Assay Method of
Ibuproen Tablet
| Sample | Dissolved a quantity of the powder containing the equivalent of 100 mg of Ibuproen as completely as possible in 60 ml of previously neutralized 96 % ethanol. Titrate with 0.1N Sodium hydroxide VS using 1 drop of 1% w/v Phenolphthalein solution as indicator.Each ml of 0.1N Sodium hydroxide s is equivalent to 20.63 mg of Ibuproden. |
| Calculation | % of content = V x F x 20.63 x 100 / W |
Assay Method of
Ibuproen Suspension
| 0.01 M Ortho phosphoric acid | Take 0.7 ml Ortho phosphoric acid (85%) in a 1000 ml Volumetric Flask & volume up to 1000 ml with Distilled water. |
| Ratio | Acetonitrile 55% : 0.01 M Ortho phosphoric acid 45% |
| Standard | Weight and transfer about 100 mg working standard of Ibuprofen in a 100 ml volumetric flask. Add 40 ml of acetonitrile and shake for 10 minutes and dilute to 100 ml with 0.01 M Ortho phosphoric acid . Take 5 ml solution in 25 ml volumetric flask & dilute to 25 ml with Mobile Phase up to the mark and mix well.Filter this solution with 0.45 m membrane filter before injection. |
| Sample | Shake the Bottle vigorously and take an amount of Suspension equivalent to 100 mg Ibuprofen in a 100 ml volumetric flask. Add 5 ml 0.01 M Ortho phosphoric acid & mix then add 40 ml acetonitrile & shake for 25 minutes and dilute with .01 M Ortho phosphoric acid & up to the mark and mix well. Filter the solution by whatman filter paper. Take 5 ml filtrate in a 25 ml volumetric flask dilute with Mobile Phase up to the mark and mix well.Finally filter this solution with 0.45 m membrane filter before injection. |
| Condition | ODS1 10 µm 4.6 x 250 mm / 40°C / 1.2 ml per minute / 220 nm / 20 µL /10 minutes / 5.5 minutes approx. |
Assay Method of
Indapamide Tablet
| Buffer | Transfer accurately weighed about 6.8045 g of Potassium dihydrogen orthophosphate in 900 ml distilled water, mix well. Adjust pH to 6.8 ± 0.05 with dilute sodium hydroxide and dilute to 1 liter with distilled water. |
| Mobile Phase | Mix 60 volume of buffer with 40 volume of acetonitrile filter through a 0.45-micron membrane filter and degas. |
| Standard | Dissolve about 25 mg of Indapamide in 30 ml of Acetonitrile and dilute to 50 ml with the same solvent. Dilute 2.5 ml to 50 ml with Acetonitrile. To 2 ml of this solution add 5 ml water and dilute to 25 ml with acetonitrile. Filter this solution with 0.45 m membrane filter before injection. |
| Sample | Shake a quantity of the powdered sample containing about 2.5 mg of Indapamide in 30 ml of Acetonitrile for 30 minutes, dilute to 50 ml with the same solvent. Transfer this solution to a 50 ml centrifuging tube & centrifuge at 2000 RPM for 15 minutes. Dilute 20 ml of supernatant to 25 ml with water. Filter this solution with 0.45 m membrane filter before injection. |
| Condition | C18 Column (4.6 mmx 25cm) & 5µm (ODS2) / UV at 242 nm |
Assay Method of
Indapamide + Perindopril Erbumine Tablet
| Buffer | Dissolve 0.92 gm of Sodium heptanesulphonate with 950 ml distilled water into 1000 ml volumetric flask & add 1 ml of Triethylamine, adjust the pH 2.5 ± 0.05 with phosphoric acid & make volume with distilled water up to 1000 ml. |
| Ratio | Mobile Phase A = Buffer pH 2.5 and Mobile Phase B = Acetonitrile |
| Solvent | Methanol : Water :: 50 : 50 |
| Standard | Accurately weigh and transfer about 25 mg of working standard of Indapamide & 80 mg of Perindopril Erbumine into 50 ml volumetric flask. Add 25 ml of Methanol & shake for 20 minutes and dilute with solvent up to the mark and mix well. Transfer 5 ml of this solution into 50 ml volumetric flask & dilute with solvent up to the mark and mix well. Filter this solution with 0.45 m membrane filter before injection. |
| Sample | Weigh and powder 20 Tablets. Take powder containing equivalent to 2.5 mg of Indapamide & 8 mg of Perindopril Erbumine into 50 ml volumetric flask. Add 25 ml of methanol & shake for 20 minutes and dilute with solvent up to the mark and mix well. Transfer this solution to 50 ml centrifuge tube & centrifuge at 2000 rpm for 15 minutes. Transfer the supernatant solution & Filter this solution with whatmann filter paper. Finally filter this solution with 0.45 m membrane filter before injection. |
