The document discusses principles and procedures for analyzing vitamins in pharmaceutical preparations and dosage forms. It provides details on several methods for analyzing Vitamin A, D, thiamine, and niacin. For Vitamin A, methods discussed include Carr-Price colorimetric method, UV spectrophotometry, and modification of Carr-Price reaction. For Vitamin D, it covers UV spectrophotometry and colorimetric reactions with glycerol-1,3-dichlorohydrin and organic aldehydes. Thiamine analysis methods include thiochrome fluorimetry, silicotungstic acid gravimetry, and colorimetric reactions with p-aminoacetophenone and 6
Determination of vitamin a in the supplied sample.Atai Rabby
Vitamin A is assayed from the biological samples by high performance liquid chromatography(HPLC) method after processing and separation of samples, as the method is specific, accurate and sensitive but it is also widely measured by calorimetrically for the last many years using the maximum intensity of color developed at 620 nm by the method of Carr and price.
Beer is an incredibly complex beverage containing more than 3000 different compounds, including carbohydrates, proteins, ions, microbes, organic acids, and polyphenols, among others.Some of the analytical methods used for quality control are presented
Determination of vitamin a in the supplied sample.Atai Rabby
Vitamin A is assayed from the biological samples by high performance liquid chromatography(HPLC) method after processing and separation of samples, as the method is specific, accurate and sensitive but it is also widely measured by calorimetrically for the last many years using the maximum intensity of color developed at 620 nm by the method of Carr and price.
Beer is an incredibly complex beverage containing more than 3000 different compounds, including carbohydrates, proteins, ions, microbes, organic acids, and polyphenols, among others.Some of the analytical methods used for quality control are presented
In this slide contains Quality control test and Analysis of Wine and Beer.
Presented by: SHAIK GOUSE UL AZAM (Department of pharmaceutical analysis ).
RIPER, anantapur
In this slide contains Quality control tests and analysis of spirits and vinegar.
Presented by: T.JAYASREE (Department of pharmaceutical analysis).
RIPER, anantapur
Carbohydrates || Food Analysis || Pharmaceutical Analysis Department || M.Pha...saimuniswetha1
Hello everyone,
Today's topic Carbohydrates in Food Analysis subject in M.pharmacy(Pharmaceutical Analysis Department) ..Don't forget to see.. please watch it... If you need explanation about Carbohydrates please click below link : https://youtu.be/aI5UnNYgufY
This presentation gives a brief introduction of Vitamin C. It Covers it's various application and uses in various industry and health care. Also, describe the main industrial process for the production of Vitamin C.
In this slide I have given brief knowledge about types of preservatives. This slide is recommended to students who are new to this particular topic or those who want notes for examination. I hope you will get benefit from this slide. Do comment for any improvement or want slides that i should prepare for you.
In this slide contains Quality control test and Analysis of Wine and Beer.
Presented by: SHAIK GOUSE UL AZAM (Department of pharmaceutical analysis ).
RIPER, anantapur
In this slide contains Quality control tests and analysis of spirits and vinegar.
Presented by: T.JAYASREE (Department of pharmaceutical analysis).
RIPER, anantapur
Carbohydrates || Food Analysis || Pharmaceutical Analysis Department || M.Pha...saimuniswetha1
Hello everyone,
Today's topic Carbohydrates in Food Analysis subject in M.pharmacy(Pharmaceutical Analysis Department) ..Don't forget to see.. please watch it... If you need explanation about Carbohydrates please click below link : https://youtu.be/aI5UnNYgufY
This presentation gives a brief introduction of Vitamin C. It Covers it's various application and uses in various industry and health care. Also, describe the main industrial process for the production of Vitamin C.
In this slide I have given brief knowledge about types of preservatives. This slide is recommended to students who are new to this particular topic or those who want notes for examination. I hope you will get benefit from this slide. Do comment for any improvement or want slides that i should prepare for you.
this presentation gives informationabout microbial assay of vitamins B2 and B12. it is based upon the guidelines of indian pharmacopoeia. this presentation highlights the principle, process and applications of microbial assay
Microbiome Identification to Characterization: Pathogen Detection Webinar Ser...QIAGEN
The research community has begun correlating the makeup of individual microbiomes with disorders and diseases such as autism, atherosclerosis, obesity and cancer. To accomplish this, researchers must first identify and characterize these microbial communities. This slidedeck will begin with a general introduction of metagenomics and an overview of experimental strategies. Following this, a comprehensive microbiome assay pipeline will be introduced. We conclude with application-based examples that demonstrate how to identify and characterize microbiome profiles.
Application of High Performance Thin Layer Chromatography with Densitometry f...iosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Nutrition for Pregnant and Lactating womanCM Pandey
These are the slides that me, Madan Pandey & my friend, Deepak Kumar Mandal has presented in our class, B. Sc. (Nutrition & dietetics) 3rd year. We have slides here about physiological changes during pregnancy & lactation; complications at these stages and nutritional requirements according to ICMR, 2010. I hope it would be useful for the friends who are studying in field of food, nutrition, health & medicine.
