A PROJECT REPORT ON INDUSTRIAL TRAINING COMPLETED AT

CHEMO TEST LABORATORIES LTD. SEWARI MUMBAI

Submitted by Mr.ROHIT MANGESH PATIL Student of M.Sc Part-II in Analytical Chemistry Of D.G.RUPAREL COLLEGE MATUNGA ROAD.

Affiliated to UNIVERSITY OF MUMBAI

ACKNOWLEDGEMENT
This project of basic chemical and instrument is required to motivate employee improve his or her standard of living, improve status in society. I student of D.G.RUPAREL COLLEGE of science heart felt thanks and appreciation to all those who have made this project see the light of the day. I would firstly like to thanks university of Mumbai for introducing new course, which give an excellent opportunity to the student to excel their specialization by caring out an assignment in that subject. My grateful thanks to Dr. Nandini Pai our H.O.D. of D.G.RUPAREL COLLEGE, for arranging my training to this esteemed organization. I would specially like to thank Mr.R.M.Patil sir (G.M. of Analytical department) for encouraging and Providing me with relevant information and material . The blend of which had made it a knowledge project.

DECLARATION

I am ROHIT MANGESH PATIL STUDENT OF M.Sc. Analytical chemistry (partII ) from D.G.RUPAREL COLLEGE, here by declare that, I have completed project on Q.C. department in academic year 2010-2011. The information submitted is true on the best of my knowledge.

DatePlace-Goregaon

Signature of the student

INDEX

Contents Sr.No. 1. 2. 3. Company Profile Quality Control Department Methods of Quality Control Analysis

Page No.

A.

Instrumental Methods 1. Ultra violet and visible spectrophotometer 2. Disintegration and Dissolution test forTablets and capsules. 3. Atomic Absorption Spectrophotometer. 4. Gas Chromatography. 5. High-Performance Liquid Chromatography. 6. Determination of moisture contain by using Karl Fischer Classical Methods of Analysis 1. LOSS OF DRYING 2.0

B.

COMPAMY PROFILE

QUALITY CONTROL DEPARTMENT

The Q.C. Dept is the heart of any industry; the quality control department deals with regular and periodic checking of raw materials, finished products, packed goods. The tests carried out include assays, related substance detection, average weight, moisture contents etc. Various Analyses of Quality Control are: 1) Raw material. 2) Finished product. 3) Stability. 4) Method validation. Following are some of the important functions of Quality Control Department: 1)To see that the product produced in company should be of consistent quality. It should be free from contaminants and also be free from environmental vectors such as humidity, micro-organisms etc. It should be well preserved while storage, to maintain its original forms intact. These are called Good Manufacturing Practices and are done by quality control department. 2)To maintain consistency in obtaining yield of the product, this is done by strictly obeying the Standard Operating Procedures (SOP) and monitoring the reactions from time to time. 3)To responsibly monitor the quality of the raw materials, the reactions going in the reactors and the finished goods produced. 4)To check the documents from stores, manufacturing, packing department before release of product. 5)To analyze the product for the market complaints, if any. The CONTROL SAMPLE from each bath dispatched, is stored for this purpose, if any customer complaints for the unsatisfactory quality of product, the control sample is analyzed for the same, and the complaint report is prepared and sent to concerned office. Control samples are stored for 2 years from the date of dispatch. 6)To check whether the new products prepared by R&D are as per validation protocol.

ULTRA VIOLET AND VISIBLE SPECTROPHOTOMETER Ultraviolet-visible spectroscopy or ultraviolet-visible spectrophotometry (UVVis or UV/Vis) refers to absorption spectroscopy in the ultravioletvisible spectral region. This means it uses light in the visible and adjacent (nearUV 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.[1] UV/Vis spectroscopy is routinely used in the quantitative determination of solutions of transition metal ions highly conjugated organic compounds, and biological macromolecules. 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).

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 or oftenAU / M * cm. The absorbance and extinction are sometimes defined in terms of the natural logarithm instead of the base-10 logarithm. The Beer-Lambert Law is useful for characterizing many compounds but does not hold as a universal relationship for the concentration and absorption of all substances. A 2nd order polynomial relationship between absorption and concentration is sometimes encountered for very large, complex molecules such as organic dyes (Xylenol Orange or Neutral Red, for example). Control of wavelengths: In liquids, the extinction coefficient usually changes slowly with wavelength. A peak of the absorbance 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.

241.15nm(Ho) 253.70nm(Hg) 287.15nm(Ho) 302.25nm(Hg) 313.16 nm(Hg) 334.15 nm(Hg) 361.50 nm(Ho)

404.66nm(Hg) 435.83 nm(Hg) 486.00 nm(D) 486.10 nm(H) 536.30 nm(Ho) 546.07 nm(Hg) 576.96 nm(Hg)

365.48 nm (Hg) 579.07 nm (Hg) Control of absorbances: check the absorbances using potassium dichromate solution uv at wavelengths indicate in table 1, which gives for each wavelengths the exact value of A(1%,1 cm) & permitted limits.