| Condition | Waters Spherisorb® S10 C8 4.6 x 250 mm / UV at 210 nm /10 minutes Indapamide : 4.5 minutes and Perindopril Erbumine : 6.0 minutes (approximately). |
Gradient Programme Ratio
| Time (min.) | Mobile phase A | Mobile phase B | Wave Length |
| 0 – 5.3 | 40 | 60 | 254 nm |
| 5.3 – 8.0 | 40 | 60 | 220 nm |
Assay Method of
Indomethacin Capsule
| Sample | Weigh and powder 20 capsules. To a quantity of mixed contents of 20 capsules containing 50 mg Indomethacin add 10 ml of water and allow to stand for 10 minutes , swirling occasionally. Add 75 ml of methanol , shake well, add sufficient methanol to produce 100 ml . Mix and filter. To 5 ml of the filtrate add sufficient of a mixture of equal volumes of methanol and phosphate buffer pH 7.2 to produce 100 ml . Measure the absorbance of the resulting solution at the maximum at 318 nm .Calculate the content of Indomethacin taking 193 as the value of A (1 % , 1 cm ) at the maximum 318 nm. |
| Calculation | % of content = Absorbance of sample x 100 / Conc. in g/100 ml x 193 |
Assay Method of
Iodine Solution
| Sample | Transfer 25 ml of sample solution, equivalent to 25 mg of Iodine, into a stoppered conical flask & rinse the pipette with distilled water. Add about 50 mL distilled water and 4-5 drops of Starch indicator into the conical flask. Titrate I2 solution immediately against the 0.1 M Thiosulphate VS solution. At the end point of titration the color of iodine become disappear. Each mL of 0.1 M SodiumThiosulphate solution is equivalent to 12.69 mg of Iodine( I2 ) |
Assay Method of
IrbesartanTablet
| Buffer | Dilute about 5.5 ml Phosphoric acid (85%) in an approximately 950 ml Distillerd water and add triehylamine, slowly and dropwise to adjust pH 3.0 ± 0.1. Then make a volume 1 Liter with distilled water. Filter this solution through 0.45 m membrane filter and Degasses it for 15 minutes by Sonicator. |
| Ratio | Acetonitrile: Buffer = 55: 45 |
| Solvent | Methanol |
| Standard | Accurately weigh and transfer 15 mg Irbesartan Working standard in a 100 ml Volumetric flask. Add 70 ml Methanol and shake for 10 minutes and make the volume up to mark with the same solvent(MeOH) & mix well. Filter this solutionwith 0.25m membrane filters before injection. |
| Sample | Take an accurately measured of 38 mg powder which contain 15 mg of Irbesartan in a 100-ml volumetric flask. Add 70 ml Methanol and shake for 30 minutes & Sonicate for 3 minutes and make the volume up to mark & mix well. Filter the solution through whatman filter paper no.1.and then filter this solution with 0.2m membrane filters before injection. |
| Condition | Waters Spherisorb ODS1 10 mm 4.6 x 250 mm /220 nm / 1 ml per minute / 20 mL / 12 minutes / 7.6 minute Irbesartan |
Assay Method of
Irbesartan + Hydrochlorhtiazide Tablet
| Buffer | Take 1000 ml of Distilled water and add 1.0 ml of Triethylamine. Adjust the pH to 3.5 with Phosphoric acid. Filter through 0.45 m membrane filter and Degasses it for 15 minutes by Sonicator. |
| Ratio | Acetonitrile(B pump 50%):Triethylamine Solution (A pump 50%)=1:1 |
| Solvent | Mobile phase |
| Standard | 6.25 mg Hydrochlorhtiazide + 37.5 mg Irbesartan ---® 100 ml (Shake 25 minutes) |
| Sample | Take an accurately measured of 95 mg powder which contain 37.5 mg Irbesartan & 6.25 mg Hydrochlorothiazide in a 100-ml volumetric flask. Add 70 ml diluents (Mobile Phase) and shake for 30 minutes and make the volume up to mark & mix well & Sonicate for 5 minutes. Cool in and Ice bath for 10 min and then filter the solution through whatman filter paper no.1.and then filter this solution with 0.2m membrane filters before injection |
| Condition | ODS2 5 mm 4.6 x 250 mm / 40°C /254 nm / 1 ml per minute / 20 mL / 15 minutes / 3 minute Hydrochlorhtiazide & 8.5 minute Irbesartan approx. |
Assay Method of
