Rifampicin PLGA nanospheres are
formulated with a specific goal in order to decrease
the dose, adverse effects and to enhance targeted
drug delivery. Rifampicin nanospheres were
prepared and evaluated by emulsion solvent
evaporation method. In vivo bio distribution studies
reveal that there was a long term accumulation of
rifampicin nanospheres in the lungs over other
organs. The increase in Cmax values confirmed that
inhalable PLGA nanospheres are suitable for
targeting and providing sustained release of antitubercular
drugs to lungs. So inhalation is a
selected administration route of Rifampicin PLGA
nanospheres. The in vivo screening of M.
tuberculosis showed good activity as well as its
activity against multidrug-resistant M. tuberculosis
and against M. tuberculosis isolates in a
potentially latent state, makes Rifampicin PLGA
nanospheres as an attractive drug dosage form
for the therapy of tuberculosis. It can be concluded
that there is a significant potential for effective
oral delivery as well as nasal delivery of the
Nanospheres for the treatment of tuberculosis.
PRECLINICAL SCREENING MODELS
In vitro methods
• Patch clamp technique in kidney cells
• Perfusion of isolated kidney tubules
• Isolated perfused kidney
In vivo methods
• Diuretic activity in rats (LIPSCHITZ test)
• Saluretic activity in rats
• Diuretic and saluretic activity in dogs
• Clearance methods
• Micro puncture techniques in the rat
• Stop-flow technique
PRECLINICAL SCREENING MODELS
In vitro methods
• Patch clamp technique in kidney cells
• Perfusion of isolated kidney tubules
• Isolated perfused kidney
In vivo methods
• Diuretic activity in rats (LIPSCHITZ test)
• Saluretic activity in rats
• Diuretic and saluretic activity in dogs
• Clearance methods
• Micro puncture techniques in the rat
• Stop-flow technique
Change of Peptides and Free -Amino Acids Contents during Nanjing Dry-Cured Du...Agriculture Journal IJOEAR
— In order to explore the relationship between the change of peptides and free-amino acid (FAA) and its unique flavour, Dry-cured duck samples of different processing phases were used to study the change of free-amino acid by High Performance Liquid Chromatography (HPLC) in this paper, meanwhile the trichloroacetic acid precipitation method for modeling use to establish the quantitative predicated peptides. The changes of small peptides and free amino acids in the process were studied. The results showed that the level and amount of proteolysis increased with the processing time at traditional technology, meanwhile the amount of peptides were positively correlated with FAA contents (R 2 =0.86).
In-Vitro and In-Vivo Assessment of Anti-Asthmatic Activity of Polyherbal Ayur...IOSR Journals
About 80% of asthmatic turn to alternative or complementary therapies typically in conjunction with their regular allopathic medication. The role of complementary and alternative medicine in adult asthma treatment is limited because these approaches have been insufficiently researched and their effectiveness is largely unproven. In the present study in –vivo and in-vitro effectiveness of a polyherbal Ayurvedic drug is evaluated for its anti-asthmatic activity. For in –vitro assessment of anti-asthmatic property of drug antiinflammatory, analgesic, immunomodulator effect, and antihistaminic, anti-cholinergic, mast cell stabilizing activity, anti-anaphylactic activity and bronchodilator effect were screen on animal models. Evaluation of Effect of Drug Distribution on Lung Mechanics is also evaluated using MATLAB. In a randomised,open, placebo controlled trial the effects of drug was compared with placebo medication (normal saline) in 60 adults with mild to moderate asthma as an adjunct to conventional treatment. Animal studies showed that drug possess significant mast cell stabilizing activity, immunomodulator activity, bronchodilator activity and anti-anaphylactic activity. Insignificant anti-cholinergic activity was found in the drug. There was significant improvement found in pulmonary function test (including FEV1, FVC and PEFR)in the group treated with polyherbal drug .Improvement remain constant in consecutive follow-ups signifies that there is no reverse bronchoconstriction after discontinuation of drug. This study signifies that polyherbal drug (Shirishadi ) may prove beneficial future alternative remedy for asthma and its effect is similar to that of modern contemporary drug when given through nasal route.
—3-Chloro-1,2-propanediol (3-chloropropanediol) is a well-known food processing contaminant found in a wide range of foods and ingredients and there has been recent concern about the levels of carcinogenic 3-chloropropanediol (3-MCPD) in some soy sauces. This paper reports on the development of an analytical method for the fast determination of 3-MCPD at trace level in commercial soy sauce using novel liquid phase extraction (LPE)/cleanup coupled with microwave-assisted derivatization (MAD) method followed by high performance liquid chromatography-ultraviolet (HPLC-UV) detection. In this method, 3-MCPD was first isolated from soy sauce sample matrix by LPE/cleanup with Extrude NT3 column cartridges and the isolated (eluent) solution was subjected to MAD with acetophenone to form 2-methyl-2-phenyl-4-(chloromethyl)-1,3-dioxolane under microwave irradiation using a specially modified domestic microwave oven, then the derivatizeddioxolane was directly analyzed with a HPLC-UV system. The optimum conditions for MAD such as the ratio of reagents, acidic catalyst, microwave irradiation power and time, as well as the chromatographic conditions were thoroughly investigated. Experimental results indicated that maximum derivatization can be achieved in 10 min under microwave irradiation at 362 watts when compared to 18 hours by conventional refluxing reaction. The proposed method provided a simple and rapid analytical procedure for 3-MCPD analysis in soy sauce with the detection limit of 80 ng mL-1. The relative standard deviations were all below 3.0 % (n = 7). Application was illustrated by the analysis of commercial sauce sample obtained from a local traditional store in central Taiwan.
Hepatoprotective Effect of Cestrum parqui L. aerial parts and Phytochemical ...Jing Zang
This study deals with the investigation of hepatoprotective effect of 70% methanolic extract from Cestrum parqui aerial parts and determination of the bioactive components of the plant. The hepatoprotective effect of Cestrum parqui methanol extract (100, 500, 1000 mg/kg) was analysed on carbon tetrachloride (CCl4)-induced acute liver injury. The administration of a single dose of 40% CCl4 (1ml/kg b.w.) causes an increase in the activities of serum alanine aminotransferase (ALT) and aspirate aminotransferase (AST) enzymes and so pretreated orally of a dose from Cestrum parqui methanol extract (100, 500, 1000 mg/kg) and silymarin (200 mg/kg) for three consecutive days prior to The administration of a single dose of CCl4 significantly prevented the increase in the activities of these enzymes. Histological analysis showed that Cestrum parqui methanol extract at doses of 500 and 1000 mg/kg and silymarin reduced the incidence of liver lesions including vacuole formation, neutrophil infiltration and necrosis of hepatocytes induced by CCl4. The extract cause a negative result on the antioxidative enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GRd) and decreased malondialdehyde (MDA) level in liver, as compared to those in the CCl4-treated group and this suggests that the hepatoprotective activity of the extract is due to the antioxidant effect of the extract. Phytochemical analysis of the methanol extract from Cestrum parqui aerial parts showed that it contained different phytoconstituents, flavonoids, tannins, saponins, alkaloids, terpenes and carbohydrates.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
The research article published in the journal RSC Advances in 2017, entitled as "One step preparation of a biocompatible,
antimicrobial reduced graphene oxide–silver nanohybrid as a topical antimicrobial agent"
Polymeric Nanoparticles of Rifampicin were prepared by emulsion solvent evaporation technique using poly methyl methacrylate as polymer matrix and Poly vinyl alcohol as surfactant. Drug entrapped free flowing nanoparticles of Rifampicin were obtained after optimization using 32 factorial design and characterized for entrapment efficiency, particle size distribution, differential scanning calorimetry (DSC), X-ray diffraction (XRD), scanning electron microscopy (SEM) and in vitro and stability studies. The PMMA nanoparticles had a small size (213 ± 0.72 nm), uniform size distribution. The effects of dependent variables drug-polymer ratio and surfactant concentration on particle size and encapsulation efficiency were studied. The drug and polymer were not interacting with each other. SEM studies revealed the spherical shape of nanoparticles and in vitro release studies showed sustained drug release. RIF-polymeric nanoparticles drug delivery system proved to be promising for anti-tubercular therapy.