Madan Pandey
Central Campus of Technology, Dharan
Tribhuvan University
Kathmandu, Nepal
Validated High-performance Liquid Chromatography Method for Degradation Study...BRNSS Publication Hub
A novel and basic reversed-phase liquid chromatographic strategy has been set up for the determination
of ursodeoxycholic corrosive and silymarin and studies its degradation pattern in pharmaceutical
dosage forms. Ursodeoxycholic acid and silymarin are used to control Type 2 diabetes. The proposed
work was performed on Young Lin (S.K) isocratic System UV Detector C18 column (150 mm ×
4.6 mm). A mixture of potassium phosphate, mobile phase in this method with a flow rate of 0.7 mL/
min (UV detection at 203 nm) and the method was validated as per the ICH guidelines. Forced
degradation studies were performed by exposing the drug ursodeoxycholic acid and silymarin to
acidic, alkaline, oxidation, and thermal stress degradations. The proposed reversed-phase-highperformance
liquid chromatography technique was observed to be powerful and particular, and
this strategy is reasonable for the measure of pharmaceutical dose frames and in addition kinetic
examinations.
In this work a new prodrug polymer was
prepared with two attachment groups (amid-ester), using di
functional spacer such as ethanol amine, which could react with
polyacrylic acid producing amide group, with remain ethanol
terminal group which could react with captopril acyl chloride,
producing ester group with extended the arm substituted drug to
improve the hydrolysis and to prevent the steric effect of polymer
chains. Many advantages enhanced the prodrug of polymer. The
prepared polymers were characterized by FTIR, 1H –NMR
spectroscopies. Controlled drug release was studied in different
pH values at 37℃, using UV. Spectra with comparing with
calibration curve. The modification percentage test was studied,and swelling percentage was calculated and all physical properties were observed.
Estimation of serum triglycerides by Dr. TehmasTehmas Ahmad
Estimation of Serum Triglycerides, Practical demonstration lecture for 2nd year MBBS students of Bannu Medical College, Bannu. Lecture delivered on 13/03/2018
Background- Chitosan is the most abundant natural amino polysaccharide. Researchers have found that chitosan is biocompatible, biodegradable and nontoxic, which have made wide applicability in the pharmaceutical field. Objectives- Aim of the study was to prepare Chitosan from chitin and characterize them. Methods- Chitosan was prepared by deacetylation of chitin and characterized by U.V Spectrophotometry, FTIR (Fourier transform infrared spectroscopy), DLS (Dynamic Light Scattering), and Scanning electron microscopy (SEM). Results- The present study showed that Chitosan was successfully prepared by deacetylation of chitin. The obtained chitosan was characterized for further study. Conclusion- Our study confirms the preparation by Chitosan from Chitin for further study. Key-words- Chitin, Chitosan, Deacetylation, DLS, FTIR, SEM
Here is brief ppt on industrial production of amino acids - glutamine, lysine, tryptophan.
Please share your feedback and queries. Constructive criticism is appreciated.
Thank you
Class 12 Chemistry Investigatory project .pdfAnonymous
Chemistry Project
Ascorbic Acid
Class 12 Chemistry Investigatory project.
Investigatory Project
Project Report File
Class 12 Project
Ascorbic Acid Project
Science Project
Vitamin C
Vitamin C Project.
Estimation of Vitamin C in fruit and vegetable juices.
Aging is a natural part of human life. However, recent discoveries indicate that pharmacological approaches used for the improvement and possibly, for the delay of the aging process, might shed a new light on this topic. This might obviously contribute to the extension of the active life of older people and maintenance of their quality of life, which could consequently reduce both social and economic burden of each country, especially the developed ones.
Gelatin-grafted N- proflavine acryl amide was synthesized through two steps; firstly the Gelatin was grafted with
acrylic acid free radically using Ammonium per-sulfate at 60℃, Then it was modified to its corresponding acyl
chloride derivation, second step included the substitution with amino group of proflavine, in this research Gelatin
was used as a natural nontoxic, water soluble polymer as a drug carrier.
The prepared pro drug polymer was characterized by FTIR and 1H-NMR spectroscopies, Controlled drug release
was studied in different pH values at 37℃. Many advantages were obtained comparing with other known
methods.
Method Development and Validation of Clopidogrel Bisulphate by Reverse Phase-...SriramNagarajan15
A new, simple sensitive, rapid, accurate and precise RP-HPLC method was developed for the estimation of Clopidogrel bisulphate in bulk drug and pharmaceutical formulation. Clopidogrel bisulphate was chromatographed on a reverse phase C18column (150 mm x 4.5 mm, i.d 5μm) in a mobile phase consisting of acetonitrile and phosphate buffer (pH: 3.0) in the ratio of 60:40 % v/v. The mobile phase was pumped at a flow rate of 1 ml/min with detection at 224 nm. The detector response was linear in the concentration of 50-150 μg /ml. The limit of detection and limit of quantitation was found to be 1.3 and 4.2 µg/ml, respectively. The intra and inter day variation was found to be less than 2%. The mean recovery of the drug from the solution was 99.79%. The proposed method is simple, fast, accurate, precise and reproducible hence, it can be applied for routine quality control analysis of Clopidogrel bisulphate in bulk drug and pharmaceutical formulation. Key words: Clopidogrel bisulphate, RP-HPLC, Validation, Accuracy, Precision.
