STUDY OF CLASSICAL AND INSTRUMENTAL METHODS USED IN ANALYSIS AT QUALITY CONTROL DEPARTMENT OF EXCEL INDUSTRIES, LOTE-PARASHURAM

STUDY OF INSTRUMENTAL AND CLASSICAL METHODS USED IN EXCEL INDUSTRIES PVT LTD. A Project Submitted To University of Mumbai As a partial fulfillment for the award of the Degree of

MASTER OF SCIENCE
(Organic Chemistry) By Mr. LISHANT PRADEEP LAD With the complements from EXCEL INDUSTRIES PVT LTD.

Department of Chemistry Dapoli Urban Bank Senior Science College, Dapoli.

2011-2012

CERTIFICATE

Certified that Mr. Lad Lishant Pradeep of M.Sc. part II has satisfactorily completed Industrial Training Programme at Excel Industries Ltd, Lote, during the period 6Th July 2011 to 20Th July 2011. This dissertation represents his bonafide work.

Date: Place: Head of Department

DECLARATION

I undersigned hereby declare that the training programme arranged at Excel Industries Ltd, Lote, Dist Ratnagiri is completed and written for the purpose fulfillment of Master of science degree in organic chemistry in academic year 2011-2012. I further declare that dissertation has not previously been formed the basis for the award of any degree, diploma or fellowship or similar other titles and this is an independent work done by me.

Date: Place: Dapoli Miss. Luhar Surekha Bhimraj M.Sc. Part- II

ACKNOWLEDGEMENT

I am grateful to each and every one who have quite and helped encouragement me directly or in directly for my success in this project. Report successfully. I am sincerely thanks to the staff & management of excel Industries for co-operation. I would like to thanks Mr. S. Y. Patankar (Manager & HRD) For giving the kind Permission of doing this Project in their reputed company. I am thankful to Mr. Khadilkar & Mr. Bajage who gave us valuable information about safety department. In QA/QC Department I owe my sincere and deep gratitude & to my resp. Sir Mr. S. A. Jadhav (HOD of QA/QC) Mr. Phuke, Mr. Joshi, Mr. Pendse and Mr. Desai They all helped me for QA Analysis. I also thankful to R & D Department I am thankful to Mr. Ketan Pendse & Mr. Aniket Barve who gave me very good information about Research & Development. I am also extremely thankful to my teachers Assistant Prof. Marathe & Assistant Prof. Gandhi for their valuable guidance, suggestions & co-operation in completion of my project work.

Index
Sr.No
1. 2. 3. 4. Introduction Company Profile Safety department Preparation of standard solution i] Ammonium thiocyanate solution Ii] Sodium hydroxide Solution Iii] Sodium thiosulphate solution Iv]Hydrochloric acid solution V]Sulphuric acid Solution 5. Raw material analysis i] Analysis of sodiumpenta chlorophenate Ii] Determination of free alkalinity Iii] Determination of O.P.A. [ orthophosphoric acid ] Iv] Text method for analysis of formaldehyde V] Text method for analysis of sodium carbonate Vi] Text method for analysis of caustic soda. Vii] Text method for analysis of acetic acid. 6. Instrumental method of analysis i]Karl Fischer Apparatus ii]Gas chromatography iii] High Performance Liquid chromatography. 7. 8. 9. R & D Department Limitation Conclusion

Title

Page No.

INTRODUCTION
The quality control department controlling the quality at each and every step of the manufacturing process of products i.e. right from raw materials to finished products. The quality control department has some objectives and responsibilities.

OBJECTIVES
1. T o establish standards of quality which are acceptable to customers. 2. To locate and identify faults and defects during the manufacturing processes. 3. To keep up quality of products during the manufacturing by taking corrective actions.

RESPONSIBILITIES
1. To prepare plant control program for the management. 2. To advice production department regarding uniformity of materials, quality aspects and specifications. 3. To assist purchasing department in the evaluation of suppliers and to minimize defects of incoming raw material. 4. To analyze customer complaints & to find solutions of those solutions. In all these aspects, mentioned above, the quality control department plays a vital role. It also maintains the quality desired and fulfils customers expectations. Therefore it is very interesting and important to study various analyses carried out in the Q.C. department, by both the methods i.e. classical and advanced methods like chromatography. It is also very necessary for an analytical chemist, to understand how the Q.C. department makes use of classical and instrumental methods to achieve and maintain the desired quality.

Company Profile
Excel Industries Ltd. Is a large scale manufacturer of agrochemicals, like pesticides, fungicides etc. The company is located at Lote- Parshuram industrial area in Ratnagiri district.

