Phytochemical Analysis, Antimalarial Properties, and Acute Toxicity of Aqueous Extracts of Trisamo and Jatu-Phala-Tiga Recipes

Drug resistance remains a significant problem that threatens antimalarial drug treatment. Hence, the challenge is to find new effective antimalarial drugs. Based on our previous study, aqueous extracts of trisamo (TSM) and jatu-phala-tiga (JPT) had good in vitro antimalarial activities, and these recipes contain multiple beneficial pharmacological effects that could be useful for malaria therapy. Therefore, this study aimed to investigate the antimalarial activity and toxicity of the aqueous extracts of TSM and JPT in mouse models. The aqueous extractions were carried out using the decoction method. Compound identification was conducted using LC-QTOF-MS analysis. The antimalarial activities of TSM and JPT at doses 200, 400, and 600 mg/kg were evaluated against Plasmodium berghei ANKA infection using a four-day suppressive test. The toxic effects of oral administration of the extracts at 2 g/kg dose were determined using an acute toxicity test. The chemical constituents of TSM contained 83 compounds, whereas JPT contained 84 compounds. All doses of the extracts exhibited a significant suppression (p < 0.05) of the parasite compared to the negative control in a four-day test. The maximum activities were observed at 600 mg/kg dose with 67.02% suppression for TSM and 79.34% for JPT, followed by 400 mg/kg dose (57.63% for TSM and 64.79% for JPT) and then 200 mg/kg dose (52.35% for TSM and 54.46% for JPT). In addition, there were no significant differences (p < 0.05) in the RBC, MCV, and MCH levels of mice receiving JPT extract compared to the uninfected control. The WBC level of mice receiving 400 and 600 mg/kg of TSM, and 200 and 400 mg/kg of JPT, was significantly (p < 0.05) lower than the infected control, and the extracts did not significantly prevent the loss of platelets. For the acute toxicity test, there were no signs of toxicity or deaths in mice, and there were no differences in the histology, weight, or enzyme biochemistry of the liver and kidney between the extract and vehicle groups. However, the platelet count in the extract-treated mice was significantly higher than that in the control group. In conclusion, this study suggests that aqueous extracts of TSM and JPT have potent antimalarial activities and could be promising as new candidates for antimalarial drug development.