Method Development and Validation of Clopidogrel Bisulphate by Reverse Phase-...SriramNagarajan15
A new, simple sensitive, rapid, accurate and precise RP-HPLC method was developed for the estimation of Clopidogrel bisulphate in bulk drug and pharmaceutical formulation. Clopidogrel bisulphate was chromatographed on a reverse phase C18column (150 mm x 4.5 mm, i.d 5μm) in a mobile phase consisting of acetonitrile and phosphate buffer (pH: 3.0) in the ratio of 60:40 % v/v. The mobile phase was pumped at a flow rate of 1 ml/min with detection at 224 nm. The detector response was linear in the concentration of 50-150 μg /ml. The limit of detection and limit of quantitation was found to be 1.3 and 4.2 µg/ml, respectively. The intra and inter day variation was found to be less than 2%. The mean recovery of the drug from the solution was 99.79%. The proposed method is simple, fast, accurate, precise and reproducible hence, it can be applied for routine quality control analysis of Clopidogrel bisulphate in bulk drug and pharmaceutical formulation. Key words: Clopidogrel bisulphate, RP-HPLC, Validation, Accuracy, Precision.
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
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Factory Supply Best Quality Pmk Oil CAS 28578–16–7 PMK Powder in Stockrebeccabio
Factory Supply Best Quality Pmk Oil CAS 28578–16–7 PMK Powder in Stock
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Hot Selling Organic intermediates
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.
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
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
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
micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
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
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.
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
1. PRINCIPLES AND PROCEDURES INVOLVEDPRINCIPLES AND PROCEDURES INVOLVED
IN THE ANALYSIS OF VITAMINS ININ THE ANALYSIS OF VITAMINS IN
PHARMACEUTICAL PREPARATIONS ANDPHARMACEUTICAL PREPARATIONS AND
DOSAGE FORMSDOSAGE FORMS.
Presented by:Presented by:
PRADIPKUMAR.L.GHORI
DEP.PHARMACEUTICSDEP.PHARMACEUTICS
M.M.C.P BELGAUMM.M.C.P BELGAUM
2. INTRODUCTION TO VITAMINS:
Vital amines/growth factors/accessory factors.
Any group of organic compounds required in
small amount to perform specific biological
functions for normal maintenance of optimum
growth and health of organism.
Classification:
Fat soluble vitamins: vit-A, vit-D, vit-E, vit-K.
Water soluble vitamins: vit-C, B-complex, vit-H.
3. Need for study:
Essential nutritional factors and potent organic compounds
require great care in analytical control and their formulations.
Analytical procedures based on chemical, physical, biological and
microbiological methods are used in the assay.
The methods of analysis of vitamins and its formulations should be
based on biological response.
Chemical, physical and microbiological methods are only valid if
they correlate directly or indirectly with biological activity.
Care should be exercised as some vitamins exhibit species
specificity in that they are required in certain species of animals
and not in others.
5. *VITAMIN-A* Chemistry:
Retinol (Alcoholic)
Retinal (Aldehydic form)
Retinoic acid (Acidic form)
β-Carotene (Provitamin-A)
RC
11
C
12
R
11-cis form
11-trans form
R -CH2OH, -CHO, -COOH.
6. TISSUE.
SYNTHESIS OF GLYCO PROTEINS AND
MEVALONATE.
CAROTENOIDS FUNCTIONS AS ANTI-OXIDANTS
AND REDUCES RISK OF CANCER.
DEFICIENCY:DEFICIENCY:
NIGHT BLINDNESS, XEROPHTHALMIA,
KERATOMALACIA, MAY ALSO AFFECT ON
REPRODUCTION AND EPITHELIAL CELLS.
PROPERTIES:PROPERTIES:
1) THE FREE VITAMIN A ALCOHOL IS READILY
OXIDIZED BY ATMOSPHERIC OXYGEN OR OXIDIZING
AGENT.
2) VERY SENSITIVE TO LIGHT.
3) VITAMINS OCCUR IN NATURE MAINLY IN THE FORM
OF ESTERS WHICH ARE CONSIDERABLY STABLE
TOWARDS OXIDATION.
7. Three methods:
Bioassay based on RAT or CHICKEN growth or on liver
storage.
Colorimetric method/ Carr-price method.
UV-spectrophotometric method.
COLORIMETRIC METHOD:COLORIMETRIC METHOD:
CARR-PRICE METHOD:
Anhydrous Antimony Tri chloride (SbCl3) in chloroform
reacts with a dilute solution of vitamin A to form a transient
blue color.
Reaction occurs between Antimony tri chloride and
unsaturated side chain of vitamin A. The carotene, polyene
acids and various other material present in natural oils
produce same color.
The color rapidly reaches maximum intensity and just as
rapidly it fades. The reading must be taken within 10-
15seconds
8. Sb Cl3+
Blue color
(λ max-
550nm)
DisadvantagesDisadvantages::
Vitamin A is a recemic mixture containing 1/4th
of cis-form
and 3/4th
of Trans form in oils. While the biologically active
form is the Trans form.
