Two novel ultraviolet spectrophotometric method for determination. Two methods for the assay of rifampicin in serum are described. The absorption or reflectance in the visible range directly affects the perceived color of the. There is a paucity of data on application of this test directly to clinical specimens, although based on a. Assay of rifampicin in bulk and its dosage forms by visible.
Uvspectrophotometric methods which are stabilityindicating were developed and validated for the determination of rifampicin in bulk drug, capsules and spiked human urine. This means it uses light in the visible and adjacent ranges. Most tests use the uv visible uv vis spectroscopy as the detection method, which usually falls into the wavelength range of 1001100 nm. Uvspectrophotometry9 for the assay of rif in formulations in. It is an aggressive disease with one third of the worlds population reportedly being infected with mtb. Jun 26, 2015 the intensity of the colorimetric assay performed on the paperbased platforms can be easily captured using a digital camera and subsequently analyzed. Spectral and absorbance measurements were made on shimadzu double beam spectrophotometer uv140 with matched 1cm quartz cells and ph measurements were carried out using on systronics ph meter 335. Ungraded products supplied by tci america are generally suitable for common industrial uses or for research purposes but typically are not. Biochemical characterization of the mycobacterium smegmatis. It is more common in developing countries, with more than half of the reported cases occurring in asia. Detection of residual rifampicin in urine via fluorescence.
Apr 28, 2017 on the other hand, simple spectrophotometric methods are available to assay rifampicin alone tella et al. Mycobacterium tuberculosis ribosomal protein s1 rpsa and. Rp hplc method development and validation for estimation of isoniazid and rifampicin in tablet dosage form dissertation submitted to the tamilnadu dr. Determination of proteinunbound, active rifampicin in. Rifampicinloaded mesoporous silica nanoparticles for the. The results showed that the addition of pva and pvp significantly improved drugloading, encapsulation efficiency and release characteristics. The mechanism of rifampicin inhibition of esche richia coli rna polymerase was studied with a newly developed steady state assay for rna chain initiation and by analysis of the products formed with several 5 terminal nucleotides. Encapsulation of zincrifampicin complex into transferrin. Rifampicin is a member of the class of rifamycins that is a a semisynthetic antibiotic derived from amycolatopsis rifamycinica previously known as amycolatopsis mediterranei and streptomyces mediterranei it has a role as an ec 2. For instance, lossoffunction mutants for a gene of interest can be tested using this assay.
Rifampicin and isoniazid were kindly donated by a pharmaceutical industry and used without further purification. The cell deathbased assay can be used to determine the involvement of a gene in pti. Experiment eight assay test for rifampicin capsules. Uvvis spectroscopy is usually applied to molecules and inorganic ions or complexes in solution. Original article formulation and characterization of rifampicinloaded p3hbco4hb nanoparticles amirah m. Obviously, the presence of rifampicin interfereswitht assay.
The antimycobacterial activity of the isolated piperine was tested against m. The chromatographic conditions were the same described on the usp xxii under rifampin assay conditions. Full text biodegradable rifampicinreleasing coating of. Development and validation of liquid chromatographymass. The stabilityindicating assay is a method that is employed for the analysis of stability of samples in pharmaceutical industry. Evaluation of the meltpro tbstr assay for rapid detection. The correction for the absorption of rifampicin at the emission wavelength trivial process or reabsorption was carried out by.
Sigmaaldrich offers a number of rifampicin products. Constant monitoring of rifampicin is important for controlling the side effects and preventing overdose caused by chronic medication. Simultaneous spectrophotometric determination of rifampicin. Simultaneous content analysis of rifampicin, isoniazid and.
In absorption spectroscopy a photon is absorbed by an atom or mol. The first is a conventional plate diffusion method, measuring concentrations down to 002. The assay results of samples 16 list of figures figure 1. An hplcuv method for determining rifampicin concentration in plasma using rifapentine as an internal standard was validated. Interpretation of the ir spectra of ethanol, acetone and. The polymeric core was synthesized in the following order.
The hplc chromatogram of uv stressed rif, inh and pza. The inhibitory effect of rifampicin on the amyloid formation of hen egg white lysozyme was assessed with both thioflavin t tht fluorescence assay and fourier transform infrared ftir difference spectroscopy. Rifampicin is active against grampositive bacteria but less active against gramnegative bacteria. Redoxreaction based spectrophotometric assay of isoniazid in. We designed a polymeric micelle to improve the delivery of antitb drugs rifampicin rf and isoniazid inh. A new approach to the use of uv spectrophotometry was used for pyrazinamide pza, rifampicin rif and isoniazid inh assay in fixeddose.
Packaging 1, 5, 25 g in glass bottle 250 mg in glass insert. A commercial line probe assay for the rapid detection of. Samples were withdrawn at predetermined time points and centrifuged at 38,000 rcf for 10 min. Interaction of rifampicin and isoniazid with large. College of pharmacy, madurai 625107, in partial fulfilment of the university rules and regulations for award of master of. To standardize a highperformance liquid chromatography hplc method for the deter.
