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A Report Submitted to SVKMs Narsee Monjee Institute of Management Studies (NMIMS) As a part of industrial training for MBA ( Pharm.

Tech)
By Ms. Shailja Shah Under the Guidance of

Mr. Bagade S. B.

School of Pharmacy & Technology Management, SVKMs Narsee Monjee Institute of Management Studies (NMIMS) V.L Mehta Road Vile Parle (W) Mumbai -400056 June 2010

Statement by the candidate


As required by University regulation, I wish to state that the work embodied in this report has not been submitted for any other degree to this or to any other University. Wherever references have been made to previous work of others, it has been clearly indicated as such and included in the bibliography.

Ms. SHAILJA SHAH Forwarded Through


Faculty Ms. VARSHA PRADHAN Asst. Professor, Pharmaceutical Sciences, Department of Pharmaceutical Chemistry, School of Pharmacy & Technology Management, SVKMs, NMIMS, Vile Parle (W), Mumbai -400 056

INTRODUCTION
Pharmacy is the health profession that links the health sciences with the chemical sciences and it is charged with ensuring the safe and effective use of pharmaceutical drugs. The word derives from the Greek (pharmakon), "drug, medicine"[1] (the earliest form of the word is the Mycenaean Greek pa-ma-ko, attested in Linear B syllabic script[2]). The scope of pharmacy practice includes more traditional roles such as compounding and dispensing medications, and it also includes more modern services related to health care, including clinical services, reviewing medications for safety and efficacy, and providing drug information. Pharmacists, therefore, are the experts on drug therapy and are the primary health professionals who optimize medication use to provide patients with positive health outcomes. An establishment in which pharmacy (in the first sense) is practiced is called a pharmacy, chemist's or drug store. These stores commonly sell not only medicines, but also miscellaneous items such as candy (sweets), cosmetics, and magazines, as well as light refreshments or groceries. There are various industries that have changed the current scenario of the pharmaceutical sector drastically. Cadilla healthcare is considered to be a pioneer in the miscellaneous segments of pharmaceutical industry. 'Cadila Healthcare' an Indian pharmaceutical company head quartered at Ahmedabad in Gujarat state is the fifth largest pharmaceutical company in India with US$290m in turnover in 2004. It is a significant manufacturer of generic drugs. Cadilla healthcare have developed a drug named Roserin which has reduced the cost of curing TB by 33%.

History
Cadila Laboratories was founded in 1952 by Shri Ramanbhai Patel (1925-2001), formerly a lecturer in the L.M. College of Pharmacy, and his business partner Shri Indravadan Modi. The company evolved over the next four decades into one of India's established pharmaceutical companies.

In 1995 the Patel and Modi families split, with the Modi family's share being moved into a new company called Cadila Pharmaceuticals Ltd. and Cadila Healthcare became the Patel family's holding company. Cadila Healthcare did its IPO on the Bombay Stock Exchange in 2000. Its stock code on the Bombay exchange is 532321.

In 2001 the company acquired another Indian pharmaceutical company called German Remedies. On June 25, 2007, the company signed an agreement to acquire 100 per cent stake in Brazils Quimica e Farmaceutica Nikkho do Brasil Ltda (Nikkho) for around 26 million dollars.

In keeping with its mission to create healthier communities globally, Zydus Cadila is actively involved in developing the community of which it forms a part.Through the Ramanbhai Foundation, it has undertaken initiatives in the field of pharmaceutical research, education and healthcare. Under the aegis of the Foundation, the group organises an International Symposium on the latest trends in Pharmaceutical Sciences, once every two years. Three International symposia have been held so far with more than 40 research scientists, academicians and research professionals from across the world sharing their insights on the latest developments in pharmaceutical research.

The Zydus School for Excellence a centre for learning where young minds are free to grow in relationship to his or her potential has also been set up Under the aegis of the Foundation. The Shri Ramanbhai Patel Memorial Scholarship Programme jointly funded by a grant from the Ramanbhai Foundation and Zydus School for Excellence funds the education of students from disadvantaged backgrounds. Shri Ramanbhai B. Patel - AMA Centre for Excellence in Education, provides a platform for parents, teachers and students to highlight the critical educational issues of the day. The centre conducts open house discussions, memorial lectures on excellence in education, progressive learning programmes for academicians and knowledge sharing forums. The centre also honours teachers for their contributions to a childs world of learning.

Dedicated to the memory of the groups founder, the IPA Shri Ramanbhai B. Patel Foundation (IRF) has been jointly set up with the Indian Pharmaceutical Association. IRF recognises and honours commitment and excellence in the field of pharmacy. As a part of its outreach programmes, the group organises rural healthcare camps annually with its team of medical advisors and employees, volunteering their services. So far, the group has organised general healthcare camps, diagnostic, dental-care, eye-care and paediatric camps. The group has also initiated 'Project Unnati' for adolescent girls at Moraiya which focusses on health and overall development. Through the Ramanbhai Foundation, the group also extends its support during times of natural calamities. In recognition of its CSR initiatives, the group bagged the Social and Corporate Governance Awards 2008 in the Best Social Responsibility Practice category presented by the BSE, NASSCOM Foundation and Times Foundation. Three of the groups facilities including the formulation manufacturing plant at Moraiya, and API plants at Ankleshwar and Dabhasa near Vadodara are approved by the USFDA. More than 900 professionals spearhead the groups research programme. Over 350 scientists are working on new molecular entity research at the Zydus Research Centre. The group has six INDs in various stages of clinical trials and two more INDs are in pre-clinical evaluation. Zydus Cadila is a partner of choice for several global pharma majors such as Boehringer Ingelheim, Bayer Schering Pharma, Madaus AG, Nycomed, Hospira, Bio Sidus of Argentina, Mallinckrodt of USA, to name a few. One of the most reputed pharma companies globally, Zydus Cadila aims to be a leading global healthcare provider with a robust product pipeline andsales of over $1 billion by 2010. It plans to achieve sales of over $3 billion by 2015 and be a global research-driven company by 2020.

OPERATIONS Formulations Business - India


With three multi-therapy divisions and eight specialty divisions, Zydus Cadila is one of the leading player in the Indian healthcare industry. It is the leading player in the cardiovascular, gastrointestinal and women's healthcare segments. The group has strong presence in respiratory, pain management, CNS, anti-infectives, oncology, neurosciences, dermatology and nephrology segments. It has been able to maintain overall position and market share through faster growing chronic / lifestyle segments. With several new product introductions and pillar brands such as Aten, Ocid, Deriphyllin, Pantodac, Atorva, Nucoxia, Mifegest to name a few, Zydus Cadila is considered a tour-de-force in therapy management and brand management. The group has several in-licensing alliances with global multinationals such as Schering AG, Boehringer Ingelheim, Viatris, etc. The portfolio of over 200 products are marketed by a specialised field force of 3,000. With one of the strongest distribution channels in the industry, the group reaches out to 1,00,000 chemists and serves 2,00,000 doctors including physicians, specialists and super specialists. Consumer Products Business - Zydus Wellness Ltd. Apart from a strong presence in the prescription market, the group also caters to the growing health and wellness segment through its subsidiary Zydus Wellness Ltd. Zydus Wellness aims to promote healthy living by anticipating the emerging and day-to-day needs in dietetic / health foods. Health and wellness have been identified as the emerging areas in consumer healthcare. The Company is focussed on empowering individuals who wish to adopt healthy eating habits and lifestyles. The Company is a pioneer, offering healthier dietary options to the consumers. The product range comprises Sugar Free Gold Indias No.1 sweetener with a market share of over 70%, Sugar Free Natura a zero calorie sucralose based sugar substitute, Sugar Free Dlite a low calorie healthy drink and Nutralite a premium cholesterol-free table spread. Nutralite has emerged as the second largest brand in the category of butter and butter substitutes. The Company also caters to the skincare segment with its Everyuth and Dermacare brands, which occupy a unique distinction of being a skincare brand of a healthcare company. Enriched with the power of natural ingredients, EverYuth has a strong presence in advanced skincare segments like soap-free, face washes, face masks, scrubs etc.

GLOBAL
Zydus Cadila has strong global opetations to market both formulations and APIs. With operations in U.S.A., Europe, Latin America, Japan and over 40 emerging markets across the world, Zydus Cadila is a global healthcare provider. The group also manufactures and markets a wide range of intermediates and APIs.

PRODUCTS

From nine pharmaceutical production operations in India as well as a major R&D operation Zydus Cadila develops and manufactures a large range of pharmaceuticals as well as diagnostics, herbal products, skin care products and other OTC products. The company also makes EverYuth Naturals Walnut Scrub & Ultra Mild Scrub -India's leading scrub brand, EverYuth Naturals Golden Glow Peel-Off-the no. 1 in the peel-off category and a face wash range. It is also the maker of Sugar Free, India's most popular artificial sweetener and Nutralite, India's most popular cholesterol-free margarine.

MANUFACTURING
Wide ranges of capabilities have helped the group create a robust manufacturing infrastructure. Spread over five states of India, the group has eight state-of-the-art manufacturing facilities, creating a strong global manufacturing hub for the group.

Active pharmaceutical ingredient plants


The company makes active pharmaceutical ingredients at three sites in India: 1. Ankleshwar plants- Zydus Cadila's plant complex at Ankleshwar in Bharuch District of Gujarat has been producing drug material since 1972. There are around 10 plants in the complex, which is ISO 9002 and ISO 14001 certified as well as FDA Approved. Total plant capacity at Ankleshwar is around 180 million tones. 2. Vadodara plant- Zydus Cadila's plant at Dhabhasa, in Vadodara District's Padra taluka (in the eastern part of the district) in Gujarat, was commissioned in 1997 by a company called Banyan Chemicals, and acquired by Zydus Cadila in 2002. The plant has a 90 million tone capacity. It is an FDA-approved facility that is also approved to WHO GMP guidelines. 3. Patalganga plant- Zydus Cadila acquired an API plant at Patalganga in Maharashtra state, 70 km from Mumbai, in the 2001 German Remedies deal. This plant operates to WHO GMP standards.