The Carr-price reaction does not differentiate between the
two isomers, so it is not a specific test for all-trans
compounds.
It does not give a stable blue color.
R
9. Applications:
This test can be used as an identification test for
vitamin A
The blue color method can be used in determining
Vitamin A in agricultural feeds and in oleomargarines.
The method extensively used in biochemical work
because of its great sensitivity.
10. METHOD2: MODIFICATION OF CARR-PRICE :
Formation of blue color by reaction with vitamin A and activated
Glycerol-1,3-dichlorohydrin.
Activation of reagent may be accomplished by distillation of
glycerol 1,3-dichlorohydrin in presence of antimony tri chloride.
Maximum absorbance at 550nm.
Color is more stable than the color produced in Carr-price
reaction.
The intensity of solution reaches 2min. and the color is stable for
additional 3min.
Disadvantages:Disadvantages:
Not as sensitive compared to carr-price method.
R
CH2Cl
CHOH
CH2Cl
blue color 550nm
Sb Cl3
+
11. SPECTROPHOTOMETRIC METHOD:
Method 3: DIRECT SPECTROPHOTOMETRIC METHOD (IP-1996 method):
Saponified sample is extracted with ether. Ether is evaporated. Residue is
dissolved in isopropanol.
Measure the absorbance at about 300, 310, 325 and 334 nm. Determine the
wavelength of maximum absorption.
If the wavelength of maximum absorption lies between 323 and
327 nm and the absorbance at about 300 nm relive to that at
about 325 nm does not exceed 0.73, a corrected absorbance is
derived from the equation.
A325 (CORR.) = 6.815 A325 – 2.555 A310 – 4.260 A334
Calculate the potency of the sample from the expression.
Vitamin A potency in Units per g = A325
1%,
1 CM) X 1830
12. If the corrected absorbance lies within +/- 3.0% of the
uncorrected absorbance, ignore the corrected absorbance
and calculate the potency from the uncorrected
absorbance.
If the wavelength of maximum absorption lies outside the
range 323 to 327 nm, or if the relative absorbance at
about 300 nm exceeds 0.73, the unsaponifiable fraction of
the sample must be further purified by chromatography.
13. Method 4: CONVERSION OF VITAMIN A TO
ANHYDRO VITAMIN A:
Vitamin A may be easily converted to
Anhydro vitamin A (II) in anhydrous solvents in the presence
of traces of mineral acids or strong acids.
Anhydro-vitamin A results in a considerable displacement of
absorption towards visible region with absorption maxima at
358nm, 377nm and 399nm in benzene. There is also an
increase in absorptivity.
R mineral/organic acid
anhydro vit A. 1,3-dihydro vitamin
14. Formula:
USP units /ml. aliquot = __ A___
0.0122
A Increase in absorbance caused by
dehydration.
0.0122 Increase in absorbance corresponding to
1USP unit of vitamin A
This method can be applied for the vit-A concentrates, fish
oils, oleomargarine, butter etc. this has been reported to be
very specific.
15. *VITAMIN D**VITAMIN D*
Steroidal vitamin/ anti-ricketic vitamin.
Two forms:
◦ Ergocalciferol (vitamin D2)
◦ Cholecalciferol (vitamin D3)
Functions:
Regulates plasma levels of calcium and phosphate.
In osteoblasts of bone, vit D stimulates uptake of calcium and
deposition of calcium phosphate.
1, 25 di-hydroxy cholecalciferol is active form also known as
calcitrol.
Diffiency:
Rickets in children and osteomalacia in adults.
16. Properties:
1) It is not affected by dilute acids, alkalis or air
2) Vitamin D3 is stored in the skin as 7-dehydro cholestrol
which is converted to activated vitamin D2 by sunshine or
ultraviolet irradiation.
Ergosterol vitamin D2. (Activated form)
7-dehydro cholestrol vitamin D3(Activated form).
3) Biological assay methods uses rats and chicks are
available for evaluation of vitamin D activity.
4) Physiochemical procedures employ the methods of uv-
spectrophotometry / colorimetry.
CH3
CH3
CH3CH2
OH
CH3
17. Method 1: UV-SPECTROPHOTOMETRY:
Both D2 and D3 have an absorption maximum at 265nm in hexane. The
A1%
1cm of D2 is 459nm that of D3 is 474.
The determination involving uv-spectrophotometry suffers from lack of
specificity.
The interference of interfering substances can be avoided by
chromatographic separation.
TWO STEP ADSORPTION CHROMATOGRAPHIC METHOD:
Passage of oil through an activated earth this removes vitamin A,
sterols and irradiation products of ergosterol other than Vitamin D.
The second adsorption step removes impurities due to first adsorbent,
unsaturated compounds of squalene type and vitamin A decomposition
products.
The eluate is collected and analyzed spectroscopically.
18. COLORIMETRIC METHOD
Many colorimetric methods have been proposed for the
quantitative determination of vitamin D.
“MODIFIED CARR-PRICE METHOD”:
The glycerol-1,3-dichloro hydrin reacts with vitamin D
producing blue color.
