Karl Fischer titration

Karl Fischer titration is a classic titration method in analytical chemistry that uses coulometric or volumetric titration to determine trace amounts of water in a sample. It was invented in 1935 by the German chemist Karl Fischer.

Advantage of analysis

The popularity of the Karl Fischer titration is due in large part to several practical advantages that it holds over other methods of moisture determination, including:

01.  High accuracy and precision

02.  Selectivity for water

03.  Small sample quantities required

04.  Easy sample preparation

05.  Short analysis duration

06.  Nearly unlimited measuring range (1ppm to 100%)

07.  Suitability for analyzing:

Ø  Solids

Ø  Liquids

Ø  Gases

08.  Independence of presence of other volatiles

09.  Suitability for automation

10.  In contrast, loss on drying will detect the loss of any volatile substance.

The major disadvantage is that the water has to be accessible and easily brought into methanol solution. Many common substances, especially foods such as chocolate, release water slowly and with difficulty, and require additional efforts to reliably bring the total water content into contact with the Karl Fischer reagents.

TRANSGENESIS

TRANSGENESIS:

Modification of germ cells by gene manipulation in vitro is called transgenesis/ transfection/ transformation.

TRANSGENIC ANIMAL:

Animal carrying new gene(s) called transgenic. Particular variants of species into which a new gene(s) has been inserted.

Every cell of the animal carries new genetic information within the genome.

Why Transformation is performed?

1. To incorporate trait to enhance the conventional animal product
2. To incorporate traits for biosynthesis of new product
3. Study of gene expression in mammalian development
4. Study of tissue specific expression
5. Study of genetic basis of various diseases
6. Gene function study through knockouts
7. Minimize the time requirement to introduce a trait that is needed in breeding

Transgenic Mice Production

Embryonic Stem (ES) Cell Method

Cells of the blastocyst stage of a developing mouse can proliferate in cell culture and still retain the capability of differentiating into all other types, including germ line cells, after they are reintroduced into another blastocyst embryo. Such cells are called pluripotent embryonic stem cells.

Another advantage: the transgenes can be targeted to introduced into a prefixed location.

Production of Chimeric Mouse :

An early embryo is isolated from the fertilized mouse of black colour. The ES cells of trophoblast stage of embryo are grown on culture medium. A small number of ES cells can be injected into blastocoel space of an embryo of a white (albino) mouse through microinjection technique. The ES cells of black mouse intermingle with that of albino. The microinjected embryo is transplanted into the uterus of a surrogate mother (another mouse of which ova we not used). The progeny born has black and white skin colour. Such mouse was called chimera or chimeric mouse.

Fig: Production of Transgenic Mice 

Application of Transgenic Mice

1. Understanding gene regulation
2. Immunological specificity understanding and antibody development
3. Drug production
4. Onset and progress of human disease e.g. Alzheimer disease, arthritis, muscular dystrophy, tumorigenesis etc.

Powdered Glass Test

Rinse thoroughly with Purified Water six or more containers selected at random, and dry them with a current of clean, dry air. Crush the containers into fragments about 25 mm in size, divide about 100 g of the coarsely crushed glass into three approximately equal portions, and place one of the portions in the special mortar. With the pestle in place, crush the glass further by striking 3 or 4 blows with the hammer. Nest the sieves, and empty the mortar into the No. 20 sieve. Repeat the operation on each of the two remaining portions of glass, emptying the mortar each time into the No. 20 sieve. Shake the sieves for a short time, then remove the glass from the Nos. 20 and 40 sieves, and again crush and sieve as before. Repeat again this crushing and sieving operation. Empty the receiving pan, reassemble the nest of sieves, and shake by mechanical means for 5 minutes or by hand for an equivalent length of time. Transfer the portion retained on the No. 50 sieve, which should weigh in excess of 10 g, to a closed container, and store in a desiccator until used for the test.

Spread the specimen on a piece of glazed paper, and pass a magnet through it to remove particles of iron that may be introduced during the crushing. Transfer the specimen to a 250-mL conical flask of resistant glass, and wash it with six 30-mL portions of acetone, swirling each time for about 30 seconds, and carefully decanting the acetone. After washing, the specimen should be free from agglomerations of glass powder, and the surface of the grains should be practically free from adhering fine particles. Dry the flask and contents for 20 minutes at 140 , transfer the grains to a weighing bottle, and cool in a desiccator. Use the test specimen within 48 hours after drying.