The organization is certified with ISO 9001, ISO 14001 ISO -18001

and

The Company was established by Champabhai Shorff in 1941 at Jogeshwari, Mumbai with the initial investment of Rs. 10,000. The Company has shown a tremendous growth & in the year 2008 the turnover of the company was 571 crores which indicates the Progress of the company and it has not stopped yet. Following are the directors of company with their respective duration.

Director 1. 2. 3. 4. C. C. Shroff G. C. Shroff K. C. Shroff A. C. Shroff

Duration 1942-1965 1966-1986 1987-1996 1997 onwards.

The company has now grown up with different plants all over India. Following are the different plants of company with their respective establishment years. Year 1941 1967 1971 1975 1983 1989 2003 Site Location Jogeshwari Amboli Bhavnagar Roha Lote (Khed) Silvasa Gajode

Company manufactures different products having different applications. Following are the companys products with their market names & applications.

Product S.P.C.P. (Sodium 1. Pentachloro Phenate) H.E.D.P. (Hexa Hthylene 2. Diphosphoric Acid.)

Market Name

Application

Biocel (SG) (Granules) Antifungal Biocel SP (Powder) generally used in Biocel SF (Flakes) paint industries. Biocel -90 As a complexing agent and it removes hardness & of Codex water at industrial level Acetyl chloride Raw material for pharmaceutical Purposes. 5. 4 To prepare 4 polymers which 4 are used in CDS.
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3. Acetyl chloride Tri Hydroxy accetophenone

THP

SAFETY DEPARTMENT Safety is the state of being "safe" (from French sauf), the condition of being protected against physical, social, spiritual, financial, political, emotional, occupational, psychological, educational or other types or consequences of failure, damage, error, accidents, harm or any other event which could be considered non-desirable. Safety can also be defined to be the control of recognized hazards to achieve an acceptable level of risk. This can take the form of being protected from the event or from exposure to something that causes health or economical losses. It can include protection of people or of possessionsNatural accidents Manmade accident They are mainly due to Unsafe Act Unsafe Condition Hazardous chemicals used in factory Examples 1. 2. 3. 4. 5. 6. Corrosive Poisonous Flammable Irritating Oxidizing chemicals Compressed gases Types of chemicals Acids and Alkali Cynide Methyl chloride Benzy 1 chloride , C12 , SO2

Thus for handling such dangerous chemicals proper precautions and use of safety equipments is must. To face such as an insanitary condition, company has set up alarm System which is daily tested. 1st Alarm Indicates to turn in safety mode. 2th Alarm - Informs the way to exit with the help of wind meter.

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Safety equipments used inside the factory Non-respiratory equipments Rubber Hand gloves PNC Hand gloves Leather Hand gloves Safety Belt Helmet Safety shower and eye washer

Respiratory equipments Dust mask Breathing air mask Pressure suit

1) To arouse & maintain the interest of managers/officers/ heads of the department/supervisors & operating personnel in the matters relating to safety of Man, Materials, Equipments, machines & instruments, Environment, society. 2) To create understanding regarding interrelatedness of safety purpose. 3) To create participative technique whereby safer design safer operation & safer partials are evolved. 4) To investigate into the past accidents & dangerous occurrence so that they do not recur. 5) To create safe working particles &strive towards commutability of understanding. 6) To keep our neighborhood trained in the in the matters relating to first aid, fire fighting, & rescue operation.

Safety:Positive organized activity or programmed based on knowdge of the reaction between man & his working environment which aids business enterprises by minimizing human, economic& sociological looses caused by injuries health impairment, fires, explosion & other occupational accidents.

First Aid:IS-11163 :-first aid & dressings. IS-4015 :-Pesticide poisoning.

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IS-2190, 2217, 884:- fire fighting. ` Aims:1) To preserve life. 2) To promote recovery 3) To prevent worsening of the causalities condition. 4) To arrange transportation to hospital.

PPEs:As the barriers between the hazards & workers which help to eliminates on injury or reduce its severity.

Types of PPEs:1) Non-respiratory PPEs. 2) Respiratory PPEs.

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Preparation of Standard Solutions

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Standard solution

A standard solution is a solution containing a precisely known concentration of an element or a substance i.e., a known weight of solute is dissolved to make a specific volume.

Uses: A solution of acid can be standardized by titrating it against a solution of alkali of known concentration. Once this has been calculated, it can in turn be used as a standard solution to find the concentration of a solution of alkali. Standard solutions are also commonly used to determine the concentration of an analyte species. By comparing the absorbance of the sample solution at a specific wavelength to a series of standard solutions at differing known concentrations of the analyte species, the concentration of the sample solution can be found via Beer's Law. Any form of spectroscopy can be used in this way so long as the analyte species has substantial absorbance in the spectra. The standard solution is a reference guide to discover the molarity of unknown species.