Simultaneous determination of rifampicin, isoniazid, pyrazinamide and ethambutol in 4fdc tablet by raman spectroscopy associated to chemometric approach article mar 2017. Isolation, identification and antimycobacterial evaluation. Two spectrophotometric methods are described for the determination of isoniazid inh in pharmaceuticals. Validation of a simple hplcuv method for rifampicin. If the light is in the visible region, meaning the wavelength of 400700 nm or more broadly 360900 nm, the color of the assay can be visibly captured by naked eyes. Simultaneous estimation of rifampicin and isoniazid in. Spectra were automatically obtained by uv probe system software. Most tests use the uvvisible uvvis spectroscopy as the detection method, which usually falls into the wavelength range of 1001100 nm. Assessment of the inhibitory effect of rifampicin on. It is the measurement of the wavelength and intensity of absorption of near ultraviolet and visible light by a sample. The decreased fluorescence intensity of bsaau ncs in the presence of rifampicin allows for the sensitive detection of rifampicin in a range from 0.
In this study, we present an easy to use, effective. The degree of resistance varies depending on the site of mutation in the rna polymerase. Rphplc,7,20 hptlc8 and uv spectrophotometry9 and for the determination of piperine and rifampicin in individually submission date. Liquid chromatography mass spectroscopy, plasma, rifampicin rifampicin 34methyl piperazinyliminomethyl rifamycin is a firstline semisynthetic agent along with isoniazid and pyrazinamide in the treatment of pulmonary and extrapulmonary tuberculosis 1, 2. Simultaneous determination of isoniazid and rifampicin by. Highly luminescent and biocompatible p and n codoped. Assay results of inh and rif determination in pharmaceutical formulations n 6. Preparation of calibration curve of metformin 67 12. Original article formulation and characterization of rifampicin loaded p3hbco4hb nanoparticles amirah m. The assay was linear over the concentration range of 0. Assay of rifampicin in bulk and its dosage forms by. On the other hand, simple spectrophotometric methods are available to assay rifampicin alone tella et al. Assay for pathogenassociated molecular pattern pamp.
The mechanism of rifampicin inhibition of escherichia coli rna polymerase was studied with a newly developed steady state assay for rna chain initiation and by analysis of the products formed with several 5terminal nucleotides. We can divide spectroscopy into two broad classes of techniques. Stabilityindicating uvspectrophotometric assay of rifampicin. In the first method fcr method, inh is reacted with folinciocalteu reagent in na 2 co 3 medium and the resulting blue colored chromogen measured at 760 nm. Approximately 50% of multidrugresistant tuberculosis and over 90% of. Increases in rifampicin concentration increased the fluorescence quenching of the imbedded bsaau ncs in the waxprinted paper sensor. The fluorescence intensity of probe is linearly proportional to rifampicin over the concentration rage from 0 to 20. Zincalginate beads for the controlled release of rifampicin. Mar 26, 2018 the fluorescence intensity of probe is linearly proportional to rifampicin over the concentration rage from 0 to 20. Qabsorbance ratio spectrophotometric method for the. Assay of rifampicin in bulk and its dosage forms by visible spectrophotometry using chloranilic acid elisha divakar tella 1.
A digimed dm21v6 ph meter was used for the determination of the end point in the titration of inh. Biodegradable rifampicinreleasing coating of surgical meshes. In addition, the liver tissue of male mice treated with rifampicin showed a decrease. An hplc uv method for determining rifampicin concentration in plasma using rifapentine as an internal standard was validated. Rifampicin or rifampin r is a common drug used to treat inactive meningitis, cholestatic pruritus and tuberculosis tb, and it is generally prescribed for longterm administration under regulated dosages.
Biodegradable rifampicinreleasing coating of surgical. The homogeneity of rifampicin microparticleinhalac 230. The assay is based on the ultraviolet uv absorbance maxima at about 202 wavelength of gentamycin using water as solvent. A typical example of a straightforward tablet assay is the analysis of furosemide frusemide tablets.
Preparation of calibration curve of ciprofloxacin 64 10. Analysis procedure identification of rifampicin standard, isoniazid standard and pyrazinamide standard by fourier transform. An integral part of the strategy to fight the disease is use of quality antitb drugs. Rifampicin is used to treat tuberculosis and tuberculosisrelated mycobacterial infections. Each colorimetric assay was performed using a total volume of 30 l prepared bsaau ncs, and the evaluation was performed with a total of 30 l spiked urine containing different concentrations of rifampicin. Since simple uv spectrophometric method for the assay of rifampicin in combined dosage form is.
Because of the interference of rifampicin,t assay cannot be used as a reliable. Encapsulation of zincrifampicin complex into transferrinconjugated silver quantumdots improves its antimycobacterial activity and stability and facilitates drug delivery into macrophages. Materials and methods instruments lab india double beam uv visible spectrophotometer uv 3092 with 1 cm matched quartz cells were used for all absorbance measurements with uv win software. Two uvspectrophotometric methods which are stabilityindicating were developed and validated for the determination of rifampicin in bulk drug, capsules and spiked human urine. Ion mass spectrometry was monitored in positive ion that has a mass 823. Research article assessment of the inhibitory effect of. A rapid, specific and accurate high performance liquid chromatographic method for the determination of rifampicin rif by uv detection in human plasma was developed and validated. The major effect of rifampicin was found to be a total block of the translocation step that would ordinarily follow. The spectrophotometry uv with auc method can be used for simultaneous determination of rfp, inh and prz mixture.
