Methods to increase sensitivity of LC/MS analysis
The present disclosure discusses a method of separating a sample (e.g., pharmaceutical drug, genotoxic impurity, biomarker, and/or biological metabolite) including coating a metallic flow path of a chromatographic system; injecting the sample into the chromatographic system; flowing the sample through the chromatographic system; separating the sample; and analyzing the separated sample using mass spectroscopy. In some examples, the coating applied to the surfaces defining the flow path is non-binding with respect to the sample—and the separated sample. Consequently, the sample does not bind to the low-binding surface of the coating of the flow path. The applied coating can increase the chromatographic peak area for the sample of the chromatographic system.
1. A method of separating a sample comprising:
providing a chromatographic system having a metallic flow path with an alkylsilyl coating on fluid exposed surfaces, the metallic flow path with the alkylsilyl coating preventing the sample from binding to underlying metal surfaces;
injecting the sample into the chromatographic system, wherein the sample comprises gentisic acid and/or 4-hydroxybenzoic acid;
flowing the sample through the chromatographic system; and
separating the sample, wherein coating the metallic flow path of the chromatographic system increases a chromatographic peak area of the gentisic acid and/or 4-hydroxybenzoic acid in the sample in comparison to an uncoated chromatographic system having metallic surfaces exposed to fluid by 1.3 × or greater.
2. The method of claim 1 , wherein the alkylsilyl coating is applied to internal portions of the column, frits and end caps.
3. The method of claim 1 , wherein the alkylsilyl coating is applied to wetted surfaces located upstream of the chromatographic column.
4. The method of claim 1 , wherein the chromatographic system comprises a mix-mode chromatographic column.
5. The method of claim 1 , wherein the chromatographic system comprises a phenyl-hexyl chromatographic column.
6. The method of claim 1 , wherein the alkylsilyl coating comprises bis(trichlorosilyl)ethane or bis(trimethoxysilyl)ethane.