Table 1

235 257 313 350

124.5 144.0 48.6 106.6

122.9 to 126.2 142.4 to 145.7 47.0 to 50.3 104.9 to 108.2

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. Resoultion power: when express in monograph,record the spectrum of 0.02% v/v solution of toluene in hexane uv,The ratio of the absorbance at the maximum about 269 nm to that at the maximum about 266 nm is not less then 1.5 unless otherwise specified in the monograph. Specrum Reagent : The temperatures of the all solution used in the test should not differ by more than 0.5 degree Resolution : Reduced to its most basic denition, resolution is dened as the ability of an instrument to separate light into nite,distinct wave lengthregions and distinguish these niteregions from each other. Resolution is primarily governed by the physical slit width of the instrument, in combination

with the inherent optical dispersion of light from the exit of the monochromator to the detector of the instrument.Reducing the physical slit width decreases the spectralbandwidth and improves the ability of the instrument to resolve closely spaced peaks.

SAMPLE NAME : KOLD TIME TABLET (PARACETAMOL)

ACETAMINOPHEN

OH

NHCOCH3 C8H9NO2 Paracetamol is 4-hydroxyactanilide Mol.Wt.151.16

TEST

: Assay (Paracetamol)

WEIGHT VARIATION : Weight of 20 t6ablets :- 13.7504gm/20 = 0.6875gm Avg of tablet :- 0.6875gm STANDARD PREPARATION :- 0.01mg/ml of paracetamol in the diluent

0.1321gm of Paracetamol 1ml of above solution

100ml with diluent 100ml with diluent

ASSAY PREPARATION :_ Weigh accurately 100mg of powder in 100ml of vol flask with 0.1N HCl and take 1ml of this solution in 100ml flask 0.1815gm eq. to 132mg 100ml with diluent 1ml of above solution 100ml with diluent WAVELENGTH :- 242 nm

STD ABSORPTION :- 0.740

SPL ABSORPTION :- 0.736 STD PURITY :- 99.405%

CALCULATION 0.736 X 132.1 X 1 X 100 X 100 X 0.6875 X 99.405 0.740 100 100 0.1815 1 150 = 98.94% ie = 98.94 X 500 =494.7mg/tab. 100

CONCLUSION :- Kold time tablet contain NLT 90.0% and NMT 110.0% of Paracetamol and by analysis got 98.94%

V.

PHENYL BUTAZONE USP

-------- PHE230501

1.0

6.63 0.5

PHENYAL BUTAZONE

STD-1

10

15

20

25

time

DISINTEGRATION AND DISSOLUTION TEST FOR TABLETSS AND CAPSULES

This test determines whether tablets or capsules disintegrate within a prescribed time when placed in a liquid medium under the prescribed experimental conditions. For the purpose of this test, disintegration does not imply complete solution of the tablet or capsule or even its active constituent. Disintegration is defined as that state in which no residue of the tablet or capsule remains on the screen of the apparatus or, if a residue remains, it consists of fragments of insoluble coating of the tablets or of capsule shells or is a soft mass with no palpable core. If discs have been used with capsules, any residue remaining on the lower surfaces of the discs consists only of fragments of shells. A rigid basket-rack assembly supporting six cylindrical glass tubes, 77.5 2.5 mm long, 21.5 mm in internal diameter and with a wall thickness of about 2 mm. The tubes are held vertically by two superimposed transparent plastic plates, 90mm in diameter and 6mm thick perforated by six holes having the same diameter as the tubes. The holes are equidistant from the centre of the plate and are equally spaced from one another. Attached to the under side of the lower plate is a piece of woven gauze made from stainless steel wire 635 m m in diameter and having nominal mesh apertures of 2.00 mm. The upper plate is covered with a stainless steel disc perforated by six holes, each about 22 mm in diameter, which fits over the tubes and holds them between the plastic plates. The holes coincide with those of the upper plastic plate and the upper open ends of the glass tubes. The plates are held rigidly in position and 77.5 mm apart by vertical metal rods at the periphery and a metal rod is also fixed to the centre of the upper plate to enable the assembly to be attached to a

mechanical device capable of raising and lowering it smoothly at a constant frequency of between 28 and 32 cycles per minute through a distance of 50 to 60 mm. The design of the basket-rack assembly may be somewhat different provided specifications for the glass tubes and the screen mesh size are unchanged. A cylindrical disc for each tube, each 20.7 0.15 mm thick in diameter and 9.5 0.15 mm thick, made of transparent plastic with a relative density of 1.18 to 1.20, and pierced with five holes, each 2 mm in diameter, one in the centre and the other four spaced equally on a circle of radius 6mm from the centre of the disc. Four equally-spaced grooves are cut in the Lateral surface of the disc in such a way that at the upper surface of the disc they are 9.5mm wide and 2.55mm deep and at the lower surface 1.6 mm square. The assembly is suspended in the liquid medium in a suitable vessel, preferably a 1000-ml beaker. The volume of liquid is such that the wire mesh at its highest point is at least 25mm below the surface of the liquid, and at its lower point is at least 25mm above the bottom of the beaker. A thermostatic arrangement for heating the liquid and maintaining the temperature at 37 2 .

