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During the Internal Standard Method process, at what stage of the sample prep should the internal...

During the Internal Standard Method process, at what stage of the sample prep should the internal standard be added to the analyte(s), and why?

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Internal standards are mostly added and used before the sample can be injected into column when many sample preparation steps are required.

when many analytes with similar properties are frequently encounters in Gas Liquid Chromatography.

When the matrix effect on an analyte is unknown or variable, the method of standard additions is useful.

because

In HPLC separation, internal standard addition compensate/adapt/adjust for evaporation losses of solvents when fractions are held in autosampler to run overnight.

internal standard addition improve data quality when added with every sample with same concentration in calibration step of Liquid Chromatography.

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