Degradation Studies Research Articles

The CombE-IDMS Alternate Potency Method for H5N1 and H5N8 Cell-Based Vaccines.

Assaying the potency of inactivated viral influenza vaccines is performed using single radial immunodiffusion, which is the globally accepted release method for potency. Under conditions of a rapidly emerging pandemic, such as the 2009 H1N1 influenza pandemic, a recognized obstacle in the delivery of vaccines to the public is the time needed for the distribution of calibrated SRID reagents (antisera and antigen standards) to vaccine manufacturers. Previously, we first described a novel streamlined MS-based assay, CombE-IDMS, which does not rely on antisera/antibodies or reference antigens, as a potential rapidly deployable alternate potency method through a comparison with SRID on adjuvanted seasonal quadrivalent vaccine cell-based (aQIVc) materials. In this report, we further demonstrate that the CombE-IDMS method can also be applied to measure the potency of pre-pandemic H5N1 and H5N8 monovalent vaccine materials, each subtype both unadjuvanted and adjuvanted, through a forced degradation study. Overall, CombE-IDMS results align with those of the gold standard SRID method on both H5N1 and H5N8 materials under conditions of thermal, pH, oxidative and freeze/thaw stress, lending further evidence for the CombE-IDMS method's suitability as an alternate assay for potency of both seasonal and pandemic influenza vaccines.

Thermal degradation studies of 2-amino 6-nitrobenzothiazole-oxamide-formaldehyde copolymer and its composites

Composites are viewed as promising advanced materials because of their superior features, including enhanced flame retardancy, density, relatively large surface areas, strength and thermomechanical/optoelectronic/magnetic capabilities. 2-amino 6-nitrobenzothiazole, oxamide, and formaldehyde has been used for the synthesis of copolymerizing polycondensation technique, and newly synthesized copolymer with activated charcoal was used to synthesize its composite. Several physicochemical techniques, and Ultraviolet-visible, SEM, TGA, and FTIR techniques, have been utilized to illustrate the produced copolymer and its composite. The non-aqueous conductometric titration was used to determine the molecular weight of the 2-amino 6-nitrobenzothiazole-oxamide-formaldehyde copolymer.based on TGA data, thermal stability of the composite is higher than copolymer, possibly because there are more carbonized residues. Based on Ea values determined by Sharp-Wentworth and Freeman-Carroll technique for copolymer and its composite, it is observed that they are in good agreement with each other. It is abundantly obvious from the results of the sequence of reaction that the decomposition reaction roughly follows a first-order kinetics. The composite's higher surface area, porous makeup, and smaller particle size may have contributed to its superior performance. The copolymer and its composite have better heat stability.

Size exclusion chromatography as green support for forced degradation study of adalimumab.

Size exclusion chromatography (SEC) has become a powerful tool for analysing size variants of biologic drugs in their native form. Modern SEC can be defined by the use of chromatographic columns packed with sub-3 µm particles, allowing an increase in method throughput compared to that of conventional SEC. We performed the forced degradation study of adalimumab, the first genetically engineered fully humanised immunoglobulin G1 monoclonal antibody, and evaluated tha possibilities of an advanced SEC column packed with sub-3 µm particles for elucidation of the degradation pathway. Our results revealed the main adalimumab degradation products to be antibody fragments. Acidic and basic conditions had the most intensive effect on the degradation of the adalimumab while the drug exhibits relative stability under thermal and photolytic stress conditions. The AGREE and AGREEprep calculators were used for the evaluation of the environmental performance of the forced degradation procedure. The results of the green score evaluation are presented as round pictograms with a circle in the centre that shows the overall score of 0.81 and 0.61, respectively. Both pictograms are highlighted in green, indicating the eco-friendly conditions.

A new stability indicating HPLC and LC-APCI-MS methods for the estimation of Flucytosine in pharmaceutical dosage forms

Flucytosine is an anti-fungal agent which acts against mycotic infections. The anti-mycotic activity of Flucytosine is due to its conversion in to 5-fluoro uracil. In the present study the authors have proposed a new stability indicating isocratic RP-HPLC as well as LC-APCI-MS methods for the estimation of Flucytosine and the methods are validated as per ICH guidelines. Thermo scientific - TSQ Quantis with Vanquish HPLC coupled with MS was employed for the study with Simpack C18 (250 mm x 4.6 mm x 5µm) column. A triple quadrupole mass spectrometer with atmospheric pressure chemical ionization (APCI) source, running in the positive mode (as well as negative mode) was used for the detection. A mixture of 0.1% Acetic acid: Methanol was used as mobile phase (50:50, v/v) (UV detection at 235 nm). A wide linearity concentration range 1.0-120 μg/ml was observed and the method is found to be precise and accurate and can be used for the estimation of marketed formulations. Stress degradation studies were performed and the method is found to be selective and specific.