Formulation plants
The company operates formulation plants at six locations: 1. Moraiya plant- Zydus Cadila's formulation plant at Moraiya in Sanand taluka on the outskirts of Ahmadabad is the largest formulation plant in India. It plant became Food and Drug Administration (FDA)-approved in 2004/2005. The plant makes tablets, capsules, and soft gel capsules as well as injectable drugs in both sterile liquid and lyophilized form. Zydus Cadila also runs a large R&D operation at Moraiya. 2. Vatwa plant- Zydus Cadila's plant at Vatwa, an industrial suburb of Ahmadabad, makes nutraceuticals. The plant was acquired Remedies. 3. Changador plant- Zydus Cadila's plant at Changodar, 20 kilometers from Ahmadabad on the city's outskirts, manufactures fine chemicals. Zydus is current constructing a facility at Changodar to make vaccines for hepatitis B and rabies. 4. Navi Mumbai plant- This operation, at Navi Mumbai in Maharashtra, is a 50/50 joint venture with Germany's Altana Pharma AG, makes intermediates of the drug pantoprazole. 5. Goa plants- The Companys plants at Ponda in the southern Indian state of Goa do formulation work as well as manufacture oncology drugs and a herbal laxative branded Agiolax based on Psyllium seeds. 6. Baddi plant- In 2004 Zydus commissioned at formulation plant at Baddi, in Himachal Pradesh state of northern India. The Baddi plant makes solid oral pharmaceuticals. Plant at Mumbai where tablets are made from hands of labours.

ANALYTICAL DEVELOPMENT LABORATORY (ADL)


It is commonly abbreviated as ADL. This is a department that requires extensive manual work. It performs the testing of formulated substances from F&D lab and yields the result with respect to the pharmacokinetic profile of the drug. The analysis and the result interpretation of each formulation is different and is symbiotically depends upon the markets they are formulated for. The F&D department of each market sends their test formulations to their respective ADL. After the result is thoroughly studied and evaluated, if a formulation is feasible for a specific market it is sent to pilot plant for batch manufacturing. The various equipments and instruments utilized are listed below.

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY


High-performance liquid chromatography (or high-pressure liquid chromatography, HPLC) is a form of column chromatography used frequently in biochemistry and analytical chemistry to separate, identify, and quantify compounds based on their idiosyncratic polarities and interactions with the column's stationary phase. HPLC utilizes different types of stationary phase (typically, hydrophobic saturated carbon chains), a pump that moves the mobile phase(s) and analyte through the column, and a detector that provides a characteristic retention time for the analyte. The detector may also provide other characteristic information (i.e. UV/Vis spectroscopic data for analyte if so equipped). Analyte retention time varies depending on the strength of its interactions with the stationary phase, the ratio/composition of solvent(s) used, and the flow rate of the mobile phase. With HPLC, a pump (rather than gravity) provides the higher pressure required to propel the mobile phase and analyte through the densely packed column. The increased density arises from smaller particle sizes. This allows for a better separation on columns of shorter length when compared to ordinary column chromatography.

OPERATION
The sample to be analyzed is introduced in small volume to the stream of mobile phase. The analyte's motion through the column is slowed by specific chemical or physical interactions with the stationary phase as it traverses the length of the column. How much the analyte is slowed depends on the nature of the analyte and on the compositions of the stationary and mobile phases. The time at which a specific analyte elutes (comes out of the end of the column) is called the retention time; the retention time under particular conditions is considered reasonably unique identifying characteristic of a given analyte. The use of smaller particle size column packing (which creates higher backpressure) increases the linear velocity giving the components less time to diffuse within the column, leading to improved resolution in the resulting chromatogram. Common solvents used include any miscible combination of water or various organic liquids (the most common are methanol and acetonitrile). Water may contain buffers or salts to assist in the separation of the analyte components, or compounds such as trifluoroacetic acid which acts as an ion pairing agent. A further refinement to HPLC has been to vary the mobile phase composition during the analysis; this is known as gradient elution. A normal gradient for reversed phase chromatography might start at 5% methanol and progress linearly to 50% methanol over 25 minutes; the gradient chosen depends on how hydrophobic the analyte is. The gradient separates the analyte mixtures as a function of the affinity of the analyte for the current mobile phase composition relative to the stationary phase. This partitioning process is similar to that which occurs during a liquid-liquid extraction but is continuous, not step-wise. In this example, using a water/methanol gradient, the more hydrophobic components will elute (come off the column) when the mobile phase consists mostly of methanol (giving a relatively hydrophobic mobile phase). The more hydrophilic compounds will elute under conditions of relatively low methanol/high water. The choice of solvents, additives and gradient depend on the nature of the stationary phase and the analyte. Often a series of tests are performed on the analyte and a number of trial runs may be processed in order to find the HPLC method which gives the best separation of peaks.

Types
Partition chromatography
Partition chromatography uses a retained solvent, on the surface or within the grains or fibres of an "inert" solid supporting matrix as with paper chromatography; or takes advantage of some additional coulombic and/or hydrogen donor interaction with the solid support. Molecules equilibrate (partition) between a liquid stationary phase and the eluent. Known as Hydrophilic Interaction Chromatography (HILIC) in HPLC, this method separates analytes based on polar differences. HILIC most often uses a bonded polar stationary phase and a non-polar, water miscible, mobile phase. Partition HPLC has been used historically on unbonded silica or alumina supports. Each works effectively for separating analytes by relative polar differences, however, HILIC has the advantage of separating acidic, basic and neutral solutes in a single chromatogram. The polar analytes diffuse into a stationary water layer associated with the polar stationary phase and are thus retained. Retention strengths increase with increased analyte polarity, and the interaction between the polar analyte and the polar stationary phase (relative to the mobile phase) increases the elution time. The interaction strength depends on the functional groups in the analyte molecule which promote partitioning but can also include coulombic (electrostatic) interaction and hydrogen donor capability.

Use of more polar solvents in the mobile phase will decrease the retention time of the analytes, whereas more hydrophobic solvents tend to increase retention times.

Normal-phase chromatography
Also known as normal-phase HPLC (NP-HPLC), or adsorption chromatography, this method separates analytes based on adsorption to a stationary surface chemistry and by polarity. It was one of the first kinds of HPLC that chemists developed. NP-HPLC uses a polar stationary phase and a non-polar, non-aqueous mobile phase, and works effectively for separating analytes readily soluble in non-polar solvents. The analyte associates with and is retained by the polar stationary phase. Adsorption strengths increase with increased analyte polarity, and the interaction between the polar analyte and the polar stationary phase (relative to the mobile phase) increases the elution time. The interaction strength depends not only on the functional groups in the analyte molecule, but also on steric factors. The effect of sterics on interaction strength allows this method to resolve (separate) structural isomers. The use of more polar solvents in the mobile phase will decrease the retention time of the analytes, whereas more hydrophobic solvents tend to increase retention times. Very polar solvents in a mixture tend to deactivate the stationary phase by creating a stationary bound water layer on the stationary phase surface. This behavior is somewhat peculiar to normal phase because it is most purely an adsorptive mechanism (the interactions are with a hard surface rather than a soft layer on a surface). NP-HPLC fell out of favor in the 1970s with the development of reversed-phase HPLC because of a lack of reproducibility of retention times as water or protic organic solvents changed the hydration state of the silica or alumina chromatographic media. Recently it has become useful again with the development of HILIC bonded phases which improve reproducibility.

Displacement chromatography
The basic principle of displacement chromatography is: A molecule with a high affinity for the chromatography matrix (the displacer) will compete effectively for binding sites, and thus displace all molecules with lesser affinities. There are distinct differences between displacement and elution chromatography. In elution mode, substances typically emerge from a column in narrow, Gaussian peaks. Wide separation of peaks, preferably to baseline, is desired in order to achieve maximum purification. The speed at which any component of a mixture travels down the column in elution mode depends on many factors. But for two substances to travel at different speeds, and thereby be resolved, there must be substantial differences in some interaction between the biomolecules and the chromatography matrix. Operating parameters are adjusted to maximize the effect of this difference. In many cases, baseline separation of the peaks can be achieved only with gradient elution and low column loadings. Thus, two drawbacks to elution mode chromatography, especially at the preparative scale, are operational complexity, due to gradient solvent pumping, and low throughput, due to low column loadings. Displacement chromatography has advantages over elution chromatography in that components are resolved into consecutive zones of pure substances rather than peaks. Because the process takes

advantage of the nonlinearity of the isotherms, a larger column feed can be separated on a given column with the purified components recovered at significantly higher concentrations.

Reversed-phase chromatography (RPC)

A chromatogram of complex mixture (perfume water) obtained by reversed phase HPLC

