Antianemic Activity Research Articles

Unveiling the Antianemic Activity, Physicochemical Aspects, Antioxidant Properties, and Mineral Profile of Petroselinum crispum L.

Anemia is a widespread global health concern necessitating effective, accessible, and natural interventions. The potential of medicinal plants to address anemia has garnered significant interest. Among these plants, parsley (Petroselinum crispum (Petroselinum crispum) L.) stands out as an edible and herbal-based option for combating anemia. Aim of the study: This study investigated the potential of P. crispum (PC-Ext) as an emerging antianemic product, focusing on its physicochemical attributes, antioxidant properties, and mineral profile. Both qualitative and quantitative analyses of the phenolic compounds in P. crispum were conducted by using high-performance liquid chromatography with a diode array detector (HPLC-DAD). Anemia was induced in rats by intravenous injections of phenylhydrazine, administered at a dose of 40 mg/kg for two consecutive days. The antianemic activity of PC-Ext was assessed at a dose of 500 mg/kg twice daily for 5 weeks by estimating blood parameters, such as serum iron and ferritin. Additionally, the osmotic fragility test measured the capacity of red blood cells to withstand osmotic shock of various concentrations of saline. Aqueous extract of P. crispum was rich in phytochemical compounds, including syringic acid, quercetin, catechin, gallic acid, and luteolin. The findings demonstrate the effectiveness of P. crispum in ameliorating phenylhydrazine-induced reductions in red blood cell count (RBCs), hemoglobin (Hb), and hematocrit (HCT) levels. Consequently, PC-Ext exhibits significant activity against phenylhydrazine-induced anemia in rats, as demonstrated by its ability to prevent hemolysis. Iron estimation within PC-Ext further confirms its utility in addressing both iron deficiency and ferritin-deficiency anemia. Therefore, PC exhibits a favorable effect against both types of anemia, iron deficiency, and hemolysis. The results of this study provide robust scientific validation for ethnomedicinal use and the potential utility of P. crispum, positioning it as a promising source for future pharmaceutical development.

Antianemic Activity of Allolobophora caliginosa Extract against Phenylhydrazine-induced Anemia in Rats

Background:: Anemia is a global hematological disorder. Earthworms have antimicrobial, antispasmodic, antihypertensive, antiallergic, anti-inflammatory, and anticoagulant properties. Objective:: Our study aimed to investigate the anti-anemic activity of the earthworm Allolobophora caliginosa extract (AcE) on phenylhydrazine-induced hemolytic anemia in rats. Methods:: Eighteen adult male albino rats were randomly divided into 3 groups (n = 6 per group) as follows: Control, PHZ, and AcE-treated groups. Induction of anemia was done by intraperitoneal administration of phenylhydrazine (40 mg/kg) for 2 consecutive days. Treatment of AcE by intraperitoneal injection (45 mg/kg) for six successive days. Results:: Treatment of anemic rats with AcE extract caused significant increases in red blood cells (RBCs), hemoglobin (HB), hematocrit (HCT), total protein (TP), albumin, glutathione reduced (GSH), catalase (CAT), and glutathione-S-transferase (GST), as compared to the PHZ group. While, mean corpuscular haemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), malondialdehyde (MDA), and nitric oxide (NO) levels significantly decreased after intraperitoneal administration of AcE, when compared to the PHZ group. Liver sections from the AcEtreated group restored the normal micromorphological features of the liver. The expression of caspase- 3 was weak and/or there was no expression in the liver sections from AcE groups. AcE caused a significant reduction in the incidence of DNA damage, which appeared as less DNA in the tails. Conclusion:: The present research sheds light on the antianemic efficacy of AcE in rats. This study reveals that AcE has antianemic activity, which can be linked to its iron and branched-chain amino acid content, as well as antioxidant and anti-apoptotic properties.

Anti-Anemic Activity of Raphanus sativus Leaves on Phenylhydrazine Induced Anemia in Wistar Albino Rats

Anti-Anemic Activity of Raphanus sativus Leaves on Phenylhydrazine Induced Anemia in Wistar Albino Rats

Investigation of Acute Toxicity and Biological Activity of an Extract obtained on the Basis of a Local Medicinal Herbal Collection

The presented article describes the results of research work on the alimentary and hemolytic anemic effect of herbal decoction of medicinal properties, which is used in folk medicine for the treatment of various diseases. The extract of the sum of decoctions of medicinal plants was administered orally to experimental rabbits for 60 days in an amount of 10 ml / kg and 25 ml / kg, and their antianemic activity was assessed by such indicators as the number of shaped blood elements, as well as the general condition of experimental animals and changes in weight. Extract of tincture of sum of medicinal plants in studies of therapeutic effect in alimentary anemia on the other hand, already on the 7th day of treatment, the effect against hemolytic anemia began to manifest itself in less than 21 days, and it becomes more intense until the end of the experiment. These effects were manifested by a positive change in the number of shaped blood elements, an improvement in the general condition of experimental animals, a significant increase in appetite, and the restoration of lost body weight. This entails extensive scientific studies of the activity and mechanism of action of this tincture against the background of various anemia.