All parts of the apparatus, including any metal that may come into contact with the sample to be tested or the dissolution medium, should be made from a chemically inert material and should not adsorb, react or interfere with the preparation or the dissolution medium. The dissolution assembly should be constructed in such a way that any vibration is reduced to a minimum. Use an apparatus that allows full visibility of all operations. The apparatus "Paddle" (Fig. 3) consists of a cylindrical vessel of suitable glass or other suitable transparent material with a hemispherical bottom and a nominal capacity of 1000 ml. The vessel is covered to prevent evaporation of the medium with a cover that has a central hole to accommodate the shaft of the stirrer and other holes for the thermometer and for devices for withdrawal of liquid. The stirrer consists of a vertical shaft with a blade at the lower end. The blade is constructed around the shaft so that it is flush with the bottom of the shaft. When placed inside the vessel, the shaft's axis is within 2mm of the axis of the vessel and the bottom of the blade is 25 2mm from the inner bottom of the vessel. The upper part of the shaft is connected to a motor provided with a speed regulator so that smooth rotation of the stirrer can be maintained without any significant wobble. The apparatus is placed in a water-bath that maintains the dissolution medium in the vessel at 37 0.5 C. The apparatus "Basket" (Fig. 4) consists of the same apparatus as described for "Paddle", except that the paddle stirrer is replaced by a basket stirrer. The basket consists of two parts. The top part, with a vent, is attached to the shaft. It is fitted with three spring clips, or other suitable attachments, that allow removal of the lower part so that the preparation being examined can be placed in the basket. These three spring clips firmly hold the lower part of the basket concentric with the axis of the vessel during rotation. The lower detachable part of the basket is made of welded-seam cloth, with a wire thickness of 0.254 mm diameter and with 0.381 mm square openings, formed into a cylinder with a narrow rim of sheet metal around the top and the bottom. If the basket is to be used with acidic media, it may be plated with a 2.5-m layer of gold. When placed inside the vessel, the distance between the inner bottom of the vessel and the basket is 25 2mm.

High-Performance Flow-Through Dissolution Manifold [model 2008-6CM]

High-Performance Flow-Through Dissolution systems represent the latest and most significant advancements in dissolution science. A modification of the USP type 4 apparatus and in-line with joint recommendations from the AAPS, FIP, USP and FDA, these systems are ideal for product development, especially for devices, films, implants, stents. These High-Performance dissolution systems can also easily be used with tablets and capsules for much greater reliability, precision and correlation to in vivo results. Either a general research or a custom designed High-Performance Flow-Through Dissolution System is available to meet general or specific pharmaceutical need.

There systems are unsurpassed in there ability to optimize convective diffusion and hydrodynamics which in turn provide vast improvements in precision, relevant sensitivities, mathematical and in vivo correlations. Additionally, these systems are designed to be integrated with HPLC components, are easily automated and have very rapid method development cycles.

TOLERANCES:An amount of ciprofloxacin Hcl to not less than 80.0% Q of lable amount of ciprofloxacin dissolve in 30 min in medium.

WAVELENGTH:STD ABSORPTION:TABLETS 1 TABLETS 2 TABLETS 3 TABLETS 4 TABLETS 5 TABLETS 6 STD PURITY CALCULATION TAB-1 = ::::::-

276nm 0.687 0.667 0.664 0.668 0.664 0.665 0.665

:-

99.465%

0.667 X 28.17 X 1 X 900 X 100 X 99.46 0.687 50 100 500 1

= 93.01%

TAB-2 TAB-3 TAB-4 TAB-5 TAB-6

- 0.664 =92.59% - 0.668 =93.15% - 0.664 =92.59% - 0.665 =92.73% - 0.665 =92.79%

MEAN=92.8% CONCLUSION:- Cipocial tablets contain not less than 80.0% of Ciprofloxacin Hcl. And by the analysis got 92.8%.

ATOMIC ABSORPTION SPECTROPHOTOMETER

GC GAS CHROMATOGRAPHY: Gas chromatography (GC) is an analytical technique for separating compounds based primarily on their volatilities. Gas chromatography provides both qualitative and quantitative information for individual compounds present in a sample. Compounds move through a GC column as gases, either because the compounds are normally gases or they can be heated and vaporized into a gaseous state. The compounds partition between a stationary phase, which can be either solid or liquid, and a mobile phase (gas). The differential partitioning into the stationary phase allows the compounds to be separated in time and space.