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1. AMMONIUM THIOCYANATESOLUTION
Aim: Preparation of standard ammonium thiocyanate solution. Requirements: A.R. grade ammonium thiocyanate solution dist. Water, A.R. grade silver nitrate, Cone. , nitrobenzene, Ferric alum indicator. Procedure: A) Preparation of ammonium thiocyante solution Weigh about 17 gms of A.R. grade ammonium thiocyanate in a beaker Add about 500 ml distilled water and stir well with glass rod to dissolve it. Transfer the solution in 2 liter volumetric flask and dilute upto mark with dist. water. Shake well to make it homogeneous. This will give 0.1 N (approx) Ammonium thiocyanate solutions. Standardize it against A.R. grade silver nitrate. Standardisation of ammonium thiocyanate solution. Weigh accurately about 0.38 g of AR grade solution in conical flask. Add about 75 ml of dist. Water & 10 ml of cone in it. Also add 2 ml of nitrobenzene.

B)

Then titrate it against prepared ammonium thiocyanate solution using Ferric alum as an indicator. End point will be milky white to reddish brown. Note down the C.B.R. CALCULATION: Normality of ammonim thiocyanate solution = = = 0.1137 N
Normality of ammonium thiocyanate thiocyanate solution = 0.1137 N
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2. SODIUM HYDROXIDE SOLUTION Aim : Preparation of 0.1 N NaOH solutions. Requirements: A.R. grade Sodium hydroxide, dist. water, KHP. (Potassium hydrogen phthalate), Phenolphthalein indicator. Procedure : a) Preparation of Sodium hydroxide solution (0.1 N)

Weigh about 8 gms of A.R. grade Sodium hydroxide flakes in a beaker and add about 500 ml. distilled water and stir well with glass rod to dissolve it. Allow it to cool upto room temperature. Transfer the solution in a 2 liter volumetric flask and dilute it upto the mark with dist. Water shake well to make it homogeneous. This will give a Solution of approx 0.1 N NaOH standardize it against potassium hydrogen phthalate. b) Standardisation of sodium hydroxide solution

Weigh accurately about 0.40 gm of A.R. grade KHP (Previously dried at 100 -120 C for one hour and cooled in desiccator) in a conical Flask. Add about 75 ml of dist. Water and shake well to dissolve it. Titrate it against prepared sodium hydroxide solution using phenolphthalein as an indicator .End point will be colourless to pink. Note down C.B.R.
CALCULATION:

Normality of NaOH solution = = = 1.023 N Normality of NaOH solution = 1.023 N

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3.SODIUM THISULPHATE SOLUTION

Aim: Preparation of 0.1 N. Sodium thiosulphate solution, dist. Water A.R. grade potassium dichromate, Sodium hydrogen Carbonate, Potassium Iodide, starch indicator Conc . HCL Procedure : a) Preparation of 0.1 N Sodium thiosulphate solution Weigh about 50 g of A.R. grade sodium thiosulphate in a beaker. Add 500 ml of distilled water and stir well with glass rod to dissolve it . Transfer the solution in 2 liter volumetric flask and dilute it upto mark with dist. Water. Shake well to make it homogeneous. This will give a solution of approx 0.1 N Sodium thiosulphate solution. Standardise it against AR grade potassium dichromate. b) Standardisation of sodium thiosulphate solution Weigh accurately about 0.11 g of A.R. grade potatssium dichromate (Previously dried at 130-150 for one hour and cooled in dessicator ) in 150 ml iodometric flask. Add about 75 ml of dist. Water. Then add 2 g of sodium hydrogen carbonate and about 3 g of potassium iodine shake well the flask to dissolve it. Then add 6 ml conc. HCL slowly by rotating the flask. Stopper the flask & allow it to stand for 5 minutes in a dark. Titrate it against prepared sodium thiosulphate solution. When solution becomes pale yellow add starch indicator End point will be greenish blue. Note down the C.B.R. (in ml)

CALCULATION:

Normality of Sodium thiosulphate solution ( = = = 1.019 N

)=

Normality of Sodium thiosulphate solution = 1.019 N.