Anti-Anemic Potential of a Herbal Recipe Against Phenylhydrazine-Induced Anemia in Rats

Anemia is characterized by an abnormal decrease in the level of hemoglobin or the number of red blood cells in the blood. It is a public health problem that mainly affects pregnant women and young children. In the present study, the objective was to evaluated the anti-anemic potential of a recipe composed of three medicinal plants, <i>Xylopia aethiopica </i>(Dunal) A. Rich<i>, Zanthoxylum leprieurii (GUILL) </i>and<i> Harungara madagascariensis (LAM), </i>used to treat sickle cell anemia in rats. Aqueous and hydroethanol extracts of recipe composed were first prepared and quantitative phytochemical tests were carried out. Then, the anti-anemic activity of the recipe extracts was assessed. The anemia was induced in animals by intraperitoneal injection of phenylhydrazine at a repeated dose of 40 mg/kg bw for two days. Treatment was carried out with the aqueous and hydroethanol extracts of the plant recipe at doses of 400 and 800 mg/kg bw. The obtained results indicated that recipe composed contains total phenols (43.31±0.92 mg GAE/g and 33.82±0.6 mg GAE /g of dry weight), total flavonoids (17.07±0.89 mg QE and 19.76±1.15 mg QE/g of dry weight) and total tannin (8.65±0.01 mg QE and 26.5±0.95 mg Cat E/g of dry weight) in decocted and hydroethanol extracts respectively. The anemia was effectively corrected after 21 days of oral administration of the extracts to the rats. The extracts significantly increased hemoglobin (P < 0.05), red blood cell count (P < 0.001), and hematocrit (P < 0.01) as early as the 7^th days of treatment compared with untreated controls. These results show that the plant recipe composed of <i>Xylopia aethiopica, Zanthoxylum leprieurii </i>and<i> Harungara madagascariensis </i>possess antianemic properties.

Assessment of the effect of iron-rich humic substances on hematological parameters in the model of acute posthemorrhagic and iron deficiency anemia

Aim. To assess the effect of iron-rich humic substances on hematological parameters in acute post-hemorrhagic and iron deficiency anemia.Materials and methods. Materials for the study were samples of iron-rich active pharmaceutical ingredients based on humic substances (Fe(III) hydroxide complexes with humic substances and polymaltose): HA-Fe3+, HA-PMFe3+, FA-Fe3+, and FA-PM-Fe3+. The anti-anemic activity of the substances was studied on 53 female Wistar rats of the conventional rat line in the model of acute posthemorrhagic and iron deficiency anemia. Anti-anemic activity was assessed by the hemoglobin level, erythrocyte count, hematocrit, and serum iron level.Results. The studied substances HA-Fe3+ and FA-Fe3+ are the most effective in correcting the consequences of both experimental acute posthemorrhagic anemia and iron deficiency anemia. Their effect is comparable to that of the positive control drug Ferrum Lek.Conclusion. Fe(III) hydroxide complexes stabilized by humic and fulvic acids exhibit anti-anemic activity.

Antianemic activity of quinoa (Chenopodium quinoa Willd) Collana Negra variety and kanihua (Chenopodium pallidicaule Aellen) Ramis variety seed flour in anemic rats