Reversed phase HPLC (RP-HPLC or RPC) has a non-polar stationary phase and an aqueous, moderately polar mobile phase. One common stationary phase is a silica which has been treated with RMe2SiCl, where R is a straight chain alkyl group such as C18H37 or C8H17. With these stationary phases, retention time is longer for molecules which are more non-polar, while polar molecules elute more readily. An investigator can increase retention time by adding more water to the mobile phase; thereby making the affinity of the hydrophobic analyte for the hydrophobic stationary phase stronger relative to the now more hydrophilic mobile phase. Similarly, an investigator can decrease retention time by adding more organic solvent to the eluent. RPC is so commonly used that it is often incorrectly referred to as "HPLC" without further specification. The pharmaceutical industry regularly employs RPC to qualify drugs before their release. RPC operates on the principle of hydrophobic forces, which originate from the high symmetry in the dipolar water structure and play the most important role in all processes in life science. RPC is allowing the measurement of these interactive forces. The binding of the analyte to the stationary phase is proportional to the contact surface area around the non-polar segment of the analyte molecule upon association with the ligand in the aqueous eluent. This solvophobic effect is dominated by the force of water for "cavity-reduction" around the analyte and the C18-chain versus the complex of both. The energy released in this process is proportional to the surface tension of the eluent (water: 7.3 106 J/cm, methanol: 2.2 106 J/cm) and to the hydrophobic surface of the analyte and the ligand respectively. The retention can be decreased by adding a less polar solvent (methanol, acetonitrile) into the mobile phase to reduce the surface tension of water. Gradient elution uses this effect by automatically reducing the polarity and the surface tension of the aqueous mobile phase during the course of the analysis. Structural properties of the analyte molecule play an important role in its retention characteristics. In general, an analyte with a larger hydrophobic surface area (C-H, C-C, and generally non-polar atomic bonds, such as S-S and others) results in a longer retention time because it increases the molecule's non-polar surface area, which is non-interacting with the water structure. On the other hand, polar groups, such as -OH, -NH2, COO- or -NH3+ reduce

retention as they are well integrated into water. Very large molecules, however, can result in an incomplete interaction between the large analyte surface and the ligand's alkyl chains and can have problems entering the pores of the stationary phase. Retention time increases with hydrophobic (non-polar) surface area. Branched chain compounds elute more rapidly than their corresponding linear isomers because the overall surface area is decreased. Similarly organic compounds with single C-C-bonds elute later than those with a C=C or C-C-triple bond, as the double or triple bond is shorter than a single C-C-bond. Aside from mobile phase surface tension (organizational strength in eluent structure), other mobile phase modifiers can affect analyte retention. For example, the addition of inorganic salts causes a moderate linear increase in the surface tension of aqueous solutions (ca. 1.5 107 J/cm per Mol for NaCl, 2.5 107 J/cm per Mol for (NH4)2SO4), and because the entropy of the analyte-solvent interface is controlled by surface tension, the addition of salts tend to increase the retention time. This technique is used for mild separation and recovery of proteins and protection of their biological activity in protein analysis (hydrophobic interaction chromatography, HIC). Another important component is the influence of the pH since this can change the hydrophobicity of the analyte. For this reason most methods use a buffering agent, such as sodium phosphate, to control the pH. The buffers serve multiple purposes: they control pH, neutralize the charge on any residual exposed silica on the stationary phase and act as ion pairing agents to neutralize charge on the analyte. Ammonium formate is commonly added in mass spectrometry to improve detection of certain analytes by the formation of ammonium adducts. A volatile organic acid such as acetic acid, or most commonly formic acid, is often added to the mobile phase if mass spectrometry is used to analyze the column eluent. Trifluoroacetic acid is used infrequently in mass spectrometry applications due to its persistence in the detector and solvent delivery system, but can be effective in improving retention of analytes such as carboxylic acids in applications utilizing other detectors, as it is one of the strongest organic acids. The effects of acids and buffers vary by application but generally improve the chromatography. Reversed phase columns are quite difficult to damage compared with normal silica columns; however, many reversed phase columns consist of alkyl derivatized silica particles and should never be used with aqueous bases as these will destroy the underlying silica particle. They can be used with aqueous acid, but the column should not be exposed to the acid for too long, as it can corrode the metal parts of the HPLC equipment. RP-HPLC columns should be flushed with clean solvent after use to remove residual acids or buffers, and stored in an appropriate composition of solvent. The metal content of HPLC columns must be kept low if the best possible ability to separate substances is to be retained. A good test for the metal content of a column is to inject a sample which is a mixture of 2,2'- and 4,4'- bipyridine. Because the 2,2'bipy can chelate the metal, the shape of the peak for the 2,2'-bipy will be distorted (tailed) when metal ions are present on the surface of the silica.

Size-exclusion chromatography
Size-exclusion chromatography (SEC), also known as gel permeation chromatography or gel filtration chromatography, separates particles on the basis of size. It is generally a low resolution chromatography and thus it is often reserved for the final, "polishing" step of a purification. It is also useful for determining the tertiary structure and quaternary structure of purified proteins. SEC is used primarily for the analysis of large molecules such as proteins or polymers. SEC works by trapping these smaller molecules in the pores of a particle. The larger molecules simply pass by the pores as they are too large to enter the pores. Larger molecules therefore flow through the column quicker than smaller molecules, that is, the smaller the molecule, the longer the retention time. This technique is widely used for the molecular weight determination of polysaccharides. SEC is the official technique (suggested by European pharmacopeia) for the molecular weight comparison of different commercially available low-molecular weight heparins.

Ion-exchange chromatography
In ion-exchange chromatography, retention is based on the attraction between solute ions and charged sites bound to the stationary phase. Ions of the same charge are excluded. Types of ion exchangers include:

Polystyrene resins These allow cross linkage which increases the stability of the chain. Higher cross linkage reduces swerving, which increases the equilibration time and ultimately improves selectivity. Cellulose and dextran ion exchangers (gels) These possess larger pore sizes and low charge densities making them suitable for protein separation. Controlled-pore glass or porous silica

In general, ion exchangers favor the binding of ions of higher charge and smaller radius. An increase in counter ion (with respect to the functional groups in resins) concentration reduces the retention time. An increase in pH reduces the retention time in cation exchange while a decrease in pH reduces the retention time in anion exchange. This form of chromatography is widely used in the following applications: water purification, pre-concentration of trace components, ligand-exchange chromatography, ion-exchange chromatography of proteins, high-pH anion-exchange chromatography of carbohydrates and oligosaccharides, and others.

Isocratic flow and gradient elution


A separation in which the mobile phase composition remains constant throughout the procedure is termed isocratic (meaning constant composition). The mobile phase composition does not have to remain constant. A separation in which the mobile phase composition is changed during the separation process is described as a gradient elution. One example is a gradient starting at 10% methanol and ending at 90% methanol after 20 minutes. The two components of the mobile phase are typically termed "A" and "B"; A is the "weak" solvent which allows the solute to elute only slowly, while B is the "strong" solvent which rapidly elutes the solutes from the column. Solvent A is often water, while B is an organic solvent miscible with water, such as acetonitrile, methanol, THF, or isopropanol. In isocratic elution, peak width increases with retention time linearly according to the equation for N, the number of theoretical plates. This leads to the disadvantage that late-eluting peaks get very flat and broad. Their shape and width may keep them from being recognized as peaks. Gradient elution decreases the retention of the later-eluting components so that they elute faster, giving narrower (and taller) peaks for most components. This also improves the peak shape for tailed peaks, as the increasing concentration of the organic eluent pushes the tailing part of a peak forward. This also increases the peak height (the peak looks "sharper"), which is important in trace analysis. The gradient program may include sudden "step" increases in the percentage of the organic component, or different slopes at different times - all according to the desire for optimum separation in minimum time. In isocratic elution, the selectivity does not change if the column dimensions (length and inner diameter) change - that is, the peaks elute in the same order. In gradient elution, the elution order may change as the dimensions or flow rate change. The driving force in reversed phase chromatography originates in the high order of the water structure. The role of the organic component of the mobile phase is to reduce this high order and thus reduce the retarding strength of the aqueous component.

Parameters
Internal diameter
The internal diameter (ID) of an HPLC column is an important parameter that influences the detection sensitivity and separation selectivity in gradient elution. It also determines the quantity of analyte that can be loaded onto the column. Larger columns are usually seen in industrial applications, such as the purification of a drug product for later use. Low-ID columns have improved sensitivity and lower solvent consumption at the expense of loading capacity.

Larger ID columns (over 10 mm) are used to purify usable amounts of material because of their large loading capacity. Analytical scale columns (4.6 mm) have been the most common type of columns, though smaller columns are rapidly gaining in popularity. They are used in traditional quantitative analysis of samples and often use a UV-Vis absorbance detector. Narrow-bore columns (12 mm) are used for applications when more sensitivity is desired either with special UV-vis detectors, fluorescence detection or with other detection methods like liquid chromatography-mass spectrometry Capillary columns (under 0.3 mm) are used almost exclusively with alternative detection means such as mass spectrometry. They are usually made from fused silica capillaries, rather than the stainless steel tubing that larger columns employ.

Particle size
Most traditional HPLC is performed with the stationary phase attached to the outside of small spherical silica particles (very small beads). These particles come in a variety of sizes with 5 m beads being the most common. Smaller particles generally provide more surface area and better separations, but the pressure required for optimum linear velocity increases by the inverse of the particle diameter squared. This means that changing to particles that are half as big, keeping the size of the column the same, will double the performance, but increase the required pressure by a factor of four. Larger particles are used in preparative HPLC (column diameters 5 cm up to >30 cm) and for nonHPLC applications such as solid-phase extraction.

Pore size
Many stationary phases are porous to provide greater surface area. Small pores provide greater surface area while larger pore size has better kinetics, especially for larger analytes. For example, a protein which is only slightly smaller than a pore might enter the pore but does not easily leave once inside.

Pump pressure
Pumps vary in pressure capacity, but their performance is measured on their ability to yield a consistent and reproducible flow rate. Pressure may reach as high as 40 MPa (6000 lbf/in2), or about 400 atmospheres. Modern HPLC systems have been improved to work at much higher pressures, and therefore are able to use much smaller particle sizes in the columns (<2 m). These "Ultra High Performance Liquid Chromatography" systems or RSLC/UHPLCs can work at up to 100 MPa (15,000 lbf/in), or about 1000 atmospheres. The term "UPLC" is a trademark of the Waters Corporation, but is sometimes used to refer to the more general technique.

Manufacturers of HPLC chromatographs


Agilent Technologies Shimadzu Scientific Instruments Beckman Coulter Hitachi PerkinElmer, Inc. Waters Corporation

Manufacturers of HPLC columns and accessories


Agilent Technologies Shimadzu Scientific Instruments AkzoNobel Beckman Coulter Thermo Fisher Scientific Tosoh Corporation Varian, Inc. Waters Corporation

GAS CHROMATOGRAPHY
Gas chromatography (GC), is a common type of chromatography used in analytic chemistry for separating and analyzing compounds that can be vaporized without decomposition. Typical uses of GC include testing the purity of a particular substance, or separating the different components of a mixture (the relative amounts of such components can also be determined). In some situations, GC may help in identifying a compound. In preparative chromatography, GC can be used to prepare pure compounds from a mixture.[1] In gas chromatography, the moving phase (or "mobile phase") is a carrier gas, usually an inert gas such as helium or an unreactive gas such as nitrogen. The stationary phase is a microscopic layer of liquid or polymer on an inert solid support, inside a piece of glass or metal tubing called a column (an homage to the fractionating column used in distillation). The instrument used to perform gas chromatography is called a gas chromatograph (or "aerograph", "gas separator"). The gaseous compounds being analyzed interact with the walls of the column, which is coated with different stationary phases. This causes each compound to elute at a different time, known as the retention time of the compound. The comparison of retention times is what gives GC its analytical usefulness.

