Stability indicating analytical method development and validation for estimation of Ceftazidime and Avibactam in bulk and pharmaceutical dosage form using RP-HPLC
Stability indicating analytical method development and validation for estimation of Ceftazidime and Avibactam in bulk and pharmaceutical dosage form using RP-HPLC
The drug or drug combination may not be official in any pharmacopoeias.
A proper analytical procedure for the drug may not be available in the literature due to patent regulations.
Analytical methods may not be available for the drug in the form of a formulation due to the interference caused by the formulation excipients.
Analytical methods for the quantitation of the drug in biological fluids may not be available.
Analytical methods for a drug in combination with other drugs may not be available.
The existing analytical procedures may require expensive reagents and solvents. It may also involve cumbersome extraction and separation procedures and these may not be reliable.
Analytical method validation as per ich and usp shreyas B R
Analytical method validation is a process of documenting/ proving that an analytical method provides analytical data acceptable for the intended use.After the development of an analytical procedure, it is must important to assure that the procedure will consistently produce the intended a precise result with high degree of accuracy. The method should give a specific result that may not be affected by external matters. This creates a requirement to validate the analytical procedures. The validation procedures consists of some characteristics parameters that makes the method acceptable with addition of statistical tools.
Introduction to Dissolution equipment's, Calibration of dissolution apparatus, Dissolution procedure development and validation, Dissolution method development for generic drug products.
To recommend acceptable amounts for residual solvents in pharmaceuticals for the safety of the patient. The guideline recommends use of less toxic solvents and describes levels considered to be toxicologically acceptable for some residual solvents.
The guideline applies to all dosage forms and routes of administration.
This guidelines does not address all possible solvents, only those identified in drugs at that time, neither address solvents intentionally used as excipients nor solvates.
The maximum acceptable intake per day of residual solvent in pharmaceutical products is defined as “permitted daily exposure” (PDE)
Previously, another terms were used like “Tolerable daily intake” (TDI) & “Acceptable daily intake” (ADI) by different organization & authorities, but now usually this new term “PDE” is used
Stability indicating analytical method development and validation for estimat...SriramNagarajan18
Stability indicating analytical method development and validation for estimation of Sacubitril and Valsartan in bulk and pharmaceutical dosage form using RP-HPLC
Traditional Kashmiri Recipe “Shangri-Kahwa” as a Stimulant Drink and Effectiv...SriramNagarajan15
The popular recipe “Shangri-kahwa” is an age old home remedy for respiratory and various other problems in almost whole of Kashmir. It is prepared from important spices like liquorice, clove, cinnamon, and cardamom, which have documented health benefits. Information about its use and method of preparation was obtained from group discussions held in some villages of Baramullah district of Jammu and Kashmir. People in these villages believe that Shangri-kahwa is cost effective, delicious, made from easily available ingredients and can be prepared easily at home. Being residents of this area, the authors are aware of the popularity of this magical drink used as a first line of treatment for various ailments at home, particularly during cold days. This recipe is extremely famous in these villages both as a refreshing and stimulant drink, as well as believed to be highly efficacious in respiratory illnesses. It is cost effective and highly palatable. The ingredients of Shangri-kahwa are being used extensively in Unani system of medicine and Ayurveda for almost same indications as the recipe is used. This study was carried out to highlight the effectiveness and focus the attention of the researchers towards this attractive and effective dosage form used as home remedy in Kashmir.
The drug or drug combination may not be official in any pharmacopoeias.
A proper analytical procedure for the drug may not be available in the literature due to patent regulations.
Analytical methods may not be available for the drug in the form of a formulation due to the interference caused by the formulation excipients.
Analytical methods for the quantitation of the drug in biological fluids may not be available.
Analytical methods for a drug in combination with other drugs may not be available.
The existing analytical procedures may require expensive reagents and solvents. It may also involve cumbersome extraction and separation procedures and these may not be reliable.
Analytical method validation as per ich and usp shreyas B R
Analytical method validation is a process of documenting/ proving that an analytical method provides analytical data acceptable for the intended use.After the development of an analytical procedure, it is must important to assure that the procedure will consistently produce the intended a precise result with high degree of accuracy. The method should give a specific result that may not be affected by external matters. This creates a requirement to validate the analytical procedures. The validation procedures consists of some characteristics parameters that makes the method acceptable with addition of statistical tools.
Introduction to Dissolution equipment's, Calibration of dissolution apparatus, Dissolution procedure development and validation, Dissolution method development for generic drug products.