The Andean grains from the Peruvian Altiplano, quinoa (Chenopodium quinoa Willd) and kanihua (Chenopodium pallidicaule Aellen) have high protein content and an optimal balance of essential amino acids and minerals such as iron (19.8 mg/100 g y 17.6 mg/100 g, respectively). The objective of this research was to evaluate the antianemic activity of extruded flour from quinoa seeds variety Negra Collana and kanihua variety Ramis in anemic Holtzman strain rats. The results of the proximal analysis showed high protein content in quinoa at 22% and kanihua at 16.2%, and the acute toxicity test showed harmlessness up to the dose of 15000 mg/Kg in both flours confirmed with the anatomopathological observation of organs such as liver, stomach, lung, kidneys, and brain. In the evaluation of the antianemic activity, a basal average of 29.3 ± 0.2% of hematocrit was observed in the group of anemic rats treated with quinoa flour, and in twelve weeks, it increased to 53.8 ± 0.3% of hematocrit (p ≤ 0.05). A group of anemic rats treated with kanihua flour had a basal average of 29.5 ± 0.3%, and in twelve weeks, it increased to 51.7 ± 0.3% (p ≤ 0.05). A group of rats without anemia treated with quinoa and kanihua flour showed a basal average of 50.2 ± 0.2% and 49.3 ± 0.3%; in twelve weeks, it increased to 55.2 ± 0.2% and 54.8 ± 0.1%, respectively. It was concluded that oral administration of 360 mg/Kg every 24 h of quinoa flour and kanihua flour increased hematocrit levels by 24.5 ± 0.5% and 22.2 ± 0.3%; weight 65.8 ± 0.3 g and 59.2 ± 0.1 g; height 6.8 ± 0.1 cm and 5.7 ± 0.5 cm, respectively (p ≤ 0.05). In rats without anemia increased hematocrit levels by 5.3 ± 0.0% and 5.5 ± 0.0%; weight 37.7 ± 0.1 g and 21.7 ± 0.05 g; height 4 ± 0.0 cm and 3.9 ± 0.0 cm, respectively (p ≤ 0.05).

Administration Routes as Modulators of the Intrahepatic Distribution and Anti-Anemic Activity of Salicylic Acid/Fe3O4 Nanoparticles.

The liver is a key organ in the pharmacokinetics of iron oxide nanoparticles (IONPs). This paper examined how the intravenous (IV) or intragastric (IG) route of administration influenced the intrahepatic distribution or therapeutic effects of IONPs. Wistar rats, some with bleeding-induced anemia, and iron oxide nanoparticles functionalized with salicylic acid (SaIONPs), with an average hydrodynamic diameter of 73 nm, compatible with rat sinusoid fenestrations, were used in this study. Light microscopy and multispectral camera analysis of Prussian blue labeled SaIONPs allowed mapping of intrahepatic nanoparticle deposits and revealed intrahepatic distribution patterns specific to each route of administration: loading of Kupffer cells and periportal hepatocytes when the IV route was used and predominant loading of hepatocytes when the IG route was used. Reducing the time to return to baseline values for hemoglobin (HGB) in rats with bleeding-induced anemia with IV or IG therapy has proven the therapeutic potential of SaIONPs in such anemias. The long-term follow-up showed that IV therapy resulted in higher HGB values. Proper use of the administration routes may modulate intrahepatic distribution and therapeutic effects of nanoparticles. These results may be beneficial in theragnosis of liver disease.

Anti-anemic Activity of Sodium, Calcium, Iron-Polygalacturonate In Vivo in Rabbits

Anti-anemic Activity of Sodium, Calcium, Iron-Polygalacturonate In Vivo in Rabbits

Antianemic activity of polyherbal formulation, Raktavardhak Kadha, against phenylhydrazine-induced anemia in rats

BackgroundOver two billion people around the world suffer from anemia. Majority of populations are using dietary supplements and herbal medicines for the management of the anemic conditions. Many polyherbal formulations such as RaktavardhakKadha (RK), are available in the Indian market as a nutritional supplement and herbal-based medicine for anemia. ObjectivesThe present study is aimed at investigating antianemic potential of polyherbal formulation, RK, against phenylhydrazine-induced anemia in rats. Materials and methodsRK was subjected to preliminary phytochemical analysis and iron estimation. Anemia was induced by phenylhydrazine administration (40 mg/kg, i.p.) for 2 consecutive days. Antianemic activity of RK was investigated at the dose of 1.8 ml/kg, twice daily for 12 days by estimating blood parameters and pathological changes in liver, heart, spleen and bone marrow. ResultsRK was found to contain saponins, steroids, flavonoids, tannins and phenolic compounds. Iron content was found to be 5 mg/100 ml in RK. Anemia induction by phenylhydrazine injections to rats caused significant decrease in red blood cells (RBCs), hemoglobin and hematocrit. These decreased levels of RBCs, hemoglobin and hematocrit in blood was significantly improved by the treatment with RK. Furthermore, RK restored pathological changes in liver, heart, spleen and bone marrow tissues near to normal. ConclusionThis study suggests antianemic activity of RK, which can be attributed to its iron content and ability to prevent hemolysis.