Procedure

Transfer 10.00 g of the prepared specimen, accurately weighed, to a 250-mL conical flask that has been digested (aged) previously with High-Purity Water in a bath at 90 for at least 24 hours or at 121 for 1 hour. Add 50.0 mL of High-Purity Water to this flask and to one similarly prepared to provide a blank. Cap all flasks with borosilicate glass beakers that previously have been treated as described for the flasks and that are of such size that the bottoms of the beakers fit snugly down on the top rims of the containers. Place the containers in the autoclave, and close it securely, leaving the vent cock open. Heat until steam issues vigorously from the vent cock, and continue heating for 10 minutes. Close the vent cock, and adjust the temperature to 121, taking 19 to 23 minutes to reach the desired temperature. Hold the temperature at 121 ± 2.0 for 30 minutes, counting from the time this temperature is reached. Reduce the heat so that the autoclave cools and comes to atmospheric pressure in 38 to 46 minutes, being vented as necessary to prevent the formation of a vacuum. Cool the flask at once in running water, decant the water from the flask into a suitably cleansed vessel, and wash the residual powdered glass with four 15-mL portions of High-Purity Water, adding the decanted washings to the main portion. Add 5 drops of Methyl Red Solution, and titrate immediately with 0.020 N sulfuric acid. If the volume of titrating solution is expected to be less than 10 mL, use a microburet. Record the volume of 0.020 N sulfuric acid used to neutralize the extract from 10 g of the prepared specimen of glass, corrected for a blank. The volume does not exceed that indicated in Table for the type of glass concerned.

Table: Test Limits for Powdered Glass Test

			Limits
Type	General Description a	Type of Test	Size, b  mL	mL of 0.020 N Acid
I	Highly resistant, borosilicate glass	Powdered Glass	All	1.0
III	Soda-lime glass	Powdered Glass	All	8.5
a  The description applies to containers of this type of glass usually available. b  Size indicates the overflow capacity of the container.

The basic requirements for quality control

The basic requirements for quality control are as follows:

(a)     adequate facilities, trained personnel and approved procedures must be available for sampling, inspecting, and testing starting materials, packaging materials, and intermediate, bulk, and finished products, and where appropriate for monitoring environmental conditions for GMP purposes;

(b)    samples of starting materials, packaging materials, intermediate products, bulk products and finished products must be taken by methods and personnel approved of by the quality control department;

(c)     qualifications and validation must be performed;  

(d)    records must be made (manually and/or by recording instruments) demonstrating that all the required sampling, inspecting and testing procedures have actually been carried out and that any deviations have been fully recorded and investigated;

(e)     the finished products must contain ingredients complying with the qualitative and quantitative composition of the product described in the marketing authorization; the ingredients must be of the required purity, in their proper container and correctly labeled;

(f)     records must be made of the results of inspecting and testing the materials and intermediate, bulk and finished products against specifications; product assessment must include a review and evaluation of the relevant production documentation and an assessment of deviations from specified procedures;

(g)   No batch of product is to be released for sale or supply prior to certification by the authorized person(s) that it is in accordance with the requirements of the marketing authorization. In certain countries, by law, the batch release is a task of the authorized person from production together with the authorized person from quality control;

(           h)    Sufficient samples of starting materials and products must be retained to permit future examination of       the product if necessary; the retained product must be kept in its final pack unless the pack is exceptionally large.

Cross-Contamination

Cross-Contamination

The term cross-contamination refers to product-to-product contamination. It can be through careless sharing of the manufacturing equipment and utensils, sharing of space without proper cleaning, poor production planning and particularly inadequate dust control. It is known that contamination by some potent drugs, even at a very minute level, can cause hyper-sensitivity reaction like anaphylaxis which may even risk the life of the patient under certain clinical circumstances. In the current GMP approach the possibility of cross-contamination with penicillin, steroids and some highly sensitive drugs has become a great concern to all. For instance a very small fraction of population may get severe allergic reaction to penicillin called anaphylactic shock with nominal amounts of penicillin i.e. as low as 0.01 IU with parenteral dosage. In case of oral administration the tolerance limit is higher.

While dealing with a highly sensitive product likes penicillin the recommended approach il having a dedicated manufacturing facilities where the objective is totally isolating such

How can cross-contamination prevent?