QbD-driven RP-HPLC method for novel chemo-herbal combination, in-silico, force degradation studies, and characterization of dual drug-loaded polymeric and lipidic nanocarriers

BackgroundIn cancer therapies, chemo-herbal combinations are receiving increased attention. A multiple tyrosine kinase inhibitor, lenvatinib (LTB) is beneficial in treating thyroid, lung, endometrial, and liver cancers. An isoflavone called biochanin A (BCA) is well known for its diverse biological properties that have been studied to potentiate the anti-cancer potential and lower the normal cell toxicities of other therapeutics. LTB and BCA can be combined for cancer treatment and may increase their therapeutic potential at lower doses. In brief, the quality by design (QbD)-driven RP-HPLC method was developed, validated, and utilized for applications employing the study of forced degradants and the successful development of LTB and BCA co-loaded nanocarriers.ResultsThe RP-HPLC method employed Box–Behnken design with peak resolution 6.70 ± 0.006, tailing factor 1.06 ± 0.05 for BCA and 1.17 ± 0.021 for LTB, and theoretical plates number > 2000. RP-HPLC applications utilized the investigation of a total of 41.17% and 70.58% degradants for LTB and BCA in contrast to in-silico predicted studies using Zeneth software. The poly (lactic-co-glycolic acid) nanoparticles (PLGA NPs) were formed with particle size 185.3 ± 12.3 nm, zeta potential − 13.3 ± 0.35 mV, and percentage entrapment efficiency (%EE) for the LTB and BCA 53.64 ± 4.81% and 61.29 ± 4.67%, respectively. However, the developed Cubosomes (CBs) exhibited 182.4 ± 16.3 nm aerodynamic particle size, − 10.8 ± 0.39 mV zeta potential, and % EE for LTB and BCA 55.62 ± 7.73% and 72.88 ± 5.52%, respectively. The percentage drug loading (%DL) of LTB and BCA from PLGA NPs was found to be 3.7 ± 0.46% and 4.63 ± 0.48%, whereas CBs exhibited higher % DL for BCA (5.42 ± 1.10%) and LTB (4.43 ± 0.77%).ConclusionThe RP-HPLC method was developed and validated according to ICH and USP guidelines. In-vitro and in-silico forced degradation studies are evident to quantify the type of degradant and its exact mechanism of degradation. In-silico toxicity assessment for LTB, BCA, and their degradants explains the necessity of conducting degradation studies during drug development. Finally, the applications of the developed RP-HPLC method explain the usefulness of analytical methods in the development of chemo-herbal drug nanocarriers (polymeric and lipidic).Graphical abstract

The role of heat treatment on the in vitro degradation and corrosion-initiated failure of the biodegradable Mg–Zn–RE alloy

ABSTRACT In the current work, magnesium (Mg) alloy containing zinc (4%) and rare earth (RE) elements (1%) was produced by stir casting and the role of heat treatment on altering the microstructure, in vitro degradation and corrosion-initiated mechanical failure. Solutionising has introduced considerable grain refinement from 122 ± 6.9 μm to 95 ± 7.1 μm and decreased the amount of intermetallic. Potentiodynamic polarization tests indicated improved corrosion resistance for the solutionised alloy as indicated by with a lower corrosion current value (0.021 × 10-4 A.cm -2) compared with base alloy (0.027 × 10-3 A.cm -2) and annealed alloy (0.168 × 10-3 A.cm -2). In vitro degradation studies conducted in simulated body fluid solutions demonstrated better degradation resistance for the solutionised alloy compared with base and annealed alloy. Tensile tests conducted before and after exposing the samples to a corroding environment for 24 h showed the promising role of microstructure modification by solutionising on enhancing the resistance to corrosion-initiated failure with improved ductility compared with the base and annealed alloys. From the results it is concluded that solutionising can be successfully adopted to enhance the corrosion and mechanical properties of Mg-Zn-RE alloy for manufacturing of degradable implants for bone fixing applications.