Change of Peptides and Free -Amino Acids Contents during Nanjing Dry-Cured Du...Agriculture Journal IJOEAR
— In order to explore the relationship between the change of peptides and free-amino acid (FAA) and its unique flavour, Dry-cured duck samples of different processing phases were used to study the change of free-amino acid by High Performance Liquid Chromatography (HPLC) in this paper, meanwhile the trichloroacetic acid precipitation method for modeling use to establish the quantitative predicated peptides. The changes of small peptides and free amino acids in the process were studied. The results showed that the level and amount of proteolysis increased with the processing time at traditional technology, meanwhile the amount of peptides were positively correlated with FAA contents (R 2 =0.86).
In-Vitro and In-Vivo Assessment of Anti-Asthmatic Activity of Polyherbal Ayur...IOSR Journals
About 80% of asthmatic turn to alternative or complementary therapies typically in conjunction with their regular allopathic medication. The role of complementary and alternative medicine in adult asthma treatment is limited because these approaches have been insufficiently researched and their effectiveness is largely unproven. In the present study in –vivo and in-vitro effectiveness of a polyherbal Ayurvedic drug is evaluated for its anti-asthmatic activity. For in –vitro assessment of anti-asthmatic property of drug antiinflammatory, analgesic, immunomodulator effect, and antihistaminic, anti-cholinergic, mast cell stabilizing activity, anti-anaphylactic activity and bronchodilator effect were screen on animal models. Evaluation of Effect of Drug Distribution on Lung Mechanics is also evaluated using MATLAB. In a randomised,open, placebo controlled trial the effects of drug was compared with placebo medication (normal saline) in 60 adults with mild to moderate asthma as an adjunct to conventional treatment. Animal studies showed that drug possess significant mast cell stabilizing activity, immunomodulator activity, bronchodilator activity and anti-anaphylactic activity. Insignificant anti-cholinergic activity was found in the drug. There was significant improvement found in pulmonary function test (including FEV1, FVC and PEFR)in the group treated with polyherbal drug .Improvement remain constant in consecutive follow-ups signifies that there is no reverse bronchoconstriction after discontinuation of drug. This study signifies that polyherbal drug (Shirishadi ) may prove beneficial future alternative remedy for asthma and its effect is similar to that of modern contemporary drug when given through nasal route.
—3-Chloro-1,2-propanediol (3-chloropropanediol) is a well-known food processing contaminant found in a wide range of foods and ingredients and there has been recent concern about the levels of carcinogenic 3-chloropropanediol (3-MCPD) in some soy sauces. This paper reports on the development of an analytical method for the fast determination of 3-MCPD at trace level in commercial soy sauce using novel liquid phase extraction (LPE)/cleanup coupled with microwave-assisted derivatization (MAD) method followed by high performance liquid chromatography-ultraviolet (HPLC-UV) detection. In this method, 3-MCPD was first isolated from soy sauce sample matrix by LPE/cleanup with Extrude NT3 column cartridges and the isolated (eluent) solution was subjected to MAD with acetophenone to form 2-methyl-2-phenyl-4-(chloromethyl)-1,3-dioxolane under microwave irradiation using a specially modified domestic microwave oven, then the derivatizeddioxolane was directly analyzed with a HPLC-UV system. The optimum conditions for MAD such as the ratio of reagents, acidic catalyst, microwave irradiation power and time, as well as the chromatographic conditions were thoroughly investigated. Experimental results indicated that maximum derivatization can be achieved in 10 min under microwave irradiation at 362 watts when compared to 18 hours by conventional refluxing reaction. The proposed method provided a simple and rapid analytical procedure for 3-MCPD analysis in soy sauce with the detection limit of 80 ng mL-1. The relative standard deviations were all below 3.0 % (n = 7). Application was illustrated by the analysis of commercial sauce sample obtained from a local traditional store in central Taiwan.
Hepatoprotective Effect of Cestrum parqui L. aerial parts and Phytochemical ...Jing Zang
This study deals with the investigation of hepatoprotective effect of 70% methanolic extract from Cestrum parqui aerial parts and determination of the bioactive components of the plant. The hepatoprotective effect of Cestrum parqui methanol extract (100, 500, 1000 mg/kg) was analysed on carbon tetrachloride (CCl4)-induced acute liver injury. The administration of a single dose of 40% CCl4 (1ml/kg b.w.) causes an increase in the activities of serum alanine aminotransferase (ALT) and aspirate aminotransferase (AST) enzymes and so pretreated orally of a dose from Cestrum parqui methanol extract (100, 500, 1000 mg/kg) and silymarin (200 mg/kg) for three consecutive days prior to The administration of a single dose of CCl4 significantly prevented the increase in the activities of these enzymes. Histological analysis showed that Cestrum parqui methanol extract at doses of 500 and 1000 mg/kg and silymarin reduced the incidence of liver lesions including vacuole formation, neutrophil infiltration and necrosis of hepatocytes induced by CCl4. The extract cause a negative result on the antioxidative enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GRd) and decreased malondialdehyde (MDA) level in liver, as compared to those in the CCl4-treated group and this suggests that the hepatoprotective activity of the extract is due to the antioxidant effect of the extract. Phytochemical analysis of the methanol extract from Cestrum parqui aerial parts showed that it contained different phytoconstituents, flavonoids, tannins, saponins, alkaloids, terpenes and carbohydrates.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
The research article published in the journal RSC Advances in 2017, entitled as "One step preparation of a biocompatible,
antimicrobial reduced graphene oxide–silver nanohybrid as a topical antimicrobial agent"
Polymeric Nanoparticles of Rifampicin were prepared by emulsion solvent evaporation technique using poly methyl methacrylate as polymer matrix and Poly vinyl alcohol as surfactant. Drug entrapped free flowing nanoparticles of Rifampicin were obtained after optimization using 32 factorial design and characterized for entrapment efficiency, particle size distribution, differential scanning calorimetry (DSC), X-ray diffraction (XRD), scanning electron microscopy (SEM) and in vitro and stability studies. The PMMA nanoparticles had a small size (213 ± 0.72 nm), uniform size distribution. The effects of dependent variables drug-polymer ratio and surfactant concentration on particle size and encapsulation efficiency were studied. The drug and polymer were not interacting with each other. SEM studies revealed the spherical shape of nanoparticles and in vitro release studies showed sustained drug release. RIF-polymeric nanoparticles drug delivery system proved to be promising for anti-tubercular therapy.
ISSN 2347-2251
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The Indo-American Journal of Pharma and Bio Sciences is an online international journal that publishes articles quarterly.It's important to note that the specific policies, guidelines, and the editorial board of IAJPB may change over time, so it's advisable to visit the journal's official website or contact the journal of the research on journaling.
To form the basis of a respiratory disease model in rats by investigating the microbial distribution and composition in the lower respiratory tracts of normal rats. Methods: DNA was extracted from the intestine, trachea, bronchus and lung samples collected from healthy rats under sterile conditions. The 16S rDNA V4-V5 region was sequenced using Illumina high-throughput technology. Results: The sequencing results showed that there was no significant difference in abundance and species diversity of microbiota between the lower respiratory and the intestine. The microbiota structure analysis showed samples from lungs and intestinal shared similarity. However, the dominant species at the levels of phylum, family, and genus diverged. The similarity analysis showed that the lung microbiota were different from the intestines. The linear discriminant analysis showed significantly different species in different tissues; function prediction also showed different microbiota function in different tissues. Conclusions: These results suggest that bacterial colonization depends on the sample’s anatomical location. The human pathogen Acinetobacter lwoffii was also detected in the rat lower respiratory tract samples.
