L.. Peptic ulcer disease. The Lancet. 2017;390(10094):613–624. https://doi.org/10.1016/S0140-6736(16DOIGOOGLE SCHOLAR
    11. 11.Mbah, C. J.; Udeh, N. E.; Okechukwu, P. N.. Ethnobotanical survey and pharmacological evaluation of Bixa orellana used in traditional medicine in Africa. African Journal of Traditional, Complementary and Alternative Medicines. 2012;9(4):500–506.GOOGLE SCHOLAR
    12. 12.Mian, S. S.; Shuaib, M.; Zehra, A.; Ansari, M. S.; Husain, I.; Begam, B. et al. Synergistic analgesic effects of ginger, colchicum, and nux vomica extracts in a Unani polyherbal combination. South Eastern European Journal of Public Health. 2025;S1:894–906. https://doi.org/10.70135/seejph.vi.3756DOIGOOGLE SCHOLAR
    13. 13.Middleton, E.; Kandaswami, C.; Theoharides, T. C.. The effects of plant flavonoids on mammalian cells: Implications for inflammation, heart disease, and cancer. Pharmacological Reviews. 2000;52(4):673–751. https://doi.org/10.1016/S0031-6997(24DOIGOOGLE SCHOLAR
    14. 14.Morris, C. J.. Carrageenan-induced paw edema in the rat and mouse. Methods in Molecular Biology. 2003;225:115–121. https://doi.org/10.1385/1-59259-374-7:115DOIGOOGLE SCHOLAR
    15. 15.Morris, S. S.; Jeyabalan, G.; Jha, A. K.; Verma, S.; Swarnkar, Y.. Phytochemical screening and in vitro anthelmintic activity of seed extracts of plants Carum carvi of family Apiaceae. Asian Journal of Research in Pharmaceutical Science. 2016;6(4):246–254. https://doi.org/10.5958/2231-5659.2016.00036.9DOIGOOGLE SCHOLAR
    16. 16.Nyiramugisha, D.; Odoma, S.. Analgesic activity of methanol leaf extract of Momordica foetida (Schumach) in albino rats. KIU Journal of Health Sciences, One(One). 2021;31–37. https://doi.org/10.59568/KJHS-2021-1-1-005DOIGOOGLE SCHOLAR
    17. 17.Paduch, R.; Kandefer-Szerszeń, M.; Trytek, M.; Fiedurek, J.. Terpenes: Substances useful in human healthcare. Archivum Immunologiae et Therapiae Experimentalis. 2007;55(5):315–327. https://doi.org/10.1007/s00005-007-0039-1DOIGOOGLE SCHOLAR
    18. 18.Pietta, P. G.. Flavonoids as antioxidants. Journal of Natural Products. 2000;63(7):1035–1042. https://doi.org/10.1021/np9904509DOIGOOGLE SCHOLAR
    19. 19.Prasad, S.; Kashyap, R. S.. Bixa orellana: A plant of multiple uses. Journal of Medicinal Plants Studies. 2013;1(5):14–20.GOOGLE SCHOLAR
    20. 20.Rivera-Mondragón, A.; Castañeda-Hernández, G.; Martínez-Cruz, M et al. Chemical composition and pharmacological activities of Bixa orellana: A review.*. Phytotherapy Research. 2017;31(8):1215–1228.GOOGLE SCHOLAR
    21. 21.Russo, E. B.. Cannabinoids in the management of difficult to treat pain. Therapeutics and Clinical Risk Management. 2008;4(1):245–259. https://doi.org/10.2147/TCRM.S1928DOIGOOGLE SCHOLAR
    22. 22.Savla, S. R.; Oza, M. J.; Laddha, A. P.; Kulkarni, Y. A.. Anti-nociceptive activity of biochanin A-an important isoflavone from natural source. Pharmacognosy research-Natural products. 2025;100157.GOOGLE SCHOLAR
    23. 23.Scholz, J.; Woolf, C. J.. Can we conquer pain?. Nature Neuroscience. 2002;5(Suppl.):1062–1067. https://doi.org/10.1038/nn942DOIGOOGLE SCHOLAR
    24. 24.Shaikh, J. R.; Patil, M.. Qualitative tests for preliminary phytochemical screening: An overview. International Journal of Chemical Studies. 2020;8(2):603–608. https://doi.org/10.22271/chemi.2020.v8.i2i.8834DOIGOOGLE SCHOLAR
    25. 25.Syamkumar, T. S.; S, G.; Augustine, A.. Study of pharmacological profile of chloroform leaf extract of Ludwigia perennis-A wetland plant. European Journal of Medicinal Plants. 2023;34(8):58–70. https://doi.org/10.9734/ejmp/2023/v34i81154DOIGOOGLE SCHOLAR
    26. 26.Vane, J. R.; Botting, R. M.. Mechanism of action of anti-inflammatory drugs. International Journal of Tissue Reactions. 1998;20(1):3–15.GOOGLE SCHOLAR
    27. 27.Vilar, D. D. A.; Rodrigues, J. A. G.; Dos Santos, S. C.; da Silva, F. M. R.. Morphological and anatomical characterization of Bixa orellana L. (Bixaceae). Revista Brasileira de Farmacognosia. 2010;20(6):889–893.GOOGLE SCHOLAR
    28. 28.Vriens, J.; Nilius, B.; Vennekens, R.. Herbal compounds and toxins modulating TRP channels. Current Neuropharmacology. 2008;6(1):79–96. https://doi.org/10.2174/157015908783769644DOIGOOGLE SCHOLAR
    29. 29.Wagner, K.-H.; Elmadfa, I.. Biological relevance of terpenoids: Overview focusing on mono-, di- and tetraterpenes. Annals of Nutrition and Metabolism. 2003;47(3–4):95–106. https://doi.org/10.1159/000070030DOIGOOGLE SCHOLAR
    30. 30.Yong, Y. K.; Zakaria, Z. A.; Kadir, A. A.; Somchit, M. N.; Lian, E. C. G.; Ahmad, Z.. Chemical constituents and antihistamine activity of Bixa orellana leaf extract. BMC Complementary and Alternative Medicine. 2013;13:1–7.GOOGLE SCHOLAR

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    Mishra, T., Kaushik, S., Bisht, P., & Bhatt, B. (2026). Anti-Inflammatory and Analgesic Effects of Bixa orellana Chloroform Extract. Pharmacognosy Research, 18(2), 548–554. https://doi.org/10.5530/pres.20260084