Here is the titration method for testing of Vitamin "C "in a supplement drug. It becomes more difficult to test Vitamin "C" in drug having Other vitamins like Vitamin "A ", Vitamin "E " Vitamin B", extracts like Ginkobiloba or Gensing and minerals like Zinc, or Seinite etc. So here is easy method to test the Vit. "C" in that drug.
PROCEEDURE:In the test , 1,2-diphenol-indophenol and metaphosphoric-acetic acid are used. first of all we are going to prepare both of these.
PREPARATION OF 1,2-diphenol-indophenolWeigh 25mg of 1,2-diphenol-indolphenol in 100ml volumetric flask. add 25 ml of water and mix the weighed quantity. Now weigh 50mg of Sodium by carbonate and add in the mixture. add more 25ml of water and dissolve the reagents by continuous stirring by any mechanical means or electromagnetic source. and in last make the volume upto the marks using water.
PREPARATION OF Metaphosphoric Acetic-AcidIn a 100ml volumetric flask, Add 3ml of conc. phosphoric acid and 8ml of glacial acetic acid and makeup the volume upto the mark using distilled water.
ASSAY OF VITAMIN "C"Weigh accurately the product equivalent to 100mg of Vitamin C in 200ml volumetric flask. add 70ml of metaphosphoric acetic-acid in flask and sonicate for 30 minutes. Then makeup the volume upto the mark using distilled water. Mix the solution for 5 minutes and filter through whattman filter paper. or centrifuge some solution. take 4ml of the solution in a 100ml conical flask and add 5ml of metaphosphoric acetic acid. Now titrate with 1,2-diphenol-indophenol until rose pink color . each ml of 1,2-diphenol-indophenol used is equivalent to 0.136mg of Vit.C. Similar Posts :
Here is the list of factors for some raw material in B.P and U.S.P. By using these factors you can remove unwanted chemical groups from active and effective one. According to your label claim and raw material. I explain, let one has Lincomycin HCl and label claim for drug is only lincomycin then one should remove the HCl group or in other words, He should add so much quantity to manufacture a batch that the active Lincomycin should gave 100% assay when compared to B.P or U.S.P. Reference standard Lincomycin. For that reason here is the list of Raw Material factors that should be multiplied with the required quantity of raw material to fulfill the requirements of active present in raw material.
|Sr. No||Product Name||Molecular Weight||Factor|
|1||Atorvastatin Calcium Trihydrate||1209.42||1.0843|
|3||Ciprofloxacin HCl Monohydrate||385.52||1.1646|
|10||Iron III Hydroxide Polymaltos Complex||449.16||3.04|
|14||Pantoprazole Sodium Sesquihydrate||432.4||1.1306|
|16||Gentamicin Sulphate||Nlt 590 µg/mg||1.6949|
ABOUT:Gentamicin Sulfate is the sulfate of a mixture of aminoglycoside substances having antibacterial activity produced by the growth of Micromonospora purpurea or Micromonospora echinospora.
PHYSICAL APPEARANCE:Gentamicin Sulfate occurs as a white to light yellowish white powder. It is highly hygroscopic.
CHEMICAL FORMULA:C21H43N5O7. xH2SO4 CHEMICAL STRUCTURE:
IUPAC NAME:(6R)-2-Amino-2,3,4,6-tetradeoxy-6-methylamino-6-methyl-α-D-erythrohexopyranosyl-(1→4)-[3-deoxy-4-C-methyl-3-methylamino-β-L-arabinopyranosyl-(1→6)]-2-deoxy-Dstreptamine sulfate
CONTENTS:It contains not less than 590 mg (potency) and not more than 775 mg (potency) per mg, calculated on the dried basis. The potency of Gentamicin Sulfate is expressed as mass (potency) of gentamicin C1
- Soluble in water.
- Typically insoluble in ethanol. (96%)
IDENTIFICATION BY PPT. FORMATION:Dissolve 50 mg of Gentamicin Sulfate in 5 mL of water, and add 0.5 mL of barium chloride TS: a white precipitate is formed.