Introduction
Malaria remains one of the most serious illnesses in tropical and subtropical regions.It is a vector-borne disease caused by obligate intracellular Plasmodium parasites and is transmitted through the bite of infected female Anopheles mosquitoes [1].Five species of Plasmodium, P. falciparum, P. vivax, P. malariae, P. ovale, and P. knowlesi, can cause diseases in humans [2].Malarial paroxysm is a distinct clinical feature of the disease caused by the rupture of infected red blood cells (RBCs) [2].Common symptoms include fever, chills, and headaches, whereas the major complications of severe malaria include cerebral malaria, pulmonary edema, acute renal failure, severe anemia, bleeding, liver injury, and death [2].According to the World Malaria Report of 2022, the estimated number of deaths globally was 619,000, and 247 million cases were reported [1].Te African region of the World Health Organization (WHO) has the highest malaria burden, accounting for approximately 95% of the global cases [1].However, the case incidence has dropped since 2000 before increasing in 2020 due to a service interruption during the COVID-19 pandemic.Although efective treatment is a key factor in combating this disease, drug resistance poses a signifcant risk to the control and elimination of malaria [3].After the emergence of chloroquine resistance in the 1960s, the WHO recommended artemisinin-based combination therapy (ACT) as the frst-line treatment for uncomplicated malaria [4], which included artemetherlumefantrine (AL), artesunate-amodiaquine (AS-AQ), artesunate-mefoquine (AS-MQ), artesunate-sulfadoxinepyrimethamine (AS-SP), dihydroartemisinin-piperaquine (DHA-PPQ), and artesunate-pyronaridine (AS-PY) combinations [3,4].Te various treatments were determined based on the areas of resistance.Currently, the emergence of partial artemisinin resistance is of great concern and has been observed in countries in the WHO African region and the Greater Mekong Subregion (GMS) [1].A treatment failure rate of >10% for AL in Burkina Faso and Uganda and for DHA-PPQ in Burkina Faso has been observed [1].In GMS, mutations associated with SP resistance have been observed; hence, the failure of AS-SP could be of concern [1].In addition, a high prevalence of mutations associated with partial artemisinin resistance was found in Myanmar and Tailand, and high rates of DHA-PPQ plus primaquine treatment failure were found in Sisaket Province, Tailand, which led the province to change its frst-line drug to AS-PY in 2020 [1].Novel strategies are needed to eradicate malarial parasites to overcome the emergence of drugresistant parasites.Utilizing traditional medicine is one of several interesting ideas, especially polyherbal or herbal recipes, owing to their positive efects as a result of synergistic interactions [5].Our previous study on the in vitro antiplasmodial properties of aqueous and ethanolic extracts of ten herbal traditional recipes reported that aqueous extracts of trisamo (TSM) and jatu-phala-tiga (JPT) exhibited good antimalarial activities [6].Both are traditional herbal recipes that have been used for several centuries in Tailand [7].Te TSM is composed of three Terminalia species: Terminalia bellirica (Gaertn.)Roxb., Terminalia chebula (Roxb.ex DC.), and Terminalia arjuna (Roxb.ex DC.).Trisamo means "three" (tri-) fruits of Terminalia species (-samo), and these plants belong to the family Combretaceae [7].Te common names of T. bellirica, T. chebula, and T. arjuna are beleric myrobalans, chebulic myrobalans, and arjuns, respectively [7].TSM is indicated for promoting good general health and relieving abdominal bloating and is also used as an antipyretic, expectorant, and rejuvenator [8].Several biological benefts of the ingredients in the TSM recipe have been reported which include antipyretic, antibacterial, antioxidant, anti-infammatory, antihyperglycemic, anticlastogenic, immunomodulatory, analgesic, radioprotective, gastrointestinal motility-promotion, cardioprotective, antiaging cytoprotective, anticancer, antidiabetic, woundhealing, and antinociceptive properties [8][9][10][11].Te meaning of JPT corresponds to the benefts of four fruits: Jatu means "four," phala means "fruits," and tiga means "benefts" or "usefulness."Te four fruit ingredients include Phyllanthus emblica Linn.(P.emblica), T. bellirica, T. chebula, and T. arjuna.Indian gooseberry or P. emblica belongs to the family Euphorbiaceae and is commonly used in Ayurvedic systems for its many benefcial characteristics such as antidiabetic, antimicrobial, anti-infammatory, and antiaging properties [7,12].JPT is well known for its antioxidant activity, and it is used as an antipyretic, laxative, stomachic, colon cleanser, detoxifying agent, health promotion agent, and rejuvenator in Tai traditional medicine [13,14].Scientifc evidence has revealed that JPT has antimutagenic, cardioprotective, radioprotective, hepatoprotective, anti-infammatory, and antiobesity properties [15,16].In addition, the TSM and JPT recipes consist of numerous secondary metabolites such as favones, alkaloids, phenols, tannins, coumarin, terpenoids, glycosides, and saponins [17][18][19].Several classes of phytoconstituents from natural products are responsible for their antimalarial activity [20].Alkaloids, including terpenoidal, quinolone, and isoquinoline alkaloids, were identifed with promising antimalarial activity [21].Te antimalarial action of plant favonoids is believed to act by inhibiting fatty acid biosynthesis and the infux of L-glutamine-myoinositol in the infected red blood cells [22].Terpene and coumarin derivatives have been reported to have potent antimalarial activities [23,24].Regarding the abovementioned, the phytoconstituents deposited in the TSM and JPT recipes may provide great potential for antimalarial activities.Tus, this study aimed to investigate the antimalarial activity and toxicity of TSM and JPT in a mouse model.

