Current Issue : April - June Volume : 2018 Issue Number : 2 Articles : 11 Articles
RP-HPLC is the most powerful and dominant analytical technique available in todays world of pharmaceutical analysis. This review presented a step-by-step approach to RP-HPLC method development and its validation. Method development, validation and its life cycle management must incorporate in it the concept of QbD and QRM. Exhaustive chromatographic literature review and collecting maximum information on sample is very first prerequisite of method development. It further requires performing various experiments with sound judgement on selection of chromatographic conditions like mobile phase, stationary phase, column temperature, flow rate, detector and detection wavelength, injection volume, etc for method optimization. Further to fasten and simplify the development work latest automated method scouting techniques can be used. Gradient scouting is a powerful technique to screen between isocratic or gradient method. Validation of analytical procedure requires addressing various validation characteristics like accuracy, precision, specificity, linearity, range, LOD, LOQ, robustness. Statistical analysis of validation data shall be used to evaluate validation characteristics against predetermined acceptance criteria. Once the validation is successful the analytical method could be transferred to laboratories for routine application....
Andrographis paniculata (Family: Acanthaceae) locally known as Kiriyattu or Kalmegh, finds its use in various Ayurvedic formulations. Andrographolide is the major constituent extracted from the leaves of the plant which is a bicyclic diterpenoid lactone. Isolation and structural elucidation by spectral analysis of any plant active constituent from ethanolic extract of Andrographis paniculata with the aid of DPPH radical scavenging assay by column chromatography. Ethanolic extract was subjected to activity guided column chromatographic fractionation using methanol as eluent. 200 fractions of 5 ml each were collected and were screened for free radical scavenging activity using DPPH (2, 2 diphenyl 1-picryl hydrazyl) radical scavenging assay. The consecutive fractions showing highest similar activity were pooled to obtain pooled fractions (Pf), concentrated and named as Pf1-Pf10. Pfs were subjected to TLC (Thin-layer chromatography) and among all of these, Pf 10 gave a single spot. UV (Ultraviolet)-Visible spectra of Pf 10 showed a single peak at 324 nm. The FT-IR (Fourier transform Infrared) and NMR (Nuclear Magnetic Resonance as HNMR) Spectra of Pf 10 were carried out and the structure was elucidated using www.nmrdb.org software. UV-Visible �» max (H2O) at 324 nm and IR (KBr) cm-1 exhibited bands at 3242, 2920, 1734 and HNMR (CDCl3) �´ : 0.896, 0.904, 1.247, 6.425, 7.226, 7.229. By combining the spectral results and with the help of the software from www.nmrdb.org, which helps in revealing HNMR results from the structure, the structure was established as 19-dehydroxy 9,11- didehydro 3,14- dihydroxy methyl andrographolide....
A simple, novel, accurate, precise, linear, rapid and economical RP-HPLC method was developed for the estimation of abacavir. The chromatographic separation was achieved using a Grace Smart RP 18 (250 x 4.6 mm, 5 �¼) column and isocratic elution, a mobile phase comprising of Methanol: 0.005 mM 1-Octanesulphonic acid at pH 5.0 was adjusted with O-Phosphoric acid in the ratio of 60:40 (v/v). The flow rate was 1.0 ml/min with detection at 280 nm using a UV detector and drug eluted with retention time of 4.157 min. The calibration curves were linear (r�²=0.9994) in the concentration range of 2-10 �¼g/ml. The limit of detection and limit of quantitation were found to be 0.6070 and 1.821 �¼g/ml respectively. Thus the simple, novel, economical, accurate, precise and rapid RP-HPLC method was developed for estimation of abacavir and validated as per ICH guidelines. Hence the proposed method was successfully applied for the quantitative estimation of abacavir in pure and pharmaceutical dosage form....
A novel, simple, accurate and precise area under curve spectroscopic method was developed and validated for the estimation of abacavir in bulk and pharmaceutical dosage forms and has an absorption maximum at 291-301 nm in 0.1 N HCl. The linearity was found to be in the concentration range of 2-16 �¼g/ml and the correlation coefficient was found to be 0.999 and it has showed good linearity, reproducibility, precision in this concentration range. The regression equation was found to be Y = 0.047 X + 0.003. The % recovery values were found to be within 99.43-100.1 % showed that the method was accurate. The LOD and LOQ were found to be 0.0640 and 0.1920 mcg/ml, respectively. The % RSD values were less than 2. The present methods were accomplish the validation parameters according to ICH guidelines like accuracy, precision, linearity, range, ruggedness, limit of detection and limit of quantization etc. Proposed method was successfully applied for the quantitative estimation of abacavir in bulk and pharmaceutical dosage forms....
A simple, rapid, sensitive, robust, RP-HPLC analytical method was developed and validated for the analysis of urapidil in bulk and in pharmaceutical formulation. The development of proposed method was achieved on a LC - GC Qualisil BDS C18 column (250 mm-4.6 mm, 5 Ã?µm) by gradient mode at ambient temperature, employing a mobile phase as Methanol: Water (70:30 v/v) pH of the aqueous mobile phase was adjusted to 5.8Ã?± 0.02 with ortho-phosophoric acid at a flow rate of 1.0 ml/min and detection at 268 nm. Design of experiments employing ââ?¬Ë?Central Composite Designââ?¬â?¢ (CCD) and ââ?¬Ë?Response Surface Methodologyââ?¬â?¢ (RSM) were applied as advancement to traditional ââ?¬Ë?One Variable at Timeââ?¬â?¢ (OVAT) approach to evaluate the effects of variations in selected factors (methanol content, flow rate) as graphical interpretation for robustness and statistical interpretation was achieved with linear regression and ANOVA. The method succeeded over the validation parameters viz linearity, precision, accuracy, limit of detection and limit of quantitation and robustness. The method was applied efficiently for analysis of urapidil....
