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- Posts: 1
- Joined: Mon Feb 19, 2018 4:14 am
I am a novice in area of LC-MS/MS. For couple weeks I am trying to learn the technique without any specialized supervision. I have optimized my compound using flow injection (automated optimization). The system was tuned by a specialized personnel from sciex. I know my parent ion and most abundant daughter ions. However, I have not bought the internal standard yet. All I am trying to do is, making a calibration plot with my standard sample. My concerns are:
1. I am running MRM scan with dwell time of 0.51 msec and running it for 450 cycles. I am running sample with autosampler but I am using a zero volume connector (no column). For the quantification, I am using the automatic qunatification tool (Analyst) and it is giving me variable peak areas for my replicate samples. Moreover, I should have observed an increase in peak area with increasing standard but I am not observing any trend.
Is this beause I am not using any internal standard? Is it necessary to run internal standard with sample in LC-MS/MS?