Analytical Method Development and Validation of Minocycline Hydrochloride in Bulk and Tablet Dosage forms using RP-HPLC Method

OBJECTIVES AND PLAN: In modern days usage of large number of drugs is increased to save the life of human beings, to control the spreading of large number of diseases around the world. A drug is having many physical and chemical properties that show variation in properties at various stages of bulk drug manufacturing, synthesis stage and formulation stage. These properties are estimated by qualitative and quantitative approach by using various number of new techniques. Modern pharmaceutical industries generally require precise analytical instruments at very low concentrations with various types of instruments like HPLC, LC-MS, UP-LC, GC, NMR and IR1. Using these instrumental techniques high resolution can be achieved. There is continuous change in techniques in the method development for various numbers of drugs for the quantification purpose are used presently. There are often a huge number of alternative methods for solving any analytical method development, but by understanding the advantages and limitations of the various tools we can choose the most appropriate instrumental method to achieve the limitations in sensitivity, precision, and accuracy. Various number of combination drug formulations are released in to the market to reduce the side effects and to minimize the drug toxicity. In these formulations resolution has to be done to determine the content of drug present in it. The advanced instruments like LC-MS and UP-LC are highly economic to develop the new methods used in industry. Many analytical techniques are used in the pharmaceutical industry to characterize a diversity of substances including active ingredients, impurities, counter-ions, excipients, extractable and process contaminants. HPLC is widely used in the pharmaceutical industry for quantitative analysis of a broad range of substances. While UV remains a primary detection technique, many analytes of interest (e.g., active pharmaceutical ingredient (API), impurities, counter-ions, excipients, extractables, and contaminants) lack a sufficient UV chromophore. Furthermore, the response obtained with UV and many other detectors is highly dependent on analyte properties. This posses significant challenges since individual standards are not always available, or are impractical, to use for calibration. Hence number of companies are mainly focusing on HPLC methods which are simple, precise, economical and efficient for the estimation purpose. The Validation of Analytical Procedures (Q2R1) issued by International Conference on Harmonization (ICH) requires that analytical test procedures should be fully validated and the analytical procedures should be investigated in order to ensure the quality of the drug substance or drug product2. The main aim of the present investigation is to develop a validated RP-HPLC-PDA method for the estimation of Minocycline Hcl in bulk, pharmaceutical dosage form, rapid, sensitive and cost effective method.7. SUMMARY: Minocycline hydrochloride is a broad-spectrum tetracycline antibiotic, chemically it is (4S,4aS,5aR,12aR)-4,7-bis(dimethylamino)-1,10,11,12a- tetrahydroxy-3,12-dioxo-3,4,4a,5,5a,6, 12,12a-octahydrotetracene-2-carboximidic acid hydrochloride. Minocycline passes directly through the lipid bilayer or passively diffuses through porin channels in the bacterial membrane. Tetracyclines like minocycline bind to the 30S ribosomal subunit, preventing the binding of tRNA to the mRNA-ribosome complex and interfering with protein synthesis. The aim of this present investigation is to develop an efficient, rapid and sensitive RP-HPLC- PDA method for the estimation of Minocycline hydrochloride in bulk, pharmaceutical dosage forms compatible for LC-MS methods and subsequent validation of the LC method as per ICH guidelines. METHOD VALIDATION:The specificity of the method was established by spiking solution of commonly used excipients. Peak purity tests were also carried out to show that the analyte chromatographic peak is not attributable to more than one component as the impurities are not available by purity index data, as there was no interference of impurities with the analyte peak. This shows that the peak of analyte was pure and excipients in the formulation did not interfere with the analyte.From the chromatograms and 3 D plots it can be inferred that there were no co eluting or interfering peaks where minocycline hydrochloride was eluted and the peak purity value greater than 0.9999 for minocycline hydrochloride this showed that the method was specific.Linearity studies were performed at 10-50μg/mL of Standard drug solution of minocycline hydrochloride by constructing graph between concentrations vs. peak areas. % RSD of peak areas were determined. Correlation coefficient was calculated, the data found within the acceptance criteria of 0.99.The result was found to be within limit so the method is linear over the concentration range of 10-50μg/mL of Minocycline hydrochloride.Precision of related substances were verified by repeatability. Repeatability was assessed by using a minimum of six determinations at 100 % of the test concentration (30μg/mL of minocycline hydrochloride) standard deviation and relative standard deviation were reported for precision.% RSD of area response of minocycline hydrochloride was 0.152 for system precision and 0.317 for method precision % RSD of retention times of minocycline hydrochloride was 0.055 for system precision and 0.061 for method precision % RSD should be NMT 2 for areas and NMT 1 for retention times. It indicates that the method is precise. CONCLUSIONS:Finally, it can be concluded that the proposed RP-HPLC-PDA method was validated fully as per the International Conference on Harmonization (ICH) Guidelines, and found to be applicable for routine quality control analysis for the estimation of Minocycline hydrochloride. The results of linearity, precision, accuracy and specificity proved to be within the limits. The method provides selective quantification of Minocycline hydrochloride without interference from blank, placebo and degradants. The proposed method is sensitive, reproducible, reliable, rapid, economical, and specific and LC-MS compatible, can be used for the estimation of Minocycline hydrochloride.