Carbonyl Iron + Vitamin B – Complex with Ascorbic Acid + Folic Acid + Zinc Sulphate Capsule
For Carbonyl Iron
| Reagent | i) Alpha-Alpha bipyridyl solution :Dissolve 100 mg of Alpha-Alpha bipyridyl in 100 ml of distilled water. ii) Hydroxylamine Hydrochloride solution :Dissolve 10 gm of Hydroxylamine Hydrochloride in 100 ml of water. iii) Sodium Acetate solution : Dissolve 24 gm of Sodium Acetate trihydrate in 100 ml of distilled water. iv) Sodium dithionite solution : Dissolve 10 gm of Sodium dithionite in 100 ml of distilled water. |
| Standard | Weigh accurately 250 mg of Ammonium ferrous sulphate working standard in 100 ml volumetric flask, add 5 ml of conc. H2SO4 and add 50 ml of distilled water, mix well. Heat on water bath for 30 minutes at 95°C , remove the flask from the water bath and allow the flask to cool to room temperature, make the volume up to mark with distilled water. Dilute 5 ml of the solution to 50 ml with distilled water. |
| Sample | Reduce about 900 mg pellets completely to a fine powder form. Weigh a sample quantity equivalent to 100 mg of Carbonyl Iron and transfer into a 100 ml volumetric flask. Add 5 ml of conc. H2SO4 and 50 ml of distilled water, mix well. Heat on water bath for 30 minutes at 95°C. remove the flask form water bath and allow the contents to cool to room temperature and make the volume up to mark with distilled water and filter. After filtration, dilute 5 ml of the solution to 50 ml with distilled water. Further dilute 10 ml of this solution to 25 ml with distilled water. |
| Procedure | Transfer 5 ml of each standard and sample solution into 50 ml volumetric flasks. To each flask add the following - i) 2 ml of hydroxylamine hydrochloride solution ii) 10 ml of Sodium Acetate Buffer solution iii) 4 ml of Alpha-Alpha bipyridyl solution iv) 10 ml of Sodium dithionite solution & v) Make up to volume with distilled water Measure the absorbance of standard and sample solutions at 525 nm against water as blank solution. Calculate the Carbonyl Iron content in 450 mg pellets by the following formula – Carbonyl Iron in mg /450 mg pellets = Sample absorbance x Std. Weight x 55.85 x 450 mg / Std. absorbance x Sample Weight x 392.14 |
For Vitamin B – Complex with Ascorbic Acid
| Buffer | Transfer 0.2 ml of Triethylamine ,7.5 ml glacial acetic acid & 175 ml methanol to a 1000 ml volumetric flask, dilute with 0.008 M Sodium Hexane –1 Sulphonic acid to volume& mix. Filter & degas for use. |
| Ratio | Buffer ---------à Isocratic |
| Standard | Weight accurately 30 mg Riboflavin . Transfer to a 500 ml amber volumetric flask and add approx 300 ml 2% acetic acid . Place in water bath at 75°for 15 minutes this solution is a completely clear yellow liquid. cool the solution in room temperature . Add 30 mg thiamine mono nitrate, 15 mg Pyridoxine HCl, 150 mg Nicotinamide & 750 mg ascorbic acid. Dissolve by shaking vigorously add 2% acetic acid to volume & mix. Filter this solution through 0.45 µm filter & use for injection. |
| Sample | Weigh accurately 1.350 gm crushed sample to content 6 mg thiamine mono nitrate, 6 mg Riboflavin, 3 mg Pyridoxine HCl, 30 mg Nicotinamide & 150 mg ascorbic acid into a 100 ml amber volumetric flask. Add approx. 60 ml 2% acetic acid & shake 20 minutes. Place in a water bath 75°C for 15 minutes. cool the solution & add 2% acetic acid to volume & filter. Filter this solution through 0.45 µm filter before injection. |
| Condition | Waters Spherisorb® S10 C8 4.6 x 250 mm / UV at 270 nm /30 minutes/1.2 ml/minutes |
For Folic Acid
| Buffer | Transfer 2 gm of Mono basic potassium phosphate & 3 gm of Tetra butyl ammonium hydrogen sulphate into a 1000 ml volumetric flask & add 900 ml of distilled water to dissolve. Adjust 7.0 with 10% Potassium Hydroxide & dilute with distilled water to volume & mix & filter. |
| Ratio | Buffer 70% : Methanol. 30% |
| Standard | Weighed accurately 25 mg of USP Folic acid WS in a 100 volumetric flask. & add approx. 75 ml mobile phase & shake it to completely dissolve, dilute with mobile phase to volume & mix. Transfer 2 ml of this solution to a 50 ml volumetric flask & dilute with mobile phase to volume and mix. Filter this solution through 0.45 µm filter before injection. |