Gas chromatography is in principle similar to column chromatography (as well as other forms of chromatography, such as HPLC, TLC), but has several notable differences. Firstly, the process of separating the compounds in a mixture is carried out between a liquid stationary phase and a gas moving phase, whereas in column chromatography the stationary phase is a solid and the moving phase is a liquid. (Hence the full name of the procedure is "Gas-liquid chromatography", referring to the mobile and stationary phases, respectively.) Secondly, the column through which the gas phase passes is located in an oven where the temperature of the gas can be controlled, whereas column chromatography (typically) has no such temperature control. Thirdly, the concentration of a compound in the gas phase is solely a function of the vapor pressure of the gas Gas chromatography is also similar to fractional distillation, since both processes separate the components of a mixture primarily based on boiling point (or vapor pressure) differences.

However, fractional distillation is typically used to separate components of a mixture on a large scale, whereas GC can be used on a much smaller scale (i.e. microscale).[1] Gas chromatography is also sometimes known as vapor-phase chromatography (VPC), or gasliquid partition chromatography (GLPC). These alternative names, as well as their respective abbreviations, are frequently found in scientific literature. Strictly speaking, GLPC is the most correct terminology, and is thus preferred by many authors

GC analysis
A gas chromatograph is a chemical analysis instrument for separating chemicals in a complex sample. A gas chromatograph uses a flow-through narrow tube known as the column, through which different chemical constituents of a sample pass in a gas stream (carrier gas, mobile phase) at different rates depending on their various chemical and physical properties and their interaction with a specific column filling, called the stationary phase. As the chemicals exit the end of the column, they are detected and identified electronically. The function of the stationary phase in the column is to separate different components, causing each one to exit the column at a different time (retention time). Other parameters that can be used to alter the order or time of retention are the carrier gas flow rate, and the temperature. In a GC analysis, a known volume of gaseous or liquid analyte is injected into the "entrance" (head) of the column, usually using a microsyringe (or, solid phase microextraction fibers, or a gas source switching system). As the carrier gas sweeps the analyte molecules through the column, this motion is inhibited by the adsorption of the analyte molecules either onto the column walls or onto packing materials in the column. The rate at which the molecules progress along the column depends on the strength of adsorption, which in turn depends on the type of molecule and on the stationary phase materials. Since each type of molecule has a different rate of progression, the various components of the analyte mixture are separated as they progress along the column and reach the end of the column at different times (retention time). A detector is used to monitor the outlet stream from the column; thus, the time at which each component reaches the outlet and the amount of that component can be determined. Generally, substances are identified (qualitatively) by the order in which they emerge (elute) from the column and by the retention time of the analyte in the column.

Physical components

Autosamplers
The autosampler provides the means to introduce a sample automatically into the inlets. Manual insertion of the sample is possible but is no longer common. Automatic insertion provides better reproducibility and time-optimization. Different kinds of autosamplers exist in the market today. Autosamplers can be classified in relation to sample capacity (auto-injectors vs. autosamplers, where auto-injectors can work a small number of samples), to robotic technologies (XYZ robot vs. rotating robot the most common), or to analysis:

Liquid Static head-space by syringe technology Dynamic head-space by transfer-line technology Solid phase microextraction (SPME)

Inlets
The column inlet (or injector) provides the means to introduce a sample into a continuous flow of carrier gas. The inlet is a piece of hardware attached to the column head. Common inlet types are: Front inlet Back inlet Micro syringe

Columns
Two types of columns are used in GC:

Packed columns are 1.5 10 m in length and have an internal diameter of 2 4 mm. The tubing is usually made of stainless steel or glass and contains a packing of finely divided, inert, solid support material (e.g. diatomaceous earth) that is coated with a liquid or solid stationary phase. The nature of the coating material determines what type of materials will be most strongly adsorbed. Thus numerous columns are available that are designed to separate specific types of compounds. Capillary columns have a very small internal diameter, on the order of a few tenths of millimeters, and lengths between 2560 meters are common. The inner column walls are coated with the active materials (WCOT columns), some columns are quasi solid filled with many parallel micropores (PLOT columns). Most capillary columns are made of fused-silica with a polyimide outer coating. These columns are flexible, so a very long column can be wound into a small coil.

The temperature-dependence of molecular adsorption and of the rate of progression along the column necessitates a careful control of the column temperature to within a few tenths of a degree for precise work. Reducing the temperature produces the greatest level of separation, but can result in very long elution times. For some cases temperature is ramped either continuously or in steps to provide the desired separation. This is referred to as a temperature program. Electronic pressure control can also be used to modify flow rate during the analysis, aiding in faster run times while keeping acceptable levels of separation. The choice of carrier gas (mobile phase) is important, with hydrogen being the most efficient and providing the best separation. However, helium has a larger range of flowrates that are comparable to hydrogen in efficiency, with the added advantage that helium is non-flammable, and works with a greater number of detectors. Therefore, helium is the most common carrier gas used.

Detectors
A number of detectors are used in gas chromatography. The most common are the flame ionization detector (FID) and the thermal conductivity detector (TCD). Both are sensitive to a wide range of components, and both work over a wide range of concentrations. While TCDs are essentially universal and can be used to detect any component other than the carrier gas (as long as their thermal conductivities are different from that of the carrier gas, at detector temperature), FIDs are sensitive primarily to hydrocarbons, and are more sensitive to them than TCD. However, an FID cannot detect water. Both detectors are also quite robust. Since TCD is nondestructive, it can be operated in-series before an FID (destructive), thus providing complementary detection of the same analytes. Other detectors are sensitive only to specific types of substances, or work well only in narrower ranges of concentrations. They include:

discharge ionization detector (DID), which uses a high-voltage electric discharge to produce ions. electron capture detector (ECD), which uses a radioactive Beta particle (electron) source to measure the degree of electron capture.

flame photometric detector (FPD) flame ionization detector (FID) Hall electrolytic conductivity detector (ElCD) helium ionization detector (HID) Nitrogen Phosphorus Detector (NPD) Infrared Detector (IRD) mass selective detector (MSD) photo-ionization detector (PID) pulsed discharge ionization detector (PDD) thermal energy(conductivity) analyzer/detector (TEA/TCD)

Some gas chromatographs are connected to a mass spectrometer which acts as the detector. The combination is known as GC-MS. Some GC-MS are connected to an NMR spectrometer which acts as a backup detector. This combination is known as GC-MS-NMR. Some GC-MS-NMR are connected to an infrared spectrophotometer which acts as a backup detector. This combination is known as GC-MS-NMR-IR. It must, however, be stressed this is very rare as most analyses needed can be concluded via purely GC-MS.

Methods
The method is the collection of conditions in which the GC operates for a given analysis. Method development is the process of determining what conditions are adequate and/or ideal for the analysis required. Conditions which can be varied to accommodate a required analysis include inlet temperature, detector temperature, column temperature and temperature program, carrier gas and carrier gas flow rates, the column's stationary phase, diameter and length, inlet type and flow rates, sample size and injection technique. Depending on the detector(s) (see below) installed on the GC, there may be a number of detector conditions that can also be varied. Some GCs also include valves which can change the route of sample and carrier flow. The timing of the opening and closing of these valves can be important to method development.

This image above shows the interior of a GeoStrata Technologies Eclipse Gas Chromatograph that runs continuously in three minute cycles. Two valves are used to switch the test gas into the

sample loop. After filling the sample loop with test gas, the valves are switched again applying carrier gas pressure to the sample loop and forcing the sample through the Column for separation.

Carrier gas selection and flow rates


Typical carrier gases include helium, nitrogen, argon, hydrogen and air. Which gas to use is usually determined by the detector being used, for example, a DID requires helium as the carrier gas. When analyzing gas samples, however, the carrier is sometimes selected based on the sample's matrix, for example, when analyzing a mixture in argon, an argon carrier is preferred, because the argon in the sample does not show up on the chromatogram. Safety and availability can also influence carrier selection, for example, hydrogen is flammable, and high-purity helium can be difficult to obtain in some areas of the world. (See: Helium--occurrence and production.) The carrier gas flow rate affects the analysis in the same way that temperature does (see above). The higher the flow rate the faster the analysis, but the lower the separation between analytes. Selecting the flow rate is therefore the same compromise between the level of separation and length of analysis as selecting the column temperature. Many modern GCs, however, electronically measure the flow rate, and electronically control the carrier gas pressure to set the flow rate. Consequently, carrier pressures and flow rates can be adjusted during the run, creating pressure/flow programs similar to temperature programs.

Stationary compound selection


The polarity of the solute is crucial for the choice of stationary compound, which in an optimal case would have a similar polarity than the solute. Common stationary phases in open tubular columns are cyanopropylphenyl dimethyl polysiloxane, carbowax polyethyleneglycol, biscyanopropyl cyanopropylphenyl polysiloxane and diphenyl dimethyl polysiloxane. For packed columns there are more options available.

Column temperature and temperature program


The column(s) in a GC are contained in an oven, the temperature of which is precisely controlled electronically. (When discussing the "temperature of the column," an analyst is technically referring to the temperature of the column oven. The distinction, however, is not important and will not subsequently be made in this article.)

The rate at which a sample passes through the column is directly proportional to the temperature of the column. The higher the column temperature, the faster the sample moves through the column. However, the faster a sample moves through the column, the less it interacts with the stationary phase, and the less the analytes are separated. In general, the column temperature is selected to compromise between the length of the analysis and the level of separation. A method which holds the column at the same temperature for the entire analysis is called "isothermal." Most methods, however, increase the column temperature during the analysis, the initial temperature, rate of temperature increase (the temperature "ramp") and final temperature is called the "temperature program." A temperature program allows analytes that elute early in the analysis to separate adequately, while shortening the time it takes for late-eluting analytes to pass through the column.