To recommend acceptable amounts for residual solvents in pharmaceuticals for the safety of the patient. The guideline recommends use of less toxic solvents and describes levels considered to be toxicologically acceptable for some residual solvents.
The guideline applies to all dosage forms and routes of administration.
This guidelines does not address all possible solvents, only those identified in drugs at that time, neither address solvents intentionally used as excipients nor solvates.
The maximum acceptable intake per day of residual solvent in pharmaceutical products is defined as “permitted daily exposure” (PDE)
Previously, another terms were used like “Tolerable daily intake” (TDI) & “Acceptable daily intake” (ADI) by different organization & authorities, but now usually this new term “PDE” is used
Stability indicating analytical method development and validation for estimat...SriramNagarajan18
Stability indicating analytical method development and validation for estimation of Sacubitril and Valsartan in bulk and pharmaceutical dosage form using RP-HPLC
Similar to Stability indicating analytical method development and validation for estimation of Ceftazidime and Avibactam in bulk and pharmaceutical dosage form using RP-HPLC
Traditional Kashmiri Recipe “Shangri-Kahwa” as a Stimulant Drink and Effectiv...SriramNagarajan15
The popular recipe “Shangri-kahwa” is an age old home remedy for respiratory and various other problems in almost whole of Kashmir. It is prepared from important spices like liquorice, clove, cinnamon, and cardamom, which have documented health benefits. Information about its use and method of preparation was obtained from group discussions held in some villages of Baramullah district of Jammu and Kashmir. People in these villages believe that Shangri-kahwa is cost effective, delicious, made from easily available ingredients and can be prepared easily at home. Being residents of this area, the authors are aware of the popularity of this magical drink used as a first line of treatment for various ailments at home, particularly during cold days. This recipe is extremely famous in these villages both as a refreshing and stimulant drink, as well as believed to be highly efficacious in respiratory illnesses. It is cost effective and highly palatable. The ingredients of Shangri-kahwa are being used extensively in Unani system of medicine and Ayurveda for almost same indications as the recipe is used. This study was carried out to highlight the effectiveness and focus the attention of the researchers towards this attractive and effective dosage form used as home remedy in Kashmir.
RP-HPLC Assay Method Validation for the estimation of new Anti-retroviral dru...SriramNagarajan15
A Reverse phase HPLC method was developed for estimation of the Lamivudine in bulk and tablet formulation by using ODS column (250mm×4.6mm, 5µm) and Acetate buffer: acetonitrile (50:50) as mobile phase, at a flow rate of 1.5ml/min. The detection was carried at the 272nm the retention time of the Lamivudine is 1.850. The developed method was validated for the various parameters as per the ICH guidelines like accuracy precision, linearity and range, Robustnes. Linearity was obtained in the concentration range of 10µg/ml to 50µg/ml with correlation coefficient of 0.999. The accuracy of the method was assessed by recovery studies at three different concentration levels. The percentage recovery of Lamivudine was found to be in the range of 98% -102%. The method was found to be precise as indicated by the repeatability, inter-day, intra-day analysis, showing %RSD less than 2. Key words: RP-HPLC, Lamivudine, Pharmaceutical dosage form.
RP-HPLC Assay Method Validation for the estimation of new Anti-retroviral dru...SriramNagarajan15
A Reverse phase HPLC method was developed for estimation of the Lamivudine in bulk and tablet formulation by using ODS column (250mm×4.6mm, 5µm) and Acetate buffer: acetonitrile (50:50) as mobile phase, at a flow rate of 1.5ml/min. The detection was carried at the 272nm the retention time of the Lamivudine is 1.850. The developed method was validated for the various parameters as per the ICH guidelines like accuracy precision, linearity and range, Robustnes. Linearity was obtained in the concentration range of 10µg/ml to 50µg/ml with correlation coefficient of 0.999. The accuracy of the method was assessed by recovery studies at three different concentration levels. The percentage recovery of Lamivudine was found to be in the range of 98% -102%. The method was found to be precise as indicated by the repeatability, inter-day, intra-day analysis, showing %RSD less than 2. Key words: RP-HPLC, Lamivudine, Pharmaceutical dosage form.