Operation of Gas Chromatograph

Instrument Details Instrument Name Make

: : Gas Chromatograph : Shimadzu

Determination of the completion of the reaction: The batch reactions being carried out in the reactors are frequently Being monitored for their completion. This is done by sampling the solution From the regular intervals of time after the reaction is started. The samples are analyzed using the gas chromatography instrument, using a suitable column and other variables. When a sample is injected into a GC apparatus, a Chromatogram is generated, which contains peaks at different distance on time axis, this indicates the retention time of the compounds, hence the compounds are identified, and distinguished as product and reactant. Then the peak area, and hence the corresponding percentage concentration is automatically calculated by the computer, thus determining the percentage concentration of each component. As the reaction progresses the concentration of the reactants is reduced, while the product increases, this is determined from the increasing peak area of the peak corresponding the product, while that of the reactant diminishes.

ANALYSIS BY GAS CHROMATOGRAPHY

SAMPLE:SAMPLE NAME:TEST :XANTHUM GUM ORGANIC VOLATILE IMPURITY

DESCRIPTION :- White Powder CHROMATOGRAPHIC CONDITION :COLUMN :- BP 624 (30m X 0.53 mm)6% cynopropylphenyl-94% Dimethylpoly siloxane OVEN TEM :- 45*c(ISOCRATIC) INJECTORE TEM :- 180*C DETECTORE TEM :- 200*C HEADSPACE TEM :AUXILLARY TEM:- 100*C TRANSFER TEM :- 110*C METHOD :For OVI test take some liquid reagent like Dichloromethane,chloroform,Trichloroethelem and 1,4 Dioxan dissolve in water.Take 5.0 ml in vial determine OVI. STANDARD PREPARATION :-

1:- DICHLOROMETHANE

0.002 X 1.342 X 100 X 10000 100 = 26.48 ppm 2:- CHLOROFORM

0.004 X 1.476 X 100 X 10000 100 = 59.04 ppm 3:- TRICHLOROETHELENE

0.004 X 1.463 X 100 X 10000 100 = 58.52 ppm 4:- 1,4 DIOXAN 0.03 X 1.033 X 100 X 10000 100 = 309. Ppm SOLN 1+2+3+4 100 ML WATER 5 ml of water

SAMPLE PREPARATION :- 0.1123 gm spl

CONCLUSION:- Ovi in Xanthum gum NMT DCM,600PPM CHCL3, 60PPM TCE,80 PPM 1,4 DIOXAN 380 PPM and by the analysis got NIL.

ATOMIC ABSORPTION SPECTROPHOTOMETRY

This technique is based on the fact that when atoms ,ions or ion complexs of an element in the ground state are atomized in a flame ,they absorb light at the characteristic wavelength of that element.If the absorption is proportional to the number of absorbing atoms.

Apparatus
In atomic absorption spectrometry, atoms in the electronic ground-state (and ions to a much lesser degree) can absorb photons at energies (wavelengths) corresponding to the energy transition from the ground to the excited state. Because atomic absorption occurs over a very narrow band of energy, a conventional monochromator cannot be used to provide the light for absorption. Instead a hollow-cathode lamp specific for the element to be measured provides the narrow-band of light (wavelength <0.001 nm) needed to measure absorption. Skoog points out that only the small fraction of atoms & ions are actually excited by the flame emission process and that simultaneously a number of concurrent processes related to the sample matrix can cause interferences with the emission process. Although generally lesser in number and amount, matrix effects also occur for atomic absorption spectrometry. One of the goals of this laboratory is to address these sample matrix effects so that they have a minimal effect on both the accuracy and precision of your measurements. In this experiment, you will use both atomic emission and absorption to measure calcium and sodium in water samples. Before beginning this experiment, you should familiarize yourself with AA and AE methods and the IL-457 instruction manual. There are many ions present in water that are suitable analytes for either AA or AE methods. For example, many water sources are referred to as being hard, i.e. the water has a

high concentration of calcium. The calcium in hard water will form deposits (scale) over time, especially in heated water, in water pipes, causes spots on glasses, bathtubs, teakettles, and insidiously inside water heaters. Another common effect is the decreased efficiency of soaps in dissolving hydrophobic materials like grease. Hard water is treated by installing a water softener in the water system. The water softener is generally nothing more than a sophisticated, rechargeable ion exchange resin system. It removes calcium ions and substitutes sodium ions. The resultant sodium-containing water is termed softened.

Atomic Absorption Spectrophotometry (AAS) When using atomic absorption spectrophotometry (AAS) as an analytical technique the absorption of light of free atoms is measured. Therefore it is one of