Green RP-HPLC methods for assay and related substances in rivaroxaban tablets

AbstractIn this study, two different ethanol-based RP-HPLC methods for assay and quantification of rivaroxaban related substances in tablets were developed, based on green analytical chemistry (GAC) principles, using the design of experiments approach. The chromatographic separation was performed on X-Bridge C18 column (250 × 4.6 mm, 5 µm particle size), using isocratic elution with ethanol : water (35:65, % v/v) for the assay and gradient elution with ethanol/water mobile phase, for related substances, with a flow rate of 1.0 mL min−1. The gradient method was optimized for the separation of three specified impurities (impurity G, impurity H, and impurity 14) and the selectivity was further confirmed using forced degradation studies. Both methods were validated in accordance with ICH guidelines. The robustness of the methods was confirmed with the Central Composite Face Design of Experiments. Analytical Eco-scale approach and AGREE metrics confirmed that both methods are in accordance with the GAC principles. The proposed ethanol-based RP-HPLC methods were applied for assay and determination of related substances in rivaroxaban 10 mg tablets obtained from three different manufacturers available on the Macedonian market.

Bioactive Fe Foam for Degradable Bone Graft Cages

Fe foams when combined with a proper surface coating can be effectively used as a biodegradable metal with improved biocompatibility and bioactivity. Herein, Fe foams prepared via powder metallurgy route coated with two different bioactive coatings namely polyvinyl alcohol (PVA)/bioglass (BG) and polylactic acid (PLA)/BG are investigated. The effect of hydrophilic PVA and hydrophobic PLA, in conjunction with BG, has been examined. The study of biomineralization, degradation, and weight loss of the composite‐coated material over a period of 28 days is compared with that of the uncoated Fe foam samples in simulated body fluid. The composite coatings on the Fe foam effectively tailor the degradation with enhanced biomineralization. The composite‐coated material exhibits viability toward the L929 fibroblast cell line. The work reveals the great potential of Fe foams to be used as a biodegradable metallic material for temporary implant applications such as bone graft cages.

Development of a Skin Scaffold for Wound Healing Using a Polymer Blend

Native skin consists mainly of epidermal and dermal layers. Here in this work, we have constructed an artificial skin scaffold mimicking the bilayered structure of the native skin using electrospinning technique for wound healing. Polyurethane (PU) and Gelatin (Ge) were used for developing the epidermal layer and the dermal layer respectively. Ciprofloxacin HCl (Cip. HCl) a fluoroquinolone antibiotic was incorporated in both the layers for rapid wound healing. Morphology of the skin scaffold was studied using scanning electron microscopy (SEM) analysis and the chemical characterization was performed using FTIR spectroscopy. Water vapor transmission rate test and oxygen transmission rate test was conducted to evaluate the barrier properties of the scaffold. Thermal stability of the skin scaffold was evaluated using DSC and TGA while an understanding of the exudate absorbing capacity and degradation behavior of the scaffold was obtained from water absorption studies and in vitro degradation studies respectively. In vitro drug release study and drug release kinetics was explored to understand the release mechanism of Cip. HCl from the scaffold. Both the layers showed nano and micropores when analyzed using SEM. The dermal layer showed comparatively more water absorption capacity and degradation, hence providing a moist environment for the wound. The skin scaffold was permeable to water vapor and oxygen, and hence will speed up the process of wound healing. In vitro release for Cip. HCl showed a non-Fickian swelling type release with zero-order kinetics. Disk diffusion test conducted on the bilayers proved the antibacterial activity of the membrane. Hence the electrospun PU-Ge skin scaffold containing Cip. HCl is a promising candidate among modern day wound healing materials.

STABILITY INDICATING HPTLC METHOD DEVELOPMENT AND VALIDATION FOR CERITINIB IN BULK AND FORMULATION

Ceritinib is an anti-cancer of drug used in treatment of non-small cell lung cancer. A high-performance thin layer chromatography (HPTLC) method has been developed for ceritinib, which is stability indicating and simple, precise, and discriminating. The stationary phase used was aluminum-backed silica gel60 F254 plates with chloroform: methanol: glacial acetic acid as the mobile phase in the ratio of 8.5:1.5:0.5 (V/V/V). The retention factor was found to be 0.34 ± 0.02. The densitometric scanning was performed at 277 nm. The linear range for analysis was 100–600 ng band-1, which gave good linear relationship with regression coefficient of 0.998. Method accuracy was proved by the recovery studies. The detection limit and quantification limit were 7.38 and 10.03 ng band-1, respectively. Stress degradation studies like hydrolysis under different pH conditions, photolytic, thermal, and oxidative degradation were carried out as per ICH Q1A (R2) and Q1B guidelines. The method established was found to be robust, and thus it can be used as a stability-indicating method.