A novel validated stability Indicating RP-HPLC Method Development for the est...Naveen Chennamaneni
Best reserch paper A novel validated stability Indicating RP-HPLC Method Development for the estimation of Certinib in its bulk and finished Dosage form as per ICH Guidelines
Development and Validation of Reversed Phase-High-Performance Liquid Chromato...BRNSS Publication Hub
A simple, accurate, precise, and robust in vitro methods developed and validated for measurement of drug release in Aminocaproic Acid tablets. High-performance liquid chromatography (HPLC) method for quantification of drug in dissolution samples of Aminocaproic Acid tablet is developed and validated. 0.1 N Hydrochloric acid is used as dissolution medium and Basket (USP-I) as apparatus at 100 rpm. The sample was withdrawn after 60 min. The developed HPLC method was used for quantitative estimation of drug release in dissolution samples of Aminocaproic Acid tablet. Chromatogram was run through Inertsil ODS 3V, (250 × 4.6 mm), 5 μm. Mobile phase containing buffer solution and methanol in the pumped through column at a flow rate of 1 ml/min. Buffer used in this method was 13.3 g sodium dihydrogen phosphate monohydrate, 500 mg of Heptane-1-sulfonic acid sodium salt, and 1.0 mL of Triethylamine buffer with pH 2.20 adjusted by orthophosphoric acid. Optimized wavelength for Aminocaproic acid was 210 nm. Retention time of Aminocaproic acid was found about 4.0 min; linearity range was 132.605 μg/ml–828.787 μg/ml. The new method was evaluated according to ICH guideline and as far as validation results are concern correlation coefficient value was 0.9999 for the very compound, percentage recovery 100.0%, repeatability results relative standard deviation 0.9 for Aminocaproic acid. The developed HPLC method was found to be a simple and rapid one for regular analysis in professional laboratory.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
A newly validated HPLC method development for simultaneous estimation of rito...SriramNagarajan19
The aim of the present work was to develop a isocratict RP-HPLC for simultaneous analysis of ritonavir and lopinavir in tablet dosage form. Method: chromatographic system was optimized using a Agilent XDB C18(150 x 4.6mm,5µm) column with potassium dihydrogen phosphate (pH 4.6) and acetonitrile in the ratio of 45;55, as a mobile phase, at a flow rate of 1.0 ml/min. detection was carried out at 215nm by a photodiode array detector. Result: ritonavir and lopinavir were eluted with retention times of 4.821 and 3.814mins respectively. Beer’s lambert’s law was obeyed over the concentration ranges of 12.5 to 50µg/ml and 50 to 200µg/ml for ritonavir and lopinavir, respectively. Conclusion: the high recovery and low coefficients of variation confirm the suitability of the method for simultaneous analysis of both drugs in a tablet dosage form. Statistical analysis proves that the method is sensitive and significant for the analysis of ritonavir and lopinavir in pure and in pharmaceutical dosage form without any interference from the excipients. The method was validated in accordance with ICH guidelines. Validation revealed the method is specific, rapid, accurate, precise, reliable, and reproducible.
Stability indicating analytical method development and Validation for Isoniaz...SriramNagarajan18
Stability indicating analytical method development and Validation for Isoniazide, Thiacetazone and Pyridoxine by RP-HPLC UV method
Similar to Stability indicating analytical method development and validation for estimation of Ceftazidime and Avibactam in bulk and pharmaceutical dosage form using RP-HPLC (20)
Stability indicating method and validation for the simultaneous estimation of...SriramNagarajan18
Stability indicating method and validation for the simultaneous estimation of metformin and empagliflozin by using RP-HPLC in a bulk and pharmaceutical dosage forms
Stability indicating method development and validation for the estimation of ...SriramNagarajan18
Stability indicating method development and validation for the estimation of Doxorubicin by using RP-HPLC method in a bulk and pharmaceutical dosage form
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
Follow us on: Pinterest
Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
There are a number of conditions that present acutely, predominantly with pain and/or swelling
A careful and detailed history and examination, and in some cases, investigations allow differentiation between these diagnoses. A prompt diagnosis is essential as the patient may require urgent surgical intervention
Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
New Drug Discovery and Development .....NEHA GUPTA
