Background:: Standardization of Triphala Juice was performed by using the WHO Guidelines. The Parameters included Preliminary Analysis, Phytochemical Identification, Heavy Metal Estimation, etc. A new simple, specific, precise and accurate UV Spectrophotometric, High-Performance Liquid Methods: Chromatography and High-Performance Thin Layer Chromatography method has been developed for the Estimation of Gallic Acid in pure form. The UV- Spectrophotometric method was developed using Schimadzu 1800 UV - Visible spec-trophotometer using methanol as a solvent. The method was shown to be linear, with a detection wavelength of 273 nm for Gallic Acid. The separation was achieved on the Schimadzu Prominence-I RP-HPLC and the column used was C18 column using mobile phase consisting of mixture of Methanol: 0.1% OPA (50:50). The detection was carried out at 280 nm with a flow rate of 0.7ml/min. The retention time for Gallic Acid was found 3.89 minutes. The calibration curve was found linear (r2 = 0.999) for RP- HPLC method. The HPTLC method was developed using Aetron Sprayline instrument, Methanol as solvent and mobile phase consisting of Toluene: Ethyl Acetate: Formic Acid: Methanol (3:3:0.9:0.2). The method was found linear and the wavelength of detection for Gallic Acid was 254nm, respectively. The percentage recoveries for both methods were found in the 98.0- 102.0% range. Results:: The methods were validated in accordance with International Conference on harmoniza-tion acceptance criteria for specificity, linearity, precision, accuracy, robustness and system suita-bility. The excipients did not interfere in the determination of Gallic acid in Triphala Juice. Conclusion:: The suggested approach was effectively implemented for the quantitative determina-tion of gallic acid in Triphala juice, which would aid in quality control
Background:: There are very few methods for simultaneously determining a combined dose of SAL and KET. Objectives:: The current study aims to explore accurate, precise, simple, and cost-effective HPLC and HPTLC techniques for the simultaneous assessment of Salbutamol (SAL) and Ketotifen (KET). Methods:: The determination of Salbutamol and Ketotifen was performed by HPLC and HPTLC methods using 280 nm and 258 nm as the determination wavelength, respectively. Methanol was used to dissolve the drug for estimation in HPLC using mobile phase methanol: 10mM di-Potassium hydrogen orthophosphate in the ratio of 55:45 v/v of pH 4 at a flow rate of 1mL/min and in chloroform: toluene: methanol (7: 2: 3 v/v/v) for the estimation in HPTLC. Moreover, a statistical comparison was made between the results obtained through HPLC and HPTLC of Salbutamol (SAL) and Ketotifen (KET) using the Student’s t-test and F-test. Results:: A linear response was observed in the range of 4-24 μg/mL and 2-12 μg/mL, respectively, for SAL and KET for HPLC. R2 was found to be 0.9998 and 0.9999, respectively. For HPTLC, the linear response was observed in the concentration range of 20-120 ng/ spot and 10 - 60 ng/ spot for SAL and KET, respectively. R2 was found to be 0.9988 and 0.9998, respectively. The limit of detection (LOD) for HPLC was estimated as 0.34 μg/ml and 0.10 μg/ml for SAL and KET, respectively, and for the HPTLC method, the LOD was estimated as 4.8 μg/ml and 1.5 μg/ml, respectively. Analysing the marketed formulation by using both methods, SAL and KET within the range of 100 ± 2% were recovered. The results obtained after the estimation of the Mastifen S tablet by applying both methods were according to nominal content. Degradation studies were performed using both methods. It was found that Salbutamol was unstable in hydrolytic, oxidative and thermal degradation, whereas stable in photolytic conditions. Ketotifen was found to be stable in thermal and photolytic conditions and unstable in hydrolytic and oxidative conditions. Conclusion:: The proposed stability indicating HPLC and HPTLC methods for SAL and KET was found to be simple, accurate, and reproducible for quantitative estimation in pharmaceutical dosage form, without interference from the excipients or degradation products from the main drug component.