Intercontinental journal of pharmaceutical Investigations and ResearchSriramNagarajan19
Anti-inflammatory activity of the ethanolic extract of Portulaca quadrifida Linn. was studied in wister rats using the carrageenan induced left hind paw edema, carrageenan induced pleurisy and cotton pellet induced granuloma model. The ethanolic extract (200 mg/kg, p.o.,) produced the inhibition of carrageenan induced rat paw edema. It also showed an inhibitory effect on leukocyte migration and a reduction on the pleural exudates as well as reduction on the granuloma weight in the cotton pellet granuloma method. The results indicated that the ethanolic extract produced significant (P<0.001) anti-inflammatory activity when compared with the standard and untreated control.
Electrochemical study of anatase TiO2 in aqueous sodium-ion electrolytesRatnakaram Venkata Nadh
In this paper, a basic electro-analytical study on the behavior of anatase TiO2 in aqueous NaOH has been presented using cyclic voltammetry technique (CV). The study has explored the possibility of using TiO2 as anode material for ARSBs in presence of 5 M NaOH aqueous electrolyte. CV profiles show that anatase TiO2 exhibits reversible sodium ion insertion/de-insertion reactions. CV studies of TiO2 anode in aqueous sodium electrolytes at different scan rate shows that the Na+ ion insertion reaction at the electrode is diffusion controlled with a resistive behavior. Proton insertion from aqueous sodium electrolytes into TiO2 cannot be ruled out. To confirm the ion inserted and de-inserted, CV studies are done at different concentration of NaOH and it is found that at lower concentrations of NaOH, proton insertion process competes with Na+ ion insertion process and as the concentration increases, the Na+ ion insertion process becomes the predominant electrode reaction making it suitable anode materials for aqueous sodium batteries in 5 M NaOH.
Validated HPLC Method for Assay and Content Uniformity Testing of Roflumilast...Ratnakaram Venkata Nadh
Roflumilast is a selective enzyme inhibitor of phosphodiesterase-4. This drug is recommended for treatment of patients suffering from
chronic-obstructive-pulmonary-disease with chronic-bronchitis. Roflumilast is not official in pharmacopoeia and the reported methods
are having high chromatographic run times. A short run time HPLC method was developed for assay and content uniformity testing to
determine the roflumilast in blend and tablets. The mobile phase consists of 10 mM sodium dihydrogen phosphate monohydrate buffer
and acetonitrile in the ratio of 45:55 v/v. The HPLC method was developed using accucore-C18 150 × 4.6 mm, 4 μm column with a flow
rate of 1.0 mL min-1, 215 nm wavelength and 10 μL injection volume with run time of 5 min. The method linearity was proved between
5.02-40.17 μg mL-1 and obtained correlation-coefficient value is 1.0000. The mean recovery of roflumilast was 100.6%. The stability
indicating nature was established and performed the validation by considering ICH Q2 (R1) recommendations.
Substrate Inhibition in Ruthenium(III) Catalyzed Oxidation of Propane-1,3-dio...Ratnakaram Venkata Nadh
Ruthenium(III) catalyzed oxidation of propane-1,3-diol by potassium periodate was studied in aqueous perchloric acid medium. Orders
of reaction with respect to concentrations of oxidant, substrate, acid and catalyst were determined. First order in oxidant and catalyst
concentrations, and inverse fractional order in acid medium were observed. In addition, substrate inhibition (i.e. a decrease in reaction rate
with an increase in substrate concentration) was observed. Effect of addition of salt and solvent was studied. Based on the studies of
temperature variation, Arrhenius parameters were calculated. Plausible mechanism was also proposed based on observed kinetics.
Ruthenium(III) Catalyzed Oxidation of Sugar Alcohols by Dichloroisocyanuric A...Ratnakaram Venkata Nadh
Kinetics of ruthenium(III) catalyzed oxidation of biologically important sugar alcohols (myo-inositol,
D-sorbitol, and D-mannitol) by dichloroisocyanuric acid was carried out in aqueous acetic acid—perchloric
medium. The reactions were found to be first order in case of oxidant and ruthenium(III). Zero order
was observed with the concentrations of sorbitol and mannitol whereas, a positive fractional order was found
in the case of inositol concentration. An inverse fractional order was observed with perchloric acid in oxidation
of three substrates. Arrhenius parameters were calculated and a plausible mechanism was proposed
Shift of Reaction Pathway by Added Chloride Ions in the Oxidation of Aromatic...Ratnakaram Venkata Nadh
Role of added chloride ions on the shift of reaction pathway of oxidation of aromatic ketones (acetophenone,
desoxybenzoin) by dichloroisocyanuric acid (DCICA) was studied in aqueous acetic acid—perchloric
acid medium. Participation of enolic and protonated forms of ketones in the rate determining steps is
manifested from zero and first orders with respect to the oxidant in absence and presence of added chloride
ions, respectively. Positive and negative effects of acid and dielectric constant on the reaction rate were
observed. The observations deduce plausible mechanisms involving (i) rate-determining formation of enol
from the conjugate acid of the ketone (SH+) in the absence of added chloride ions and (ii) rapid formation of
molecular chlorine species from HOCl (hydrolytic species of DCICA) in the presence of added chloride ions,
which then interacts with SH+ in a rate-determining step prior to the rapid steps of product formation. The
order of Arrhenius parameters substantiate the proposed plausible mechanisms based on order of reactants
both in presence and absence of added chloride ions.
Kinetics of Ruthenium(III) Catalyzed and Uncatalyzed Oxidation of Monoethanol...Ratnakaram Venkata Nadh
Kinetics of uncatalyzed and ruthenium(III) catalyzed oxidation of monoethanolamine by N-bromosuccinimide
(NBS) has been studied in an aqueous acetic acid medium in the presence of sodium acetate
and perchloric acid, respectively. In the uncatalyzed oxidation the kinetic orders are: the first order in NBS,
a fractional order in the substrate. The rate of the reaction increased with an increase in the sodium acetate
concentration and decreased with an increase in the perchloric acid concentration. This indicates that free
amine molecules are the reactive species. Addition of halide ions results in a decrease in the kinetic rate,
which is noteworthy. Both in absence and presence of a catalyst, a decrease in the dielectric constant of the
medium decreases the kinetic rate pointing out that these are dipole—dipole reactions. A relatively higher
oxidation state of ruthenium i.e., Ru(V) was found to be the active species in Ru(III) catalyzed reactions. A
suitable mechanism consistent with the observations has been proposed and a rate law has been derived to
explain the kinetic orders.
A novel reversed-phase liquid chromatographic method for the simultaneous det...Ratnakaram Venkata Nadh
In the present study 12 impurities of bisoprolol fumarate (BISO) and hydrochlorothiazide (HCTZ) were
separated simultaneously in a single HPLC method. Out of these 12 impurities, five are potential
degradants, which are validated as per The International Council for Harmonisation of Technical
Requirements for Pharmaceuticals for Human Use (ICH) guidelines. As the two active drug substances
BISO and HCTZ have different solubilities and polarities, the most critical parameters in resolving the
components from each other are pH, temperature, and solvents. The method is precise (RSD < 1.0%),
accurate, linear (r2 > 0.999), robust, and stability indicating in the range of limit of quantification (LOQ)
to 150%. The HPLC method is then migrated to ultra-performance liquid chromatography (UPLC) to
further reduce the run time and solvent consumption, and increase the sample throughput
The emergence of multidrug-resistant TB (MDR-TB) against first-line drugs and extensively drug resistant TB (XDRTB)
due to misuse of second-line anti tubercular drugs (ATDs) is a further concern. Recommended treatment involves
long term and multiple drug therapy with severe side effects. Due to this concern nanoparticle-based systems
have significant potential for treatment and prevention of tuberculosis (TB) to overcome the need to administer
ATDs at high and frequent doses, would assist in improving patient compliance and circumvent hepatotoxic ity
and/or nephrotoxicity/ocular toxicity/ototoxicity associated with the prevalent first-line chemotherapy.