The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
These lecture slides, by Dr Sidra Arshad, offer a quick overview of physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar leads (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
Stability indicating analytical method development and validation for estimation of Ceftazidime and Avibactam in bulk and pharmaceutical dosage form using RP-HPLC
1. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
70
International Journal of Farmacia
Journal Home page: www.ijfjournal.com
Stability indicating analytical method development and validation for
estimation of Ceftazidime and Avibactam in bulk and pharmaceutical dosage
form using RP-HPLC
Syeda Saniya Fatima1*
, R. Vani2
Deccan School of Pharmacy, Aghapura, Dar-Us-Salam, Hyderabad, Telangana
Corresponding Author: Syeda Saniya Fatima
Email: ainas_tremendous.queen@yahoo.co.in
ABSTRACT
A new simple, rapid, specific, accurate and precise stability indicatind RP-HPLC method has been developed for the
estimation of Ceftazidime and Avibactam in bulk & pharmaceutical dosage form. From UV spectrophotometric
method selected wavelength for estimation of drugs were 231 nm as isobestic point and 256 nm, 210 nm as λ max of
CAZ and AVY respectively by using methanol as a solvent. RP-HPLC method was developed by using Hypersil
ODS (150×4.6mm) 5µ. The samples were analyzed by using mixed phosphate buffer (PH
adjusted to 4 using
orthophosphoric acid): Acetonitrile (60:40 % v/v) as the mobile phase at the flow rate of 1.0 ml/min and detection
wavelength is 231 nm. Both the drugs were eluted within 5 minutes and gave sharp peaks with high theoretical plate
count and low tailing factor. The retention time for Ceftazidime and Avibactam was found to be 2.523 and 4.410
min respectively. Calibration curve was linear with correlation coefiicient of 0.996 and 0.999 over a concentration
range of 240-560 µg/ml and 60-120 µg/ml for Ceftazidime and Avibactam respectively. The percent recovery was
100.10 and 99.75 for Ceftazidime and Avibactam respectively indicating accuracy and reliability of the method.
Forced degradation studies were carried out and drug peaks were well resolved without any significant degradation
products when subjected to stress conditions. So the developed stability indicating method could be utilized for
routine analysis of Ceftazidime and Avibactam in bulk and pharmaceutical dosage form.
Keywords: Ceftazidime, Avibactam, Stability indicating, UV, RP-HPLC.
INTRODUCTION
Avibactam (AVIB) is chemically named as
(1R,2S,5R)-2-Carbamoyl-5-methyl-7- oxo-1,6-
diazabicyclo [3.2.1]octan-6-yl hydrogen sulphate.
Avibactam is a non β lactam β-mlactamase inhibitor
antibiotic used for treating complicated urinary tract
and complicated intraabdominal infections caused by
antibiotic resistant-pathogens, includind those caused
by multi-drug resistant gram negative bacterial
pathogens.[4,5] Ceftazidime (CEF) is chemically
named as (7R,Z)-7-(2-(2-aminothizol-4yl)-2- (2-
carboxypropan-2-yloxyimino) acetamido)-8-oxo-3-
(pyridinium-1-ylmethyl)-5-thia-1aza bicycle[4.2.0]oct-
2-ene-2-carboxylate. [1-3]
Ceftazidime is a semi synthetic broad spectrum
beta lactam antibiotic for parentral administration.
Ceftazidime is bactericidal in action exerting its effect
by inhibition of enzymes responsible for cell-wall
synthesis, primarily penicillin binding protein 3
(PBP3). It is a third generation cephalosporin. As a
class cephalosporin’s have activity against gram-
positive and gram-negative bacteria. The balance of
activity tips toward gram positive organisms for earlier
generations; later generations of cephalosporin’s have
2. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
71
more gram-negative coverage. Ceftazidime is one of
the few in this class with activity against
pseudomonas. It is not active against methicillin-
resistant Staphylococcus aureus. [4-6] The review of
literature revealed that several analytical methods have
been reported for ceftazidime in spectrophotometry,
HPLC, HPTLC, LC/MS individually and in the
combination with other drugs. [6-12] To date there
have been no published reports about the stability
indicating method for estimation of avibactam and
ceftazidime by HPLC in bulk and in pharmaceutical
dosage forms. This present study report for the first
time stability indicating simultaneous estimation of
avibactam and ceftazidime by RP-HPLC in bulk drug
and in pharmaceutical dosage form.
Fig 1: Structure of Ceftazidime Fig 2: Structure of Avibactam
MATERIALS AND METHODOLOGY
Chemicals
Avibactam and ceftazidime were obtained as gift
sample from Chandra Pharma Research Laboratory in
Hyderabad and marketed formulation was purchased
from local market.