Background: Kutki, the dried rhizome of Picrorhiza kurroa Royle ex Benth belonging to the family of Scrophulariaceae, has been utilized globally for liver ailments. Objective: Comprehensive use of kutki needs to evaluate its role as a quality control tool for dis-crimination of kutki samples, and therefore an effective HPLC fingerprinting method was estab-lished. Methods: Reverse-phase high-performance liquid chromatography with photodiode array (RP-HPLC-PDA) detection method coupled with multivariate analysis was developed, which was mod-est, consistent and, accurate for classification of 11 kutki samples including authentic Picrorhiza kurroa rhizomes from the market of Ahmedabad and Gandhinagar in Gujarat, India. Results: The method was validated for various parameters like precision, reproducibility, and stabil-ity. The lowest value of the % relative standard deviations (RSD) was 1.31%. Chromatographic fin-gerprint profiles of 11 kutki samples, including authenticated samples, were obtained by this meth-od, which showed a total of 28 peaks, and 9 peaks were important among them. Chemometric tech-niques like PCA and HCA were applied to identify the kutki samples.The Samples of kutki could be exquisitely differentiated into two clusters. Conclusion: HPLC-PDA method coupled with multivariate analysis divulged that chromatographic fingerprint analysis was reliable and effective for quality assessment and discrimination of kutki samples.
Background: Abiraterone acetate is a derivative of steroidal progesterone, used as a first-line therapy for metastatic castration of prostate cancer. Objective: The present study encompasses the design of an experiment approach for developing a simple, reliable, and rapid RP-HPLC method for the estimation of abiraterone acetate. Method: The chromatographic separation was efficiently conducted on a Hypersil Gold C18 (50 x 4.6 mm, 5 µm) HPLC column, using the mobile phase composition of acetonitrile: dibasic potassium phosphate (0.01 mM) in the ratio of 80:20 (%v/v) at pH 6.5 with an isocratic elution mode. Furthermore, the different force degradation study including hydrolysis, oxidation, thermal, and photolytic was performed for abiraterone acetate. Result: The dynamic linearity was established in the concentration range of 0.5-10 µg/mL with r2 of 0.9998. Furthermore, the limit of detection and the limit of quantitation were 0.0978 µg/mL and 0.3260 µg/mL. The degradation of abiraterone acetate was shown in both acidic (54.16 ± 0.247 after 24 hrs) and basic conditions (35.06 ± 0.458 after 24 hrs). Furthermore, the developed method was successfully employed to quantify abiraterone acetate in bulk powder and the solid dispersion did not show any change in the retention time. Conclusion: The developed method was validated according to the ICH Q2 (R1) specification, which was found to be sensitive, accurate, precise, robust, linear, and selective compared to the reported chromatographic method.
Background: The 6-mercaptopurine is an active ingredient used to treat certain types of leukemia. This drug is an immunosuppressive and antineoplastic agent that belongs to the thiopurine class. In Brazil, 6-MP is currently available only in the form of 50 mg tablets, sold as Purinethol® and manufactured by Glaxo Smith Kline. The lack of the liquid formulation’s production impedes treatment, assuming that one of its advantages is through its applicability in pediatric patients, who show the highest incidence among others. Objective: The purpose of this work was to evaluate the development and application of a reversed phase high performance liquid chromatography (HPLC) method using an Agilent 1220 Infinity® G4294B chromatograph with photodiode array detector. Methods: HPLC assays were performed on an Eclipse plus® C18 column (4.6 x 150 mm, 3.5 μm particle size) using a gradient mode mixture of acetonitrile and aqueous acetic acid solution as a mobile phase, with a flow of 1 mL.min-1 and detection at 324 nm. The method was validated by determining its selectivity, linearity, precision, accuracy, and robustness. Results: Retention time for 6-mercaptopurine was 5.12 minutes. The detector’s response was linear at concentrations from 1.6 to 2.4 μg/mL. The results of the method’s accuracy evaluation showed mean recovery of the amount of substance added to the samples of between 99.88 and 100.5%. For precision, repeatability and intermediate precision were evaluated. The repeatability showed a standard deviation of 0.0737. The intermediate precision was assessed on three different. For three days of the studies, the values of the standard deviations were less than 3%, showing repeatability and intermediate precision adequate for the analytical method in question. The limit of detection was determined as 3.6 ng/mL. The limit of quantification was determined as 12 ng/mL. The chromatographic method was robust. Conclusion: The proposed method can be applied to control the quality of 6-MP oral suspension to ensure that the required content is delivered to pediatric oncology patients.