COLOR OF SOLUTION:Dissolve 1.0g of Gentamicin Sulfate in 10 mL of water, the solution is clear and colorless to pale yellow.
IDENTIFICATION BY THIN LAYER CHROMATOGRAPHY:Dissolve 50 mg each of Gentamicin Sulfate and Gentamicin Sulfate Reference Standard in 10 mL of water, and use both of these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under thin layer-Chromatography. Spot 20 µL of the sample solution and standard solution on a plate of silica gel for thin layer-Chromatography. Separately, shake a mixture of chloroform, ammonia solution and methanol in ration of (2:1:1) in a separator, and allow the mixture to stand for more than 1 hour. To 20 mL of the lower layer so obtained add 0.5 mL of methanol, and use this as the developing solvent. Develop the plate with the developing solvent to a distance of about 17cm in a developing container with a cover, having an opening of about 20 mm2, and without putting a filter paper in the container, and air-dry the plate. Allow the plate to stand in iodine vapors: three principal spots obtained from the sample solution are the same with the corresponding spots obtained from the standard solution in color tone and the Rf value, respectively.
pH:The pH of a solution obtained by dissolving 400 mg of Gentamicin Sulfate in 10 mL of water is between 3.5 and 5.5
LOSS ON DRYING:Not more than 18.0% when 0.15g of Gentamicin Sulfate at pressure not exceeding 0.67 kPa and temperature 110°C for 3 hours. Handle the sample avoiding absorption of moisture.
RESIDUE ON IGNITION:Should be not more than 1.0% when 1.0 gram is ignited.
SPECIFIC OPTICAL ROTATION:+107° to +121° When 100mg of Gentamicin Sulfate is dissolved in 10ml of water.
QUANTITATIVE ASSAY (BIO-ASSAY):Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics according to the following conditions. Test organism Staphylococcus epidermidis ATCC Agar media for seed and base layer
- Glucose 1.0 g
- Peptone 6.0 g
- Meat extract 1.5 g
- Yeast extract 3.0 g
- Sodium chloride 10.0 g
- Agar 15.0 g
- Water 1000 mL
REFERENCES:Japan Pharmacopia (JP XVII) Official monographs Page 983. United State Pharmacopia (USP 41) Page 938.
DESCRIPTION:Diclofenac Sodium is white or slightly yellowish, slightly hygroscopic, crystalline powder.
MOLECULAR WEIGHT:318.1 Dalton
USE AS:Cyclo-oxygenase inhibitor; analgesic; anti-inflammatory.
IUPAC NAME:Sodium [2-[(2,6-dichlorophenyl)amino]phenyl]acetate.
CONTENTS:It contains not less than 99.0% and not more than 101.0% of C14H10Cl2NNaO2.
- Sparingly soluble in water.
- Freely soluble in Methanol.
- Soluble in ethanol.
- Slightly Soluble in acetone.
MELTING POINT:Determine the melting point of sample by using M.P instrument. It decomposes at 280°C.
IDENTIFICATION BY FTIR SPECTROPHOTOMETER:Measure the FTIR spectra of sample and compare with standard.
IDENTIFICATION BY SODIUM REACTION:Dissolve 60mg in 0.5mL of methanol and add 0.5 mL of water R. The solution gives reaction of sodium.
IDENTIFICATION BY COLOR OF SOLUTION:Dissolve 1.25 gm in 25 mL methanol. And check its color & measure absorbance at 440 nm its absorbance should not be greater than 0.05
pH:Dissolve 1.0 gm in Distilled water & dilute to 100ml and determine the pH. It should be in between 7.5 to 9.0.
LOSS ON DRYING:Take 1.0gm of sample & dry in oven at 105-110oC for 3.0 hours. It should not be greater than 0.5%.
QUANTITATIVE ASSAY: (By Non-Aqueous TITRATION)Dissolve 450 mg of Diclofenac Sodium in 25mL Glacial acetic acid. Add 2 drops of crystal violet indicator. Titrate with 0.1N Perchloric acid till change in color of indicator. If necessary, carry out blank titration. Each ml of 0.1N HClO4 is equivalent to 31.81 mg of C14H10Cl2NNaO2.