The color is stable for several hours.
The reaction is specific:
Ergosterol gives pink-orange color and slowly turns to a
fluorescent green.
7-dehydro cholesterol produce no color but after hours produce
faint pink color.
Cholesterol does not show reaction or absorption.
19. REACTION WITH ORGANIC ALDEHYDE:
This is based on reaction between Vitamin D and organic
aldehyde (vanillin, furfural or anisaldehyde) and
sulphuric acid or perchloric acid.
CH3
CH3
CH3CH2
OH
CH3
CHO
OMe
OH
H2SO4/HCLO4
Green Color (625nm)+
CH3
CH3
CH3CH2
OH
CH3
H2SO4/HCLO4
Green Color (625nm)
OH
CHO
+
CH3
CH3
CH3CH2
OH
CH3
O CHO
H2SO4/HCLO4
Green Color (625
nm)
+
20. MODIFIED CARR-PRICE METHOD:
The modified method has improved sensitivity and
reproducibility by addition of acetyl chloride to SbCl3.
Mixture of vitamin A and vitamin D2 is dissolved in benzene
after passing through a column of bentonite activated with
acid.
The treatment selectively destroys vitamin A and the eluate
is then analyzed using antimony trichloride (SbCl3) reaction.
CH3
CH3
CH3CH2
OH
CH3
Green Color (625nm)CH3COClSb Cl3 ++
21. VITAMIN DVITAMIN D
CONTD…CONTD…
Vitamin D gives color with glycerol-1,3-
dichlorohydrin in presence of acetyl chloride
CH3
CH3
CH3CH2
OH
CH3
Green Color (625nm)CH3COCl
CH2 Cl
CH OH
CH2Cl
++
22. *THIAMINE*
Chemistry:Chemistry:
Contains pyridine and thiazole ring structures.
Functions:Functions:
Prosthetic group in decarboxylation. and transketolase reactions.
Deficiency disorders:Deficiency disorders: Beri beri.
Properties:Properties:
1) White crystalline powder with slight characteristic odour.
2) It is hygroscopic.
3) Soluble in water, glycerol, alcohol. Practically insoluble in ether, benzene,
hexane, and chloroform
Chemical, biological and microbiological methods are available for the analysis of
thiamine.
◦ The bio-assay is based on prevention and cure of polyneuritis and weight gain
in animals. Microbiological methods using Phycomyces blakesleeanus and
Lactobacillus ermenti are available. A yeast fermentation method is also
NN
N SCH3
NH3+ CH2 CH2 O H
CH3
. 2 Cl
+
-
2
23. FLOURIMETRIC METHOD (Thiochrome)
1. Thiamine is quantitatively isolated from the foods, biologicals,
and pharmaceuticals usually by boiling with dilute acids and
treatment with enzyme preparations containing phosphatases
which will free thiamine from its natural complexes.
2. Protein substances must be digested with proteolytic enzyme like
papain.
3. Purification of extract is done by passing through zeolite, an
inorganic ion exchanger and thiamine is retained in the zeolite.
4. Add acidified potassium chloride to elute thiamine.
5. Further it is oxidized with alkaline potassium ferricyanide to
produce thiachrome. This compound is having blue fluorescence.
NN
N SCH3
NH3+ CH2 CH2 O H
CH3
NN
N SCH3
CH2 CH2 O H
CH3
NK3 Fe (C N)6
+
2 Cl-
24. SILICO TUNGSTIC ACID METHOD (Gravimetry):
{H4Si(W3O10)4 nH2O }
Prepared by One mole of Silicic acid for every twelve moles of
Sodium Tungstate.
Thiamine in tablets and solutions may be determined by
precipitation with silicotungstic acid.
The sample is dissolved in acidified water and heated to boiling.
The precipitate is filtered washed with acid and water finally
with acetone. Then its weighed to constant weight.
Each gram is equivalent to 0.1936 g of thiamine hydrochloride.
25. COLORIMETRIC METHOD:
Method 1: with p-Amino acetophenone:
The extraction is done as given in thiochrome method.
Thiamine couples with diazotized p-Amino Acetophenone
which has an absorption maximum at 520nm.
This method can be used to determine thiamine in presence
of phosphorylated thiamine and is useful in urine analysis.
This method is not recommended for assay of materials rich
in protein content and low in thiamine.
Results of this method agrees with the biological method.
NN
N SCH3
NH3 CH2 CH2 O H
CH3
N
C O C H3
N H
complex
diazotised p-amino acetophenone
+
520 nm
26. Method 2: using 6-Aminothymol.
A color reaction between thiamine and diazotized 6-
Amino thymol is seen. This method is simple fast and
no interference is seen between the degraded thiamine
and 6-Aminothymol.
NN
N SCH3
NH3 CH2 CH2 O H
CH3
OH
CH (CH3)2
CH3
N NH
color complex+
27. Miscellaneous Methods:Miscellaneous Methods:
Method 3: Non-aqueous titration:
Thiamine HCl can be titrated with perchloric acid in glacial acetic acid solution, if an
excess of mercuric acetate is added. Both the nitrogen are titrated. p-
Naphthol-benzein and quinaldine red are suitable indicators.