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4. HYDROCHLORIC ACID SOLUTION

Aim : Prepartion of 0.5 N HCL solution. Requirement: Conc.HCL, dist. Water , A.R. grade sodium carbonate, Methyl orange indicator. Procedure: a) 8 Preparation of0.5 N HCL solution Take about 500 ml o dist water in a beaker and add 90 ml conc. HCL slowly with constant stirring. Allow it to cool to room temperature. Transfer the Solution in a 2 liter volumetric flask and dilute it up to the mark with dist. Water. Shake well to make it homogeneous. This will give a solution of approx 0.5 N HCL. Standardize the Solution using A. R. grade sodium carbonate. Standardisation of HCL solution Weight accurately about 0.6 gram of A.R. grade solution sodium carbonate (previously dried at 250-2700 c for 1hr and cooled in dessicator) in a conical flask. Add about 75 ml of dist. Water and shake well to dissolve it This will against prepared HCL solution using methyl orange as an indicator .End point will be red. Note down C.R.B. CALCULATION : Normality of HCL solution = = =0.507 N

b)

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5. SULPHURIC ACID SOLUTION

Aim: Preparation of 0.1 N Requirements: Conc. Procedure: a) Preparation of sulphuric acid solution (0.1 N) Take about 500 ml of dist water in a beaker and add 6 ml of conc. slowly with constant stirring. Allow it to cool upto room temperature. This will give a solution of approx 0.1 N Solution. Titrate it against standard NaOH solution (0.1N) b) Standardization of Sulphuric acid solution Take 25 ml std. NaOH solution by pipette in a conical flask. Titrate it against prepared sulphuric acid solution using phenolphthalein indicator. End point will be pink to colourless. Note down C.B.R. Std. NaO solution. Phenolphthalein indicator. solution

CALCULATION: = (Std. NaOH) 0.1 Normality of Normality of Soln , ( 24.9 =

n)

solution = 0.1004 N

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Raw Material Analysis

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1. ANALYSIS OF SODIUMPENTA-CHLOROPHENATE
Aim : To determine the chlorophenate content of given sample. Principle:
Acidimetric titration of aqueous solution of sodium pentachlorophenate using bromophenol blue indicator.

Reagents : 1) 2) 3) 4) Procedure: Weigh accurately about 1 g of sample in 250 ml conical flask. Add 50 ml of water & 2/3 drops of phenolphthalein indicator & titrate it with std. HCL to Remove of free alkanity take this reading as A. Then add 30 ml of diethylether & then titrate it with std. HCL using Bromophenol as an indicator. Endpoint will be blue to greenish. Take this reading as B. Std. HCL 0.5 N Phenolphthalein indicator Bromophenol blue indicator Diethylether

CALCULATION: A = 8.7 ml, B = 14.4 ml Chlorophenates as Where N = Normality of HCL M = Mass in gm of material taken. =
( )

ONa =

= 81.17
(Sodium pentachlorophenate % should be max of 85% , as per Q.C. Department )

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2. DETERMINATION OF FREE ALKALINITY

Aim: To determine free alkalinity of the given sample. Principle: Acidimetric titration of aqueous solution of sodium pentachlorophenate using thynolphthalein indicator. Reagents: Weigh accurately about 1 g of sample in 250 ml conical flask and add 75ml of water. Shake well to dissolve it and then add thymolpthalein indicator 2-3 Drops of titrate with std. solution. End point will be blue to colourless. CALCULATION: Free alkalinity = Where V = Volume in ml of Std. N = Normality of M = Mass in gms of material taken = = 0.156 % (Free alkalinity should be between 0.1 0.25 % as per specifications of Q.C. dept. ) required for titration

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3. DETERMINATION OF ORTHOPHOSPHORIC ACID (O.P.A)

Aim: Determination of orthophosphoric acid (O.P.A) impurity. Reagents: 1) 2) 3) Procedure: Weigh accurately 1 gm of sample in a flask. Containing about 50 ml of distilled water. Add 10 gm of sodium chloride in it. Mix well to dissolve it. Titrate the Solution with 0.1 N NaOH Solution using phenolphthalein as an indicator. End point will be colourless to pink. Sodium choloride. Standard sodium hydroxide solution of 0.1 N . Phenolphthalein indicator.

CALCULATION: O.P.A. content = Where V = Volume in ml of Std. NaOH solution required for titration. N = Normality of Std. NaOH solution. M = Mass in gm of material taken for test. = = 2.009% (O.P.A. content should be 1.5 2.5 % per specifications of Q.C. department.)

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4.TEST METHOD FOR ANALYSIS OF FORMALDEHYDE Subject: Test method for analysis of formaldehyde Determination of assay Reagents: 1) 2) 3) 4) Thymolphthalein 0.1 N NaOH solution Ammonium sulphate solution 0.5 N HCL

Procedure: 1) Weigh accurately 1 gm of sample and dissolve it in 50 ml water, in a conical flask. Add 2-3 drops of thymolphthalein indicator & 0.1 N NaOH solution till solution turns faints blue. Weigh 2 gms of ammonium sulphate, add to it thymolphthalein indicator and NaOH solution till blue solution turns colourless. Add ammonium sulphate solution to the first & then titrate with 0.5 N HCL.