Cross-contamination should be avoided by taking appropriate technical or organizational measures, for example:

(a) Carrying out production in dedicated and self-contained areas (which may be required for products such as penicillin's, live vaccines, live bacterial preparations and certain other biological s);

(b) Conducting campaign production (separation in time) followed by appropriate cleaning in accordance with a validated cleaning procedure;

(c) Providing appropriately designed airlocks, pressure differentials, and air supply and extraction systems;

(d) Minimizing the risk of contamination caused by re-circulation or re-entry of untreated or insufficiently treated air;

 (e) Wearing protective clothing where products or materials are handled;

(f) Using cleaning and decontamination procedures of known effectiveness;

(g) Using a “closed system” in production;

(h) Testing for residues;

(i) Using cleanliness status labels on equipment.

Quality Assurance

Quality Assurance

Quality assurance is a wide-ranging concept covering all matters that individually or collectively influence the quality of a product. It is the totality of the arrangements made with the object of ensuring that pharmaceutical products are of the quality required for their intended use. QA therefore includes GMP and other factors such as product design and development

QA=Product design + GMP + QC + quality goal activities

Criteria of Quality

Safety

Potency

Efficacy

Stability

Acceptability

Regulatory compliance

1.1 Principle. “Quality assurance” is a wide-ranging concept covering all matters that individually or collectively influence the quality of a product. It is the totality of the arrangements made with the object of ensuring that pharmaceutical products are of the quality required for their intended use. Quality assurance therefore incorporates GMP and other factors, including those outside the scope of this guide such as product design and development.

1.2 The system of quality assurance appropriate to the manufacture of pharmaceutical products should ensure that:

(a) Pharmaceutical products are designed and developed in a way that takes account of the requirements of GMP and other Good manufacturing practices for pharmaceutical products, Part One. In: WHO Expert Committee on specifications for Pharmaceutical Preparations. Thirty-second report. Geneva, World Health Organization, 1992, Annex 1 (WHO Technical Report Series, No. 823).

Associated codes such as those of good laboratory practice (GLP) and good clinical practice (GCP);

(b) Production and control operations are clearly specified in a writ­ten form and GMP requirements are adopted;

(c) Managerial responsibilities are clearly specified in job descriptions;

(d) Arrangements are made for the manufacture, supply and use of the correct starting and packaging materials;

(e) All necessary controls on starting materials, intermediate prod­ucts, and bulk products and other in-process controls, calibra­tions, and validations are carried out;

(f) The finished product is correctly processed and checked, accord­ing to the defined procedures;

(g) pharmaceutical products are not sold or supplied before the au­thorized persons (see also sections 9.11 & 9.12) have certified that each production batch has been produced and controlled in ac­cordance with the requirements of the marketing authorization and any other regulations relevant to the production, control and release of pharmaceutical products;

h) Satisfactory arrangements exist to ensure, as far as possible, that the pharmaceutical products are stored by the manufacturer, dis­tributed, and subsequently handled so that quality is maintained throughout their shelf-life;

(i) There is a procedure for self-inspection and/or quality audit that regularly appraises the effectiveness and applicability of the qual­ity assurance system;

(j) Deviations are reported, investigated and recorded;

(k) There is a system for approving changes that may have an impact on product quality;

(l) Regular evaluations of the quality of pharmaceutical products should be conducted with the objective of verifying the consis­tency of the process and ensuring its continuous improvement.

1.3 The manufacturer must assume responsibility for the quality of the pharmaceutical products to ensure that they are fit for their in­tended use, comply with the requirements of the marketing authoriza­tion and do not place patients at risk due to inadequate safety, quality or efficacy. The attainment of this quality objective is the responsibil­ity of senior management and requires the participation and commit­ment of staff in many different departments and at all levels within the company.

This is a code governing the testing of chemicals to obtain data on their properties and ensuring safety with respect to human health and the environment. It is different from that described in “Good laboratory practices in governmental drug control laboratories” in the Thirtieth report of the WHO Expert Committee on specifications for Pharmaceutical Preparations (WHO Technical Report Series, No. 748, 1987, Annex 1).

Company, the company’s suppliers, and the distributors. To achieve the quality objective reliably there must be a comprehensively de­signed and correctly implemented system of quality assurance in­corporating GMP and quality control. It should be fully documented and its effectiveness monitored. All parts of the quality assurance sys­tem should be adequately staffed with competent personnel, and should have suitable and sufficient premises, equipment, and facilities.

