Answer Posted / sushant

1.HPLC method development start with literature servey to
get rough idea abt development
2.then find out functionallity, molecular structure,
Polarity of particular API or molecule. from which we can
decide method development phase RPC or NPC.
3.Find out Solubility, Max UV absorbace so that we be
assure about respective diluent & wavelegh for method
development.
if we will not get UV absorbace then there is dervatisation
method.
4.Mostly many oragnic drug molecules are polar or mid polar
so we usually go for RPC.
5.satrt with mobile phase composition Use water & MEthanol
or ACN according to sollubillity in 50:50 ratio.
6.Select Column -start with C-18 column make HYpersil or
Inertsil(3 microne x 4.6 ID )10-15 cm
7. feed other parameter like wavelength,ambient Temp,
1ml/min Flow rate
8.About sample preparation for assay preparation use conc
in between 100-500 ppm as per peak response.
9.If there are more than 10 molecule in particular API go
for gardient method for better resolution & short run time
while developing any method always follow ICH parameter abt
Tailing factor, Capacity factor, Assymetry, thereotical
plates.Change MP composition ,Column.
Once you get better result then for sharpness or to save
time adjust flowrate & increase Temp.
10.If our desired peak isnot retain on water /ACN/Methanol
composition MP then go for Buffer. Phosphate buffer is
widly use buffer for its PKa value,easily Available, gives
more Hydrophillic effect, stable or non degradable for 2-3
days.
...............Before starting any method development on
HPLC Flush the system with water, check pressure, saturate
column with MP,check callibration status, lamp Hours.
Always keep data about whatever trials you have done during
method development.
Method Development is very challenging task so always be
logical before any changes in any parameter.

Mostly For HPLC method Development 10 gm sample is enough
but for overall development max 100 gm sample is sufficient.
Its a tentative quantity.

After method development method validation must be done.

in which situation require to take incident in validation?

1742

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Why acetonitrile and water are used as extraction solvent when analysis melamine? I thought they are miscible and won't be able to separate...

1700

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how require to select dissolution media? what is discrimination?

802

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What is the difference between chromatographic purity and related substances?

1387

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How can we confirm the HPLC column is end-capped or not? Is it possible to identify by physical appearance?

3233

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what is mean by extactable and leachable study?

1185

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[3/30, 13:29] Manoj P Venkatpurwar: How hplc column selection according to structure? How mobile phase buffer selection on molecule structure?

813

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1)What's the meaning of Absorption,give a example. 2)What's the meaning of Adsorption,give a example. 2)what is the difference between Absorption and Adsorption.

2896

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how to selecet an exact coloumn for an new molecule development by hplc how to select exact salt as buffer for new molecule development by hplc what is the the process to select the mode of saparation of compoundes by hplc what is the use of ph of buffer what is use of buffer,ph,organic phase,ans methods how the molecules get saparated in coloumn,

2613

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How to decide assay range for non pharmacopeial API analysis by HPLC? Could you give me any reference for same e.g Guidelines or Paper publications?

3370

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In Dissolution Test why limit is define Q+5% what is the role of +5%.

3114

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How do we get end points and how many end points are possible for citric acid and di-acid not theorotically answer should be given practically.

2591

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How to do regeneration of Metacarb Pb plus column?

2586

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Why we select scan range from higer wavelength to lower wavelength in uv visible spectroscopy and ftir spectroscopy ? .

921

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Difference between the quantitative analysis and qualitative analysis?

1193

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