Management and Preparation of TSM and JPT Recipes.
T. bellirica, T. chebula, T. arjuna, and P. emblica were bought at a Tai pharmacy store in the southern Tai province of Nakhon Si Tammarat's Muang District.Te morphological identifcation of plants was confrmed by a botanist, and the deposited specimens SMD074002003 (T.bellirica), SMD070006007 (T.chebula), SMD070006002 (T.arjuna), and SMD209003007 (P.emblica) were at Walailak University in Tailand's School of Medicine's Department of Medical Sciences.Te fruits were dried for 3 days in an oven (Memmert Model SFE 600, Schwabach, Germany) after being washed with tap water.Each fruit was ground using a herb grinder (Taizhou Jincheng Pharmaceutical Machinery Co., Ltd., Model: SF, Jiangsu, China).Te TSM and JPT recipes were prepared according to Tai herbal pharmacopeia [7,25].Te TSM recipe was prepared by mixing T. bellirica, T. chebula, and T. arjuna, in a 1 : 1 : 1 ratio, and the JPT recipe was prepared by mixing T. bellirica, T. chebula, T. arjuna, and P. emblica in a 1 : 1 : 1 : 1 ratio.

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Advances in Pharmacological and Pharmaceutical Sciences 2.2.Aqueous Extraction Method.Aqueous extractions of TSM and JPT were performed using the decoction method [26,27].For each recipe, 60 g of plant material suspended in 600 mL of water was extracted by boiling for 30 min.Ten, fltration through flter paper (Whatman, Buckinghamshire, England) was used to separate the liquid from the marc.Subsequently, the marc was re-extracted twice by boiling in 600 mL of water for 30 min.Te rotary evaporator (Rotavapor, Buchi, China) was used to concentrate the combined fltrate at 45 rpm and 45 °C.Ten, the extract was dried at −89 °C in a freeze-drying apparatus (Martin Christ, Germany).Te crude extract was weighed, and the yield was determined as follows: Percentage yield � weight of crude extract initial weight of herbal recipe × 100.
(1)  [29] according to a previously described method [6,26,30].Tree groups of fve mice each were formed from a total of ffteen mice.PBS was used as the vehicle control for Group I. TSM and JPT were administered at a dose of 2 g/kg to Groups II and III, respectively.All mice were weighed before receiving the extract or PBS.To assess the toxicity of the extracts, 2 g/kg of the extract was administered after the mice had fasted for 3 h.Te mice were observed immediately after feeding and then carefully observed for 30 min.Behavioral changes, signs of toxicity, and mortality were observed twice daily for 14 days.On day 14, all mice were weighed, anesthetized with 2%

Compound Identifcation
Advances in Pharmacological and Pharmaceutical Sciences isofurane, and euthanized through cardiac puncture.Blood samples were collected from the heart for hematological and biochemical analyses.Te liver and kidneys were removed and weighed to determine the relative organ weights.Relative organ weight was calculated using the following formula; thereafter, the organs were used for histopathological analysis.

Hematological, Biochemical, and Histopathological Assessment in Acute Toxicity
Test.Blood was collected into two types of tubes including EDTA and serum clot activator tubes.Blood in EDTA tubes was used for the hematological analysis (RBC, HGB, HCT, MCV, MCHC, MCH, PLT, and WBC), whereas serum were used for evaluation of renal and hepatic functions, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), blood urea nitrogen (BUN), and creatinine.Blood samples were analyzed using an AU480 chemistry analyzer (Beckman Coulter, USA).For histopathological assessment, the liver and kidneys were fxed in 10% formalin, and hematoxylin and eosin staining was performed as previously described [31][32][33].

Statistical Analysis. SPSS for Microsoft Windows
(version 17.0; IBM, Armonk, NY, USA) was used to conduct the statistical analyses.Te mean ± standard error of the mean (SEM) is used to express all data.Te data were examined for normality of distribution before being subjected to a one-way analysis of variance (ANOVA), with a significance level of p < 0.05.