Method 4: Argentometric Method:.
Total chlorine in thiamine HCl can be determined by dissolving in acidified water
(HNO3 ) added with excess of silver nitrate. The precipitate is filtered and washed.
The filtrate is then titrated with 0.1N Ammonium Thiocyanate.
1ml of 0.1N AgNO3 is equivalent to 0.003546 g of chlorine.
Method 5:
Chloride as hydrochloride is determined by titration with 0.1 N sodium hydroxide to
pH7 using bromothymol blue as indicator.
Each ml of 0.1 N NaOH is equivalent to 0.003546 g of chlorine
Method 6
The nitrate in thiamine mononitrate is determined by precipitation with nitron
(1,4-Diphenyl-3-phenylamino-1,2,4-triazolium hydroxide inner salt)
from an acidified solution.
Nitron is a compound C20H16N4 used in the qualitative and quantitative determination
of nitric acid with which it forms an insoluble nitrate
28. *NIACIN (NICOTINIC ACID)*
Chemistry:Chemistry:
◦ It’s a heterocyclic. 3-pyridine carboxylic acid.
Functions:Functions:
◦ It functions as oxidising co-enzymes of many dehydrogenases
Deffiency disordersDeffiency disorders:
◦ Pellagra, characterised by dementia, dermatitis and diarrhoea.
PROPERTIES:PROPERTIES:
1. Solubility: Soluble in boiling water and in boiling ethanol (95%). Sparingly
soluble in water. Very slightly soluble in chloroform. Practically insoluble in
ether. It dissolves in dilute solutions of alkali hydroxides and carbonates.
2. Non-hygroscopic and stable in air.
3. It has absorption maximum in UV at 262nm.
4. Identification test:
Thiamine is dissolved in water, and neutralized to litmus paper with 0.1M
sodium hydroxide, add 3 ml of copper sulphate solution; a blue precipitate is
N
COOH
29. ACID BASE TITRATION:
Nicotinic acid can be titrated with NaOH using
phenolphthalein solution as indicator.
COLORIMETRIC METHOD USING CYANOGEN
BROMIDE:
This is based on a color reaction of pyridine and
λ,β− unsubstituted derivatives. Cyanogen bromide breaks one
carbon-nitrogen linkage and produces a color compound upon
addition of amine or ammonia.
N
COOH
NaOH
N
COONa
H2 O
+
+
N
COOH
SO3H
NH2
CNBr
cleavage of CN bond of pyridine by CNBr red color measured spectroscopically+ +
30. DETERMINATION OF NICOTINAMIDE
ACID-BASE TITRATION:
◦ Nicotinamide when boiled with sodium hydroxide solution, it
releases the nitrogen of the amido group in the form of ammonia,
which can be collected in sulphuric acid and determined by
titration.
The liberated ammonia collected in sulphuric acid and
determined by titration with NaOH.
2NH3 + H2SO4----------- (NH4)2
(NH4)2 + 2 NaOH----- Na2SO4 + 2 H2O + 2NH3
End- point being determined using phenolphthalein
indicator.
N
CONH2
alkaline hydrolysis
N
COOH
NH3
Nicotinic acid
+
31. HOFFMAN REARRANGEMENT REACTION:
Nicotinamide undergoes a Hoffmann rearrangement to form a 3-amino pyridine
which reacts as an aromatic amine.
It can be diazotized and coupled with N-(1-naphthyl)- ethylene di-amine to produce
red colored azo compound.
This is the basis of USP assay of nicotinamide in capsules, injectables and tablets.
N
CONH2 hoffman -co
rearrangement
N
NH2
3-Amino Pyridine
NaNO2/HCl
HNO2 <8*C
N N Cl
diazotised compound
NH-CH2-CH2-NH2
. HCl
N N NH-(CH2)2NH3
red colored azo-compound
+
+ +
32. VITAMIN C (ASCORBIC ACID)
Chemistry:Chemistry:
◦ It’s a heterocyclic furan-2-one
derivative
Functions:Functions:
◦ Anti-oxidants, helps in synthesis of collagen and
hydroxylation reactions.
Defiency disorders:Defiency disorders:
◦ Scurvy, swollen joints, hemorrhages in various tissues
and delayed wound healing.
Properties:Properties:
1. It occurs as a white or slightly yellow crystal or powder.
2. In dry state it is stable to air but degradation seen in presence of some
metals and it is unstable to light.
3. Many chemical methods have been reported and this is based on the
reducing properties of ascorbic acid.
OO
OHOH
CHOH
CH2OH
33. VOLUMETRIC METHODS
Iodometric method:
Ascorbic acid content of pure solutions or the purity of the
substance can be determined by titration with 0.1 N Iodine
solution.
Modifications of this method using instruments like
potentiometric titration or polarized platinum-platinum
electrodes and a dead-stop end point, have been applied to
pharmaceutical products.