FORCED DEGRADATION STUDIES OF VINCAMINE BY HPTLC

Vincamine is an alkaloid with vasodilator properties. Vinpocetine, a semi-synthetic vincamine derivative alkaloid, is used to treat cerebrovascular diseases like stroke and dementia. Being an important molecule, the current research work examines the forced degradation studies of vincamine using planar chromatography, HPTLC using stability studies parameters such as degradation by acid and base hydrolysis, oxidative stress degradation, hydrolytic induced degradation, photolytic degradation and dry heat degradation. The compound was found to be stable to oxidative stress, but significant degradation occurred under acid hydrolysis, base hydrolysis, water hydrolysis, and to a lesser extent, under thermal stress and photolytic stress. The percentage recovery of vincamine was found to be lower in acidinduced degradation (31.8 %), and base-induced degradation (8.2 %) than in UV-induced degradation (94.9 %) and dry heat-induced degradation (93.1 %). In pharmaceutical research and development, forced degradation experiments are essential for predicting long-term stability.

A eco-friendly, low cost green synthesis of Ag@Sm2O3/rGO nanocomposites with enhanced UV light photocatalytic and antimicrobial activity

In this research, we investigated the potential of a leaf extract from the Punica Granatum peel as a reducing and capping agent in the manufacture of a nanocomposite consisting of Ag-doped Sm2O3 nanoparticles encased in reduced graphene oxide (rGO). XRD, SEM, TEM, FTIR, XPS, PL, and DRS were all used to confirm the performance and characterisation of nanocomposite materials. Photocatalytic degradation studies of 4-nitrophenol (4-NP) and methylene blue (MB) provided interesting insights into the photocatalytic activity of Ag@Sm2O3/rGO. The results show that Ag@Sm2O3 particles between 10 nm and 20 nm in size are uniformly distributed across the RGO surface. Maximum photodegradation efficiency towards MB (96.7 %), high apparent constant (0.8716 min−1), and long-term stability under UV light exposure are all achieved when rGO is combined with Ag@Sm2O3 as the photocatalyst. It was confirmed that certain types of bacteria and fungi responded differently to the antibacterial properties of doped and undoped nanoparticles. The surface surface plasmon resonance (SPR) of Ag NPs had a role in the relationship between Sm2O3 nanoparticles and rGO sheets, which led to this finding. A novel Sm2O3-based photocatalyst with high solar absorption and higher catalytic activity is proposed, and a viable mechanism for developing such a material is shown in this study.

Comparative in vitro evaluation of five Commelina forage species on ruminal fermentation and methanogenesis

In vitro gas measurement study was known to be a helpful tool for investigating the nutritional quality of feed for ruminant animals. This research was conducted to explore the ruminal fermentation and methane emission potential, and their variability pattern with changing seasons and altitudes for Commelina species, using in vitro test. Samples of the species were collected from different altitudes and seasons and used for the in vitro degradability study. Five species (C.africana, C. albescence, C. benghalensis, C. imberbis and C. diffusa), two altitudes (low and mid), and two seasons (wet and dry) were arranged in a 5 × 2 x 2 factorial manner in a completely randomized design, with three repeats of each treatment. In vitro gas and methane production as well as methane to total gas ratio were all meaningfully (P < 0.001) affected by season and altitude with the highest values observed in wet season and mid altitudes. At 24 h incubation, 44.76 and 37.82 ml/200 mg DM of total gas production was noted for wet and dry seasons respectively, while 39.77 and 47.38 ml/200 mg DM was recorded for low and mid altitudes respectively. Average gas production from immediately fermentable fractions (a) for wet season (4.21 ml) was reasonably (P < 0.001) higher than those for dry season (2.16 ml). Midlands (5.04 ml) had higher (P < 0.001) ‘a’ value than lowlands (2.77 ml). Both methane production and methane to total gas ratio exhibited a tangible decrease (P < 0.001) from wet season to dry season. 4.88 ml/200 mg DM and 6.52 ml/200 mg DM methane records were recorded for lowlands and midlands respectively. It appears that Commelina species contain nutrients that degrade in vitro, highlighting their potential supplement value for animals. The promising gas production potential added to its low methanogenic coefficient relative to the other species makes C. diffusa to the preferred supplement for poor-quality roughages followed by C. benghalensis and C. imberbis and the rest species being least preferred. But further nutrient analysis, minerals, secondary metabolites and the like, was required. And, in vivo trials must be conducted to strengthen the implications of this study.