Antianemic Activity of Coconut (Cocos nucifera) Haustorium Waste Filtrate in Mice Induced by Sodium Nitrite

This study aims to determine the antianemic activity of coconut haustorium (Cocos nucifera) waste against mice induced by sodium nitrite orally for 43 days. The condition of anemia was determined by calculating the hemoglobin level using the Point of Care Test (POCT) method with the easytouch® GCHb and the resulting Hb level was 7.96 ± 0.05 g/dL. After the anemia condition is achieved, the test treatment is given for 21 days by dividing the mice into four groups, each consisting of three mice. The blank group (I) was administered distilled water, the control group (II) was administered Inbion® supplements at a dose of 0.65 mg/20 g BW, groups III and IV were administered coconut haustorium filtrate at a dose of 5 and 15 g/kg, respectively. Then measured the hemoglobin level and the number of erythrocytes. Based on the results of the study, the average hemoglobin levels of the blank, control, I and II groups were 13,9; 15,4; 16,0; and 16.8 g / dL, respectively. The number of erythrocytes during anemia was 1,750,000 / mm3 and after giving coconut kentos was 8,260,000 / mm3. From this study it can be concluded that the highest anti-anemia activity showed by coconut haustorium waste filtrate at dose 15 g/kgBW and had a significant difference in hemoglobin levels (p &lt;0.05) with the blank group.

Research antianemic action of the iron (IV) clathrochelate complexes on the sickling piglets

A lot of antianemic drugs for animals are presented on the modern pharmaceutical market of veterinary medicine, but they have several drawbacks. So finding new drugs with goal to treat ill animals by anemia and to prevent the iron deficiency anemia is a pressing issue. We previously reported the results of preclinical studies of iron in rare high valence IV. This allowed us to determine, supplement, and generalize the data on clinical studies of the new drugs with the active substance iron (IV) clathrochelate. Therefore, the article presents the results of studies of its antianemic effect on sickling piglets. Experiments were carried out on piglets-analogues neonates, which were divided into three groups: control and two experimental groups. Piglets were kept with sows on suckling. For the purpose of prevention iron deficiency anemia the traditional solution of iron dextran was administered once intramuscularly to piglets of I control group. The aqueous solution of iron (IV) clathrochelate complexes was administered once intramuscularly to piglets of II experimental group. Iron (IV) clathrochelate complexes was dissolved in a solvent of rheopolyglucin and administered once intramuscularly to piglets of III experimental group. 1 mL of test solutions contained 100 mg of active substance. The investigative material were the samples of blood and serum of piglets, their liver and spleen. The experiment lasted during a 30-day period since birth of piglets. According to the results of the experiments, iron (IV) clatrochelate complexes which was dissolved in a water for injection and rheopolyglucin had higher antianemic activity compared to the control. This is evidenced by the dynamics of probable changes in the number of erythrocytes, hemoglobin content and hematocrit, Iron content in serum and its mass fraction in the blood, liver and spleen of piglets. The effectiveness of the action of iron (IV) clatrochelate complexes is explained by the full supply of piglets with iron and its higher bioavailability.

Evaluation of the anti-anemic activity of date syrup in Wistar rats

The main objective of this study is to determine the effect of food intake of date syrup on the treatment of anemia.First, we used 3 samples of dates from different regions for the preparation of the syrup. We have studied certain criteria (physical, chemical, biochemical, microbiological); where the results showed that the prepared date syrup has a good hygienic quality, and of remarkable nutritional quality due to its ideal content with many properties, such as carbohydrate and protein content.To evaluate the anti-anemic properties of date syrup in rats, anemia is induced by a food having an iron deficiency. Date syrup, prepared from Djamaa dates, was administered by gavage of anemic rats at doses of 1000 mg / kg / day and 2000 mg / kg / day resulting in an increase in hemoglobin, red blood cell count, hematocrit and serum iron. Thanks to the results obtained, date syrup can be classified among the foods that help fight iron deficiency anemia.

