what is method development parameter in HPLC method
development
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Answer / abdulkalim
Parameters involved during initial stage of Analytical
Method development:
1. Optimization of Mobile Phase
2. Selection of Buffers
3. Selection of Ion pair reagent
4. pH of buffer/mobile phase
5. Composition of Mobile Phase
6. Selection of Column
7. Optimization of Analyte signal
8. Objectives of Seperation
8.1 Analysis time
8.2 Resolution
8.3 k factor (Capacity factor)
8.4 Peak height
8.5 Asyymetry
8.6 Theoritical Plates
9. Flow rate
| Is This Answer Correct ? | 90 Yes | 7 No |
Answer / ramana
Parameters involved during initial stage of Analytical
Method development:
1. Solubility
2. Pka Value
3. Selection of Buffers
4. pH of buffer/mobile phase
5. Composition of Mobile Phase
6. Selection of Column
7. Optimization of Analyte signal
8. Objectives of Seperation
8.1 Analysis time
8.2 Resolution
8.3 k factor (Capacity factor)
8.4 Peak height
8.5 Asyymetry
8.6 Theoritical Plates
9. Flow rate
| Is This Answer Correct ? | 54 Yes | 2 No |
Answer / yojana
1. Check solubility,if sample dissolved in water then use
reverse phase,othervise use normal phase.
2.wavelength selection
3.selection of mobile phase,buffer.
4. pH OF BUFFER &MOBILE PHASE
5. selection of column
6. check retention time
7 .Flow rate
8. check system suitability parameter like
i)theoritical plates
ii) Assymetry
iii) Resolution
iv) capacitor fator
| Is This Answer Correct ? | 31 Yes | 2 No |
Answer / vinod kumar, nektar therapeuti
1.If molecular weight less than 2000. based on their
solubility follow as below
2.If water soluble and non ionic follow reverse phase.
3.If water soluble and ionic follow reverse phase ion pair
method.
4.If organic soluble follow normal phase.
5.If molecular weight is more than 2000.Based on their
solubility follow as below.
6.If organic soluble follow gel permeation chromatography
7.If water soluble ion exchange or gel filtration.
8.Once the technique is fixed optimise column.
9.Optimise buffers
10.optimise pH of buffers
11.optimise concentration of buffers.
12.optimise type of organic solvent.
13.optimise column temperature
14.optimise gradient programme to improve resolution and to
elute out late eluting peaks.
| Is This Answer Correct ? | 19 Yes | 11 No |
Answer / sivannarayana
1.Solubility
2.Find out the UV scan For wave length
3.optimaize Buffer & pH Value Optimaize
4.Optimaize Column Temparature & optimaize sample cooler Temparature.
5.optimaize the GP For better Resolution
| Is This Answer Correct ? | 4 Yes | 0 No |
Answer / anand patel
as per my knowledge system suitability test is the method
delopment parameter
| Is This Answer Correct ? | 10 Yes | 19 No |
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Dear Sir, This is Thirunavukkarasu am working in Ordain Health Care AR&D, Am Having one problem While doing method development my question Metadoxine and Pyridoxine same chemical nature and structural activity i tried lot of methods but Analyte eluted same Retantion time i cant separate both peaks. solubility, pH both are same only only difference melting point metadoxine 210 C Pyridoxine 102 C please do the help needful for me