2) 3)

CALCULATION: Purity of formaldehyde = = = 71.59 % (Formaldehyde purity should be above 70% as per Q.C. dept.)

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5.TEST METHOD FOR ANALYSIS OF SODIUM CARBONATE Subject: Test method for analysis of sodium carbonate. Determination of assay Principle: Acid base titration + HCL Reagents: 1) 2) 3) Standard hydrochloric acid solution of 0.5 N Methyl orange indicator Sample of sodium carbonate. 2 NaCL + O+

Procedure: Dry the sodium carbonate sample in oven for 1 hr. at 260 C. Weigh accurately about 1 g of dried sample in conical flask. Dissolve it in 75 ml of distilled water. Titrate with std. hydrochloric acid solution using methyl orange as an indicator. End point will be yellow to orange red. CALCULATION: Sodium carbonate purity i.e. %=

Where, V = Volume in ml of Std. HCL required for titration N = Normality of Std. HCL solution M= Mass in gms of material taken % = = 98.41 %
(Sodium carbonate purity should be very high i.e. above 98% as per Q.C. dept.)
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6.Methods for the analysis of caustic soda.

Subject: Test method for analysis of Caustic soda Determination of assay Principle: Acid base titration. NaOH + HCL Reagents : 1) 2) Standard hydrochloric acid solution Phenolphthalein indicator NaCL + O

Procedure: Weigh accurately 0.5 g of NaOH on 100 % basis in a conical flask containing 75 ml of dist water stir well to dissolve it. Titrate it with Std. HCL Solution using phenolphthalein indicator. End point will be pink to colourless. CALCULATION: Sodium hydroxide % = Where, V= Volume in ml of std. HCL required for titration N= Normality of Std hydrochloric acid. M = Mass in gms of material taken. Purity of NaOH i.e. NaOH % = = 48.73 % (Purity of NaOH should be around 49% as per Q.C. dept. )

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7. Methods for the analysis of Acetic Acid .

Subject: Test method for analysis of acetic acid. Part I: Determination if assay [purity] Principle: Acid base titration. CH3COOH + NaOH

CH3COONa + H2O

Reagents:1) Standard NaOH solution of 0.1 N. 2) Phenolphthalein indicator. Procedure: Weigh accurately 1.0 g of sample in 250 ml conical flask Add about 75 ml of dist. Water. Titrate it with std. solution of NaOH using phenolphthalein indicator. End point will be colourless to pink. CALCULATION: Acetic acid % = Where, V = Volume in ml of Std. NaOH solution required for titration N = Normality of NaOH solution M = Mass in gms of material taken F = 6 (acetic acid factor) Acetic acid % = = 99.79 % (Acetic acid purity should be very high i.e. above 99% as per Q.C. dept.)

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Instrumental Method of Analysis

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Karl Fisher instrument.

Determination of moisture percentage of the sample using Karl Fisher apparatus

Reagents: 1] Karl Fisher reagent 2] Menthol 3] Distilled water 4] Commercial grade menthol for cleaning the dispensing system Procedure: 1] Add about 150 ml of K/F regent in dry reservoir bottle fitted with the adaptor. 2] Add K/F grade menthol to the reaction breaker are fully immersed. 3] Switch on the instrument the 1st reading obtained shows the volume of K/F reagent required for removal of moisture from methanol in beaker. Confirm the end point 2-3 times pressing start key to neutralize the traces of moisture in beaker. Determination of titre factor: Confirm the end point by pressing START key and wait for audio signal. In the mean time take reweighed water ready for addition. When buzzer stop, add accurately weighed water into the methanol. Press TART key. The digital display show message inject for 10 second display will show volume of K/f
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reagent being added. As before the operating will stop on reaching the end point the green lamp will go OFF and red lamp will become ON with audio signal.
Read the display and note it down. This reading indicates the volume of reagent required to react with neutrals the reweighed water. Repeat the procedure 2 to 3 times and find out overage consistent results.

Titre Factor

Liquid samples are usually injected by graduated micro syringe through a self sealing rubber septum. A solid sample can be dissolved in suitable solvent and then injected with the help of a micro syringe. The advantage of the Karl Fischer method
The capability to accurately measure small amounts of moisture. The volumetric method is preferred when moisture levels get high, when complex solvents are needed to release bound water in the sample or when other complex chemical reactions affect the combined reagent/solvents use in the econometric methods

Testing sample:
Keep the weighed sample ready for introducing into titration beaker and repeat procedure as above. The reading appearing on digital display shows volume of K/F reagent required to neutralize the moisture in sample. CACULATION: Moisture content % == Where, V = Volume in mi of std. K/F reagent required to neutralize the moisture present in sample.