Common pharmacopeial calculations in usp monographs. Uvspectrophotometry with its variants constitutes an evident alternative to the already existing methods. The potential of polymeric micelles constructed by coalescing natural and synthetic polymers for tuberculosis tb treatment was evaluated in this work. Chemical structure of rifampicin which absorbance of an aqueous solution. Chemometric assisted spectrophotometric methods for the. Unbound rifampicin was analyzed using ultra performance liquid chromatography with a beh c18 column, isocratic elution and ultraviolet uv detection. Evaluation of the meltpro tbstr assay for rapid detection of. Ultraviolet visible spectroscopy or ultraviolet visible spectrophotometry uv vis or uv vis refers to absorption spectroscopy or reflectance spectroscopy in part of the ultraviolet and the full, adjacent visible spectral regions.
We reveal that tht fluorescence assay gives a false positive result due to rifampicin interference, while ftir difference spectroscopy provides a reliable assessment. Each inh and rif can be assayed by ultraviolet spectrophotometry. Assessment of the inhibitory effect of rifampicin on amyloid. On the mechanism of rifampicin inhibition of rna synthesis. Two uv spectrophotometric methods which are stabilityindicating were developed and validated for the determination of rifampicin in bulk drug, capsules and spiked human urine. A validated highperformance liquid chromatography method for the determination of rifampicin and desacetyl rifampicin in plasma and urine a. Spectral and absorbance measurements were made on shimadzu double beam spectrophotometer uv 140 with matched 1cm quartz cells and ph measurements were carried out using on systronics ph meter 335. The method, which requires a simple protein precipitation procedure as sample preparation, was performed to compare venous and arterial plasma kinetics after a single therapeutic dose of rifampicin 8. These methods can provide high selectivity and good detection limits. Assay of formulations and % recovery data sample labelled amount mg amount found mg in method % recovery proposed method proposed reported rifampicin capsule tablet 150 150 148. The supernatant was diluted in methanol and quantified for rifampicin by uvvis spectroscopy as described above.
Determination binary mixture of isoniazide and rifampicin. Rifampicin in urine was detected by a simple plate diffusion assay using as the test organism a strain of staphylococcus aureus made resistant to other antituberculosis drugs. Multiwavelength spectrophotometric determination of. Besides, this probe can be also used to assay rifampicin in serum samples with satisfactory recoveries. Tuberculosis tb is one of the most important human bacterial diseases caused by mycobacterium tuberculosis mtb commonly affecting the lungs. Line probe assay has been shown to be highly sensitive and specific in the detection of rifampicinresistant tb when used on culture isolates. Uv spectrophotometric methods which are stabilityindicating were developed and validated for the determination of rifampicin in bulk drug, capsules and spiked human urine. The deficiency in delivery of proper dose of rifampicin has serious implications as it is known that doses of rifampicin. Rifampicin is a bactericidal antibiotic drug used to treat inactive meningitis and tuberculosis. Certificate this is to certify that the dissertation entitled rphplc method development and validation for estimation of isoniazid and rifampicin in tablet dosage form submitted by mr.
A validated highperformance liquid chromatography method for. Development and validation of high performance liquid. Either test a alone or any two of tests b, c and d may be applied. Five milligrams of rifampicinloaded msnp were suspended in 10 ml of pbs sink conditions and agitated at 37 c. Simultaneous determination of isoniazid and rifampicin by uv spectrophotometry article pdf available in farmacia 652. Wahab 1school of pharmaceutical sciences, universiti sains malaysia, 11800 penang, malaysia.
Common calculation for determination of the assay result chromatography. The assay results for the capsule by proposed method were. The blend of microparticles with inhalac 230 was prepared in ratio of 1. It interferes with the synthesis of nucleic acids by inhibiting dna dependent rna polymerase. In one class of techniques there is a transfer of energy between the photon and the sample. Versatile phresponsive chitosangpolycaprolactonemaleic. Ironii, formed as a result of reaction between inh and ironiii, is made to react with ferricyanide, and the. A shimadzu model 1700 japan double beam uv visible spectrophotometer with spectral width of 2 nm, wavelength accuracy of 0.
Three antituberculosis medications are investigated in this work consist of rifampicin, isoniazid and pyrazinamide. Pdf simultaneous determination of isoniazid and rifampicin. Medical university, chennai in partial fulfilment for the award of the degree of master of pharmacy in pharmaceutical analysis department of pharmaceutical analysis k. Rifampicin was extracted from plasma utilizing liquidliquid extraction process using a 70. Analysis procedure identification of rifampicin standard, isoniazid. Pdf simultaneous determination of isoniazid and rifampicin by uv. The purity of the compound was ascertained by uv, ftir studies and by differential scanning calorimetry dsc. Negative control reagent was used as notemplate control. Pdf one of the most effective antituberculosis treatments is the. Assay of rifampicin in serum journal of clinical pathology.
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