O O
OH OH
HOHC
HOH2C
I2
O O
O O
HOHC
HOH2C
2
HI+ +
Dehydro ascorbic acidAscorbic acid
34. 2) Titration with 2,6-DICHLORO PHENOL-INDOPHENOL:
A typical procedure for eliminating interfering substances
consists of
a) The conversion of total ascorbic acid to dehydro ascorbic acid by
passing it through Norit or by using ascorbic acid oxidase.
b) The reduction of dehydro ascorbic acid to ascorbic acid with
hydrogen sulphide at pH 4 to 7
c) The titration of ascorbic acid with dichloro phenol –indophenol.
OO
OHOH
CHOH
CH2
OH
NaO N O
Cl
Cl
OO
OO
CHOH
CH2OH
2
,6
- dichloro phenol indo phenol (pink or blue color) dehydro ascorbic acid
NaO N
Cl
OH
Cl
H
reduced form (color less)
+ +
35. METHOD 3: Titration With N-BROMO
SUCCINIMIDE:
N-Bromo succinimide in aqueous solution readily
oxidizes an aqueous solution of ascorbic acid to dehydro
ascorbic acid, while N-Bromo succinimide is irreversibly
reduced to succinimide with the formation of Hydrogen
Bromide.
After all the ascorbic acid has been oxidized, the
slightest excess of N-Bromo succinimide, In presence of
potassium Iodide, liberates iodine.
The end point is being determined using starch solution
added in the end.
OO
OHOH
CHOH
CH2OH
CH2-CO
CH2CO
N Br
OO
OO
CHOH
CH2OH
CH2-CO
CH2CO
NH H Br+ + +
36. COLORIMETRIC METHODS
METHOD 1: Using 2,4-DINITRO PHENYL
HYDRAZINE:
Dehydro ascorbic acid couples with 2,4-dinitro phenyl
hydrazine to form an osazone which develops a red color in
strong sulphuric acid.
In this reaction, de-hydro ascorbic acid is used so the
titrimetric impurities like –SH, etc. are degraded previously,
so pure ascorbic acid react with DNP and quantized the
exact result.
OO
OO
CHOH
CH2OH
NH-NH2
NO2
O2N
H2SO4
OO
NN
CHOH
CH2OH
N
HN NO2
O2N
NO2
NO2
osazone (pale brown to red color)
H
+
37. METHOD 2: Using 4-METHOXY-2-NITROANILINE:
This is based on ability of ascorbic acid to couple with
Diazonium compounds.
Ascorbic acid with diazotized 4-methoxy-2-nitro aniline forms
a deep blue compound.
This method has an advantage as it is specific for ascorbic acid.
This method has been applied to pharmaceutical preparations,
natural juices, powdered milk and fortified feed.
OO CHOH
OHOH
CH2OH
HNO2
< 8*, NaNO2/HCl
N
NO2
OMe
N-Cl
OO CHOH
OH OH
CH2OH
Deep blue color complex
N
NO2
OMe
N-Cl
diazotised 4-methoxy 2-nitroaniline
+
38. RIBOFLAVIN (VITAMIN B2)
It is also called as lactoflavin.
Properties:
Yellow to orange-yellow, crystalline powder with slight. Odour
Deffiency:
Solubility:
Very slightly soluble in water; more soluble in saline solution than
in water; practically insoluble in chloroform, in ethanol (95%)
and in ether.
Aqueous solutions exhibit an intense yellow color-green fluorescence at pH6.
Specific rotation is Between –115o
and –135o
, determined in a 0.5% w/v solution in
carbonate-free 0.05M sodium hydroxide .
Riboflavin shows absorption maxima at 224, 267, 373, 445, and 475nm.
Irradiation either with uv or visible light, of alkaline solutions, produces luminoflavin,
irradiation of acid or alkaline solutions produces luminochrome, a blue fluorescent
substance.
Reducing agents such as, sodium hydrosulphite, reduce riboflavin to a dihydro
compound, leucoflavin which is not fluorescent. But this is reversible and leucoflavin is
readily oxidized back to riboflavin by atmospheric oxygen.
N
CH2
N
N
NCH3
CH3
O
O
C
H
OH
C HOH
C HOH
CH2
OH
39. METHOD 1: FLUORIMETRIC METHOD
There are three methods are in use in
fluorimetric determination of riboflavin;
Direct determination
Direct additive determination
Adsorptive additive determination
40. FLUORIMETRIC METHOD: DIRECT
DETERMINATION:
This method is employed for the mixtures which are free of interfering pigments
or substances and contain relatively high concentration of riboflavin.
An appropriate quantity is weighed and add with boiling distilled water and
shaken for few minutes, if required, boil the solution. It is then centrifuged.
Sample solution: a suitable aliquot of clear liquid is diluted appropriately to
yield concentration of 0.2mcg/ml.
Blank solution: add a few granules of sodium hydrosulphite to the aliquot of
sample solution. This shows indication of purity of solution.
Measurement and calculation: Readings must be taken as rapidly as possible.
Mcg of riboflavin = A – C * dilution factor
B – C wt of sample (g.)
A reading of unknown concentration.
C unknown blank
B reading of standard
D reading of standard blank.
41. FLUORIMETRIC METHOD: DIRECT ADDITIVE
METHOD
The interference of other substances can be avoided by this method. In this method the
addition of the known quantity of riboflavin to the assay solution is used to compensate
for interfering substance which may absorb the incident or fluorescent light.