Good Manufacturing Practice (GMP)

GMP refers to the Good Manufacturing Practice Regulations promulgated by the US Food and Drug Administration under the authority of the Federal Food, Drug and Cosmetic Act. These regulations require that manufacturers, processors, and packagers of drugs, medical devices, some food, and blood and service providers take proactive steps to ensure that their products are safe, pure, and effective. GMP regulations require a quality approach to manufacturing, enabling companies to minimize or eliminate contamination, mix-ups, and errors. Failure of industries to comply with GMP regulations can result in very serious consequences including recall, seizure, fines, and jail terms.

GMP regulations address issues including recordkeeping, personnel qualifications, sanitation, cleanliness, equipment verification, process validation, and complaint handling. Most GMP requirements are very general and open-ended, allowing each manufacturer to decide individually how to implement the necessary controls. This provides much flexibility, but also requires that the manufacturer interpret the requirements in a manner which makes sense for each individual business.

GMP is sometimes referred to as "cGMP". The "c" stands for "current," reminding manufacturers that they must employ technologies and systems which are up-to-date in order to comply with the regulation. Systems and equipment used to prevent contamination, mix-ups, and errors, which may have been "top-of-the-line" 20 years ago, may be less than adequate by today's standards.  GMP is a good business tool which helps to refine both compliance and performance at company level.

Main Parameters of GMP

i.Quality Assurance

ii. GMP for pharmaceutical products

iii. Sanitation and hygiene

iv. Qualification and validation

v. Complaints

vi. Product recalls

vii. Contract Manufacturing and analysis

viii. Self inspection and quality audits

ix. Personnel

x. Training

xi. Personal hygiene

xii. Premises

xiii. Equipment

xiv. Materials

xv. Documentation

xvi. Good practices in production

xvii. Good practices in quality control

Prescriptions

DEFINITION

Prescription is an order written by a physician, dentist, veterinarian or a registered medical practitioner to a pharmacist to compound and dispense a specific medication for the patient.

Important features of a prescription:

·         Directions are given to the pharmacist about what type of preparation (tablet, power, mixture etc.) is to be prepared.

·         It contains directions for the patients, the dose of the drug and the dose interval, and how it is to be taken.

·         Prescriptions are generally written in Latin language, so that the prescription remains unknown to the patients to avoid self-medication.

PARTS OF A PRESCRIPTION

A typical prescription consists of the following parts:

1. Date

Date on the prescription helps the pharmacists to know when the medicines were last dispensed if the prescription is brought for redispensing of the prescription. In case of habit forming drug  the date prevents the misuse of the drug by the patient.

2. Name, age, sex and address of the patient

By name and address the patient and the prescription can be identified. Age and sex of the patient is especially required for child patient to check the prescribed dose.

3. Superscription

It is represented by a Latin symbol ℞, an abbreviation of Latin term ‘recipe’ which means ‘take thou’ or ‘you take’.

[N.B. In olden days, the symbol was considered to be originated from the sign of Jupiter, the Greek God of healing. This symbol was employed by the ancient in requesting God for the quick recovery of the patient.]

4. Inscription

This is the main part of the prescription.  It contains the names and quantities of the prescribed medicaments. The medicament may be official preparation or nonofficial preparation. If is official preparation (i.e. from pharmacopoeia or formulary) then only the name of the preparation is written e.g. Piperazine Citrate Elixir IP.

If it is nonofficial preparation then the quantity of each ingredient will be given. The type of preparation will also be given e.g.               Sodium bicarbonate           3g

                                Simple Syrup                        6ml

                                Purified Water q.s.               100ml

The inscription of prescriptions containing several ingredients are divided into the following parts:

(a)     Base: The active medicaments those are intended to produce the therapeutic effect.

(b)     Adjuvants: These are included either to enhance the action of the drug or to make the preparation more palatable.

(c)     Vehicle: It is the main carrier of the drug. In liquid preparations drugs are either dissolved or dispersed in the vehicle.

5. Subscription

In this part the prescriber gives direction to the pharmacist regarding the dosage form to be prepared and the number of doses to be dispensed.