Data reduction and analysis


Qualitative analysis: Generally chromatographic data is presented as a graph of detector response (y-axis) against retention time (x-axis), which is called a chromatogram. This provides a spectrum of peaks for a sample representing the analytes present in a sample eluting from the column at different times. Retention time can be used to identify analytes if the method conditions are constant. Also, the pattern of peaks will be constant for a sample under constant conditions and can identify complex mixtures of analytes. In most modern applications however the GC is connected to a mass spectrometer or similar detector that is capable of identifying the analytes represented by the peaks. Quantitative analysis:

The area under a peak is proportional to the amount of analyte present in the chromatogram. By calculating the area of the peak using the mathematical function of integration, the concentration of an analyte in the original sample can be determined. Concentration can be calculated using a calibration curve created by finding the response for a series of concentrations of analyte, or by determining the relative response factor of an analyte. The relative response factor is the expected ratio of an analyte to an internal standard (or external standard) and is calculated by finding the response of a known amount of analyte and a constant amount of internal standard (a chemical added to the sample at a constant concentration, with a distinct retention time to the analyte). In most modern GC-MS systems, computer software is used to draw and integrate peaks, and match MS spectra to library spectra.

Application
In general, substances that vaporize below ca. 300 C (and therefore are stable up to that temperature) can be measured quantitatively. The samples are also required to be salt-free; they should not contain ions. Very minute amounts of a substance can be measured, but it is often required that the sample must be measured in comparison to a sample containing the pure, suspected substance. Various temperature programs can be used to make the readings more meaningful; for example to differentiate between substances that behave similarly during the GC process. Professionals working with GC analyze the content of a chemical product, for example in assuring the quality of products in the chemical industry; or measuring toxic substances in soil, air or water. GC is very accurate if used properly and can measure picomoles of a substance in a 1 ml liquid sample, or parts-per-billion concentrations in gaseous samples. In practical courses at colleges, students sometimes get acquainted to the GC by studying the contents of Lavender oil or measuring the ethylene that is secreted by Nicotiana benthamiana plants after artificially injuring their leaves. These GC analyses hydrocarbons (C2-C40+). In a typical experiment, a packed column is used to separate the light gases, which are then detected with a TCD. The hydrocarbons are separated using a capillary column and detected with an FID. A complication with light gas analyses that include H2 is that He, which is the most common and most sensitive inert carrier (sensitivity is proportional to molecular mass) has an almost identical thermal conductivity to hydrogen (it is the difference in thermal conductivity between two separate filaments in a Wheatstone Bridge type arrangement that shows when a component has been eluted). For this reason, dual TCD instruments are used with a separate channel for hydrogen that uses nitrogen as a carrier are common. Argon is often used when analysing gas phase chemistry reactions such as F-T synthesis so that a single carrier gas can be used rather than 2 separate ones. The sensitivity is less but this is a tradeoff for simplicity in the gas supply.

GCs in popular culture


Movies, books and TV shows tend to misrepresent the capabilities of gas chromatography and the work done with these instruments. In the U.S. TV show CSI, for example, GCs are used to rapidly identify unknown samples. "This is gasoline bought at a Chevron station in the past two weeks," the analyst will say fifteen minutes after receiving the sample. In fact, a typical GC analysis takes much more time; sometimes a single sample must be run more than an hour according to the chosen program; and even more time is needed to "heat out" the column so it is free from the first sample and can be used for the next. Equally, several runs are needed to confirm the results of a study - a GC analysis of a single sample may simply yield a result per chance (see statistical significance). Also, GC does not positively identify most samples; and not all substances in a sample will necessarily be detected. All a GC truly tells you is at which relative time a component eluted from the column and that the detector was sensitive to it. To make results meaningful, analysts need to know which components at which concentrations are to be expected; and even then a small amount of a substance can hide itself behind a substance having both a higher concentration and the same relative elution time. Last but not least it is often needed to check the results of the sample against a GC analysis of a reference sample containing only the suspected substance. A GC-MS can remove much of this ambiguity, since the mass spectrometer will identify the component's molecular weight. But this still takes time and skill to do properly. Similarly, most GC analyses are not push-button operations. You cannot simply drop a sample vial into an auto-sampler's tray, push a button and have a computer tell you everything you need to know about the sample. According to the substances one expects to find the operating program must be carefully chosen. A push-button operation can exist for running similar samples repeatedly, such as in a chemical production environment or for comparing 20 samples from the same experiment to calculate the mean content of the same substance

ULTRA-VIOLET SPECTROSCOPY
Ultraviolet-visible spectroscopy or ultraviolet-visible spectrophotometry (UV-Vis or UV/Vis) refers to absorption spectroscopy in the ultraviolet-visible spectral region. This means it uses light in the visible and adjacent (near-UV and near-infrared (NIR)) ranges. The absorption in the visible range directly affects the perceived color of the chemicals involved. In this region of the electromagnetic spectrum, molecules undergo electronic transitions. This technique is complementary to fluorescence spectroscopy, in that fluorescence deals with transitions from the excited state to the ground state, while absorption measures transitions from the ground state to the excited state.

Applications
UV/Vis spectroscopy is routinely used in the quantitative determination of solutions of transition metal ions and highly conjugated organic compounds.

Solutions of transition metal ions can be colored (i.e., absorb visible light) because d electrons within the metal atoms can be excited from one electronic state to another. The colour of metal ion solutions is strongly affected by the presence of other species, such as certain anions or ligands. For instance, the colour of a dilute solution of copper sulfate is a very light blue; adding ammonia intensifies the colour and changes the wavelength of maximum absorption (max). Organic compounds, especially those with a high degree of conjugation, also absorb light in the UV or visible regions of the electromagnetic spectrum. The solvents for these determinations are often water for water soluble compounds, or ethanol for organicsoluble compounds. (Organic solvents may have significant UV absorption; not all solvents are suitable for use in UV spectroscopy. Ethanol absorbs very weakly at most wavelengths.) Solvent polarity and pH can affect the absorption spectrum of an organic compound. Tyrosine, for example, increases in absorption maxima and molar extinction coefficient when pH increases from 6 to 13 or when solvent polarity decreases. While charge transfer complexes also give rise to colours, the colours are often too intense to be used for quantitative measurement.

The Beer-Lambert law states that the absorbance of a solution is directly proportional to the concentration of the absorbing species in the solution and the path length. Thus, for a fixed path length, UV/Vis spectroscopy can be used to determine the concentration of the absorber in a solution. It is necessary to know how quickly the absorbance changes with concentration. This can be taken from references (tables of molar extinction coefficients), or more accurately, determined from a calibration curve. A UV/Vis spectrophotometer may be used as a detector for HPLC. The presence of an analyte gives a response assumed to be proportional to the concentration. For accurate results, the instrument's response to the analyte in the unknown should be compared with the response to a standard; this is very similar to the use of calibration curves. The response (e.g., peak height) for a particular concentration is known as the response factor.

Beer-Lambert law
The method is most often used in a quantitative way to determine concentrations of an absorbing species in solution, using the Beer-Lambert law: ,

where A is the measured absorbance, I0 is the intensity of the incident light at a given wavelength, I is the transmitted intensity, L the pathlength through the sample, and c the concentration of the absorbing species. For each species and wavelength, is a constant known as the molar absorptivity or extinction coefficient. This constant is a fundamental molecular property in a given solvent, at a particular temperature and pressure, and has units of 1 / M * cm

Practical considerations
The Beer-Lambert law has implicit assumptions that must be met experimentally for it to apply. For instance, the chemical makeup and physical environment of the sample can alter its extinction coefficient. The chemical and physical conditions of a test sample therefore must match reference measurements for conclusions to be valid.

Spectral bandwidth
A given spectrometer has a spectral bandwidth that characterizes how monochromatic the light is. If this bandwidth is comparable to the width of the absorption features, then the measured extinction coefficient will be altered. In most reference measurements, the instrument bandwidth is kept below the width of the spectral lines. When a new material is being measured, it may be necessary to test and verify if the bandwidth is sufficiently narrow. Reducing the spectral bandwidth will reduce the energy passed to the detector and will, therefore, require a longer measurement time to achieve the same signal to noise ratio.

Wavelength error
In liquids, the extinction coefficient usually changes slowly with wavelength. A peak of the 3absorbance curve (a wavelength where the absorbance reaches a maximum) is where the rate of change in absorbance with wavelength is smallest. Measurements are usually made at a peak to minimize errors produced by errors in wavelength in the instrument, that is errors due to having a different extinction coefficient than assumed.

Stray light
Another important factor is the purity of the light used. The most important factor affecting this is the stray light level of the monochromator [2] . The detector used is broadband, it responds to all the light that reaches it. If a significant amount of the light passed through the sample contains wavelengths that have much lower extinction coefficients than the nominal one, the instrument will report an incorrectly low absorbance. Any instrument will reach a point where an increase in sample concentration will not result in an increase in the reported absorbance, because the detector is simply responding to the stray light. In practice the concentration of the sample or the optical path length must be adjusted to place the unknown absorbance within a range that is valid for the instrument. Sometimes an empirical calibration function is developed, using known concentrations of the sample, to allow measurements into the region where the instrument is becoming non-linear. As a rough guide, an instrument with a single monochromator would typically have a stray light level corresponding to about 3 AU, which would make measurements above about 2 AU problematic. A more complex instrument with a double monochromator would have a stray light level corresponding to about 6 AU, which would therefore allow measuring a much wider absorbance range.

Ultraviolet-visible spectrophotometer
The instrument used in ultraviolet-visible spectroscopy is called a UV/Vis spectrophotometer. It measures the intensity of light passing through a sample (I), and compares it to the intensity of light before it passes through the sample (Io). The ratio I / Io is called the transmittance, and is usually expressed as a percentage (%T). The absorbance, A, is based on the transmittance: A = log(%T / 100%) The basic parts of a spectrophotometer are a light source, a holder for the sample, a diffraction grating or monochromator to separate the different wavelengths of light, and a detector. The radiation source is often a Tungsten filament (300-2500 nm), a deuterium arc lamp, which is continuous over the ultraviolet region (190-400 nm) or more recently, light emitting diodes (LED) and Xenon arc lamps for the visible wavelengths. The detector is typically a photodiode or a CCD. Photodiodes are used with monochromators, which filter the light so that only light of a single wavelength reaches the detector. Diffraction gratings are used with CCDs, which collects light of different wavelengths on different pixels.