Evaluation of the anti-anemic activity of date syrup in Wistar rats

Evaluation of the anti-anemic activity of date syrup in Wistar rats

Sıçanlarda fenilhidrazin ile indüklenmiş anemi üzerine iki Murva türünün in vivo antianemik etkileri

Aim: The present study was carried out to evaluate the antianemic effects of methanol extracts of Marsdenia tenacissima and Sansevieria roxburghiana on Phenylhydrazine (PHZ) induced anaemia in rats.Materials and Methods: PHZ (40mg/kg, i.p) injection was administered to all rats on Day 0 and Day 1 day except the group I. Group II was disease control, group III to IV treated with 100, 200 mg/Kg of methanol extract of M. tenacissima whereas group V &amp;amp; VI received 50, 100 mg/kg of methanol extract of S. roxburghiana. Group VII received the higher doses of both extracts continuously from the day 3 to 28. RBC, Haematocrit and Haemoglobin were analysed on day 0,2,7,14,21 and 28.Results: Continuous 28 days oral administration extracts raised the RBC, haemoglobin, and Haematocrit periodically in rats which were treated with PHZ.Conclusion: These plants possess antianemic activity and can be used for rapid recovery and management of anaemia.

Antianemic action of the iron (IV) clathrochelate complexes

Anemia is one of the most common non-contagious diseases of pigs. Modern antianemic drugs have several drawbacks, so finding new drugs is a pressing issue. We previously reported the results of preclinical studies of iron in rare high valence IV. This allowed us to determine, supplement, and generalize the data on clinical studies of the new drugs with the active substance iron (IV) clathrochelate. Therefore, we studied its antianemic effect on piglets. Experiments were carried out on piglets-analogues neonates, which were divided into three groups: control and two experimental groups. Piglets were kept with sows on suckling. For the purpose of prevention of iron deficiency anemia, the traditional solution of iron dextran was administered once intramuscularly to piglets of I control group. The aqueous solution of iron (IV) clathrochelate complexes was administered once intramuscularly to piglets of II experimental group. Iron (IV) clathrochelate complexes were dissolved in a solvent of rheopolyglucin and administered once intramuscularly to piglets of III experimental group. 1 mL of test solutions contained 100 mg of active substance. The investigative material were the samples of blood and serum of piglets, their liver and spleen. The experiment lasted during a 30-day period since the birth of the piglets. According to the results of the experiments, iron (IV) clatrochelate complexes which were dissolved in water for injection and rheopolyglucin had higher antianemic activity compared to the control. This is evidenced by the dynamics of probable changes in the number of erythrocytes, hemoglobin content and hematocrit, iron content in serum and its mass fraction in the blood, liver and spleen of piglets. The effectiveness of the action of iron (IV) clatrochelate complexes is demonstrated by the full supply of piglets with iron and its higher bioavailability.

Anti-Anemic and Haemopoietic Evaluation of Trigonella foenum-graecum (Fenugreek) in Rodent Model

The health benefits and medicinal properties of herbal food products are known since antiquity. Fenugreek, Trigonella foenum-graecum Linn (T. foenum-graecum, Fabaceae), a seed spice used to enhance flavor, color and texture of food is employed for medicinal purposes in many traditional systems. Ethno botanical survey of T. foenum-graecum revealed the seeds of the plant to be useful in anemia. The objective of this study was to study the ant-anemic effect of hydroalcoholic extract of seeds of T. foenum-graecum against phenylhydrazine induced anemic rat model. The hydroalcoholic extracts of seeds were prepared by soxhlation. Phytochemical analysis of the extracts was performed using standard testing procedures. Hemolytic anemia was induced in male Wistar rats by intraperitoneal administration of phenylhydrazine HCl (PHZ) at doses of 40 mg/kg of body weight during two successive days then one day after the animals were treated orally by the hydroalcoholic extracts with the amounts of 200 mg/kg and 400 mg/kg of body weight and Dexorange (reference drug) up to 13 days. The rats were analyzed for hematological parameters such as hemoglobin (Hb), red blood cell count (RBC) and white blood cell count (WBC) on day 2 and 13. Phytochemical screening of the extracts indicated the presence of carbohydrates, saponins, sterols, polyphenols, tannins and flavonoids. Anemia was induced successfully in Groups II, III, IV and V which was indicated by a mean reduction of 51.6% in RBC count; 52.85% in Hb content and 54.9% in WBC. Analysis of hematological parameters on day 13 showed that extract significantly (p&lt;0.05) improved Hb, RBC and WBC count at a dose of 400 mg/kg body weight. This study, not only substantiates the folklore use of the seed of T. foenum-graecum, but also suggests its inclusion in the treatment of anemia as it exhibited significant anti-anemic activity&#x0D; Keywords: Trigonella foenum-graecum, Anemia, Phytochemical analysis, Phenyl hydrazine, Dexorange