Percentage Yield of TSM and JPT Recipes.
Te percentage yield of the aqueous extracts of JPT (40.12) was slightly higher than that of TSM (39.62).Te TSM appeared as a dark brown solid, and JPT was a light brown crumbly solid.

Compound Composition of TSM and JPT Detected by LC-QTOF-MS Analysis.
Te compounds in the TSM and JPT extracts were tentatively identifed using LC-QTOF-MS analysis.Tables 1 and 2 show the compounds found in the aqueous extracts of TSM and JPT, respectively.Te chemical constituents of TSM contained 83 compounds, whereas JPT contained 84 compounds.Figures 1 and 2 show the peak chromatograms of TSM and JPT, respectively.

Efects of the Extracts on Percentage Parasite Suppression.
Percentage parasitemia and efects of crude extracts on the percentage suppression of P. berghei infection are shown in Table 3. Te standard drug administered at a concentration of 25 mg/kg eliminated 100% of the blood-stage parasites.Administration of TSM and JPT extracts exhibited signifcant (p < 0.05) dose-dependent percentage parasite suppression compared to the negative control, with mean suppression percentage ranges 52.35-67.02%for TSM and 54.46-79.34%for JPT.In addition, percentage suppression at all doses of TSM was signifcantly lower than that in the chloroquine group (p < 0.05), whereas the percentage suppression at 600 mg/kg JPT showed no signifcant diference (p < 0.05) compared to chloroquine administration.Hematological alterations between normal and infected controls showed signifcant diferences in RBC, MCV, and MCH levels (p < 0.05), while hemoglobin, hematocrit, and MCHC levels were not signifcantly diferent among all groups.Mice administered a standard drug demonstrated signifcantly higher RBC levels (p < 0.05), but MCV and MCH levels were signifcantly lower (p < 0.05) than those in the infected controls.TSM administered at a dose of 400 mg/kg revealed a signifcant decrease in RBC (p < 0.05), but JPTadministration did not show a signifcant decrease in RBC compared with uninfected mice and those administered chloroquine.Te diferences between MCV and MCH levels showed that mice receiving JPT at all doses exhibited signifcantly (p < 0.05) lower levels than the infected control, and no diference was observed when compared to the chloroquine group.Te MCV and MCH levels in the TSM group were signifcantly (p < 0.05) higher than those in the uninfected and chloroquine groups.Administration of 400 mg/kg TSM resulted in a signifcant increase in MCV, compared to 400 and 600 mg/kg of JPT, and all doses of TSM resulted in a signifcant diference in the MCH level compared to 400 mg/kg of JPT.Te platelet counts of infected mice and mice that received the extracts were signifcantly (p < 0.05) lower than those of the normal control.Compared to the infected control, only the positive control group showed a signifcant increase in platelet counts (p < 0.05).In addition, chloroquine also produced a signifcant (p < 0.05) decrease in the WBC count compared to that in infected mice.Te extracts did not prevent a signifcant (p < 0.05) loss of WBC compared with chloroquine.TSM (400 and 600 mg/kg) and JPT (200 and 400 mg/kg) were signifcantly decreased when compared to the infected control.9 1 0 1 1 1 2 1 3 1 4 1 5 1 6 1 7 1 8 1 9 2 0 2 1 2 2 2 3 2 4 2 5 2 6 2 7 2 8 2 9 3 0 3 1 3 2 3 3 3 4 3 5 3 6 3 7 3 8 3   Data are presented as mean ± SEM (n � 5 per group).Diferences were considered statistically signifcant at p < 0.05.a compared with the negative control group receiving PBS, b compared with the positive control group receiving CQ, c compared with TSM 200 mg/kg, d compared with TSM 400 mg/kg, e compared with TSM 600 mg/kg, f compared with JPT 200 mg/kg, g compared with JPT 400 mg/kg, and h compared with JPT 600 mg/kg.Advances in Pharmacological and Pharmaceutical Sciences mice administered that the extracts showed no signifcant diference (p > 0.05) (see in Table 4).