| Sample | Weigh accurately 0.450 gm crushed sample containing equivalent to 0.5 mg Folic Acid into a 50 ml volumetric amber glass flask & add 35 ml mobile phase & shake for 20 minutes. dilute with mobile phase to volume & mix. Filter this solution through whatmann filter paper & finally filter this solution through 0.45 µm filter before injection. |
| Condition | m Bondapak C18 125 A° 10 mm 3.9mm x 300 mm HPLC Column / UV at 280 nm /10 minutes |
For Zinc Sulphate
| Standard | Prepare the Zinc standard solution of 0.2, 0.4, 0.6, 0.8, 1.0 ppm using 3% v/v Nitric Acid solution as solvent. |
| Sample | Weigh accurately 225 mg crushed sample to content 11.26 mg Zinc into a 100 ml volumetric flask. Add 5 ml of water & 5 ml of Nitric Acid & shake 5 minutes. Then add 50 ml of water & heat in a water bath for 10 minutes and Shake 10 minutes. Cool the solution & add water to volume & filter. Transfer 10 ml of this solution to a 100 ml volumetric flask & dilute with 3% v/v Nitric Acid. Further transfer 4 ml of this solution to a 100 ml volumetric flask & dilute with 3% v/v Nitric Acid and mix. Filter this solution through 0.45 µm filter before injection. |
| Procedure | Follow the SOP(QC/SOP/o69) of Atomic Absorption Spectrophotometer & all the safety instruction. Carry out the test following SOP & Safety instruction using 213.9 nm wavelength & Air Acetylene method. The analytical result is obtained in ppm. Calculation : % of Elemental Zinc = Result (in ppm) x 100 / Conc. of Zn (in ppm) |
Assay Method of
Isoniazid Tablet
| Sample | Weigh and powder 20 tablets. Dissolve a quantity of the powder containing 0.40 g of Isoniazid as completely as possible in water , filter and wash the residue with sufficient water to produce 250 ml . Mix . To 50 ml of the resulting solution add 50 ml of water , 20 ml of hydrochloric acid and 0.2 g of potassium bromide and titrate with 0.0167M potassium bromate VS determining the end point electrometrically.Each ml of 0.0167M potassium bromate VS is equivalent to 3.429 mg of Isoniazid . % of Isoniazid = No. of ml of 0.0167M potassium bromate VS x F x 3.429 X 100 / Amount Isoniazid taken |
Assay Method of
Isosorbide Dinitrate Tablet
| Mobile Phase | Mix 15 volumes of Absolute ethanol and 85 volumes of trimethylpentane (iso Octane), filter through a 0.45-micron membrane filter and degas. |
| Solvent | Mobile Phase |
| Standard | 25 mg ---® 25 ml ---® take 1 ml ---® 10 ml |
| Sample | Same as standard. |
| Condition | 250 x 4.6 mm aminopropylmethylsilyl silica gel column (10 µm) / UV at 230 nm / 15 minutes |
Assay Method of
Isosorbide Mononitrate Tablet
| Mobile Phase | Mix 15 volumes of Absolute ethanol and 85 volumes of trimethylpentane (iso Octane), filter through a 0.45-micron membrane filter and degas. |
| Solvent | Mobile Phase |
| Standard | 25 mg ---® 25 ml ---® take 1 ml ---® 10 ml |
| Sample | Same as standard. |
| Condition | 250 x 4.6 mm aminopropylmethylsilyl silica gel column (10 µm) / UV at 230 nm / 15 minutes |
Assay Method of
Itopride HCl Tablet
| Standard | Take 50 mg of working standard of Itopride HCl in a 100 ml volumetric flask. Add 70 ml of 0.1 N HCl and shake for 30 minutes. Adjust the volume to100 ml with 0.1 N HCl . Take 2 ml of the solution in a 50 ml volumetric flask. Make the volume up to the mark (50 ml) with 0.1 N HCl . Take absorbance at 257 nm. Use 0.1 N HCl as blank. |
| Sample | Weigh and make fine powder of 20 tablets. Take an accurately weighed portion of powder equivalent to 50 mg of Itopride HCl in a 100 ml volumetric flask. Add 70 ml of 0.1 N HCl and shake for 30 minutes. Adjust the volume to100 ml with 0.1 N HCl & mix. Filter the solution by Whatman filter paper. Take 2 ml of the filtrate in a 50 ml volumetric flask. Make the volume up to the mark (50 ml) with 0.1 N HCl. Take absorbance at 257 nm. Use 0.1 N HCl as blank |