In Vitro and In Vivo Hydrolytic Degradation Behaviors of a Drug-Delivery System Based on the Blend of PEG and PLA Copolymers.

This paper presents the in vitro and in vivo degradation of BEPO, a marketed in situ forming depot technology used for the formulation of long-acting injectables. BEPO is composed of a solution of a blend of poly(ethylene glycol)-block-poly(lactic acid) (PEG-PLA) triblock and diblock in an organic solvent, where a therapeutic agent may be dissolved or suspended. Upon contact with an aqueous environment, the solvent diffuses and the polymers precipitate, entrapping the drug and forming a reservoir. Two representative BEPO compositions were subjected to a 3-month degradation study in vitro by immersion in phosphate-buffered saline at 37 °C and in vivo after subcutaneous injection in minipig. The material erosion rate, as a surrogate of the bioresorption, determined via the depot weight loss, changed substantially, depending on the composition and content of polymers within the test item. The swelling properties and internal morphology of depots were shown to be highly dependent on the solvent exchange rate during the precipitation step. Thermal analyses displayed an increase of the depot glass transition temperature over the degradation process, with no crystallinity observed at any stage. The chemical composition of degraded depots was determined by 1H NMR and gel permeation chromatography and demonstrated an enrichment in homopolymers, i.e., free PLA and (m)PEG, to the detriment of (m)PEG-PLA copolymers in both formulations. It was observed that the relative ratio of the degradants within the depot is driven by the initial polymer composition. Interestingly, in vitro and in vivo results showed very good qualitative consistency. Taken together, the outcomes from this study demonstrate that the different hydrolytic degradation behaviors of the BEPO compositions can be tuned by adjusting the polymer composition of the formulation.

Novel stability indicating HPLC method for the quantification of Nirmatrelvir in bulk drugs

Nirmatrelviris anantiviral drug that acts asa 3C-likeprotease inhibitor. It is used with the combination of Ritonavir for the treatment of COVID-19. The World Health Organization strongly recommended this medicine to moderate and mild COVID patients, which is now the best option for COVID-19 therapeutics against SARS-CoV-2. As a result, there is an urgent need for quick and low-level quantitative analytical techniques for Nirmatrelvir and its products by focusing on the mass-compatible buffer with a shorter run time as the target. Accordingly, we developed a procedure with the following optimized conditions; mobile phase: 0.10 % formic acid in water and acetonitrile (55:45 %v/v), column oven temperature: 45 °C, flow rate: 1.0 mL/min with 15 min run time in isocratic elution mode. The developed procedure is linear and accurate from 0.20 to 500 µg/mL. Forced degradation studies in all solid and liquid degradations were executed to demonstrate the method's stability indicative capacity. To demonstrate the peak purity of the desired compound orthogonally, all degradation samples were then subjected to peak purity analysis using an HPLC PDA (High-performance liquid chromatography photodiode array) detector and LCMS (Liquid chromatography-mass spectrometry) analysis. The literature methods were developed with highly concentrated buffers which cause ion suppression in LCMS and increase backpressure in HPLC systems but the proposed method is conducive and more sensitive (limit of quantitation: 0.20 µg/mL and limitation of detection: 0.0 6 µg/mL) than the reported methods and has a wide linearity range as well. The developed technique has been fully validated by ICH (International Council for Harmonisation) guidelines. Further, it has been evaluated for greenness character using the AGREE (Analytical GREEnness) statistical tool and the obtained score is 0.61 out of 1.

Bio-based polyester-polyurethane foams: synthesis and degradability by Aspergillus niger and Aspergillus clavatus.

In this article, the degradability by Aspergillus niger and Aspergillus clavatus of three bio-based polyurethane (PU) foams is compared to previous degradability studies involving a Pseudomonas sp. bacterium and similar initial materials (Spontón et al. in Int. Biodet. Biodeg. 85:85-94, 2013, https://doi.org/10.1016/j.ibiod.2013.05.019 ). First, three new polyester-polyurethane foams were prepared from mixtures of castor oil (CO), maleated castor oil (MACO), toluene diisocyanate (TDI), and water. Then, their degradation tests were carried out in an aqueous medium, and employing the two mentioned fungi, after their isolation from the environment. From the degradation tests, the following was observed: (a) the insoluble (and slightly collapsed) foams exhibited free hydroxyl, carboxyl, and amine moieties; and (b) the water soluble (and low molar mass) compounds contained amines, carboxylic acids, and glycerol. The most degraded foam contained the highest amount of MACO, and therefore the highest concentration of hydrolytic bonds. A basic biodegradation mechanism was proposed that involves hydrolysis and oxidation reactions.