Nanostructured delivery systems constitute a wide range of systems varying from liposomes, micelles, micro- and
nanoemulsions, to polymeric nanoparticles (PNPs ) and solid lipid nanoparticles (SLNs). Pulmonary administration
of inhaled nanoparticles in the form of dry powder inhalers offer particular advantages for pulmonary administration
of anti tubercular drugs (ATDs). Present review comprehensively about different approaches of nanobased
drug delivery, devises and techniques for pulmonary delivery of nanoparticle encapsulated ATD.
Kinetic, isotherm and thermodynamics investigation on adsorption of divalent ...Ratnakaram Venkata Nadh
Three novel and distinct agricultural waste materials, viz., Casuarinas fruit powder (CFP), sorghum stem powder
(SSP) and banana stem powder (BSP) were used as low cost adsorbents for the removal of toxic copper(II) from
aqueous solutions. Acid treated adsorbents were characterized by SEM, EDX and FTIR. Different factors effecting
adsorption capacity were analyzed and the effi ciency order was BSP>SSP>CFP. Based on the extent of compatibility
to Freundlich/Langmuir/D-R/Temkin adsorption isotherm and different models (pseudo-fi rst and second order,
Boyd, Weber’s and Elovich), chemisorption primarily involved in the case of CFP and SSP, whereas, simultaneous
occurrence of chemisorption and physisorption was proposed in the case of BSP. Based on the observations, it was
proposed that three kinetic stages involve in adsorption process viz., diffusion of sorbate to sorbent, intra particle
diffusion and then establishment of equilibrium. These adsorbents have promising role towards removal of Cu(II)
from industrial wastewater to contribute environmental protection.
Kinetic, thermodynamic and equilibrium studies on removal of hexavalent chrom...Ratnakaram Venkata Nadh
Removal of Cr(VI) by biosorption on two agro waste materials, casuarinas fruit powder (CFP) and sorghum
stem powder (SSP), has been investigated. The prepared adsorbent materials were characterized by SEM, EDX,
FTIR and BET. These biomaterials effectively removed Cr(VI) with a maximum removal of 93.35% and 63.75% using
15 gL−1 and 5 gL−1 of CFP and SSP, respectively, at 60 oC with 20mgL−1 initial Cr(VI) concentration in solution. In both
cases of adsorbents, kinetic data of adsorption fitted well in pseudo-second-order in terms of correlation coefficient
(R2). This helps in proposing the process of adsorption as chemical coordination, which is correlated with the thermodynamic
study results conducted at different values of temperature. Langmuir, Freundlich and D-R models were evaluated
for description of metal sorption isotherms. Values of coefficients of intra-particle diffusion and mass transfer have
also been determined at different values of temperature.
Novel coumarin isoxazoline derivatives: Synthesis and study of antibacterial ...Ratnakaram Venkata Nadh
A highly efficient and mild protocol for the syntheses of ethyl-3-
[7-benzyloxy-4-methyl-2-oxo-2H-8-chromenyl]-5-aryl-4,5-dihydro-4-
isoxazole carboxylates and ethyl-3-[7-benzyloxy-3-chloro-4-methyl-2-
oxo-2H-8-chromenyl]-5-aryl-4,5-dihydro-4-isoxazole carboxylates in
good yields via [3 þ 2] cycloaddition of in situ–generated nitrile
oxides from 7-benzyloxy-4-methyl-coumarin hydroxymoylchlorides
and 7-benzyloxy-3-chloro-4-methyl-coumarin hydroxymoylchlorides
respectively with ethyl-3-aryl prop-2-enoate has been developed.
The new compounds are screened for antibacterial activity.
Ultra performance liquid chromatographic method for simultaneous quantificati...Ratnakaram Venkata Nadh
Plerixafor (PLX) injections are administered to patients with cancers of lymphocytes
(non-Hodgkin’s lymphoma) and plasma cells (multiple myeloma). The main
objective of the current study was to develop a short reverse phase chromatographic
method for the simultaneous quantification of PLX and its impurities, in an injection
formulation, to reduce the time required for these quality tests. Furthermore, the
present work describes the role of nonalkyl branched nonquaternary ion pair reagent
in improving the peak shape and reducing column equilibration time. The separation
of PLX and its related substances is pH dependent (optimum pH = 2.50) and was
achieved on an octadecylsilyl (C18) column. The method was validated for its intended
purpose in accordance with the current regulatory guidelines for validation. The
proposed method can be applied for quality control, release, and stability analyses of
active pharmaceutical ingredient, PLX, as well as finished products, PLX injections
Caralluma lasiantha: A review on it’s vital role in Indian Traditional MedicineRatnakaram Venkata Nadh
Caralluma is a genus used as traditional medicine. Caralluma lasiantha is medicinally important due
to existence of pregnane glycosides, which may possess various biological activities. This article thoroughly
reviewed about the usage of C. lasiantha in traditional medicinal system, phytochemicals present in it, profile
identification studies, anti-hyperglycemic effect, antibacterial, antifungal, cytotoxic and antioxidant activities
Phytochemical Investigation of Caralluma lasiantha: Isolation of Stigmasterol...Ratnakaram Venkata Nadh
Stigmasterol, a phytosteroid was isolated for the first time from C. lasiantha using n-hexane as a solvent. Stigmasterol was characterized on the basis of physical, chemical and spectral data (IR, 1H NMR, 13C NMR, 1HNMR, DEPT-45, 90 & 135, and MS) analysis as well as by comparing them to their literature data. A sequence of steps was adopted like saponification, fractional crystallization and gradient elution column chromatography to isolate stigmasterol because some phytosterols possess identical physical properties which makes it difficult to isolate the constituents.
Phytochemical Screening of Caralluma lasiantha Isolation of C21 Pregnane SteroidRatnakaram Venkata Nadh
Phytochemical screening of Caralluma lasiantha was carried out and one C21 pregnane steroid was isolated from chloroform extract. Based on spectroscopic studies (IR, 1H NMR, 13C NMR and ESI-MS) the isolated compound is 3b,14b-dihydroxy-14b-pregn-5-en-20-one which was earlier isolated from other species.
Supercritical fluid (CO2) chromatography for quantitative determination of se...Ratnakaram Venkata Nadh
In the present study, two cancer therapeutic drugs (docetaxel and bortezomib) were separated from their
potential impurities on a chromatographic platform by utilizing CO2 gas (supercritical state) and quantified.
The chromatographic separations were achieved on two short columns BEH-2EP (100mm 3mm, 1.7 mm)
and CHIRALPAK AD-3 (100 mm 4.6 mm, 3 mm) for docetaxel and bortezomib, respectively. The present
work describes the role of organic modifiers in the separation of polar compounds by supercritical fluid
chromatography. The two new methods were fully validated in accordance with the current ICH
(International Council for Harmonization of technical requirements for pharmaceuticals for human use)
guidelines. The stability indicating power of the methods was demonstrated from the stress studies
conducted on the injection formulations of the two compounds. The methods are precise with % RSD of
0.4, linear with the correlation coefficient of r2 $ 0.999 and accurate in the range of 50–150% of the
target assay concentration. The two methods can be equally employed for the assay determination of
docetaxel and bortezomib APIs as well.