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1] Moistures content of Biocel -90 = 1) Titre Factor 2) Titre Factor Mean = = = =7.95 =8.04 =7.995

moisture content % = 1) moistures content % = =0.9667 Moisture content of Biocel -90 = 0.9667 %

2] Moistures content of codex calculated

1) Titre Factor 1) Titre Factor Mean of Titre Factor Mean of Titre Factor Moisture content % = = = = = = =0.9665 Moisture content in Codex= 0.9665 % =7.16 =7.26 =7.21 =7.21

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3] Moisture content of acetyl chloride as calculated 1) Titre Factor = = 2) Titre Factor Mean of Titre Factor Moisture content % = = = =1.20 % Moisture content of acetyl chloride =1.20 % = =8.65 =8.61 =8.63

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Chromatography
Chromatography is the most accepted separation tool in modern analytic Laboratories. The acceptance of Chromatography as an analytical tool for separation, identification and quantization of mixtures is due to simplicity of the equipment required and the ease of interpretation of the data produced. This has been due to rapid technological advancement over the past 25 years. There are several types of Chromatography technique depending upon the Nature of the two phases and the mechanism of distribution of the substance between them. The Chromatography methods are basically developed to separate complex mixture of the component which is otherwise difficult to separate by existing chemical technique. The Chromatography technique has proved to be a very important tool in the separation and quantitative analysis gasses and organic compounds including pesticides and pollutants. Gas Chromatography: Gas Chromatography is a physical method of separation accomplished by Distribution of sample solution in vaporized phase between moving phase and stationary phase. When stationary phase is solid adsorbent then technique gas solid Chromatography (GSC) when stationary phase is nonvolatile liquid held in column the technique is called Gas liquid Chromatography. This technique is widely used than the former. GSC has limited application because of sampler anent retention of active or polar molecules and severe tailing of elution peats. (a consequence of the non-linear character of adsorption process.) Thus, this technique has not found application except for the separation of certain lowmolecular weight gaseous species.

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Principles of gas Chromatography: Gas liquid Chromatography (GLC) or Gas Chromatography (GC) is a separation of a mixture in microgram quantity by passage of vaporized sample in gas stream (mobile phase) through a column containing fixed liquid coated on inert solid phase (stationary phase). During the process, the components of vaporized sample are fractionated as a consequence of being partitioned between mobile gaseous phase and stationary phase in the column.

Instrument for Gas Liquid Chromatography

1. 2. 3. 4. 5. 6. 7. 8. 1. A gas cylinder a carrier gas Pressure regulator and flow control. Sample injection system. The column. The thermal compartment. The detection system. The recorder. Amplifier.

A gas cylinder containing a carrier gas: The most commonly used carrier gasses are helium, hydrogen, nitrogen, organ, and CO2. Chaise of carrier gas depends uponi) Nature of the sample ii)The type of detector being employed. iii) Column efficiency. Generally nitrogen is used as a carrier gas because ofi) High thermal conductivity. ii) Not reactive with analyte. iii) Not explosive. iv)Not expensive.

Hydrogen is explosive and reactive with sample components hence it is not used.Carrier gas function is to transport the analytic through column.
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2.

Pressure regulator and flow control : It is attached to the Carrier gas cylinder whose function is to regulate the pressure and flow rate respective to desired level. 3. Sample injection system : The function of sample injection system is to vaporize the sample instantaneously because the injectors temperature is maintained above the boiling component in the sample. Liquid samples are usually injected by graduated micro syringe through self sealing rubber septum. A solid sample can be dissolved in suitable solvent and then injected with the help of micro syringe. Gaseous samples can be introduced into the column by means of special gas sampling valves. 4) The column: The columns used in GC are of two types. i) ii) Packed column. Open tabular (or capillary) column.

Packed column are made from glass or metal tubing. The length is typically 2 to 3 m long and diameter of 2-4 mm, column is in the form of coils. Capillary columns which are made up of glass fused silica, of length 25 to 50 inside diameters of 0.25-0.50 mm. In Gas solid chromatography the column are packed with solid stationary phase such as activated charcoal, silica gel and alumina. In gas liquid chromatography (GLC) the stationary phase is coated on same inert support material such as diatomaceous earth or glass of uniform size. The most widely used liquid stationary phases for both packed & open tabular column in G.C. with their maximum temperature limit are polydimethyl siloxane (623K), 50% phenyl polydimethyl siloxane (523K).