The objective of the work was to develop UV spectroscopic method for simultaneous estimation of pitavastatin calcium and metformin hydrochloride. Pitavastatin calcium and metformin hydrochloride have absorbance maxima (�»max) at 249 nm and 233 nm respectively. Beerâ��s law was obeyed in the concentration range 0.25-1.25 �µg/ml and 12.5-62.5 �µg/ml for pitavastatin calcium and metformin hydrochloride, respectively. Results of methods were validated statistically. Novel, simple, sensitive, rapid, accurate and economical spectrophotometric methods have been developed for simultaneous estimation of pitavastatin calcium and metformin hydrochloride....
A simple, sensitive and acute UV spectrophotometric method has been developed for the determination of alogliptin in bulk and pharmaceutical tablet dosage formulation. This method obeys Beerâ��s law in the concentration range of 3-18 �µg/ml with correlation coefficient of 0.9994 and absorption maximum at 272-282 nm in Distilled water. The percentage recovery of alogliptin ranged from 100.05 to 100.40 %. The developed method was validated for accuracy, precision, LOD, LOQ as per ICH guidelines. Statistical analysis proved that the developed method shows the % RSD within the acceptance limit. It was concluded that the developed method was simple, accurate, precise, reproducible, selective and it can be employed for the routine analysis of alogliptin in pure and tablet dosage form....
Cosmetics, products are repeatedly applied directly to the human skin, mucous membranes, should be safe for health. However, recently there has been increasing concern about their safety. Unfortunately, using these products in some cases is related to the occurrence of negative effects resulting from intentional or the accidental presence of chemical substances, including toxic metals. In present study, heavy metals like lead (Pb), cadmium (Cd), chromium (Cr), arsenic (As), mercury (Hg), zinc (Zn), nickel (Ni), manganese (Mn), copper (Cu) were quantitatively estimated by using Inductive couple plasma atomic emission spectroscopy in 21 samples of cosmetic products (like lipsticks, face wash, Face cream, eyeliners, kajal, mascara, foundation etc.) purchased from local market. The outcome results were indicating that the quantity of heavy metals in cosmetic preparation is out of limit or either in limit. The comparison was done with World Health organization, Food Drug Administration, International Purity and Specification guidelines and the permitted limits....
A novel, simple, accurate and precise area under curve spectroscopic method was developed and validated for the estimation of tenofovir disoproxil fumarate in bulk and pharmaceutical dosage forms and has an absorption maximum between 253-263 nm in 0.1 N H2SO4. The Linearity was found to be in the concentration range of 5-40 �¼g/ml and the correlation coefficient was found to be 0.9995 and it has showed good linearity, reproducibility, precision in this concentration range. The regression equation was found to be Y = 0.0246X + 0.0032. The % recovery values were found to be within 99.36 -100.28 % showed that the method was accurate. The LOD and LOQ were found to be 0.316 and 0.948 �¼g/ml, respectively. The % RSD values were less than 2. The method has been validated according to ICH guidelines for linearity, accuracy, precision, robustness, ruggedness. Limit of detection and limit of quantitation. Proposed method was successfully applied for the quantitative estimation of tenofovir disoproxil fumarate in bulk and pharmaceutical dosage form....
A simple, novel, accurate, precise, linear, rapid and economical RP-HPLC method was developed for the estimation of tenofovir disoproxil fumarate. The chromatographic separation was achieved using a phenomenex gemini-NX-5 C18 (250 x 4.6 mm, 5 �¼) column and isocratic elution, a mobile phase comprising of Methanol: 0.05mM Phosphate buffer (60:40 v/v) pH 5 was adjusted with O-Phosphoric acid. The flow rate was 1.0 ml/min with detection at 260 nm using a UV detector and drug eluted with retention time of 5.082 min. The calibration curves were linear (r�²=0.9990) in the concentration range of 10-50 �¼g/ml. The limit of detection and limit of quantitation were found to be 0.0616 and 0.1848 �¼g/ml respectively. Thus the simple, novel, economical, accurate, precise and rapid RP-HPLC method was developed for estimation of tenofovir disoproxil fumarate and validated as per ICH guidelines. Hence the method holds good for routine analysis tenofovir disoproxil fumarate in pure and pharmaceutical dosage form....
Herbal formulations are popular in worldwide due to safer and not causing any toxicity. Recently there have been reported herbal medicines adulterated with undeclared synthetic drugs. A simple, specific, accurate and isocratic RP-HPLC method was developed and subsequently validated for determination of adulterant. Separation was achieved with a Phenomax Luna CÃ?¬18 (250 Ã?â?? 4.5 mm) 5 Ã?µm column with a mobile phase comprising phosphate buffer: Acetonitrile pH4 adjusted with 0.1% ortho phosphoric acid in the volume ratio of 45:55% v/v was developed. The detection was carried out using a photo diode array detector set at a wavelength of 254 nm. The method was validated as per ICH guideline demonstrating the accuracy and precision within corresponding linearity range of 10 Ã?µg/ml to 60 Ã?µg/ml and get the correlation coefficient of paracetamol, diclofenac sodium and aceclofenac were found to be 0.995, 0.997 and 0.995, respectively. The developed method was validated with respect to linearity, accuracy, precision, limit of detection and quantification and robustness. The method can be used for quality control of herbal formulation....
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