Quality-by-design-based development and validation of a stability-indicating ...Ratnakaram Venkata Nadh
A systematic design-of-experiments was performed by applying quality-by-design concepts to determine
design space for rapid quantification of teriflunomide by the ultraperformance liquid chromatography
(UPLC) method in the presence of degradation products. Response surface and central composite
quadratic were used for statistical evaluation of experimental data using a Design-Expert software. The
response variables such as resolution, retention time, and peak tailing were analyzed statistically for the
screening of suitable chromatographic conditions. During this process, various plots such as perturbation,
contour, 3D, and design space were studied. The method was developed through UPLC BEH C18
2.1 � 100 mm, 1.7-μ column, mobile phase comprised of buffer (5 mM K2HPO4 containing 0.1%
triethylamine, pH 6.8), and acetonitrile (40:60 v/v), the flow rate of 0.5 mL min 1 and UV detection at
250 nm. The method was developed with a short run time of 1 min. Forced degradation studies revealed
that the method was stability-indicating, suitable for both assay and in-vitro dissolution of a drug product.
The method was found to be linear in the range of 28–84 μg mL 1, 2.8–22.7 μg mL 1 with a correlation
coefficient of 0.9999 and 1.000 for assay and dissolution, respectively. The recovery values were found in
the range of 100.1–101.7%. The method was validated according to ICH guidelines.
A convenient new and efficient commercial synthetic route for dasatinib (Spry...Ratnakaram Venkata Nadh
A new and efficient synthetic route for dual-Src/Abl kinase inhibitor
dasatinib (Sprycel®), an anticancer drug, is described. This commercially
viable process yields dasatinib monohydrate free of potential impurities
with consistent yield of 68% in route A and 61% in route B with HPLC
purity >99.80% over four stages.
Utilization of agro-waste for removal of toxic hexavalent chromium: surface i...Ratnakaram Venkata Nadh
Abundantly available agricultural waste materials
(banana bunch, sorghum stem and casuarinas fruit) are
processed with negligible cost and are found to be highly
suitable as biosorbents for chromium(VI) removal from
aqueous environment due to high surface area and functional
groups of adsorbents. The equilibrium data have
been analyzed for the adsorbate–adsorbate/adsorbent
interactions and found to be fitted to the data in the order,
Hill–de Boer C Fowler–Guggenheim % Frumkin[Kiselev.
To determine the characteristic parameters for process
design, mass transfer studies have been carried out using
two-parameter isotherm models (Harkins–Jura, Halsey,
Smith, El-Awady and Flory–Huggins) and three-parameter
isotherm models (Redlich–Peterson and Sips) which are
applied to the experimental data. The fitness of the isotherms
describes that both mono- and multilayer adsorptions
occur in the present studied three biosorbents in
preference to the latter. The mechanism of adsorption has
been studied using diffusion kinetic models (viz. liquid film
diffusion, Dunwald–Wagner intra-particle diffusion model
and moving boundary model) and described the possibility
of diffusion in the order of banana bunch–stem powder[
sorghum stem powder[casuarinas fruit powder in
terms of diffusion coefficients. In essence of all the results,
the selected adsorbents can be used as a potential adsorbent
for the removal of Cr(VI) from aqueous solutions.
Evaluation of in vitro antibacterial activity of Caralluma lasiantha for scie...Ratnakaram Venkata Nadh
Caralluma lasiantha is used as a traditional medicine in India to heal body
heat and inflammations. In order to find out a scientific validation for the Indian
traditional knowledge, antibacterial activity of C. lasiantha extracts was studied
against inflammation causing bacteria (viz., Staphylococcus aureus, Escherichia coli,
Streptococcus Sp., Bacillus subtilis, Enterobacter aerogenes, Klebsiella pneumoniae)
along with other Gram-positive and Gram-negative bacteria. Solvents with different
polarity were used for extraction from dry roots and stems. Minimum inhibitory
concentrations (MIC) were also studied. Differential antibacterial activity was
exhibited by extracts and higher inhibition potential against Gram-positive bacteria
was explained. The observed antibacterial activities were correlated with the chemical
structures of phytochemicals present in C. lasiantha. Anti-inflammation activities
are related to C. lasiantha extracts through their antibacterial activities.
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
Basavarajeeyam is an important text for ayurvedic physician belonging to andhra pradehs. It is a popular compendium in various parts of our country as well as in andhra pradesh. The content of the text was presented in sanskrit and telugu language (Bilingual). One of the most famous book in ayurvedic pharmaceutics and therapeutics. This book contains 25 chapters called as prakaranas. Many rasaoushadis were explained, pioneer of dhatu druti, nadi pareeksha, mutra pareeksha etc. Belongs to the period of 15-16 century. New diseases like upadamsha, phiranga rogas are explained.
New Drug Discovery and Development .....NEHA GUPTA
The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
The Gram stain is a fundamental technique in microbiology used to classify bacteria based on their cell wall structure. It provides a quick and simple method to distinguish between Gram-positive and Gram-negative bacteria, which have different susceptibilities to antibiotics
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
Basavarajeeyam is a Sreshta Sangraha grantha (Compiled book ), written by Neelkanta kotturu Basavaraja Virachita. It contains 25 Prakaranas, First 24 Chapters related to Rogas& 25th to Rasadravyas.
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Local Advanced Lung Cancer: Artificial Intelligence, Synergetics, Complex Sys...Oleg Kshivets
Overall life span (LS) was 1671.7±1721.6 days and cumulative 5YS reached 62.4%, 10 years – 50.4%, 20 years – 44.6%. 94 LCP lived more than 5 years without cancer (LS=2958.6±1723.6 days), 22 – more than 10 years (LS=5571±1841.8 days). 67 LCP died because of LC (LS=471.9±344 days). AT significantly improved 5YS (68% vs. 53.7%) (P=0.028 by log-rank test). Cox modeling displayed that 5YS of LCP significantly depended on: N0-N12, T3-4, blood cell circuit, cell ratio factors (ratio between cancer cells-CC and blood cells subpopulations), LC cell dynamics, recalcification time, heparin tolerance, prothrombin index, protein, AT, procedure type (P=0.000-0.031). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and N0-12 (rank=1), thrombocytes/CC (rank=2), segmented neutrophils/CC (3), eosinophils/CC (4), erythrocytes/CC (5), healthy cells/CC (6), lymphocytes/CC (7), stick neutrophils/CC (8), leucocytes/CC (9), monocytes/CC (10). Correct prediction of 5YS was 100% by neural networks computing (error=0.000; area under ROC curve=1.0).
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
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In-Vivo Evaluation of Rifampicin Loaded Nanospheres: Biodistribution and Mycobacterium Screening Studies
1. Current Trends in Biotechnology and Pharmacy
Vol. 12 (2) 169-176 April 2018, ISSN 0973-8916 (Print), 2230-7303 (Online)
169
In-Vivo Evaluation of Rifampicin Loaded Nanospheres
Abstract:
Rifampicin PLGA nanospheres are
formulated with a specific goal in order to decrease
the dose, adverse effects and to enhance targeted
drug delivery. Rifampicin nanospheres were
prepared and evaluated by emulsion solvent
evaporation method. In vivo bio distribution studies
reveal that there was a long term accumulation of
rifampicin nanospheres in the lungs over other
organs. The increase in Cmax
values confirmed that
inhalable PLGA nanospheres are suitable for
targeting and providing sustained release of anti-
tubercular drugs to lungs. So inhalation is a
selected administration route of Rifampicin PLGA
nanospheres. The in vivo screening of M.
tuberculosis showed good activity as well as its
activity against multidrug-resistant M. tuberculosis
and against M. tuberculosis isolates in a
potentially latent state, makes Rifampicin PLGA
nanospheres as an attractive drug dosage form
for the therapy of tuberculosis. It can be concluded
that there is a significant potential for effective
oral delivery as well as nasal delivery of the
Nanospheres for the treatment of tuberculosis.
Keywords: Rifampicin Nanospheres, M.
tuberculosis, In vivo Bio distribution studies.