Evaluation of the in vitro antiplasmodial activity of Millettia zechiana and its action on the evolution of anemia in albino rats

Background: Malaria is a parasitic infection that leads to anaemia and death. Unfortunately, the upsurge of chemo-resistance prompted researchers to focus on new antimalarial drugs. Objectives: The aim of this work was to evaluate the antiplasmodial and antianemic activity of Millettia zechiana. Methods: the In vitro activity was assessed on clinical isolates and on the standard strain of Plasmodium falciparum K1, using the SYBR green I test. Moreover, the antianemic activity was evaluated in phenyl hydrazine induced anemic albino rats. Results/discussion: The ethyl acetate and hydroethanolic extract exhibited an antiplasmodial activity with IC50s of 6.14 and 12.14 μg/mL respectively on Plasmodium falciparum K1 strain. As for the in vivo antianemic activity, the ethyl acetate extract was the most active with better hematological reconstitution percentages. The presence of chemical compounds such as alkaloids, terpenoids and quinonic substances in both extracts, could be responsible for their activities. Conclusion: Millettia zechiana could be a potential source for novel antimalarial drugs and might be used as an improved traditional medicine on account of its availability.

Synthesis, properties, and antianemic activity of new metal complexes of sodium pectinate with iron and calcium

A new water-soluble complex based on sodium pectinate with iron and calcium (P-NaFeCa) was synthesized. The regularities of the complexation of sodium pectinate (with a 35% content of COONa groups) with biogenic metals Ca and Fe were studied. The content of macro- and microelements was determined by atomic emission spectrometry; the product was identified by IR spectroscopy. Using atomic force microscopy, it was found that the average particle size of P-NaFeCa is ~170 nm. The efficiency of P-NaFeCa (per os) was studied for the first time in animals with hemolytic anemia modeled by administration of phenylhydrazine. It was experimentally shown that in vivo P-NaCaFe at a dose of 60 mg kg–1 containing 50% of the recommended therapeutic dose of iron helps to recover the number of erythrocytes and mean concentration hemoglobin in erythrocyte, which indicates the efficiency of P-NaCaFe as an agent restoring the blood parameters after exposure to hemolytic poisons.

Multiwavelength UV-metric and pH-metric determination of the dissociation constants of the hypoxia-inducible factor prolyl hydroxylase inhibitor Roxadustat

The dissociation constant pKa of the Roxadustat with UV-VIS spectra, molar absorption coefficients and protonation equilibria are the key physicochemical parameters influencing many biopharmaceutical characteristics. Roxadustat is an orally bioavailable, hypoxia-inducible factor prolyl hydroxylase inhibitor with potential anti-anemic activity. It belongs to Active Pharmacenutical Ingredients, which have acidic/basic functionalities, their ionization state is controlled by solution pH and acid dissociation constants. Nonlinear regression of the pH-spectra with programs REACTLAB and SQUAD84 and of the pH-titration curve with ESAB determined four multiple consecutive dissociation constants with the protonation scheme. A sparingly soluble neutral molecule LH3 of Roxadustat was dissociated to the soluble anions LH2−, LH2− and L3− or protonated to the cation LH4+ in an aqueous medium. The graph of molar absorption coefficients of variously protonated species according to wavelength shows that the spectra of two anions LH2− and LH2− are nearly the same in colour. The Roxadustat spectrum exhibited five sharp isosbestic points, which were related to the LH2−/L3− equilibrium. Four consecutive thermodynamic dissociation constants were estimated using UV-metric data pKTa1 = 3.60(04), pKTa2 = 5.62(14), pKTa3 = 7.66(16), pKTa4 = 9.08(02) at 25 °C and pKTa1 = 3.60(04), pKTa2 = 5.73(10), pKTa3 = 7.52(10), pKTa4 = 8.99(02) at 37 °C and using pH-metric data pKTa1 = 4.33(09), pKTa2 = 6.57(11), pKTa3 = 8.88(05), pKTa4 = 9.03(04) at 25 °C and pKTa1 = 4.25(09), pKTa2 = 6.49(10), pKTa3 = 8.80(06), pKTa4 = 9.00(05) at 37 °C The positive values of the enthalpy ΔH0 showed that the dissociation process is endothermic and the positive values of the Gibbs free energy ΔG0 at 25 °C indicated that the dissociation process was not spontaneous, which also was confirmed by a negative value of the entropy ΔS0. Four macro-dissociation constants of Roxadustat and six protonation locations were predicted by MARVIN.