A spectrophotometer can be either single beam or double beam. In a single beam instrument (such as the Spectronic 20), all of the light passes through the sample cell. Io must be measured by removing the sample. This was the earliest design, but is still in common use in both teaching and industrial labs. In a double-beam instrument, the light is split into two beams before it reaches the sample. One beam is used as the reference; the other beam passes through the sample. The reference beam intensity is taken as 100% Transmission (or 0 Absorbance), and the measurement displayed is the ratio of the two beam intensities. Some double-beam instruments have two detectors (photodiodes), and the sample and reference beam are measured at the same time. In other instruments, the two beams pass through a beam chopper, which blocks one beam at a time. The detector alternates between measuring the sample beam and the reference beam in synchronism with the chopper. There may also be one or more dark intervals in the chopper cycle. In this case

the measured beam intensities may be corrected by subtracting the intensity measured in the dark interval before the ratio is taken. Samples for UV/Vis spectrophotometry are most often liquids, although the absorbance of gases and even of solids can also be measured. Samples are typically placed in a transparent cell, known as a cuvette. Cuvettes are typically rectangular in shape, commonly with an internal width of 1 cm. (This width becomes the path length, L, in the Beer-Lambert law.) Test tubes can also be used as cuvettes in some instruments. The type of sample container used must allow radiation to pass over the spectral region of interest. The most widely applicable cuvettes are made of high quality fused silica or quartz glass because these are transparent throughout the UV, visible and near infrared regions. Glass and plastic cuvettes are also common, although glass and most plastics absorb in the UV, which limits their usefulness to visible wavelengths.[5] A complete spectrum of the absorption at all wavelengths of interest can often be produced directly by a more sophisticated spectrophotometer. In simpler instruments the absorption is determined one wavelength at a time and then compiled into a spectrum by the operator. A standardized spectrum is formed by removing the concentration dependence and determining the extinction coefficient () as a function of wavelength.

KARL-FISCHER
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.

Coulometric titration
The main compartment of the titration cell contains the anode solution plus the analyte. The anode solution consists of an alcohol (ROH), a base (B), SO2 and I2. A typical alcohol that may be used is methanol or diethylene glycol monomethyl ether, and a common base is imidazole. The titration cell also consists of a smaller compartment with an (anode) immersed in the anode solution of the main compartment. The two compartments are separated by an ion-permeable membrane. The Pt anode generates I2 when current is provided through the electric circuit. The net reaction as shown below is oxidation of SO2 by I2. One mole of I2 is consumed for each mole of H2O. In other words, 2 moles of electrons are consumed per mole of water. BI2 + BSO2 + B + H2O 2BH+I + BSO3 BSO3 + ROH BH+ROSO3 The end point is detected most commonly by a bipotentiometric method. A second pair of Pt electrodes are immersed in the anode solution. The detector circuit maintains a constant current between the two detector electrodes during titration. Prior to the equivalence point, the solution contains I- but little I2. At the equivalence point, excess I2 appears and an abrupt voltage drop marks the end point. The amount of current needed to generate I2 in order to reach the end point can then be used to calculate the amount of water in the original sample.

Volumetric titration
The volumetric titration is based on the same principles as the coulometric titration except that the anode solution above now is used as the titrant solution. The titrant consists of an alcohol (ROH), base (B), SO2 and a known concentration of I2. One mole of I2 is consumed for each mole of H2O. The titration reaction proceeds as above, and the end point may be detected by a bipotentiometric method as described above.

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:

High accuracy and precision Selectivity for water Small sample quantities required Easy sample preparation Short analysis duration Nearly unlimited measuring range (1ppm to 100%) Suitability for analyzing: o Solids o Liquids o Gases Independence of presence of other volatiles Suitability for automation

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

PRODUCTION
The production department of Zydus Cadilla is very extensive and it comprises of various sub-departments. These departments are listed below:-

1. Warehouse o Raw material o Packaging material o ASRS 2. CENTRAL PHARMACY o Shifting o Milling o Dispensing pharmacy 3. Tablet manufacturing o Granulation o Compression o Coating 4. Table packaging o Bulk packaging o Blister packaging o Strip packaging 5. Hard Gelatin capsule 6. Soft Gelatin capsule 7. Lyophilization 8. Parenteral. 9. Aerosol manufacturing 10.Transdermal manufacturing, Pilot plant & Q.C.

WAREHOUSE

Warehouse is a primary storage for raw material and packaging material. The material from the supplier first enters in this area. Warehouse stores raw materials that are under test till manufacturing department need for processing. Warehouse is used by exporter, importer, manufacturer, researcher and employee.

Storage Areas according to temperature required:-

MATERIAL Active pharmaceutical ingredients Excipient Empty capsule shells

STORAGE TEMPERATURE Storage in A.C.room (below 25 C temp.) Stored at room temperature Stored in an A.C. room (below 25 C)

RELATIVE HUMIDITY for all the material is 40-60%. AIR FLOW: - reversed.

Steps for entry of material in warehouse:1. 2. 3. 4. 5. 6. 7. 8. Vehicle checking Unloading of raw material & packaging material Dedusting Weighing Sampling Q.C. of raw material Approval of raw material Labeling (under test-yellow label)

Departments:

1. 2. 3. 4. 5. 6.

Storage for active pharmaceutical ingredient (API) Storage for Excipient Storage for packaging material Quality control for packaging material Sampling of raw material. Hold area

The stored materials are labeled with different color of labels as per testing done in Q.C department

COLOUR OF LABEL Yellow Green Red

POSITION OF TESTING under test Approved Rejected

COLOUR OF LABEL Yellow Green Red

POSITION OF MATERIAL In ware house In A.S.R.S. Discarded

1. Raw material: In warehouse, we can also store raw material. For this purpose the calibration or checking of the weighing balance is required

Quarantine area: For in coming of raw materials and packaging materials its necessary and encloused quarantine area must be provided for raw materials and packaging materials bulk product and finished product and goods that are rejected for failure while compare with various standard. It includes:

a. Raw material receipt area:-responsible for receipt of raw material e.g. API, excipient etc. b. Packaging material receipt area:-e.g. closures and containers c. Lock and key area: storage of discarded material, failure batches, approved batches with appropriate label occurs. Various labels are used to identify the materials and products:

I. II. III. IV.

Under test label Approved label Dispensing label Identification label

Other main terms are used for notification i. ii. iii. MRN No.: Material Receipt Note A.R : Analytical Report B.M.R : Batch Manufacturing Record

The difference between raw material storage and packaging material storage are:The main difference is temperature difference. Packaging material is stored only at 25 C, where as raw material is stored according to. 1. Ambient (room temp) but temperature not exceeding 39 C 2. below 25 C 3. 2 to 8 C (cooling temp) EXAMPLES of the APIs present: Topiramate Losartan Riboflavin Simvastatin Paroxetine Meloxicam

If material is stored for more than one year & then used, it must be rechecked.

AUTOMATED STORAGE AND RETRIEVABLE SYSTEM (ASRS):

In this area the material stored are approved by Q.C. The material which are checked from ware house after getting the approval, are sifted to the A.S.R.S. No FINISHED GOODS are stored in this area. The condition at which materials are kept is almost about 25 C and at relative humidity 4065%.The capacity of this area to store the materials are about 500 pallets. The fully auto mated machine is working in this area, where to get aby of the product we need to just enter the specific number at which the material is kept and the machine it self gives the material at the unloading space. And the same procedure follows for the material to store at a specific position. In the 1st ,2nd and 4th storage raw, the raw materials are kept and in the 3rd raw, packing materials are stored. The materials which are costly or are highly important are kept in a locker or cup board. This area is known as UNDER HOLD AREA. The examples of this type of substances are morphine, dehydrated alcohol. In the cup-board the materials like hexamethazone sodium, uresamide, salmetriol are stored. Godrej crane is used to transfer the raw material. It is operated under the command of computer .SRP (excel file)is used for running of godrej crane automatically ,In which location entered in system &then crane goes to that place and takes it out. A fire management system is also placed inside this area, which is active even for small dust or small fire or spark.

CENTRAL PHARMACY

The role of central pharmacy department in any pharmaceutical industry is to dispense required ingredients in required quantity. The material to the whole production area is only supplied by this area.

Main goal of the program:


To ensure that safe; effective; high-quality essential drugs are available at all times in our health facilities, and appropriately used by the patients. Aiming to close the gap between the need of essential drugs and access to them, in addition to another gap between availability of drugs and their rational use. The department is divided into various sections as:1. Raw material staging room 2. Liquid dispensing booth 3. Sifting rooms 4. Washing area 5. Sifting material staging room 6. Dispensing booth (total 5) A. One booth for dispensing API B. Four booths for dispensing excipients.

TABLET MANUFACTURING

Tablet manufacturing is one of the important areas as tablets form the major bulk of solid dosage form. Tablets can be made in virtually any shape, although requirements of patients and tableting machines mean that most are round, oval or capsule shaped. More unusual shapes have been manufactured but patients find these harder to swallow, and they are more vulnerable to chipping or manufacturing problems. Tablet diameter and shape are determined by the machine tooling used to produce them - a die plus an upper and a lower punch are required. This is called a station of tooling. Tablets need to be strong enough to resist the stresses of packaging, shipping and handling by the pharmacist and patient.

The various areas for tablet manufacturing are:1. 2. 3. 4. 5. 6. 7. 8. 9. Granulation area Granule quarantine area Approved granule store Tablet compression area Tablet quarantine area IPQC Tablet inspection area Coating area Approved tablet area

The various tablet defects are:1. Capping 2. Lamination 3. Picking 4. Sticking. 5. Chipping 6. Mottling 7. Black spots 8. Weight variation.