Forced Degradation Studies for Glycopyrrolate, Formoterol &amp; Budesonide By UPLC

Forced degradation studies include the degradation of new drug substance and drug product at conditions more severe than accelerated conditions. These studies illustrate the chemical stability of the molecule which further facilitates the development of stable formulation with suitable storage conditions. ICH guidelines demonstrate certain degradation conditions like light, oxidation, dry heat, acidic, basic, hydrolysis etc.

Degradation study for 18650 NMC batteries at low temperature

This research focuses on the identification and quantification of lithium-ion battery degradation indicators at low temperatures. We studied the cycling aging of 18,650 commercial NMC lithium-ion batteries at 10 °C. For this purpose, we carried out life cycle tests and performed Galvanostatic Intermittent Titration Technique (GITT) tests in the voltage range for the charge and discharge processes for different States of Health (SoH). The diffusion time constant and the ohmic overpotential were determined from GITT for different SoH. As the batteries degrade both parameters increase. A degradation mechanism associated with faradaic effects (increase in ohmic drop overpotential) and a thermodynamic effect associated with changes in the active material (diffusional time constant) is observed. Furthermore, we performed Electrochemical Voltage Spectroscopy studies through Incremental Capacity (IC) curves. IC curves peaks and valleys are associated with battery phase transformations due to aging phenomena. Each peak has a unique peak height, area, and position associated with a degradation mode. This research focuses on the IC curves derived from the discharge capacities at 3A and OCP (Open Circuit Potential) to study the thermodynamic and faradaic effects separately. The peak at 3.6 V and 3.4 V for the 3A and OCP studies, respectively, is the main feature for the detection of the degradation mechanism. The valley in the OCP curves intensifies with the decrease in SoH and is shifted to lower potentials, an effect that is not observed in the 3A curves. So, we could associate this with thermodynamic effects, Loss of Lithium Inventory (LLI) and Loss of Active Material (LAM) of both the positive electrode and the negative electrode.

Determination of Nifedipine by Validated RP-HPLC Method in Bulk and Pharmaceutical Dosage Form

The present paper deals with the development and validation of reverse phase HPLC method for the determination of Nifedipine on Nucleosil 100, 5 μm, C8, 250 x 4.0 mm column. A mobile phase consisting of 40 ml 2-propanol: 60 ml phosphoric acid 0.85% was employed in this study. The flow rate was kept at 0.8 ml/min and the injection volume was 10 µl. The separation was performed at 40°C. Eluents were monitored by UV detector set at 237 nm. The developed method was statistically validated for the linearity, precision, robustness, specificity and solution stability. The specificity of the method was ascertained by force degradation studies by acid and alkali hydrolysis, oxidation, heat and photo degradation. The degraded products were well resolved from the analyte peak with significant differences in their retention time values.

Development and validation of stability indicating high‐performance thin‐layer chromatographic method for estimation of Rifapentine in pharmaceutical formulation

AbstractA simple, accurate, and precise method is described for stability indicating a high‐performance thin‐layer chromatographic (HPTLC) method for development and validation in the pharmaceutical dosage form. The method was based on HPTLC separation of the drug by measurement of the spot at 478 nm. Separation was carried out on silica gel 60GF254 by using mobile phase methanol:toluene:acetic acid (3:7:0.1, v/v/v). The linearity range was between 500–3000 ng/spot with a coefficient of determination 0.999, respectively. The retardation value found was 0.55 ± 0.02 and the % recovery was found to be 100.610 ± 0.035%–101.0178 ± 0.035%. The limit of detection value for Rifapentine was 92.901 ng/spot and the limit of quantitation for Rifapentine was found to be 281.52 ng/spot. Forced degradation studies were carried out under various conditions. In acidic conditions, the % degradation was found to be 20.07%. In alkaline and oxidative conditions, the % degradation was found to be 61.27% and 11.12%. Photolytic and thermal conditions the % degradation was found to be 49.75% and 8.35%. Maximum degradation was found in alkaline condition with 61.27%, an assay for the marketed formulation was carried out and the amount of drug was found to be 149 ± 0.05 mg with a % of 99.25% ± 0.005%.