Sample treatment: samples of natural origin should be subjected for enzymatic
hydrolysis to digest starchy substances and to “free” any “bound” riboflavin.
Standard solution I: 40mcg/ml of USP reference standard in 20% of ethanol.
Standard solution II: dilute solution I such that it contains 1.6mcg of riboflavinin
distilled water.
Measurement and calculation:
Mcg of riboflavin per gram= A – (1.07C) * 1.6 * 1 dilution factor
B- (0.94A) 16 wt of sample (g)
A fluorescence of sample solution
B fluorescence of standard II
C blank solution ( obtained by adding HYDROGEN SULPHITE to standard solution II and
this should be repeated until successive additions shows no deflections)
1.07 and 0.94 constants due to change in volume of measurement.
42. USP method: in this an additional purification step is
involved by treating with potassium permanganate
and this produces oxidation of interfering
substances. Excess permanganate is removed by
treating with hydrogen peroxide.
Applications:
This method can be used with vitamin mixtures of
wafers, flour enrichment mixtures and simple
pharmaceutical preparations, provided they are
substantially free from coloring or fluorescencing
matter.
43. FLUORIMETRIC METHOD: ADSORPTIVE ADDITIVE
DETERMINATION:
In this method most of the interfering substances are eliminated by an
adsorption step and those not eliminated are compensated by addition of a
known quantity of riboflavin to the assay solution. . this method can be
applied to universally to all samples.
Extraction: the extraction procedure is same as the direct additive
determination.
Adsorption and elution:
An aliquot of sample solution or suitable dilution of the sample extract is
measured accurately and passed through an adsorption column. The column
is prepared by using fluorosil. After elution the column is washed with hot
distilled water and the excess water is drained using vacuum.
The riboflavin is being retained in the column is then eluted using hot acetic
acid-pyridine eluent is collected and mixed well.
44. FLUORIMETRIC METHOD: ADSORPTIVE
ADDITIVE DETERMINATION:
Standard solution I: 40mcg/ml of usp reference standard in 20% of
ethanol.
Standard solution III: dilute solution I such that it contains 1.6mcg of
riboflavinin in acetic acid pyridine mixture.
Measurement and calculation:
Mcg of riboflavin per gram= A – (1.07C) * 1.6 * 1 dilution factor
B- (0.94A) 16 wt of sample (g)
A fluorescence of sample solution
B fluorescence of standard III
C blank solution ( obtained by adding hydrogen sulphite to standard
solution II and this should be repeated until successive additions shows no
deflections)
1.07 and 0.94 constants due to change in volume of measurement.
45. METHOD 2: SPECTROPHOTOMETRIC METHOD (IP 1996):
Procedure:
Carry out the procedure in subdued light.
Weigh accurately about 65 mg and transfer to an amber-glass 500-
ml volumetric flask, suspend in 5 ml of water, ensuring that it is
completely wetted.
Dissolve in 5 ml of 2M sodium hydroxide. As soon as dissolution is
complete add 100 ml of water and 2.5 ml of glacial acetic acid and
dilute to 500.0 ml with water.
To 20.0 ml of this solution add 3.5 ml of a 1.4% w/v solution of
sodium acetate and dilute to 200.0 ml with water.
Measure the absorbance of the resulting solution at the maximum at
about 444 nm.
Calculate the content of C17H20N4O6 taking 328 as the value of A(1%, 1
cm) at the maximum at about 444 nm.
46. METHOD 2:
Riboflavin has a characteristic absorption spectrum in water with a
maximum at 267nm. This is the basis of the method. This method is
based on the assumption that extraction with chloroform will remove
impurities from an aqueous solution.
Procedure:
Carry out in subdued light.
Weighed about 20mg of riboflavin transferred to a 1000ml volumetric
flask, dilute the solution and add few drops of 1N NaOH.
Shake gently until the solution is complete, then add few drops of 5N
acetic acid and dilute upto the mark.
A 20ml of aliquot of solution is taken and shaken with 25ml of chloroform
for 1min. separate the chloroform layer and discard. The extraction is
repeated twice.
47. METHOD 2 CONTD…
The absorbance of clear aqueous layer is
determined at 267nm.with water as a
reference.
Repeat the procedure with reference
sample.
% Riboflavin= 100 (As/Ar) . (Wr/Ws)
Ws and As weight in mg and absorbance
of sample respectively.
Wr and Ar corresponding values of
standard and reference sample of riboflavin.
48. REFERENCES:
Pharmaceutical analysis by Takeru Higuchi et.al. page no. 649-707.
Bentley and Drivers textbook of pharmaceutical chemistry. 8th
edition.
Vogel’s textbook of quantitative chemical analysis.
IP-1996.
Instrumental methods of chemical analysis by G R Chatwal and Sham K
Anand. Enlarged edition. 2005. p no. 2.413-2.414
Guyton textbook of medical physiology. 11th
edition.
Biochemistry by Leninger 4th
edition 2005.
Biochemistry By Jeremy M Berg, John L Tymoczko And Lubert Stryler. 5th
Edition