Granulation
Granulation may be defined as a size enlargement process which converts small particles into physically stronger & larger agglomerates. Granulation method can be broadly classified into three types:

Wet granulation Dry granulation Direct compression

The effectiveness of granulation depends on the following properties i) Particle size of the drug and excipients ii) Type of binder iii) Volume of binder iv) Wet massing time v) Amount of shear applied to distribute drug, binder and moisture. vi) Drying rate For granulation, total 5 areas are available. It is grouped based on the capacity difference. Granulation area 1 Capacity 60 kg. Granulation area 2 Capacity 100 kg. Granulation area 3 Capacity 300 kg. Granulation area 4 Capacity 400 kg. Granulation area 5 Capacity 500 kg.

Wet granulation
Important steps involved in the wet granulation:

Mixing of the drugs and excipients Preparation of binder solution Mixing of binder solution with powder mixture to form wet mass.

Coarse screening of wet mass using a suitable sieve Drying of moist granules. Screening of dry granules through a suitable sieve Mixing of screened granules with disintegrant, glidant, and lubricant.

Special wet granulation techniques:


High shear mixture granulation Fluid bed granulation Spray drying

Dry granulation Steps in dry granulation


Milling of drugs and excipients Mixing of milled powders Compression into large, hard tablets to make slug Screening of slugs

General scheme of manufacturing includes:1. 2. 3. 4. 5. 6. 7. 8. Milling Sieving Mixing Granulation Granule drying Sieving Lubrication Compression

The various equipments used are: Rapid mixer granulator High speed mixer granulator Turbo sifter:-The Turbo Sifter is effectively used after the degerminator to recover the fines and separate the high oil content grit/meal Fluidized bed dryer:- Fluid bed dryer is most suitable for pharma equipment and can be used in chemical, food, dyestuff, and polymer industries for fast and efficient drying Rotary press:- Rotary press is also known as tablet compression machine. Ganscoater 1500 (Gansons):- IIPMC is mainly used as the coating polymer.

Compression:-

Here cavin & cadmach machine are available in the compression machine. Generally, in this process first material is passed from the hopper after in the feeder powder filled in die then going in roller and compress the material. Now tooling is decided according to the product.

Tooling is divided in the 3 parts like:


1. Upper punch 2. Dye 3. Lower punch

Function of dye cavity:In the dye cavity the material is filled and compressed the both punch & ejected in outside and according to shape product is outside and according to shape product is outside the dye cavity.

I.P.Q.C. of the compression:1. Balance To check the weight of the tablets. 2. Friabilator To check the friability and if it breaks or not. 3. Disintegration tester (USP) Checks the disintrigation rate. 4. Hardness test Checks the hardness of the tablets. 5. Weight variation

Coating:Coating is a covering that is applied to the surface of an object, usually referred to as the substrate. In many cases coatings are applied to improve surface properties of the substrate, such as appearance, adhesion, wetability, corrosion resistance, wear resistance, and scratch resistance. In other cases, in particular in printing processes and semiconductor device fabrication, the coating forms an essential part of the finished product. I.P.Q.C of the coating: 1. To check the roughness of the tablet 2. Orange peel 3. Sticking & picking

TABLET INSPECTION:In this area the tablets made after compression are came here for the visual inspection of the tablets. The tablets are checked for the uniformity of the coating, any black particle or any other deformity in the tablet. The vibrating or rolloing belt is present on the machine so that the tablets are checked uniformly and a predetermined speed.

TABLET FACILITY 4
In the tablet facility-4 domperidone, liprofloxaline, minoxine and noclate like tablets are manufactured.The temperature maintained here is 20-30 C and the relative humidity is 40 60%.

TABLET PACKAGING:-

In the pharmaceutical industry it is vital step that the package selected adequately preserve the integrity of the product. The selection of a package therefore begins with a determination of the products physical and chemical characteristics.

The materials selected must have the following: 1. 2. 3. 4. 5. 6. They must protect the preparation from the Environment Condition. They must not reactive with product. They must not impart to product tastes or odors. They must be nontoxic. They must be a FDA approved. High speed packaging material.

The whole packing department is divided into main four areas. 1. Primary packaging storage 2. Approved tablet store 3. Tablet inspection store There are mainly three type of tablet packaging: 1. Packaging of tablet into strips 2. Packaging of tablet into blister 3. Packaging of tablet into plastic bottle (bulk packaging) Blister Packaging: - It is the type of packing in which the tablet is filled into plastic pockets covered with aluminum foil. Working steps:The steps of packaging in are:
o o o o o o o

Formation of cavity into PVS foil (temp = 100-180C) Filling of tablet into cavity Coding of aluminum foil Sealing of two film (temp = 120-230c) Cooling Cutting of blister Packing into final paper box.

Strip packaging:Strip packing is same as blister packing but instead of the plastic foil both the films are of aluminum. This equipment is also known as ALU-ALU packaging machine.

The step of packaging are:1. 2. 3. 4. 5. 6. 7. Filling of tablet between to foil Coding of aluminum foil Sealing of two film (temp=125-200c) Cooling Cutting of strip Packing into final paper box Printing on paper box (mfg & exp date)

Packaging area are of two type :1. Primary packaging area 2. Secondary packaging area

PACKAGING:The containers used are mainly made up from glass and plastic
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Plastic containers :The principal ingredient if the various materials used for container is the thermoplastic polymer : Although most of the plastic materials used in the medical field have a relatively low amount of added ingredients, some contain a substantial amount of plasticizers, fillers, antistatic agents, antioxidants and other ingredients added for special purposes.

Glass containers :The glass that is most resistant chemically is composed almost entirely of silicon dioxide, but it is relatively brittle and can only be melted and molded at high temperatures.

Advantages are visibility and cheap and easily made by tubing and molding. The disadvantages are that they are easily breakable.
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Blister packaging :The vials are packed in the blister and for that the machine is automatic in which one side the PVC roll came and then by means of vacuum the sufficient size of pocket is made and then from the other side the aluminum roll came and then by means of heat the sealing is done and finally at downwards the cutting of the strip occur.

EVALUATION PARAMETERS FOR THE PRODUCTS:1 Leak test by means of vacuum is done. if any crack or not properly sealed ampoules easily breaks and removed. 2 For checking the alkalinity of the glass the leaching test is performed. 3 The BREWITY INSPECTION MACHINE was there for checking the glass particles, fibers, black particle and white particle in the vials. There are 3 cameras, two of them used for same purpose for glass and fiber particle checking while another with red LED used to check black particle and volume determination. 4 Black particles and white particles are checked against the white and black background respectively manually for ampoules.

HARD GELATIN CAPSULE

Capsule is an important oral dosage from and has properly to mast the unpleasant taste of drug. It is one of the leading dosage forms after tablet. Advantage of Capsule over other dosage formulation:o o o o

Good elegance Ease and convenient of use Smooth slippery and easily swallowed Can be used for enteric coating

Departments:1. 2. 3. 4. 5. 6. Empty capsule storage area Filling area Coating area I.P.Q.C (In Process Quality Control ) Packaging Dispensed material storage area

Manufacturing Process:

Production of pellets: - Equipments used to produce extruder and spherodiser. Coating of Pellets :- Solace aero coater is used for Pellet coating

Five coatings are done on sugar seed to produce enteric coated pallets.

Name of coating material Hydroxyl propyl methyl cellulose Drug (omeprazole) Hydroxyl propyl methyl cellulose Sodium alginate Endragit

Use of the material provide hardness to sugar seed Active ingredient Barrier coating Separating layer Enteric coating.

Capsule filling:Equipment :- AF-40 and zanasi filling machine. Empty capsule shells are separated in caps and body which are then passed through filling line where drug is filled in the body. Then cap is inserted on body and filled capsules are ejected out. These capsules are then passed through polishing machine for finishing. The condition maintained in this area is, temperature 22 - 24 C and relative humidity 40 -60 %. I.P.Q.C (In Process Quality Control). Some filled capsules are randomly selected for Q.C testing. Weight variation, Content uniformity and Disintegration time are performed. Any deviation from standard value leads to rejection of whole batch.

Packaging:1. Blister packaging:It is the type of packing in which the CAPSULE is filled into Plastic pockets covered with aluminum foil. The instrument used this principal is PAM PAC 240 MACHINE.

Packaging steps:1. 2. 3. 4. 5. Formation of cavity into PVC foil. (Temp = 100-180 C) Filling of capsule into cavity. Coding of aluminum foil Cooling Cutting of blister

Type of rejections:1. 2. 3. 4. 5. 6. 7. Defective filling Concentrated or Empty capsule Colom strain Defective coding Blister with cut pockets Defective cutting Blister with joints

Examples of products:1. Pirocan 2. Fluoxetin 3. Propranalol Hcl

SOFT GELATIN CAPSULE


Product name:1. 2. 3. 4. 5. globac z depin 10 depin 5 globac PM calcit SG

Where the Z stands for 200mg weight and b stands for 200mg weight of API in the capsule.

Manufacturing:Manufacturing is divided into 3 units: 1. Gelatin preparation:- In this unit, preparation of gelatin paste which is released from raw material staging area until form uniform gelatin paste. The maintained conditions are, temp 60-65C. 2. Medicine preparation Here, API and EXIPIENT mixing is taking place. 3. feeding zone 1 and 2 It is an important part of preparation of soft gelatin capsule, where encapsulation done by pumping both medicine and gelatin with the help of lobe pump. Actual capsule drug made by 2 machine 1. Jumbo machine (70-80 thousand capsule/hour) 2. standard machine(30-40 thousand capsule/hour) 3. For benzonatate capsule(a table made as per U.S.P.) For gelatin preparation, 3 equipments are used. 1. Gelatin preparation tank. 2. Colloid mill 3. Gelatin paste storage tank.

Material added and their uses, used for gelatin solution are:-

Glycerin water mixer Sorbitol Methyl & propyl paraben Titanium dioxide Colour Flavor

To make the outer layer of capsule As plasticizer As preservative Anti oxidant To give good appearance To give a good taste

Manufacturing procedure for soft gel capsule:Stage 1:- preparation of gelatin paste Stage 2:- preparation of medicine Stage 3:- Encapsulation In this first Medicine loading takes place then formation of gelatin belt takes place in the bochang machine then the Lubrication of gelatin belt takes place. After that loading of medicine on gelatin belt takes place and then sealing of gelatin belt takes place.