the branches of atomic spectroscopy, together with flame photometry (see Standardbase techniques: Flame Photometry that measures the intensity of light emitted by free atoms when their electrons return to ground state after the excitation by light). However - unlike flame photometry - AAS is based on the first half of the excitation process, while atoms absorb light getting their electrons from the ground state to a higher energy level. Although the atomic absorption spectrophotometer (fig. 1.) is quite expensive, the technique is very wide-spread, thank to the fact that by AAS it is possible to determine about 70 elements (mainly metals) at very low concentrations. The sample is atomised at a very high temperature (2500-3000 C) and the free atoms have line spectrum. It means that they can only absorb the energy of light at discrete energy levels according to the excitations of electrons. Excitation energies in this case are determined by the difference between the energy level of the ground state and one of the excitation states of their electrons. Only a light with a concrete wavelength belongs to each of these excitation energies and when this light is absorbed it is missing from the continuous spectra of the electromagnetic radiation: a black line appears in the absorption spectrum of the atom. There are no vibration or rotation energy levels that would widen the lines to brands in the spectrum (like it happens in the case of UV-Vis spectrophotometry, when molecules and ions are measured, see Standardbase technique: UV-Vis Spectrophotometry). Using AAS free atoms are lit by monochromatic light (called resonance radiation that has got a special wavelength) that belongs to one line of their spectrum and therefore it has the suitable excitation energy mentioned above. Only the examined atoms can absorb it. As a result of absorption, the intensity of light decreases, which is proportional to the number of the examined atoms being present. That makes very sensitive quantitative measurements possible. To produce the proper monochromatic light necessary for the AAS, so called hollow cathode lamps are used. The cathode of this sort of lamp is made of the metal under investigation (or its alloy). It means that different lamps are used for the determination of each element. It is named after the cylindrical shape of the cathode that gives direction to emerging beam, and helps re-deposit sputtered atoms back on cathode. The anode is made of tungsten and the electrodes are surrounded by noble gases. At high voltage the cathode produces electrons that speeding up in the electric filed cause the ionisation of noble gas atoms. These high-speed noble gas ions bombard the cathode and therefore sputtering occurs, dislodging atoms from the surface of cathode. These free atoms are excited by the high-speed electrons and then emit the line spectrum characteristic of the particular element that is the cathode made of. AAS called a destructive technique, because only solutions containing the

investigated element can be used. Solid samples should be accurately weighed and then dissolved, often using strong acids (e.g. in cases when soil samples contaminated with heavy metal ions are measured). However only a very small amount of sample is enough, because of the high sensitivity of the technique. The solvent of the solution is evaporated and all materials present in the sample are vaporised and dissociated to atoms at the very high temperature. (The process in the reality is a bit more complicated, since ions and oxides are also produced, decomposition and association reactions take place too.) The following atomisation methods are known: Flame atomisation Graphite furnace atomisation Mercury hydride atomisation (this is only mentioned here, but not used while doing Standardbase experiments). The source of atoms is usually flame (flame atomisation). Metals could be measured at ppm concentration (part per million, that is mg kg -1 or mg dm-3 in case of dilute solutions). The sensitivity could be increased when the light travels for longer in the flame. Therefore most of the burners are about 5-10 cm long. The accuracy is very good, about 1-2%. The sample solution is sprayed (nebulized) continuously into the flame (similarly to the flame photometer). The graphite furnace AAS (GFAAS), a more recent technique is even more sensitive than the traditional, cheaper AAS using flame. Measuremnts could be done at ppb level (part per billion, ppb = 10

-3 ppm, that is g kg -1 or g dm-3 in case of dilute solutions!). The accuracy is still about 20% in this latter case. The drying, combustion, vaporisation and atomisation of sample happen in a heated graphite tube that is placed in the way of light. This graphite furnace is protected against oxidation by an inert gas (e.g. argon). The other parts of the atomic absorption spectrophotometer are similar to the one used in the case of UV-Vis spectrophotometer. The monochromator that selects the proper wavelength of the emitted spectra is the usual prism or optical filter. The detector is a photo-electron multiplier that produces an electric sign proportional to the intensity of emitted light. (It contains diodes /electrodes with increasing potential/ between the photocathode and anode, where multiplied .

ANALYSIS BY ATOMIC ABSORPTION SPECTROPHOTOMETER

SAMPLE:SAMPLE NAME :- TALCUM BP TEST :DETERMINATION OF ALUMINUM BY

DESCRIPTION :- white powder. METHOD:TEST SOLUTION :weight 0.5030 gm of sample take in polytetra fluotoethelene dish & add 5ml of lead free nitric acid .Stire gently & then add 35ml of Hydrofluoric acid. And evaporate slowly to dryness on a hot plate add to residue ml of Hcl acid.and teansfar in 50 ml Dilute with water. TEST PREPARTION:- Take 5ml of above solution add 10 ml of 25.34 Gm/1 solution of cesium chloride solution & 10 ml of Dilute & 10 ml of Hcl & Dilute to 100ml with water. STANDARD PREPARATION :- (2.0 ppm) From the stock soln(1000ppm) of ALUMINIUM From stock solution 10ml of above soln 10ml of above soln 2ml of above soln 100ml with water 100ml with water 100ml with water

CALCULATION:Weight of sample :Std concentration :Spl concentration :Splconcentration :0.5030 2.0ppm 0.314 0.02894

0.314 X 50 X 100 0.05030 X 5 = 624.24 ppm ie 624.24 10000 =0.0624 % CONCLUSION:- Talcum powder contain not more than 2.0% of Aluminium By the analysis got 0.0624%.