Introduction
Tuberculosis is a contagious disease that
transmits through air by the bacterium
Mycobacterium tuberculosis (MTB) (1). Despite
the pathogen being pulmonary targeted, it’s
pathogenicity spreads over entire body. In addition,
tuberculosis displays a vibrant range from non
infectious to hazardous ailment (2, 3).
Development of specific delivery system
with sustained release of drug can able to maintain
the adequate therapeutic concentration in the site
of action (4). Use of bio decomposable and
biocompatible polymers to develop nanospheres
is remarkable achievement in controlled drug
delivery system.
Poly lactic co glycolic acid (PLGA) polymer
its co-polymers have achieved a perspective in
preparing an array of delivery systems incorporated
with various drugs for sustained release, being
their biodegradable and biocompatible characters
with minimum toxicity (5) in particular as anti
tubercular drug (ATD) carrier (6).
Current research was aimed to investigate
pre clinically for efficacy release of pulmonary
targeted ATD rifampicin from PLGA nano carrier
in comparison with intravenous route therapy of
the same using the conventional formulation.
Materials and Methods
Materials
Rifampicin and polyvinyl alcohol (PVA) were
obtained from Sigma Chemical Co. Poly (lactide-
co-glycoside) was purchased from Boehringer
Ingelheim, Germany. Remaining chemicals used
in this study are of analytical grade.
In-Vivo Evaluation of Rifampicin Loaded Nanospheres:
Biodistribution and Mycobacterium Screening Studies
Vishnu Vardhan Reddy Beeram1
, Krupanidhi S1*
&Venkata Nadh R2
1
Department of Biotechnology, Vignan’s University, Vadlamudi, Guntur-522213, India.
email: srivyshu.pharma@gmail.com; Mobile: +91-9705070901
2
GITAM University – Bengaluru Campus, Karnataka- 561 203, India,
email: doctornadh@yahoo.co.in; Mobile: +91-9902632733
*
Corresponding Author email: krupanidhi.srirama@gmail.com
2. Current Trends in Biotechnology and Pharmacy
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170
Vishnu Vardhan Reddy et al
Formulation of Rifampicin PLGA Nanoparticle
by Emulsion Solvent Evaporation Method :
Different variants of rifampicin stacked
nanoparticles (NPs) were prepared by changing
the formulation variables (like polymer and
surfactant) and process variables (like sonication
time). In the formulations, 100 mg of rifampicin
was dissolved with various concentrations of PLGA
and surfactant in 300 ml ethanol. Then the organic
phase was emulsified by adding different
concentrations of PVAusing ultrasonicator for 35
seconds. The emulsion was included drop wise
into beaker containing different concentration of
PVAsolution which acts as continuous phase and
mixture was maintained for continuous stirring.
The emulsion was left on gentle stirring for 3 hrs
to allow for solvent evaporation. Then, suspended
nanoparticles were collected by ultracentrifugation
at 30,000 rounds per minutes (RPM) for 15 min
at 3° C. The NPs were washed three times with
cold double-deionized water and then freeze-dried
for 38 h (7).
Pharmacokinetic Studies : Rodents like Wistar
albino rat of male gender (160-180g) were selected
for investigation. Rats were resided in
polypropylene cages in a proper aerated room
under atmospheric circumstances of 22±2°C and
45-65% relative humidity, with on and off light with
equal duration in a day. Selected rats were
adjusted to laboratory conditions one week prior
to the date of experimentation. Standard diet food
and water were given at sufficient levels. Rats were
fasted overnight prior to the day of experiment
with a provision of water.
On the next day, Rifampicin nanospheres
were administered to rats (n= 6) by intravenous
route through tail vein at the dose of 2.4 mg/rat.
Rats are bled at time periods 5,15,30
min,1,2,4,6,8,12 and 24 h following administration.
After centrifugation, the plasma was divided and
frozen at -20°C for further studies (7).
Bio-distribution studies : Male Wistar rats (n=6)
weighing 160–180g were used for the bio
distribution studies. The experimental proposal
was accepted and performed as per the guidelines
of Institutional Animal Ethical Committee (IAEC)
(MIP/IAEC/2015-16/M1/07) of the Institute.
Nanospheres at a dose of 25mg/rat were
administered once by inhalational route. About
required dose of drug loaded nanospheres were
charged and nebulized for 30 sec using an in
house apparatus to obtain inhaled dose of 25mg/
rat. Before dosing, the rats were trained for 3 days
to accept or restraint the application of an infant
inhalation mask attached to our in-house
apparatus.
After inhalation, rats were bled at different
time intervals selected for Bio distribution studies.
After blood collection, animals were sacrificed by
using CO2
euthanasia and organs like lungs, liver
and kidney were collected. The organ weights were
recorded. Lungs were kept in saline prepared with
phosphate buffer at a slight acidic pH and stored
at -20ºC until analysis. The collected organs are
sliced and homogenized at 6000 RPM for 20 min.
Centrifugation was done to the collected tissue
samples at 4000 RPM for 10 min and the collected
supernatant was analyzed by HPLC.
Collection of bronchi -alveolar ravage : From
the sacrificed rats lungs were isolated in
conjunction with trachea by dissecting thoracic
region. The lungs were frequently lavaged with ice
cold phosphate buffer saline (PBS) (with 0.5M
EDTA) through cannulated trachea. Broncho
alveolar fluids were pooled, made centrifuge and
the collected macrophages were numbered and
stored at -20ºC till the analysis.
The analysis of drug was done by HPLC and
the concentration of drug was obtained from
calibration graphs. Inspecting the data visually,
maximum plasma concentration (C max
) and time
to achieve it time to maximum plasma
concentration (T max
) were determined.
The plasma concentration values were
transformed logarithmically and by applying linear
regression T1/2
was estimated. The plasma
concentration versus time curve (zero moment)
and the first moment curve area under the moment
curve (AUMC) were estimated. The area under
the curve (AUC), area under the concentration-
3. Current Trends in Biotechnology and Pharmacy
Vol. 12 (2) 169-176 April 2018, ISSN 0973-8916 (Print), 2230-7303 (Online)
171
time curve from time zero to time of last sample
intake (AUC0b
t) and area under the moment curve-
time curve from time zero to time of last sample
intake (AUMC0b
t) were calculated as per the
trapezoid rule. The first moment was calculated
as concentration times time (Cp x t). The AUMC
is the area under the (Cp x t) versus time curve.
TheAUC determines the bioavailability of the drug
for the given same dose in the formulation. Total
calculations were done by software Phoenix
WinNonlin non-compartmental analysis program.
Sample preparation for analysis
Blood sample: The blood samples collected at
respective time intervals during pharmacokinetic
and bio distribution studies were taken in
heparinised micro-centrifuges. Blood samples
were centrifuged at 4000 rpm for 10 min to
separate plasma and it was maintained at -20ºC
until analysis.
Aliquots of 150μl of plasma were mixed with
methanol (300μl) as de-proteinizing agent and the
obtained dispersal is whirl pooled around 2 min.
The samples were centrifuged at 15000 RPM for
10min at 4°C and the supernatant is collected.
Rifampicin was extracted with 3 ml of
chloroform-butanol (70:30%v/v) and 3 ml portions
of chloroform-butanol (70:30% v/v) and vortexed
for about 1 min followed by centrifuging at 4000
RPM of 10 min duration. The superficial layer was
decanted and the process was repeated for in
triplicate and superficial liquid was collected. The
collected supernatants were diluted and analyzed
by HPLC.
Tissue sample : Tissue homogenates (20% w/v)
with aqueous medium were prepared in cold 150M
KCL. Supernatant liquid collected from
homogenates by centrifugation at 15000 RPM for
10 min at 4°C was kept aside for further studies.