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5. The thermal compartment: The coiled column is ordinarily housed in a thermo stated oven. The optimum temperature depends upon the points of the sample components. A temperature is roughly equal to slightly above point of the sample. For samples with a broad boiling rang, it may be necessary to employ temperature programming, whereby the column temperature is increased either continuously or in step ass the separation proceeds.

6. The detection system:

The function of a detector is to sense the components in the gas by providing an electrical signal. The mainly used detectors in the G.C. in Excel industries are TCD & FID.

a) TCD: (Thermal conductive Detector)

It has one reference side and one sampling side. The carrier gas is passed through the reference side and comes out of the sampling side with the sample component. A change in composition in gas flowing through the sensing cell cause a heat loss and therefore a change in resistance with can be recorded. b) FID: (Flame ionization Detector) FID (Flame ionization Detector) is based on the fact that most organic compounds, when paralyzed in hot flame produce ionic intermediates that conduct electricity through the flame. The current flowing through the circuit is proportional to no. of ions striking the collector, is proportional to the concentration of cognizable sample components that enter the flame. FID response only to substance that can be ionized in an air hydrogen flame. It does not respond to inorganic compound (N2, O2, CO2, H2O) FID is sensitive than TCD, but TCD response to both organic and inorganic spices.

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Quantitative analysis: In excel industries, a variety of samples containing organic compounds as well as finished good are quantitatively analyzed by gas chromatography. There are three methods of quantitative analysis which are: 1] 2] External standard method Internal standard method

3] Area normalization method (this is the most widely used method in Excel industries)

1] External standard method:

The method includes comparative study of standard solution of analyst having known concentration with solution of sample containing analytic having known amount of sample diluted to some definite volume. Chromatogrames are obtained of the both solution. On the basis of peak area of the analyte component the percentage purity of the analyte can be found out. Here the sample matrix is kept identical to for both the solutions. Also for particular analysis the method of chromatography i.e. conditions, such as selection of column, temperature, pressure, sample conc. is developed such that there is a good resolution of analyte and other components present in sample. 2] Internal standard method: The highest precision for quantitative Chromatography is obtained by use of internal standards because the uncertainties introduced by sample injection are avoided. In this procedure, a carefully measured quantity of a standard substance is introduced into each standard and sample, and the ratio of analyte to internal standard peak areas serves as analyte parameter. For this method to be successful, it is necessary that internal standard peak be well separated from peaks of all other components of the sample the standard peak should, on the other hand, appear close to the analyte peak. With a suitable internal standard, precisions of better than 1% relative can usually be achieved.

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3] Area normalization method: In this method complete elution of all components of the sample is required. In the areas of eluted peaks are computed; after correcting these areas for differences in the detector response to different compound types, the concentration of the analyte is round from the ratio of its area to the total area of all peaks.

Standardization of Gas chromatograph

Activities for calibration of Gas Chromatography 1] Prepare fresh, standard calibration mixture of 5 % benzene in toluene. 2] Clean flame ionization detector (FID) the roughly. 3] Connect 5 % SE 30 ss column to GC which is to be calibrated. 4] Measure nitrogen (N2) hydrogen (H2) and air flow rates and set 30 ml/min, 30ml/min and 300 ml/min respectively using flow meter. 5] Set GC condition as given below. Oven temperature Injector temperature Detector temperature Attenuation Range 6] 7] 256 1000

What to achieve all temperature and stability of GLC. Inject 10 micro liter calibration of 5% benzene in Toluene and take results. Measure heights of benzene and toluene.

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High performance Liquid Chromatography

The Q.C. lab in excel department is equipped with Schimatzu high performance liquid chromatography. Various samples are analyzed using HPLC. Also samples from Research and Development (R & D) department are analyzed here. Instrument is frequently calibrated by expert chemists (Specially assigned) & also it is maintained in a good condition for its longer life & better performance. Scope of HPLC High performance liquid chromatography is the most widely used out of all the analytical separation techniques, with annual sales of HPLC equipment approaching the billion- mark. the reasons for popularity of the method is its sensitivity, its ready adaptability to accurate quantitative determinations, its suitability for separating nonvolatile species or thermally fragile ones, and above all, its widespread applicability to substances that are of prime interest to industry, to many fields of science and to the public. Examples of such Materials include: amino acids, hydrocarbons, carbohydrates, drugs, terpenoids, pesticides, antibiotics, steroids, metal organic species and a variety of inorganic substances. Principle: In this method sample is introduced into column stationary liquid phase by means of pressurized flow of liquid mobile phase. The components in the sample are fractioned during their passage through the column. The detector system senses these components as they are eluted from column and generates a signal proportional to amount of solutes passing through the system.