Instrument and chromatographic condition
All solvent used in this work are HPLC & AR
grade. Instrument and chromatographic condition RP-
HPLC Agilent 1220 infinity series separation model
equipped with UV Detector was employed in this
method. The EZchrom software was used for LC peak
integration along data acquisition and data processing.
The column used for separation of analyte is Hypersil
ODS (150×4.6mm) 5µ. Mobile phase consisting of
phosphate buffer: Acetonitrile in the ratio of 60:40 %
v/v at flow rate 1 ml/min. it was filter through 0.45 μm
nylon filter and sonicated for 5 min in ultrasonic bath.
Sample was analyzed at 231 nm at an injection volume
of 20μl.
Preparation of phosphate buffer
1.625 gm of Potassium Di Hydrogen ortho
phosphate and 0.3 gms of Di Potassium Hydrogen
ortho phosphate was weighed and dissolve in 100 ml
of water and volume was made up to 550 ml with
water. Adjust the PH
using ortho phosphoric acid. The
buffer was filtered through 0.45µ filters to remove all
fine particles and gases.
Preparation of Solutions
Preparation of Mixed Standard Solution:
(400µg/ml & 100µg/ml)
Weigh accurately 400 mg of Ceftazidime & 100
mg of Avibactam and dissolve in 100 ml of diluents.
From the above stock solution 400 µg/ml of
Ceftazidime and 100 µg/ml of Avibactam is prepared
by diluting 1 ml to 10 ml with mobile phase. This
solution (400 µg/ml of Ceftazidime & 100 µg/ml of
Avibactam) is used as working standard concentration
for estimation of IV infusion.
Preparation of Sample Solution
5 bottles were weighed & calculate the average
weight of each bottle then the weight equivalent to 1
bottle was transferred into a 500 ml of volumetric
flask. Add sufficient quantity of diluents & sonicated
for 25 minutes further the volume is made up with
diluents and filtered. From the filtered solution 1 ml
was pipette out into a 10 ml with diluents.
3. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
72
Table 1: Summary of Chromatographic conditions
S. No Parameter Description
1. Column ODS Hypersil C-18 (150×4.6mm) 5µ
2 Mobile Phase & PH
Phosphate Buffer : Acetonitrile PH
4, 60:40 v/v
3 Flow rate 1 ml/min
4 Column & sample temperature Room temperature (20-25o
C)
5 Detection Wavelength 231 nm
6 Detector UV
7 Injection Volume 20 µl
8 Retention time Ceftazidime – 2.523 Min
Avibactam - 4.410 Min
9. Run time 10 mins
Fig 3: Typical chromatogram of Avibactam and Ceftazidime
Method Validation
The validation of method was carried out as per
ICH guideline. The parameters assessed were
specificity, linearity, precision, accuracy, stability,
LOD and LOQ. Specificity is the ability of the
analytical method to measure the analyte response in
the presence of interferences including degradation
product and related substances.
Accuracy
The accuracy was determined by calculating %
recoveries of avibactam and ceftazidime. It was
4. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
73
carried out by adding known amount of each analyte
corresponding to three conc. Levels (80, 100, and 120)
of the label claims to the excipients. At each level, six
determinations were performed and the accuracy
results were expressed as percent analyte recovered by
proposed method.
Precision
Method precision
Precision of an analytical method is usually
expressed as the standard deviation. Method precision
was demonstrated by preparing six samples as per test
method representing single batch and were
chromatographed. The precision of the method was
evaluated by computing the %RSD of the results. The
individual values and the low % RSD observed on the
values are within acceptance criteria and indicates that
method is precise.
Linearity
The purpose of the test for linearity is to
demonstrate that the entire analytical system
(including detector and data acquisition) exhibits
linear responses and directly proportional over the
relevant conc. Range for the target conc. of the
analyte. The linear regression data for the calibration
plot is the indicative of a good linear relationship
between peak and concentration over wide range. The
correlation coefficient was indicative of high
significance.
Robustness
Robustness of method was investigated under a
variety of conditions including changes of
composition of buffer in the mobile phase, flow rate
and temperature. This deliberate change in the method
has no effect on the peak tailing, peak area and
theoretical plates finally the method was found to be
robust.
Ruggedness
The ruggedness of the method was studied by the
determining the analyst to analyst variation by
performing the Assay by two different analysts. %
RSD Assay values between two analysts not greater
than 2.0%, hence the method was rugged.
