Summary
In a chemical analysis, the internal standard method involves adding the same amount of a chemical substance to each sample and calibration solution. The internal standard responds proportionally to changes in the analyte and provides a similar, but not identical, measurement signal. It must also be absent from the sample matrix to ensure there is no other source of the internal standard present. Taking the ratio of analyte signal to internal standard signal and plotting it against the analyte concentrations in the calibration solutions will result in a calibration curve. The calibration curve can then be used to calculate the analyte concentration in an unknown sample. Selecting an appropriate internal standard accounts for random and systematic sources of uncertainty that arise during sample preparation or instrument fluctuation. This is because the ratio of analyte relative to the amount of internal standard is independent of these variations. If the measured value of the analyte is erroneously shifted above or below the actual value, the internal standard measurements should shift in the same direction. The earliest recorded use of the internal standard method dates back to Gouy's flame spectroscopy work in 1877, where he used an internal standard to determine if the excitation in his flame was consistent. His experimental procedure was later reintroduced in the 1940s, when recording flame photometers became readily available. The use of internal standards continued to grow, being applied to a wide range of analytical techniques including nuclear magnetic resonance (NMR) spectroscopy, chromatography, and inductively coupled plasma spectroscopy. In NMR spectroscopy, e.g. of the nuclei 1H, 13C and 29Si, frequencies depend on the magnetic field, which is not the same across all experiments. Therefore, frequencies are reported as relative differences to tetramethylsilane (TMS), an internal standard that George Tiers proposed in 1958 and that the International Union of Pure and Applied Chemistry has since endorsed.
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