CALCULATIONS:Calculate the Percentage of Diclofenac Sodium by using following Expression. %age assay = ((R x 31.81) / (W-(100-X)) x 100 Where R = Reading at Burette (Vol. of 0.1N HClO4) W = Weight of sample taken X = Water contents
STORAGE CONDITION:It should be packed in airtight containers. Containers should not be transparent to light.
REFERENCE:British Pharmacopia 2016. Volume-I, Page 740
White Crystalline powder, Slightly hygroscopic. On dried basis, It contains 98.0% to 102.0% of Pregabalin.
- Soluble in Water
- Slightly Soluble in Methanol
- Very slightly soluble in ethanol
- Practically insoluble in acetone, ethyl acetate, heptane and in methylene chloride
IDENTIFICATION BY FTIR SPECTROPHOTOMETERMeasure the FTIR spectra of sample and compare with standard.
IDENTIFICATION BY HPLCThe retention time of peak of Pregabalin in sample solution is similar to the retention time of peak of pregabalin in standard solution
APPEARANCE OF SOLUTIONDissolve 0.5 grams in CO2 Free water and dilute 20ml with same solvent (solution S) The solution is clear and not more intensely colored than GY5
WATER CONTENTSNMT 0.5%, determined on 0.200 gram
QUANTITATIVE ASSAY BY HPLC
- Mobile Phase:
- Standard solution:
- Sample Solution:
- Mode : LC 100
- Detector : UV 210 nm
- Column : A stainless steel column 0.25 m x 4.6mm x 5 µm C18 packing; Octadecylsilanized silica gel (5µm particle diameter).
- Column temperature : Ambient
- Flow rate : 35 ml/min
- Inject 10µl of mobile phase (blank) and record the chromatograph
- Inject 10µl of standard solution two times and record the chromatograph
- Inject 10µl of sample solution and record the chromatograph
- Inject 10µl of standard solution and record the chromatograph
- Inject 10µl of mobile phase (blank) and record the chromatograph
CALCULATIONSCalculate the Percentage of Pregabalin sample by using following formula %age Assay = (At/As)× (Cs/Ct) × P Where At = Av. Area of the peaks in sample chromatograms As = Av. Area of the peaks in standard chromatograms Cs = Concentration (mg/ml) of standard Ct = Concentration (mg/ml) of sample P = Potency of reference standard
Physical Appearance :Tranexamic Acid occurs as white, crystals or crystalline powder. It belongs to antifibrinolytic family. Tranexamic Acid, when dried, contains not less than 98.0% and not more than 101.0% of tranexamic acid (C8H15NO2)
Solubility:It is freely soluble in water, and practically insoluble in ethanol (99.5).
Identification : BY FTIRDetermine the infrared absorption spectrum of Tranexamic Acid as directed in the potassium bromide disk method under Infrared Spectrophotometry, and compare the spectrum with the Reference Spectrum or the spectrum of Tranexamic Acid RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
pH:The pH of a solution prepared by dissolving 1.0 g of Tranexamic Acid in 20 mL of water is between 7.0 and 8.0.
Color of solution:Dissolve 1.0 g of Tranexamic Acid in 10 mL of water: the solution is clear and colorless.
LOD: (LOSS ON DRYING)Not more than 0.5% (1 g, for 2 hours).
Residue on ignition :Not more than 0.1%
ASSAY : ( HPLC-Method)
Mobile Phase:Dissolve 11.0 g of anhydrous sodium dihydrogen phosphate in 500 mL of water, and add 5 mL of triethylamine and 1.4 g of sodium lauryl sulfate. Adjust the pH to 2.5 with phosphoric acid or diluted phosphoric acid (1 in 10), add water to make 600 mL, and add 400 mL of methanol. Standard Preparation: Weigh accurately about 50 mg of Tranexamic Acid RS, previously dried, dissolve in water to make exactly 25 mL, and use this solution as the standard solution.
Sample Preparation:Weigh accurately about 50 mg of Tranexamic Acid, previously dried, dissolve in water to make exactly 25 mL, and use this solution as the sample solution.