6. Signatura

It is usually written as ‘Sig.’. The instructions given in the prescription should be written in the label of the container so that the patient can follow them. The instructions may include:

                (a) The quantity to be taken             (b) The frequency and timing of administration of the preparation

                (c) The route of administration         (c) The special instruction (if any)

7. Renewal instructions

The prescriber indicates in every prescription, whether it should be renewed, and if renewed, for how many times. It is very important particularly for the case of habit forming drugs to prevent its misuse.

8. Signature, address and registration number of the prescriber

The prescription must be signed by the prescriber by his / her own hand. His/her address and registration number should be written in the case of dangerous and habit forming drugs.

An example of a typical prescription is given as follows:

SHARMA NURSING HOME New Delhi

Name: Mr. N. Anand           Age: 42 years                       Sex: Male Address: 32, Azad Nagar, new Delhi             ℞ (Superscription)                                 Sodium bicarbonate                           3g Inscription               Compound tincture of cardamom   2ml                                 Simple Syrup                                        6ml                                 Purified Water q.s.                               90ml Fiat misture. (Subscription) Sig. Cochleare magnum ter in die post cibos sumenda. (Signatura) Refill: ________                                                                 Sd/-                                                                                 Dr. Aswini Sharma                                                                                 Regn. No. 14328

HANDLING OF PRESCRIPTION

The following procedures should be adopted by the pharmacist while handling the prescription for compounding  and dispensing:

                (i) Receiving         

(ii) Reading and checking

(iii) Collecting and weighing the materials

(iv) Compounding, labeling and packaging

(i) Receiving

·         The prescription should be received by the pharmacist himself / herself.

·         While receiving a prescription from a patient a pharmacist should bot change his/her facial expression that gives an impression to the patient that he/she is confused or surprised after seeing the prescription.

(ii) Reading and checking

·         After receiving the prescription it should be screened behind the counter.

·         The prescription is a hospital slip or from a nursing home or from a private practitioner and their authenticity should be checked. The signature of the prescriber and the date of prescription is checked.

·         The pharmacist should read all the lines and words of the prescription. He/she must not guess any word. If there is any doubt, the pharmacist should consult with the other pharmacist or the prescriber over telephone.

3. Collecting and weighing the material

Before compounding a prescription all the materials required for it should be collected from the shelves or drawers and kept in the left hand side of the balance. After measuring each material should be kept on the right hand side of the balance. After compounding the prescription the materials are replaced back to the shelves / drawers where from they were collected.

While compounding the label of every container of material should be checked thrice in the following manner:

(i)                   When collected from the shelves/drawers.

(ii)                 When the materials are measured.

(iii)                When the containers are replaced back to the shelves/drawers.

4. Compounding, labeling and packaging

·         Only one prescription should be compounded at a time.

·         Compounding should be done on a clean table.

·         All equipment required should be cleaned and dried.

·         The preparation should be prepared according to the direction of the prescriber or as per methods given in pharmacopoeia or formulary are according to established pharmaceutical art of compounding.

·         The compounded preparations should be filled in suitable containers.

Round vials	For tablets and capsules
Oval prescription bottles Narrow mouthed	For liquid of low viscosity e.g. mixtures, oral emulsions etc.
Wide mouthed bottles	For filling liquids of high viscosity, large quantities of tablets or capsules and bulk powders.
Colored fluted bottles	For external preparations e.g. liniment and lotions.
Ointment jars and collapsible tubes	For ointments, creams or any other semisolid dosage forms.
Paper wrappers and envelops	For oral powders in divided doses.
Dropper bottles	For eye drops and ear drops.
Sifter top containers	For dusting powders.

·         The containers are labeled as per the prescriber. If required some additional instructions may also be given.

The following information should be written on the label:

Type of preparation:	The Emulsion, The Mixture, The Powder etc. Its quantity should also be mentioned.
For:	Name, Age and Sex of the patient.
Date of dispensing:	Date on which the prescription is dispensed.
Expiry date if any:	e.g. ‘Must be taken within 7 days of dispensing.’
Directions for use:	e.g ‘One teaspoonful thrice daily.’
Storage condition:	e.g. ‘Keep in a cool place’
Secondary labeling:	e.g. ‘SHAKE THE BOTTLE BEFORE USE’       ‘FOR EXTERNAL USE ONLY’ etc.
Name and signature	of the pharmacist who dispensed
Name and Address	of the Pharmacy

·         The container should be polished to remove any finger print.

·         While delivering the preparation to the patient the pharmacist should explain the mode of administration, direction for use and storage.