Limit of Detection and Limit of Quantitation
The LOD can be defined as the smallest level of
analyte that gives a measurable responses and LOQ
was determined as the lowest amount of the analyte
that was reproducibly quantified. These two
parameters were calculated using formula based on
standard deviation of the response and slope. LOD and
LOQ were calculated by equation, LOD=3.3 x ϭ/s and
LOQ= 10 x ϭ/s, where s = standard deviation, S =
slope of calibration curve.
Assay of avibactam and ceftazidime in
pharmaceutical dosage form
Assay of marketed product was carried out by
using the developed method. Sample solutions were
prepared and injected into RP –HPLC system. The
sample solution was scanned at 231 nm. The
percentage drug estimated was found to be 100.23%
and 101.21% respectively as ceftazidie avibactam. The
chromatogram showed two single peaks of ceftazidime
and avibactam was observed with retention time 2.523
and 4.410 min respectively.
Forced degradation studies
Stress studies are performed according to ICH
guidelines under following conditions.
Acid degradation
To 5 ml of sample solution add 1 ml of 0.1 N HCL
and sonicate place it aside for 3 hrs, then neutralize the
solution with 1ml of base and then transfer above
solution into 10 ml volumetric flask dilute with mobile
phase and record the chromatogram.
Alkaline degradation
To 5 ml of sample solution add 1 ml of 0.1 N
NaOH place it aside for 3 hrs, then neutralize the
solution with 1ml of acid and then transfer above
solution into 10 ml volumetric flask dilute with mobile
phase and record the chromatogram.
Peroxide degradation
To 5 ml of sample solution add 1 ml of 3% H202
and sonicate place it aside for 3 hrs, then transfer
above solution into 10 ml volumetric flask dilute with
mobile phase and record the chromatogram.
Photolytic degradation
Expose about 100 mg of sample in UV light
chamber at 231 nm for 3 hrs. weigh accurately this
5. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
74
power equivalent to 40 mg of Ceftazidime and 10 mg
of Avibactam into a 100 ml volumetric flask and make
up the volume and sonicate for 30 minutes with
intermittent shaking and volume is made up to the
mark with mobile phase and record the chromatogram.
Thermal degradation
Expose about 100 mg of sample in to dry heat at
800
C for 3 hrs. weigh accurately this power equivalent
to 40 mg of Ceftazidime and 10 mg of Avibactam into
a 100 ml volumetric flask and make up the volume and
sonicate for 30 minutes with intermittent shaking and
volume is made up to the mark with mobile phase and
record the chromatogram.
Records the peak area of stressed samples then
compare it with peak area of unstressed sample to
determine the % degradation.
Response of unstressed sample – response of stressed sample
% degradation = ------------------------------------------------------------------------------ X 100
Response of unstressed sample
RESULTS & DISCUSSION
Ceftazidime/avibactam (AVY-CAZ) is a recent
combination in the market used in the treatment of
complicated intra-abdominal infections (cIAI),
complicated urinary tract infections (cUTI), acute
pyelonephritis (AP). Literature survey reveals that
various methods for the estimation of Ceftazidime
individually and in combination with other drug is
reported, but no method has been reported for the
estimation of the Ceftazidime and Avibactam in
combine dosage form by RP-HPLC method. So here
an attempt has been made to develop simple, accurate,
sensitive, rapid and economic method for estimation of
Ceftazidime and Avibactam from combine dosage
form using HPLC. Specific objectives includes
The conditions in HPLC were optimized in order
to obtain the drugs separation of eluted compounds.
Initially various composition of mobile phase & PH
range were tried in order to have a good separation of
the titled ingredients. The composition of Mobile
phase & PH selections were based on peak parameters
like height, tailing, capacity factor, symmetric factor,
theoretical plates, run plates, run time & resolution.
The system with phosphate buffer: acetonitrile of
70:40 was found to be robust with PH 4. The optimum
wavelength for detection was 231 nm at which both
drugs have good response. CAZ was eluted at 2.523
min and AVI was eluted at 4.410 min.
System suitability test were carried out on a freshly
prepared stock solution % RSD of peak area of five
replicated injections of standards were taken & was
found to be less than 2%. Specificity of the method
developed was performed & it was found that there is
no interference of excipients with the analyte which
indicate that the method is specific for the analysis of
the analytes in their dosage form. Assay was
determined for both Standard & sample solution & the
% assay was found to be 100.23% & 101.21% and
respectively.