Chromatographic conditions:Sample Volume: 20µL Detector: UV ʎmax=220nm Column: A stainless steel column 6.0 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: 25°C Flow rate: Adjust so that the retention time of tranexamic acid is about 20 minutes.
Calculations:Contents of Tranexamic acid can be calculated by given formula %age Assay = Hs/Ha × Ca/Cs × 100 Hs = Peak height ratio from the Sample solution Ha = peak height ratio from the Standard solution Ca = concentration of Alclometasone Dipropionate in the Standard solution (mg/ml) Cs = concentration of the Sample solution (mg/ml)
Assay: (BY TITRATION) (non-aqueous )Weigh accurately 10mg of tranexamic acid in a round 250ml conical flask and add 50ml of glacial acetic acid. Dissolve in magnetic stirrer. Now simply titrate it 0.1M perchloric acid. Each ml of 0.1M perchloric acid is equivalent to 15.72 mg of tranexamic acid.
Calculation:Contents of Tranexamic acid can be calculated by given formula %age of tranexamic acid = (volume of 0.1M perchloric acid × 15.72) ÷ weight of sample taken.
ALTERNATIVE METHOD:(BY UV/vis Spectrophotometer)
STANDARD PREPARATION:Weigh accurately about 50mg of Tranexamic acid of working standard in to 50ml volumetric flask and dilute it with distilled water. Take 1ml of this solution and dilute upto 100ml with the same solvent. SAMPLE PREPARATION: Weigh accurately about 50mg of Tranexamic acid of sample in to 50ml volumetric flask and dilute it with distilled water. Take 1ml of this solution and dilute upto 100ml with the same solvent. Take Reading at 210nm Contents of Tranexamic acid can be calculated by given formula %age of tranexamic acid= At/As × Cs/Ct × 100 At = Absorbance Of Tranexamic acid in Sample Solution. As = Absorbance Of Tranexamic acid in Standard Solution. Cs = Concentration of standard solution (mg/ml) Ct = Concentration of sample solution (mg/ml)
Alclometasone dipropionate is a synthetic glucocorticoid corticosteroid and a corticosteroid ester. At is an Active raw material for different products in pharmaceutical industry. CHEMICAL FORMULA C23H37ClO7 CHEMICAL STRUCTURE DEFINITION Alclometasone Dipropionate contains not less than 97.0% and not more than 102.0% of C23H37ClO7 calculated on the dried basis. ASSAY (HPLC PROCEDURE) MOBILE PHASE: Methanol and Solution A in ratio (2: 1) SOLUTION A: 6.80 mg/ml of monobasic potassium phosphate (0.05 M) INTERNAL STANDARD SOLUTION: 2 mg/ml of betamethasone dipropionate in methanol STANDARD STOCK SOLUTION: 1.2 mg/ml of Alclometasone Dipropionate reference standard in methanol STANDARD SOLUTION: 4.0 ml of Standard stock solution and 4.0 ml of Internal standard solution. Dilute with methanol to 25 ml. ( this solution contains 0.2 mg/ml of Alclometasone Dipropionate reference standard.) SAMPLE STOCK SOLUTION: 1.2 mg/ml of Alclometasone Dipropionate in methanol SAMPLE SOLUTION: 4 ml of Sample stock solution and 4 ml of Internal standard solution. Dilute with methanol to 25 ml. CHROMATOGRAPHIC SYSTEM Mode: LC Detector: UV 254 nm Column: 4-mm x 30-cm; packing L 1 Flow rate: 1.2 ml/min Injection size: 10 µL RELATIVE STANDARD DEVIATION Relative Standard Deviation should not be more than 2%. CALCULATION %age Assay = Hs/Ha × Ca/Cs × 100 Hs = Peak height ratio from the Sample solution Ha = peak height ratio from the Standard solution Ca = concentration of Alclometasone Dipropionate in the Standard solution (mg/ml) Cs = concentration of the Sample solution (mg/ml) ACCEPTANCE CRITERIA: 97.0% - 102.0% on the dried basis IMPURITIES
- OPTICAL ROTATION :
- Loss on Drying: Dry a sample in a vacuum at a pressure not exceeding 5 mm of mercury at 105° for 3 h: