Articles
Importance of Mobile Phase Buffer Selection for HPLC to LC-MS
by Rohit T. Veeprho Laboratories Pvt.LtdBackground:
Considering
the more complex products analytical scientist needs to develop the specific,
accurate, robust stability indicating analytical method. At the time of method
development analytical scientist need to be focus on the development of mass
compatible method. At the start of method development, it is important to have
systematic information about sample.
Selection
of Buffers for HPLC:
It
is essential to separate all probable impurities from the degradation sample
with minimum run time. Samples are categorized as neutral and ionic while ionic
includes acid, base, ampholytic and organic salts. Acidic and basic nature of
samples requires buffer content mobile phase and for neutral buffer usually not
required. If there is no resolution between 2 closely eluted peak then need to
vary solvent strength of mobile phase. Solvent strength and solvent type impact
the selectivity of sample peaks. Following are the solvents which control the
retention.
During HPLC method development selection of proper
buffer is important for the separation of peaks and its symmetry. Following are
the common different types of organic/inorganic buffers:
1. Phosphate Buffers:
Potassium phosphate, Di-Potassium phosphate, Monosodium phosphate, Disodium phosphate,
Phosphoric acid etc.
2. Acetate Buffers:
Ammonium acetate, Sodium acetate, etc. (Phosphate and acetate buffers are most
common because it can be used at wavelengths below 220 nm)
3. Di-ethyl
amine/Tri-ethyl amine buffers
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About Rohit T. 
Veeprho Laboratories Pvt.Ltd
2 connections, 0 recommendations, 28 honor points.Joined APSense since, August 6th, 2020, From Navi Mumbai, India.
Created on Feb 11th 2021 00:33. Viewed 310 times.