Efects of the Extracts on Hematological and Biochemical
Changes in Acute Toxicity Test.Hematological results revealed that the platelet counts of the TSM and JPT groups were signifcantly higher than that of the vehicle control group (see Figure 4).Biochemical parameters of the liver and kidney function tests are presented in Table 5.No signifcant diferences in BUN, CREA, AST, ALT, or ALP levels were observed among the groups.

Efects of TSM and JPT on Histopathology in Acute
Toxicity Test. Figure 5 shows the histopathological examination of the liver and kidneys in the acute toxicity test at a dose of 2 g/kg.Figures 5(a

Discussion
As antimalarial drug resistance has been a major problem in malaria control, efective vaccines are unavailable.Terefore, new treatments are urgently needed.Our previous report showed that aqueous extracts from TSM and JPT have potent antiplasmodial activity against P. falciparum [6].As a result, the current study sought to assess the antimalarial properties and acute toxicity of aqueous extracts of TSM and JPT in mouse models.
For in vivo antimalarial testing, the antimalarial activities of TSM and JPT were investigated at 200, 400, and 600 mg/kg using a 4-day suppressive test.Te highest average percentage parasite suppression of 600 mg/kg JPT was 79.34%, and the extracts at all doses signifcantly suppressed parasite growth compared with the infected control.However, only JPT at 600 mg/kg showed no signifcant diference (p < 0.05) compared to chloroquine, which may imply that the efect of JPT at 600 mg/kg is similar to that of chloroquine.Te biological properties of plant extracts are known to be mediated by phytocomponents [34].Based on the LC-MS analysis, our fndings are consistent with those of previous studies.Chebulinic acid, gallic acid, ellagic acid, quinic acid, and luteolin were found in the fruits of T. bellirica, T. chebula, and T. arjuna extracts [8,35,36].In addition, the phytocomponent of P. emblica was reported to have tannins such as chebulic acid, gallic acid, and punicalagin; favonoids such as luteolin and quercetin derivatives; polyphenolics such as ellagic acid; and phenolics such as chebulinic acid [12,[37][38][39].Ellagic acid has been reported to possess antioxidant, anti-infammatory, antimutagenic, antiproliferative, and antimalarial properties [40,41].Chebulinic acid has been reported to have numerous biological activities, including antidiabetic, antifbrotic, antiinfammatory, antitumor, antiatherogenic, antioxidant, antiulcer, hepatoprotective, and antiviral properties [42].Quinic acid has an important antibacterial efect [43].Accordingly, the compound described above, or the other compounds present in TSM and JPT might exert antimalarial activities through individual or synergistic efects.In addition, the antimalarial activity of the extracts in this study showed that the percentage parasite suppression of JPT was higher than that of TSM.Te reason for this could be the extra ingredient from P. emblica in the JPT recipes.P. emblica is important in traditional medicinal systems, and its various pharmacological benefts have been reported, including antimicrobial, antioxidant, antiinfammatory, antipyretic, antitusive, antiatherogenic, anticancer, antidiabetic, antiaging, cardioprotective, gastroprotective, nephroprotective, neuroprotective, chemopreventive, analgesic, and immunomodulatory properties [44].Furthermore, JPT and its components have been reported to exhibit strong antioxidant activity [45].Based on previous evidence, we suggest that P. emblica improves the antimalarial property of the extract.
Furthermore, this study investigated the efects of the extracts on hematological parameters during malaria infection because hematological abnormalities are considered a characteristic of malaria, especially RBCs [46].Te differences in RBC, MCV, and MCH between the uninfected and infected groups were signifcant, and chloroquine improved these blood parameters to normal ranges compared to the uninfected control.Te reduction of RBCs in infected mice may be caused by the destruction or sequestration of RBCs, or reduction of RBC production in the bone marrow [9].Te signifcant increases in MCV and MCH were indicative of malaria-induced macrocytic anemia [47,48].Te RBC count in the 400 mg/kg TSM group was signifcantly lower than that in the uninfected control.Te MCV and MCH of all TSM extract doses difered signifcantly from   Advances in Pharmacological and Pharmaceutical Sciences those of the uninfected control.However, the RBC count, MCV, and MCH of mice that received JPT showed no signifcant diferences compared to the uninfected control.Tis fnding implies that only the JPT extract can prevent malaria-induced macrocytic anemia.Te ability to improve RBC and related parameters may be due to the inhibitory efects of the drug and its extracts on parasite growth.WBC and related parameters play an important role in infectious diseases [46].WBC responds to infectious agents, and thrombocytopenia is a common feature of Plasmodium infection, which is associated with several mechanisms, such as endothelial damage and isolated platelet consumption [46].Te platelet count of mice in the infected control group was signifcantly reduced, but the WBC count was increased, when compared with the uninfected control group.Te extracts from TSM and JPT showed a signifcant and dramatic decrease in platelet count compared to the normal control and chloroquine.Tis could mean that extracts at doses 200, 400, and 600 mg/kg did not maintain platelet homeostasis during malaria infection.In terms of WBC parameters, TSM reduced the WBC count in a dosedependent manner.Tis fnding may indicate that a decrease in the WBC count is associated with a reduction in the percentage of parasites.Interestingly, 600 mg/kg JPT resulted in the highest WBC count and the highest percentage parasite suppression among the extracts.Tis result implies that the antimalarial activity of JPT at this dose may be attributed to the activation of immune cells, which is consistent with a previous study [12].P. emblica, an ingredient in the JPT recipe, has been reported to exhibit the ability to enhance immunity [12].Consequently, JPT extract not only exhibited stronger antimalarial activity but also exhibited greater maintenance of RBC parameters than the crude extract from TSM.Although natural products have been used to treat several diseases since ancient times, the negative efects of plant products must be considered.According to the results of the toxicity study, there were no deaths or physical or 12 Advances in Pharmacological and Pharmaceutical Sciences behavioral changes for 14 days, thereby indicating that the LD 50 of TSM and JPT was greater than 2 g/kg.TSM and JPT were classifed as relatively low acute toxicity hazards in Category 5 according to the international system of chemical classifcation [49].Te efects of the extracts on changes in body weight and organ weight are sensitive indicators for general health status and organ damage in animals [50,51].We found no signifcant changes in body weight or organ weight when compared with the vehicle control at the endpoint.In addition, this study focused on the negative efects of the extracts on the hematological markers, biochemical enzymes, and pathology of the liver and kidney.For all blood parameters, only the platelet count was signifcantly increased in mice treated with TSM and JPT compared to the vehicle control.Tis result implies that the extract at a dose of 2 g/kg may have an efect on platelet enhancement activity, suggesting that this efect may be benefcial for improving thrombocythemia in blood diseases such as malaria.To ensure the safety of the extracts, the biochemical enzymes and pathology of the liver and kidney were assessed.Te liver and kidneys play a dominant role in drug metabolism and elimination after ingestion.Te liver is well known as the primary organ for parasite development in the pre-erythrocytic stages, resulting in stifness of the infected liver cells [52].Acute kidney injury is a well-known signifcant organ dysfunction caused by malaria infection, and hematological abnormalities are regarded as a key feature of malaria infection [46,53].Tere were no signifcant changes in biochemical enzymes and histology of the liver and kidneys.Tis fnding implies that the extracts were not associated with nephrotoxicity or hepatotoxicity caused by herbal medicine.Regarding the results of the toxicity test, aqueous extracts of TSM and JPT showed clear evidence that the extracts were considered safe in mice when administered at 2 g/kg, and these extracts may provide great choices for antimalarial drug candidates because they are safe for major cells that can be damaged by the Plasmodium parasite, such as hepatocytes and RBCs.

Conclusions
Tis study demonstrates that aqueous extracts of TSM and JPT exert potent antimalarial activities against P. berghei and are considered safe for oral administration.Terefore, TSM and JPT should be considered as an alternative treatment for malaria.Further experiments should be conducted to test the antimalarial activity in nonhuman primates and in clinical trials.
Changes in 4-Day Suppressive Test.Te results of the hematological changes are shown in Figure 3. Te indices of uninfected mice were used to represent hematologic reference values at the normal levels.
) and 5(b) show the normal structure of the liver and kidney histology, respectively, which were obtained from mice in the control group.In comparison, liver and kidney sections showed no diferences between the control and mice treated with TSM and JPT.Liver sections (Figures5(a), 5(c), and 5(e)) revealed normal hepatocytes without hepatic congestion, infammatory cell infltration, or sinusoidal dilatation.Kidney sections (Figures 5(b), 5(d), and 5(f )) showed unchanged glomeruli and renal tubules without vascular congestion.

Figure 4 :
Figure 4: Efects of the extracts on hematological changes in acute toxicity test: (a) RBCs and related parameters, (b) WBC count, and (c) platelet count.All values are expressed as mean ± SEM (n � 5 per group).Tere were statistically signifcant diferences at p < 0.05, a compared with negative control, b compared with 2 g/kg of TSM extract, and c compared with 2 g/kg of JPT extract.

Figure 5 :
Figure 5: Histopathological micrograph of the liver and kidney of mice in the acute toxicity test.All images were acquired at 20× magnifcation.Bar � 20 μm; CV: central vein; H hepatocyte; T renal tubule; G glomerulus.(a) Liver histology of control mice; (b) kidney histology of control mice; (c) liver histology of TSM-treated mice; (d) kidney histology of TSM-treated mice; (e) liver histology of JPTtreated mice; (f ) kidney histology of JPT-treated mice.

Table 1 :
Tentative identifcation of the chemical constituents from TSM by LC-QTOF-MS analysis.
tremors, convulsions, ataxia, or unusual behaviors, were not observed throughout 14 days.No mortality occurred during the experiment; therefore, the mean lethal dose (LD 50 ) of the aqueous extract of TSM and JPT administered via the oral route was higher than 2 g/kg.When compared to the vehicle control, the actual body weight, percentage of body weight change, and relative organ weights of the liver and kidney of 4Advances in Pharmacological and Pharmaceutical Sciences

Table 2 :
Tentative identifcation of the chemical constituents from JPT by LC-QTOF-MS analysis.

Table 3 :
Percentage parasitemia and suppression of TSM and JPT recipes.
Figure3: Efects of TSM and JPT on hematological parameters in the 4-day suppressive test: (a) RBC count, (b) hemoglobin levels, (c) hematocrit levels, (d) MCV levels, (e) MCH levels, (f ) MCHC levels, (g) WBC count, and (h) platelet count.Data are presented as the mean ± SEM (n � 5 per group).Tere were statistically signifcant diferences at p < 0.05, a compared with uninfected mice, b compared with negative control group receiving PBS, c compared with the positive control group receiving CQ, d compared with TSM 200 mg/kg, e compared with TSM 400 mg/kg, f compared with TSM 600 mg/kg, g compared with JPT 200 mg/kg, h compared with JPT 400 mg/kg, and i compared with JPT 600 mg/kg.

Table 4 :
Body weight and organ weight in acute toxicity test.All values are expressed as mean ± SEM.Tere were no statistically signifcant diferences at p < 0.05.

Table 5 :
Biochemical profles of the liver and kidney function in acute toxicity test.
All values are expressed as mean ± SEM.Tere were no statistically signifcant diferences at p < 0.05.