Then, 300μl of the methanol was admixed to 150μl
of the clear homogenates and the dispersal was
vortexed for 2 min. The samples were then
centrifuged at 15,000 RPM for about 10 min at 4
ºC. An equal volume of water is added to the
obtained supernatant. The samples were further
filtered using 0.2μm nylon filters and were instilled
to the HPLC system (8-11).
Bio-analytical HPLC method : The collected
serum and tissue samples were analyzed by
HPLC (Analytical technologies Ltd) comprising
C-18 column & UV detector. The mobile phase
consists of Triethanolamine acetate: acetonitrile
(97: 3% v/v) eluted by isocratic method and
detected at 262 nm, analysed at 30ºC by injecting
20μl by maintaining a flow rate of 0.9ml/min. A
wash method program which increased the %
methanol was included at the end of drug
elution to ensure washout of all interfering
exciepents. Spectral purity analysis of the
drugs peak over a range of 200–400 nm was
performed. The accuracy and precision of the
developed method for determination of drug
was comparable to the isocratic methods
described in United State Pharmacopoeia (USP).
Data Analysis
The area under the total plasma
concentration time curve from zero time to infinity
was calculated by equation AUC0–”
= area under
the plasma concentration-time curve extrapolated
to infinity
AUC0–” =
AUC0–t
+Ct
/Ke
Where Ct
is the rifampicin concentration observed
at last time and Ke
is the apparent elimination
rate constant obtained from the terminal slope of
the individual plasma concentration (12-18).
Durations of serum drug concentration following
inhalational route were analyzed by software
Phoenix Win Nonlin non-compartmental analysis
program of version 5.1.
Mycobacterial infection study in rat
MTB H37RvATCC 27294, a strain sensitive to all
the standard antimycobacterial agents, was used
for all animal infection in the experiments.
Bacterial cultures were prepared as described
previously published article (8). Wistar rats were
infected via the respiratory route to obtain low-
grade bacillary lung infection (100 bacilli) using a
modified Madison aerosol chamber (9). Bacterial
lung loads were estimated to determine suitable
infection conditions for drug efficacy experiments.
After infection, the animals were housed for the
duration of the study in a bio-safety level 3
facilities. By using microbial enumeration,
In-Vivo Evaluation of Rifampicin Loaded Nanospheres
4. Current Trends in Biotechnology and Pharmacy
Vol. 12 (2) 169-176 April 2018, ISSN 0973-8916 (Print), 2230-7303 (Online)
172
dependent variable, the number of animals
required, per treatment group was three in
experiments for drug evaluation. The course of
mycobacterial infection was monitored by
enumeration of colony forming units (CFU) from
excised lungs at 1, 2, 3, 4, 6, and 12 weeks of
post infection.
Statistical analysis : All the experiments were
performed in triplicate. Results were collected as
mean and standard deviation (mean ± SD).
Significance in difference was measured with p
value as p d” 0.5.
Results
The mean bio distribution pharmacokinetic
study parameters of F13 rifampicin nanospheres
formulation administered through Intravenous (IV)
and Inhalation are summarized in the Table 1,
Figure 1 & 2. The C-max, T-max and clearance of
F13 through Inhalation route were 42.34 ìg/mL,
14 hr and 0.82(ml/hr) respectively, similarly those
of IV route were 20.46 ìg/mL,14 hr and 3.44
respectively.
In short term study (Table-3),the potency
of rifampicin nanospheres at 25 mg/kg body weight
was compared with those of rifampicin pure drug.
rifampicin nanospheres slightly more active than
pure drug rifampicin. Long-term treatment (Table-
3) with rifampicin nanospheres at 25 mg/kg Shown
statistically very significant when compared to
negative control in the lungs and spleen (p<0.05).
The results elucidated that there was a significant
decrease in colony forming unit (CFU) of lungs
and spleen in all treated groups during both period
in contrast to negative control group.After 18 days
of treatment, rifampicin nanospheres minimized
the bacterial content by 0.54 log 10
CFU. The
activity of rifampicin nanospheres and rifampicin
pure drug in the lungs and spleen was statistically
not significant (p = ns) when compared with
negative control after 18 days of treatment. After
42days of treatment, rifampicin nanospheres
minimized the bacterial content 4.62log 10
CFU.
(9.68 14.30-9.68 log 10 CFU). The efficacy of
rifampicin nanospheres in the lungs and spleen
differs statistically from those of rifampicin pure
drug (P > 0.001) after 42days of treatment. The
results are shown in (Table 2&3 and Figure no 3).
Discussion
By comparing the Cmax
results between IV
and Inhalation administration of Rifampicin
Nanospheres it shows that more concentration of
drug was accumulated in the lungs while
Fig. 1 : XY plot for Rifampicin Nanoparticle –IV
administration Fig. 2: XY plot for Rifampicin Nanoparticle –
Inhalation administration
Vishnu Vardhan Reddy et al
5. Current Trends in Biotechnology and Pharmacy
Vol. 12 (2) 169-176 April 2018, ISSN 0973-8916 (Print), 2230-7303 (Online)
173
Fig. 3: Determination of Bacterial numbers in lungs after 6 weeks of treatment
Table 1: Bio distribution studies of Rifampicin Nanospheres (F13) – IV & Inhalation administration
Tissue T+(hr) Cmax
(μg/ml) AUC 0-”
Vd
(ml) Clearance
(μg/ml/hr) (ml/hr)
IV Administration
Lungs 1 20.46±1.05 802.3±5.29 244.8±3.95 3.44±0.31
Liver 2 24.98±1.17 1120.16±8.46 184.61±2.10 2.82±0.56
Kidney 3 22.02±0.94 690.39±5.93 218.43±42 3.02±0.14
Inhalation
Lungs 14 42.34±2.46 1996±8.32 136.33±2.48 0.82±0.04
Liver 20 10.84±1.8 484±4.26 489±5.73 2.84±0.19
Kidney 24 4.32±0.86 118±3.16 756.6±6.38 2.34±0.23
Table 2: Growth of M.tuberculosis in culture medium and their control in different conditions
Compound Concentration Log10
CFU±SD % growth of
(per ml) microorganism
compared
with controls
Only culture - 11.62±0.22 100
Pure Rifampicin 25 mg/kg 9.40±0.12** 78.40
Rifampicin NP Equivalent to 25 mg/kg 6.42±0.26*** 68.40
Values are expressed as mean ± SD. n=3; Values are statistically significant at p<0.05;
Significant-p<0.05;** Very significant -p<0.01;***-p<0.001; ns-non-significant;
In-Vivo Evaluation of Rifampicin Loaded Nanospheres
6. Current Trends in Biotechnology and Pharmacy
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174
administering the formulation through Inhalation.
From the Tmax
comparison data between IV and
Inhalation administration it shows the sustainability
time of rifampicin nanospheres in lungs i.e., it
confirms the sustained action of rifampicin
nanospheres in lungs. By comparing the AUC0-á
from Table 1, it concludes that maximum
concentration of drug was present in lungs through
inhalation than any other organ. The organ
clearance ratio of drug from lungs through
inhalation was less than the IV administration,
which confirms the sustained release of rifampicin
from nanospheres in the lungs. From the above
pharmacokinetic distribution data it shows that
rifampicin nanospheres shows more accumulation
of drug in lungs through inhalation administration
than IV, which indicates nanospheres is having
targeted and sustained release of drug results in
lungs. By this it can be confirmed that inhalable
nanospheres are suitable for targeting with
negligible toxicity and providing sustained release
of anti-tubercular drugs especially rifampicin in
lungs. The result showed the rifampicin
nanospheres leads to maximum deposition of drug
in lungs through inhalation which leads to maintain
high therapeutic concentration by improving good
pulmonary tuberculosis chemotherapy.
In vivo mycobacterium screening studies
show that on long term therapy rifampicin
nanospheres shows better control of growth of
microorganism i.e. CFU when compared to short
term therapy i.e. for 18 days after administration.
After 6 weeks of treatment the bacterial counts in
the lungs were reduced to very low numbers in all
treatment groups (range, 14.30 to 9.68 log10 CFU
Rifampicin Nanospheres Vs untreated controls),
as was the case for the spleens (p<0.001). In
summary, its good activity in vivo models, as well
as its activity against multidrug-resistant MTBand
against MTB isolates in a potentially latent state,
makes Rifampicin Nanospheres an attractive drug
dosage form for the therapy of tuberculosis. These
data indicate that there is significant potential for
effective inhalational delivery of Rifampicin
Nanospheres for the treatment of tuberculosis.
Conclusion
In vivo bio distribution studies show that
nanospheres form is the best formulation for
Rifampicin, Nanospheres accumulates maximum
Table 3: M.tuberculosis number in lungs and spleen of albino rat
Treatment batch M.tuberculosis number (Log10
CFU±SD) after
different treatment schedules
18days 42 days
Lungs Spleen Lungs Spleen
Control group 6.40 ± 0.12 2.16 ± 0.12 9.42 ± 0.42 4.12 ± 0.10
Negative control 7.42± 0.32a**
2.74 ± 0.10a**
14.30± 0.54a***
5.68 ± 0.22a***
Positive control
with Rifampicin
25 mg/kg 7.40 ± 0.20b ns
2.54 ± 0.14b ns
10.40 ± 0.30b***
3.64 ± 0.24b***
Test group with
Rifampicin Nanospheres
Equivalent to 25 mg/kg 7.82 ± 0.22b ns
2.48 ± 0.10bns
9.68 ± 0.22b***
2.28 ± 0.22b***
Values are expressed as mean ± SD. n=3; Values are statistically significant at p<0.05;
Significant-**-p<0.01;Very Significant-***-p<0.001; ns-non-significant; a-Group compared to control;
b-Groups compared to negative control;
Vishnu Vardhan Reddy et al
7. Current Trends in Biotechnology and Pharmacy
Vol. 12 (2) 169-176 April 2018, ISSN 0973-8916 (Print), 2230-7303 (Online)
175
dose in the lungs than other organs over prolonged
period of time. The plasma levels are more for
inhalable PLGA Nanospheres and are suitable for
targeting and providing sustained release of anti-
tubercular drugs to lungs. So inhalation can be
selected as administration route of Rifampicin
PLGA Nanospheres. From the in vivo screening
of M.tuberculosis, it shows good activity in vivo
models, as well as its activity against multidrug-
resistant MTBand against MTBisolates in a
potentially latent state, makes Rifampicin PLGA
Nanospheres an attractive drug dosage form for
the therapy of tuberculosis. These data indicate
that there is significant potential for effective
intravenous as well as nasal delivery of
Nanospheres for the treatment of tuberculosis.
REFERENCES
1. World Health Organization (WHO, 2015).
Global Tuberculosis Report 2015.
2. Barry, C. E. (2009). The spectrum of latent
tuberculosis: rethinking the biology and
intervention strategies. Nat. Rev. Microbiol.
7: 845–855.
3. Esmail, H., Barry, C. E., Young, D. B.
and Wilkinson, R. J. (2014).The ongoing
challenge of latent tuberculosis. Phil. Trans.
R. Soc. B 369: 387-422.
4. Anderson J.M. and Shive M.S. (1997).
Biodegradation and biocompatibility of PLA
and PLGAmicrospheres,Adv. Drug. Delivery
Rev, 28: 5–24.
5. Dutt M. and Khuller G.K. (2001). Chemo-
therapy of Mycobacterium tuberculosis
infection in mice with a combination of
isoniazid entrapped in poly (DL lactide-co-
glycolide) microparticle. J. Antimicrob.
Chemother, 4: 829-835.
6. AitMoussa L., Khassouani C.E., Hue B.,
Jana M., Begaud B. and Soulaymani R.
(2002). Determination of the acetylator
phenotype in Moroccan tuberculosis
patients using isoniazid as metabolic probe.
Int J Clin Pharm Ther, 40: 548-53.
7. MaikeLohrmann, Michael Kappl., Hans-
Juergen Butt., Nora Anne Urbanetz. and
Bernhard Christian Lippold. (2007).Adhesion
forces in interactive mixtures for dry powder
inhalers – Evaluation of a new measuring
method.ýEur. J. Pharm. Biopha, 67: 579 -
586.
8. SaniIbnYakubua., Khaled H.Assi. and Henry
Chrystync. (2013). Aerodynamic dose
emission characteristics of dry powder
inhalers using an Andersen Cascade
Impactor with a mixing inlet. ýInt. J. Pharm,
455: 213– 218.
9. Francesco Martinelli., Anna Giulia
Balducci., Alessandra Rossi., Fabio
Sonvico. and Paolo Colombo. (2015). Pierce
and inhale design in capsule based dry
powder inhalers: Effect of capsule piercing
and motion on aerodynamic performance of
drugs. Int. J. Pharm, 487:197–204.
10. Yoen-JuSon., Worth Longest P. and Michael
Hindle. (2013). Aerosolization
characteristics of dry powder inhaler
formulations for the excipient enhanced
growth (EEG) application: Effect of spray
drying process conditions on aerosol
performance. Int. J. Pharm, 443:137– 145.
11. Sarah Zellnitz., Jakob Dominik
RedlingerPohna. and Michael Kapplb.
(2003). Characterization and deposition
studies of engineered lactose crystals with
potential for use as a carrier for aerosolised
salbutamol sulfate from dry powder inhalers.
Eur. J. of Pharm. Sci, 19: 211–221.
12. Cordula Weiss., Peter McLoughlin. And
Helen Cathcart. (2015). Characterization of
dry powder inhaler formulations using atomic
force microscopy. Int. J. Pharm, 494: 393 -
407.
13. FlorisGrasmeijer., Paul Hagedoorn.,
Henderik W Frijlink. And Anne H de Boer.
(2012). Characterization of high dose
aerosols from dry powder inhalers. Int. J.
Pharm, 437: 242– 249.
In-Vivo Evaluation of Rifampicin Loaded Nanospheres
8. Current Trends in Biotechnology and Pharmacy
Vol. 12 (2) 169-176 April 2018, ISSN 0973-8916 (Print), 2230-7303 (Online)
176
14. Ehab F Elkady. andMarwaAFouad. (2011).
Forced degradation study to develop and
validate stability-indicating RP-LC method
for the determination of ciclesonide in bulk
drug and metered dose Inhalers.Talanta, 87:
222– 229.
15. De Boer A.H., Winter H.M.I. and Lerk C.F.
(1996) Inhalation characteristics and their
effects on in vitro drug delivery from dry
powder inhalers Part 1. Inhalation
characteristics, work of breathing and
volunteers’ preference in dependence of the
inhaler resistance. Int. J. Pharm, 130: 231-
244.
16. De Boer A.H., Gjaltema D, Hagedoorn P.
(1996). Inhalation characteristics and their
effects on in vitro drug delivery from dry
powder inhalers Part 2: Effect of peak flow
rate (PIFR) and inspiration time on the in
vitro drug release from three different types
of commercial dry powder inhalers. Int. J.
Pharm, 138: 45-56.
17. Tan Suwandecha., Wibul Wong poowarak.,
Kittinan Maliwan. and Teerapol Srichana.
(2014). Effect of turbulent kinetic energy on
dry powder inhaler performance. Powder
Technology, 267: 381–391.
18. Nora Y.K Chewa., Hak Kim Chana., David F
Bagsterb. and Jay Mukhraiyab. (2002).
Characterization of pharmaceutical powder
inhalers: estimation of energy input for
powder dispersion and effect of capsule
device configuration. Aerosol Science, 33:
999–1008.
Vishnu Vardhan Reddy et al