Stage 4:- Drying of capsule


Semidrying, this process takes about one hour. After this ipa washing takes place and then tray drying under one way air flow for 42-72 hours takes place. Stage 5:- Inspection of capsule. This is done manually under vibration principal.The capsule defect (physical) like uneven shape,
twins, black particles, joints are rejected.

Stage 6:- Quality Control of capsule Leak test Disintegration test Weight variation

Packaging:Taking example of product Amlodipine, for which blister packaging takes place, the sealing temp for it is 210 C, in the temp on the top is 122 C and the bottom is 123 C.

SUPPOSITORY

A Suppository is a medicated semi solid dosage form generally intended for use in rectum, vagina, and lesser extent the urethra. Rectal and urethral suppositories usually employ vehicles that melt or soften at body temperature and thus release the medicament in the body cavity and provides the direct release of medicament and thus gives the better bio-avability.

Formulation of suppository:1. Suppositories bases


Cocoa butter Hydrophilic suppository base Water soluble base/ Water dispersible base

2. Medicament :- used as per requirement. Formoress form Filling Suppositories machine is used for preparation of suppositories. Manufacturing process for suppository preparation:Stage 1:- Preparation of suppository base The suppository Base is melted at required temperature. Stage 2:- Mixing of medicine with suppository base The medicine is mixed with the suppo. Base, which is already in melted condition. Stage 3:- Loading of mixer on aluminium strip The mixer is loaded to the machine which contains alluminium strip. Stage 4:- Cooling of mixer The suppository is then cooled to 2 8 C. Stage 5:- sealing of strip The cooled suppository is then sealed with the help of some heat & taking care that suppository doesnt melt. Stage 6:- cutting of strip The suppositories are cut into five pieces. Stage 7:- Inspection area. The product is inspected for the defect, if any.

Rejection of Suppository:The suppositories which are not packed perfectly or not cooled or conataning any of the defects are rejected. Packing Suppository:The packing usually takes place as blister packing and from this the each of five suppository containing groups are separated and are packed in boxes. The condition maintained here in all areas are abound 25 C, temperature.

OINTMENT

Ointment is a semi solid dosage form in which medicament is emulsified in ointment base which are applied external on the skin or any other area other then wounded area. Ointment are normally seen like a lotion, nut they must have good penetration power and thus drugs with the same penetration power can only be used by this way.

Formulation:Ointment base Water soluble base Water miscible base Hydrocarbon base

Medicament

used as per required.

Example: - Oxalgen nano gel

Preparation of Ointment:Stage 1:- Preparation of ointment base blend. The ointment base is prepare by blending it for 1-2 hr. Stage 2:- Emulsification of medicament and Mixing of base with the medicine takes place. - Homogenization: - Homogenizer machine is used to make solution homogenised -storage:- The prepared ointment is stored till it is bring to the packing area. Stage 3:- Packing of ointment in collapsible tube. loading of mixing blend & Empty collapsible tube The empty tubes and ointment are put in their hoopers. - Filling of ointment base:- When the empty tube comes to correct position the ointment is filled in to it. -sealing of collapsible tube :- The filled tubes are then sealed with aid of heat. - printing of collapsible tube:- The printing of specific code on the tube takes place. The machine used for filling the tubes containing the ointments is Auxomatic pentecnica which is brought away from ITALY.

LYOPHILIZATION

Lyophilization is a process more commonly known as freeze-drying. Its a dehydration process typically used to make the material more convenient for transport. It works by freezing the material and then reducing the surrounding pressure and adding enough heat to allow the frozen water in the material to sublime directly from the solid phase to the gas phase.

Manufacturing process:Stage 1:- Vials are unloaded. Stage 2:- Washing of vials by use of WFI. Stage 3:- Vials are depyrolated. Stage 4:- Filling of vials takes place. Stage 5:- Lyophilization takes place.

LYOPHIZISATION PROCEDUE (FROM LIQUID STATE TO SOLID):The product in the liquid form is first prepared and then the product is filtered from the appropriate filters and then the filling of the vials takes place. During Lyophilization the liquids in the vials are formed in to solid by initially heating and then freezing the vials at -45 C.

PARENTRAL

It is sterile dosage forms which are use intended inside the body, give fast onset of action than other dosage form. But the most important thing is when it is given once its effect cannot be minimized. The various types of parentrals roots:1. 2. 3. 4. Subcutaneous injection Intradermal injection Intravenous injection Intra-arterial injection

Manufacturing process for parental preparation:Stage 1:- Vial washing with purified water & then water for injection Stage 2:- Dried with compressed air when fed the drying zone of tunnel. Stage 3:- Depyrogenation at 330 C for minute. Stage 4:- Then fed to a cooling zone. Stage 5:- Then move to grade area for aseptic filling. Stage 6:- Half stoppered vials fully loaded in lyophilyser. Stage 7:- After completion of process vials fully stoppered. Quality control test

Leakage test Clarity test Pyrogen test Sterility test

Parentral packaging:Blister Packaging is the type of packing in which the tablet is filled into plastic pockets covered with aluminum foil. Steps:1 Formation of cavity into PVS foil (temp 100-180C) 2 Filling of tablet into cavity 3 Coding of aluminum foil 4 Sealing of two film (temp = 120-230C) 5 Cooling 6 Cutting of blister 7 Packing into final paper box.

AEROSOL

Aerosol spray is a type of dispensing system which creates an aerosol mist of liquid particles. This is used with a can or bottle that contains a liquid under pressure. When the container's valve is opened, the liquid is forced out of a small hole and emerges as an aerosol or mist. As gas expands to drive out the payload, only some propellant evaporates inside the can to maintain an even pressure. Outside the can, the droplets of propellant evaporate rapidly, leaving the payload suspended as very fine particles or droplets. Typical liquids dispensed in this way are insecticides, deodorants and paints. An atomizer is a similar device that is pressurised by a handoperated pump rather than by stored gas.

Working
As gas expands to drive out the payload, only some propellant evaporates inside the can to maintain an even pressure. Outside the can, the droplets of propellant evaporate rapidly, leaving the payload suspended as very fine particles or droplets.

Aerosol propellants
If aerosol cans were simply filled with compressed gas, it would either need to be at a dangerously high pressure and require special pressure vessel design (like in gas cylinders), or the amount of gas in the can would be small, and would rapidly deplete. Usually the gas is the vapor of a liquid with boiling point slightly lower than room temperature. Chlorofluorocarbons (CFCs) were once often used, but since the Montreal Protocol came into force in 1989, they have been replaced in nearly every country due to the negative effects CFCs have on Earth's ozone layer. The most common replacements are mixtures of volatile hydrocarbons, typically propane, nbutane and isobutene. Dimethyl ether (DME) and methyl ethyl ether are also used.

Packaging
A typical paint valve system will have a "female" valve, the stem being part of the top actuator. The valve can be preassembled with the valve cup and insta can as one piece, prior to pressurefilling. The actuator is added later. Aerosol Evaluation: Flammability &combustibility Flash point Flame extension, Physicochemical characterization vapour pressure

Density Moisture content Performance Aerosol valve discharge rate Spray pattern

Dosage with metered valve Biological characterization

TRANSDERMAL MANUFACTURING

Transdermal patch is a medicated adhesive patch that is placed on the skin to deliver a specific dose of medication through the skin and into the bloodstream. Often, this promotes healing to an injured area of the body. A wide variety of pharmaceuticals can be delivered by transdermal pressure. An advantage of a transdermal drug delivery route over other types such as oral, topical, etc is that it provides a controlled release of the medicament into the patient. A disadvantage to development however, stems from the fact that the skin is a very effective barrier.

Popular uses
The highest selling transdermal patch in the United States was the nicotine patch which releases nicotine to help with cessation of tobacco smoking. The first commercially available vapour patch to reduce smoking was approved in Europe in 2007. Fentanyl, an analgesic for severe pain Other skin patches administer estrogen for menopause. This also seems to prevent osteoporosis after menopause. Nitroglycerin patches for angina are available. Clonidine has also been administered transdermally. Buprenorphine, marketed as BuTrans, as analgesia for moderate to severe chronic pain

Components
The main components to a transdermal patch are: Liner- Protects the patch during storage. The liner is removed prior to use. Drug- Drug solution in direct contact with release liner Adhesive- Serves to adhere the components of the patch together along with adhering the patch to the skin

PILOT PLANT
A pilot plant is a small chemical processing system which is operated to generate information about the behavior of the system for use in design of larger facilities. Pilot plants are used to reduce the risk associated with construction of large process plants. They do this in two ways:

They are substantially less expensive to build than full-scale plants. The business does not put as much capital at risk on a project that may be inefficient or unfeasible. Further, design changes can be made more cheaply at the pilot scale and kinks in the process can be worked out before the large plant is constructed. They provide valuable data for design of the full-scale plant. Scientific data about reactions, material properties, corrosiveness, for instance, may be available, but it is difficult to predict the behavior of a process of any complexity. Engineering data from other process may be available, but this data cannot always be clearly applied to the process of interest. Designers use data from the pilot plant to refine their design of the production scale facility.

If a system is well defined and the engineering parameters are known, pilot plants are not used. For instance, a business that wants to expand production capacity by building a new plant that does the same thing as an existing plant may choose to not use a pilot plant. Additionally, advances in process simulation on computers have increased the confidence of process designers and reduced the need for pilot plants. However, they are still used as even state-of-the-art simulation cannot accurately predict the behavior of complex systems.

CONCLUSION

I am highly thankful to ZYDUS CADILA Pharmaceutical & entire staff to have given me an opportunity to work. This training period during my educational period was very important for me to improve myself as a pharmacist. It has helped me a lot in gaining practical knowledge with regards to what are the latest inventions and researches going on. Also this training has been beneficial to me because it has given me a chance to imply my theoretical knowledge, practically. For me it has been a great experience to take training in such a renowned & well established pharmaceutical company. This training shall help me a lot in future as a Professional pharmacist.