A calibration curve was found to be linear in the
concentration range of 240-560 µg/ml and 60-140
µg/ml for CAZ & AVI. Accuracy studies of the
method developed wad determined by spiking the
known amount of analyte to sample solution and the
percentage mean recovery was found to be 100.10%
and 99.75% for CAZ & AVI respectively. Precision of
the method & system were found to be within limits.
Limit of detection was found to be 11.2 and 2.5 µg/ml
for CAZ & AVI respectively and Limit of
Quantification was found is to be 33.8 and 7.69 µg/ml
for CAZ & AVI respectively. Robustness of the
method was determined by varying flow rate and
wavelength & Ruggedness of the method was
determined by carrying out the determination by two
different analyst.
Samples containing Ceftazidime and Avibactam
were subjected to various stress conditions and it was
found that overall net degradation was within the
limits without any significant degradation products.
A RP-HPLC method developed for Ceftazidime
and Avibactam shows that the results obtained for RP-
HPLC are promising with better resolution in set
chromatographic conditions. The developed methods
were statistically validated which suggested that the
methods were within the acceptable limits hence these
methods can be used for the routine determination of
Ceftazidime and Avibactam in bulk drug and
pharmaceutical formulation.
6. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
75
Fig 4: Linearity curve of Ceftazidime Fig 5: Linearity curve of Avibactam
Table: 2 Results of Linearity
CEFTAZIDIME AVIBACTAM
S.No. Conc.(µg/ml ) Area Conc.(µg/ml ) Area
1 240 1344.606 60 108.783
2 320 1849.853 80 159.306
3 400 2338.421 100 204.849
4 480 2563.186 120 222.682
5 560 3106.591 140 243.873
Table: 3 Precision method of proposed RP- HPLC method
CEFTAZIDIME AVIBACTAM
S.No. Rt Area Rt Area
1 2.510 2192.417 4.397 267.545
2 2.523 2322.573 4.410 211.442
3 2.523 2321.138 4.413 202.102
4 2.523 2333.196 4.413 200.853
5 2.507 2350.119 4.397 202.888
6 2.497 2341.355 4.390 198.551
Avg 2.513833 2310.133 4.403333 213.8968
Stdev 0.010926 6.248003 0.009893 2.001001
%RSD 0.433746 0.270460 0.224216 0.935498
Table: 4 Recovery data for Ceftazidime
Recovery
level
Amount
taken
(mcg/ml)
Accuracy of Ceftazidime
Area Average
area
Amount
recoverd
%Recovery Average %
Recovery
80% 80+320=400 2608.241 2607.639 10.8 111.3
2609.160
2605.517
100% 80+400=480 2194.643 2244.257 13.25 95.85
y = 1.538x + 33.44
R² = 0.999
0
50
100
150
200
250
300
0 50 100 150
7. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
76
2211.816
100.10
2326.313
120% 80+480=560 3109.681 3109.702 22.5 93.16
3112.744
3106.682
Table: 5 Recovery data for Avibactam
Recovery
level
Amount
taken
(mcg/ml)
Accuracy of Avibactam
Area Average
area
Amount
recoverd
%Recovery Average %
Recovery
80% 20+80=100 236.388 237.6637 9,56 92.65
99.75
234.158
242.445
100% 20+100=120 224.953 254.704 15.03 99.34
269.243
269.916
120% 20+120=140 280.051 275.7967 21.07 107.35
280.286
267.053
Table: 6 Robustness data
Parameter
CEFTAZIDIME AVIBACTAM
Retention time(min) Tailing factor Retention time(min) Tailing factor
Flow Rate
0.8 ml/min
1.2 ml/min
3.130
2.090
1.258
1.036
5.443
3.663
1.167
0.943
Wavelength
229nm
233nm
2.513
2.517
1.222
1.179
4.380
4.380
1.088
1.125
Table: 7 Result of Ruggedness
Ceftazidime % Assay Avibactam % Assay
Analyst 01 99.3 Analyst 01 96.28
Analyst 02 99.30 Analyst 02 95.97
% RSD 1.82% % RSD 1.67%
Table: 8 Result of LOD and LOQ
DRUG LOD(μg/ml) LOQ(μg/ml)
Ceftazidime 11.2 33.8
Avibactam 2.5 7.69
8. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
77
Table: 9 Results of Forced Degradation studies
STRESS CONDITION PEAK AREA % DEGRADATION
CAZ AVI CAZ AVI
Unstressed sample 2192.417 267.545 0 0
Acid 3981.699 391.641 0.81 0.46
Alkaline 3934.512 396.009 0.79 0.48
Oxidation 3943.709 391.379 0.79 0.46
Photolytic 3951.770 397.622 0.80 0.49
Thermal 3971.642 385.227 0.81 0.43
CONCLUSION
Ceftazidime and Avibactam are essential for
treatment of complicated intra-abdominal infections