DETERMINATION OF PH VALUE The pH is determine conventially represent the acidity or alkalinity of an aqueous solution.in pharmacopeia ,standards and limits of pH have been provided for thopharmacopia

HPLC

HPLC: a method of chromatographic separation based on the difference in the distribution of species between two non-miscible phases,in which the mobile phase is a liquid which percolates through a stationary phase contained in a column mainly based on mechanisms HPLC of - adsorption - mass distribution - ion exchange - size exclusion - stereochemical interaction

Instrument Details : Instrument Name : High Pressure Liquid Chromatograph Make : Younglin

APPARATUS a pumping system an injector, a chromatographic column (a column temperature controller may be used) a detector a data acquisition system ( an integrator or chart recorder) The mobile phase is supplied from one or several reservoirs and flows through the column usually at a constant rate , and then through the detector.

SAMPLE:1

SAMPLE NAME:

PHENYL BUTAZONE USP

C19H20N2O2

Mol.Wt.308.38

PHENYL BUTAZONE

is 4-butyl-1, 2-diphenyl-3, 5-pyrazolidinedione.

TEST :- ASSAY DESCRIPTION :- WHITE POWDER METHOD ACETATE BUFFER :- Transfer 2.72 gm of sodium acetae to a 1000ml and dissolve in about 700 ml of water.Adjust with glacial acetic acid to pH of 4.1 filter through a 0.5u filter.Dilute with filtered water to 1000ml. MOBILE PHASE :- Prepare a filtered and degassed mixture of acetonitrile with 560ml of acetic acid buffer (400;560) make adjustment if necessary. CHROMATOGRAPHIC CONDITION :- The liquid chromatographis equipped with a 254 nm detectore and a 4.6 mm X 25 cm column that contain packing L7(C8).The flow rate 2.4 ml per minut. STANDERD PREPARATION:- Dissolve an accurately weight quantity of USP phenyl Butazone RS in acetonitrile.With add of sonicate and ilute quantity of acetonitril to obtaind the concentration about 1.4 mg per ml .pepette out 10 ml of this solution 50 ml of flask.0.0142gm 10mlwith ACN 10ml of above solution 50ml with CAN

ASSY PREPARATION ;- Transfer about of 140mg of phenyl butazone accuretly weight to a 100ml volumetric flask.add 75ml of ACN and sonicate to dissolve. Dilute with ACN to volume. And pipette out 10ml of this solution in 50 ml with ACN . 0.1430gm 10mlof above solution 100ml with ACN 50ml with ACN

STD PURITY:STD AREA 1 :-12716.269 2 :-12773.408 3 :-12761.542

MEAN :-12750.406 SDV :-30.153250 RSD :-0.2236488

99.24% oanhydrous basis

CLCULATION:12843.697 X 14.2 X 5 X 100 X 50 X 99.24

12750.40 = 99.27

10

50

0.1430 5

CONCLUSION:- Phenyl butazone contain not less than 98.0% and not more
than 102.0% on dried basis.And but the analysis got 99.44% on dried basis.

V.

PHENYL BUTAZONE USP

-------- PHE230501

1.0

6.63 0.5

PHENYAL BUTAZONE

STD-1

10

15

20

25

time

Ret.times areamV.s Hight min mV 6.633 12716.28 630 9 Total 12716.28 630 9
V.

Area% 100.00 100.00

Height% W05(min) 100.00 100.00 0.31

PHENYL BUTAZONE USP

-------- PHE230502

1.0

6.62 0.5

PHENYAL BUTAZONE

STD-2 0.0 5 10 15 20 25 time

Reten.Time [min]

Area [mV.s]

Height [mV]

Area

[%]

Height [%]

WOS [min]

6.623 Total

12773.408 12773.408

617 617

100.000 100.000

100.0 100.0

0.32 0.32

V.

PHENYL BUTAZONE USP

-------- PHE230503

1.0

6.64 0.5

PHENYAL BUTAZONE

STD-3 0.0 5 10 15 20 25 time

Reten.Time [min]

Area [mV.s]

Height [mV]

Area

[%]

Height [%]

WOS [min]

6.640 Total

12761.542 12761.542

602 602

100.000 100.000

100.0 100.0

0.33 0.33

V.

PHENYL BUTAZONE USP

-------- PHE230504

1.0

6.64 0.5

PHENYAL BUTAZONE

SPL 0.0 5 10 15 20 25 time

Reten.Time [min]

Area [mV.s]

Height [mV]

Area

[%]

Height [%]

WOS [min]

6.657 Total

12843.697 12843.697

589 589

100.000 100.000

100.0 100.0

0.4 0.4

Determination of Moisture by Karl Fischer Titration:

Determination of the water content in the compounds is an important factor for many chemical reactions, in order to check the product quality and durability. Earlier loss on drying was one of the methods used for the determination of the moisture, but it has limited use as it also measures the loss of volatile components. Karl Fischer titration serves a moisture specific method for the determination of trace amounts of water in a sample that uses colorometric or volumetric titration. Preparation of the K F Reagent: The German chemists KARL FISCHER prepared the reagent containing Pyridine, SO2 and Iodine, which react according to the Bunsen reaction, hence can be used for the determination of the amount of water in the sample as; one mole of I2 is consumed for each mole of H2O 2H2O + SO2 + 2Py + I2 H2SO4 + 2 HI*Py Now a days a pyridine free KF reagent is used which is odour free and has high rate of reaction. The KF reagent contains base like Imidazole or Secondary amine instead of pyridine, which not catalyses but participates in the reaction. Pyridine has following drawbacks, which can be overcome by using the pyridine free reagent . o It has unpleasant odour, where as Pyridine free reagent is odourless o It is carcinogenic. o The rate of reaction is slow, hence the end point is determined with low accuracy, where as the pyridine free reagent has high rate of reaction hence gives a sharp end point, with more accuracy and reproducible.

First Generation of Pyridine-free Karl Fischer Reagents: As Anders Cedergren noticed the pyridine was just a buffering agent and not a reactive part According to the results it was possible to replace the unpleasant pyridine with the odourless imidazole. 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. The titration cell also consists of a smaller compartment with an (anode) immersed in the anode solution of the main compartment. 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. Reaction: When the KF reagent comes in contact of methanol it forms methyl sulphite (active species of KF Titration). The reaction is as follows, SO2 + MeOH + B MeSO-3 + HB

Methyl sulphite gets oxidized to methyl sulphate, with quantitative oxidation of I2 to 2I- per molecule of H2O as shown in the following reaction. MeSO-3 + H2O + I2 +2B MeSO -4 +2HB + 2I B= Base The end point is detected most commonly by a bipotentiometric method. A second pair of Pt electrodes is 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.

GENERAL PROCEDURE TO DETERMINE THE MOISTURE CONTENT IN SAMPLES BY KARL-FISCHER TITRATION METHOD Reagents Karl Fischer`s reagent Methanol ( AR Grade ) Distilled Water Apparatus > Karl Fischer`s apparatus >Standard volumetric glassware >Analytical balance Operation of Karl-Fischer Apparatus A Instrument Details Instrument Name Make : : Karl-Fischer Apparatus : Analab

1 2 3 4 5 6 7

DETERMINATION OF KF FACTOR AquaCalModel : 10 Switch ON the main and fill the reaction vessel with fresh dried methanol (approx. 50 ml). Ensure the connection of electrodes. Switch ON the stirrer switch to start stirrer. Neutralize methanol by K.F. solution by pressing START button. Gradually addition of K.F. reagent has been started and when the vessel solution becomes neutralized, it will give beep sound. Press ESCAPE button. Standardization of K.F. reagent a) Weigh accurately about 5 mg of distilled water. b) Record the weight on approved format.

c) Transfer the weight in the beaker, containing neutralized methanol. d) Press the START button, the titration will automatically start. e) After the neutralization with K.F. reagent, it will give beep sound. f) Record the volume of required of K.F. reagent and that will display on the screen. g) Calculate the Factor by using formula Weight of water in mg K.F. Factor = -----------------------------------Volume use of K.F. in ml 8 Repeat the same operation for two more readings and report the results in the approved format.

Water equivalence factor is always in milligrams/ ml acceptable range = 5.5 to 7.5 (factor may change as per manufacturer

Weight of water in mg K.F. Factor = -----------------------------------Volume use of K.F. in ml

Procedure for Estimation: Ensure that the reservoir is filled with Karl Fischer Reagent. Add sufficient quantity of methanol (specially dried for KF) to the beaker (mounted on a magnetic stirrer) Switch on the instrument and adjust the speed and proper rotation of magnetic needle. Neutralize the methanol by adding KF reagent When end point is reached, add requisite quantity of the test sample (solid or liquid) to the beaker. Now, titrate the sample till end point is indicated by beep. Note down the end point. Calculate the Percentage of the moisture content as below. 1) DPTDTA (DI-PARA-TOLUOYL-D-TARTARIC ACID) Determination of factor Wt.of water T.R Factor Mean

Factor = Wt. of sample T.R Factor Moisture

Percentage of Moisture = Water Equivalent Factor x T.R. Weight of sample x 10 = = %

Result : % moisture in the sample is %

2.SAMPLE Wt. of sample T.R Factor Moisture

Percentage of Moisture = Water Equivalent Factor x T.R. Weight of sample x 10 = = %

Result : % of moisture in the sample = %

3.

SAMPLE T.R Factor Moisture

Wt. of sample

Percentage of Moisture = Water Equivalent Factor x T.R. Weight of sample x 10 = = %

Result : % moisture in the sample is %

Precautions: 1. Before assembling the parts ensure that, all parts are perfectly free from moisture. 2. During the moisture content determination, the sample should be added without touching the sides of the beaker or the electrode, dont spill drop of KF on the cabinet, it fails it should wiped off with soft clothe or moistened with acetone.