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The important components of HPLC 1. Mobile phase reservoirs & solvent treatment systems. A modern HPLC apparatus is equipped with one or more glass or stainless steel reservoirs, each of which contains 200-1000 ml of a solvent. The reservoirs are often equipped with a means of removing dissolved gases usually oxygen and nitrogen that Vacuums pumping system, devices for heating and stirring the solvents or systems for sparging in which the dissolved gases are swept out of the solution by fine bubbles of on inert gas of low solubility. A separation that employs a single solvent of composition is termed as isocratic elution. Frequently sedation efficiency is greatly enhanced by greatly elution. Here two or three solvent systems that differ significantly in polarity are employed. 2] High Pressure Pump: Pumps are designed to operate at a constant pressure or at constant flow. The requirement for an HPLC pumping system are: 1] The generation of pressure of up to 6000 psi. 2] Pulse free output 3] Flow rates ranging from 01. to10 ml/min. 4] Flow control & flow reproducibility of 0.5 % relative or better. 5] Corrosion resistant component. 3] Precolumn: The purpose of precolumn is to remove impurities from solvent , thus it prevents contamination of analytical column. 4] Sample injection system: The sample is injected in to a sample loop by means of hypodermic syringe through self sealing septum of silicon or teflon. Sample injection system takes only the required injection system takes only the required quantity & remaining is rejected.

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5] Column 7 column packing: The column are ordinary constructed from smooth bore stainless tabing although heavy walled glass tubing is occasionally uncounted .The latter is restricted to pressure that are lower than 600 psi. Variety of packed columns differing in size & packing are available from several manufactures .Their costs range from Rs 10000 25000. The majority of liquid chromatographic columns range in length from 1030 cm normally the columns are straight, with added length, where needed, being gained by coupling two or more columns together. the inside diameter of column is 4 to 10 mm. the most common particle size of packing is 5 10 mm. Column packing material: i ] Porous beads : Porous solid adsorbents like silica gel and alumina are used for sepration of compounds with low molecular and low polarity. ii] Pellicular beads: These are made up of glass beads coated with thin layers of porous material like slices gel, alumina or iron exchanges resin.

6] Detectors: The most common detector for HPLC is UV detector which is based upon absorption of ultraviolet or visible radiation. They have a wide range .they can be used for gradient elution. The most powerful UV spectrophotometric detectors are diode array instruments which permit collection of data for on entire spectrum in about one second. 7] Quantitative work: The quantitative work of HPLC is also done by area normalization method as in case of GC analysis.
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REASERCH & DEVELOPMENT DEPARTMENT

The R&D department consists of three main units. There aims & objectives are as below:1) Micro lab Aim: - Synthesis or development of specified product with great yield of purity. Objectives:1) Set up the process for synthesis or development of product. 2) Obtain desired purity & yield. 3) Work on micro scale up to 100gm.

2) Kilo lab:Aim: - To obtain product of kilo level with purity & yield. Objectives:1) Obtain desired product at kilo level up to 10 kg to 50 kg. 2) Set up process for product obtained from micro lab.

3) Pilot plant:Aim: - To obtain desired product on large scale & set process which is finalized for production plant. Objectives:1) Set up SOP for desired product with finalized level. 2) Set up proportion, time& conditions which is required for finalized product during production.

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Limitations
The Project work out at Q.C. department was very helpful to me for
the future carrier. Also I gained a lot of information about the analyses, Instruments during my project, however it has got few limitations such as: 1] Since the analysis work at Excel Industries is confidential, certain Information about the analysis (e.g. certain parameters about calculation) were kept confidential. 2] The analysis work using G.S. & H.P.L.C. had to be done under observation Of the expertise & it was limited. 3] Company do not give information about reaction mass i.e. reactance, Products & reagents etc. 4] Certain instruments for e.g. Karl Fischer have calibration frequencies once In a six months. Still I have tired you make the best of the work & for that I am grateful to Q.C. department of Excel Industries Ltd.

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Conclusion
The project work at Excel Industries, Lote, was a very important experience for me. It has been realized that how the use of classical & instrumental methods for the analyses is done. Further the Q.C. department is very important department of the organization which involves a number of activities regarding to the quality & certain specification of the products being analyzed. The department plays on important role in manufacturing of quality products & in achieving customer satisfaction. The theoretical knowledge about the classical methods, instruments like GC, HPLC etc. was tested during the project work. The study of how the instruments are handled, & operated, also the parts of the instrument, their roles etc. were very important & it strengthens the theoretical knowledge. Thus project work at Excel industries was a very important & knowledgeful experience.

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