(CIAI), complicated urinary tract infections (cUTI),
acute pyelonephritis (AP) so it is therefore necessary
to know the quality of these drugs which is possible
through the use of simple, sensitive and cost effective
analytical methods so that the compounds can be
rapidly, routinely and consistently assessed. Literature
survey reveals that there are methods to estimate the
ceftazidime individually and with other combinations,
but not a single method is reported for estimation of
Ceftazidime and Avibactam in bulk and
multicomponent formulation. So, an attempt was made
to develop RP-HPLC method. A RP-HPLC method
proposed as a suitable method for the simultaneous
estimation of Ceftazidime and Avibactam. The
chromatographic conditions included mixed phosphate
buffer (PH
4): Acetonitrile in the ratio of 60:40 % v/v
as mobile phase. The optimum wavelength for
detection was 231 nm where as CAZ was eluted at
2.523 Min and AVY was eluted at 4.410 min. The
calibration curve of Ceftazidime and Avibactam is
linear in the range of 240-560 µg/ml and 60-140 µg/ml
respectively.
Accuracy study showed the percentage mean
recovery of Ceftazidime and Avibactam is 100.10%
and 99.75% respectively. The amount of Ceftazidime
and Avibactam present in the taken formulation was
found to be 100.23% and 101.21% respectively.
Forced degradation studies were carried under various
stress conditions. It was found to be stable and the net
degradation was within the limits without any
significant degradation products. From the above
experimental results and parameters it was concluded
that, this newly developed methods for the estimation
of Ceftazidime and Avibactam was found to be
simple, rapid, economic, precise, accurate and
reproducible. Forced degradation studies carried out
are helpful to determine stability of this drug in
combination. The analytical technique showed reliable
method hence it can be effectively applied for routine
analysis in research institutions & quality control
department in industries.
REFERENCES
[1]. Amareswari S, Nandakishore A, Aasif M, Khan S. Stability indicating RP-HPLC method for the estimation of
ceftazidime pentahydrate and tazobactam sodium in bulk and dosage forms. Ind J of RES Pharm and biotech,
1(4), 2013, 543-548.
[2]. Marianna Z, Anna J, Agnieszka S and Wojciech M stability of ceftazidime pentahydrate in medicinal
preparations biotm and ceftium. Acta poloniae pharmaceutical and drug research, 62(1), 2005, 11-15.
9. Syeda S F et al / Int. J. of Farmacia, 2016; Vol-(2) 1: 70-78
78
[3]. Cinzia A, Monica B, Roberto V, Pierangelo B, Rta P Determination of Ceftazidime concentration in Mueller
Hinton Agar by High Performance Liquid Chromatography. J Chromatogr A, 742, 1996, 121-126.
[4]. Nanda R, Shelke A. Development and validation of RP_HPLC method for the simultaneous estimation of
ceftazidime sodium and tazibactam sodium in marketed formulation. Int J Pharm Res., 5(3), 2013, 983-999.
[5]. Reddy J, Ganapaty S. A validated stability indicating RP-HPLC method for simultaneous determination of
tobramycin and ceftazidime in pharmaceutical formulation. Int J Pharm., 5(3), 2015, 976-984.
[6]. Masoom RS et al. Development and validation of high performance liquid chromatographic method for the
simultaneous determination of ceftazidime and sulbactam in spiked plasma and combined dosage form.
American J applied Sci., 6(10), 2009, 1781-1786.
[7]. International conference on harmonization, (ICH) “Q2A: Text on Validation of analytical procedure,” Federal
Register (notices), 65(40), 1995, 11260 11262.
[8]. International conference on harmonization, “Q2B- Validation of analytical Procedures: Methodology”, US
Food and Drug Administration, 1996.
[9]. International conference on harmonization, (ICH) “Q1B stability testing: Photostability testing of new drug
substances and products, 2006.
[10]. www.drugbank.ca/drugs/DB00438/Ceftazidime.
[11]. www.drugbank.ca/drugs/DB09060/Avibactam.
[12]. www.scbt.com/datasheet-205243-ceftazidime.