mobile phase: methanol/ACN (90:10, v/v) + 9 μM TEA.
Can I use n-hexane as the injection solvent?
Will this cause column clogging or damage?
I know that acetonitrile and n-hexane are immiscible.
You did not provide any information about the HPLC column or sample you selected. The column type, name, dimensions, sample and full method info would be needed to understand what you are really doing first. However, hexane is not miscible with either methanol or acetonitrile.
Your injection solution should be the same as your mobile phase. This is a basic fundamental of chromatography. *The sample and mobile phase solution should be fully miscible (100%). Chromatography is not possible if the sample can not be fully dissolved in the mobile phase. Hexane is NOT miscible with methanol or acetonitrile so would not be used with those solvents alone. Also, running pure ACN and Methanol on a column does not sound like an appropriate mobile phase so you really need to stop what you are doing and speak to a local chromatographer (or your teacher) at your school to obtain help in using the HPLC system. You may damage one of the columns or instrument.
Using n-hexane as the injection solvent with the given mobile phase composition of methanol/acetonitrile (90:10, v/v) + 9 μM triethylamine (TEA) is not recommended, as n-hexane is immiscible with the polar mobile phase components, which could lead to precipitation and clogging of the column frit or inlet. Additionally, n-hexane is a strong eluting solvent that may disrupt the retention mechanism and cause peak distortion or analyte breakthrough, and it may not be compatible with the HILIC (hydrophilic interaction liquid chromatography) stationary phase of the Polaris NH2 column, potentially causing damage. Instead, it is advisable to use a more polar solvent that is miscible with the mobile phase, such as methanol or acetonitrile, as the injection solvent, and to perform a small-scale test injection to evaluate the compatibility before proceeding with a large number of samples.
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