LOD

LOSS ON DRYING

Loss on drying
The classic laboratory method of measuring high level moisture in solid or semisolid materials is loss on drying (LOD). In this technique a sample of material is weighed, heated in an oven for an appropriate period, cooled in the dry atmosphere of a desiccator, and then reweighed. If the volatile content of the solid is primarily water, the LOD technique gives a good measure of moisture content. Procedure: Weigh accurately dry and cleaned glass stopper shallow weighing dish of weight (w1) .Then add accurate weight of sample and reweight dish with sample (w2). Distribute the sample evenly in weighing dish by sidewise shaping. Place to bottle & uncovered at given temperature for given hours under vacuum. After specific hrs take out the dish from the oven and immediately cover if with stopper & place it in a desicatar to get it cooled to Room Temperature. weight the dish along with sample & lid, Record the weight (w3). Calculate LOD by formula W2-W3 % LOD = ---------- X 100 W2-W1

Determination of Assay (Purity)

Sample Name : STRUCTURE

Molecular weight Molecular formula IUPAC Name Appearance Assay by titration Aim :. Theory : Standard Operating Procedure for determination of Assay Observation 1) Solution in burette : 2) Solution in conical flask : 3) Indicator : 4) End point : For Lot 1) (500 kg) Reading Initial Final Difference

I cm

C.B.R.

For Lot2 Reading Initial Final Difference Calculation For Lot 1)

I cm

C.B.R. cm

Percentage purity = Burette Reading Normality Wt. of sample = = %

For Lot 2) Percentage purity = Burette Reading Normality Wt. of sample

= Result : Passes Analysis Aim :

Reagents : Procedure to find out purity

Observation .Weight of sample taken = g 1)Solution in burette 2)Solutioninconicalflask 3)Indicator 4)End point Reading Initial Final Difference Calculation Percentage purity = Burette Reading Normality 63.54 Wt. of sample10 = : : : I cm 0.0 C.B.R. cm

()

Result : The purity of given Cu powder is %

LOSS ON DRYING

Loss on drying
The classic laboratory method of measuring high level moisture in solid or semisolid materials is loss on drying (LOD). In this technique a sample of material is weighed, heated in an oven for an appropriate period, cooled in the dry atmosphere of a desiccator, and then reweighed. If the volatile content of the solid is primarily water, the LOD technique gives a good measure of moisture content. Procedure: Weigh accurately dry and cleaned glass stopper shallow weighing dish of weight (w1) .Then add accurate weight of sample and reweight dish with sample (w2). Distribute the sample evenly in weighing dish by sidewise shaping. Place to bottle & uncovered at given temperature for given hours under vacuum. After specific hrs take out the dish from the oven and immediately cover if with stopper & place it in a desicatar to get it cooled to Room Temperature. weight the dish along with sample & lid, Record the weight (w3). Calculate LOD by formula W2-W3 % LOD = ---------- X 100 W2-W1 WEIGHT CHEMISTRY SAMPLE:1

SAMPLE NAME:-

DITHRANOL IP Anthralin; Dioxyanthranol

C14H10O3

Mol.Wt.226.23 Dithranol is 1,8-dihydroxyanthrone

TEST:- ASSAY DESCRIPTION:- Yellow coloured powder. METHOD :-

Weight accurately about 0.2 gm of substance.And dissolve in 50ml of anhydrous pyridine.Titrate with 0.1 M tetra butyl ammonium hydroxide.Determine the end point potentiometrically carry out the blank titration.1.0ml of TBAH equivalent To 0.02262 gm of Dithronol. Sample weight :- 0.2022gm Factor :0.02262gm

Actual molarity:- 0.1 M TBAH is 0.10065 M Titration Teading:- 8.8ml Calculation:8.8 X 0.02262 X 0.10065 X 100

99.08% AS IT IS

LOSS ON DRYING:Weight of empty :Weight of dish +Sample :Weight of sample :29.3436gm 30.3462gm 1.0026gm

After drying at 105degree :Weight of dish + Sample :Loss in Weight :30.3434gm 0.0028gm

CALCULATION :Loss in weight Sample taken 0.0028 1.0026 =0.28% ASSY ON DRIED BASIS:99.08 X 100 (100-0.28) X 100 X 100 X 100

=99.36% CONCLUSION:- Dthranol contain not less than 98.5% and not more than 101.0% on dried basis.And by the analysis got 99.36%.

SAMPLE :1

Sample name :

Molecular Formula: Molecular Weight:

Weight of LOD dish (W1) = g Weight of dish + sample (W2) =g ( Before drying) Weight of sample = g Weight of dish + sample (W3)=g ( After drying ) LOD =g W2-W3 % LOD = ---------- X 100 W2-W1 = =%

Result =Passes

Sample name :

Molecular formula Molecular weight

Weight of LOD dish (W1) = g Weight of dish + sample (W2) =g ( Before drying) Weight of sample = g Weight of dish + sample (W3)= g ( After drying ) LOD =g W2-W3 % LOD = ---------- X 100 W2-W1 = =%

Result =Passes