Instrumax CAD-piperidine-Test report

1, instruments and materials

Instruments:Agilent1200 liquid chromatograph+Charged Aerosol Detector (CAD)(INSTRUMAX, Model: CADetector a1)

Reagents: pure water (Watson's distilled water), acetonitrile (chromatography grade), trifluoroacetic acid (analytical grade)

Samples: standards and samples (provided by user teachers)

2, methods and results

2.1 Chromatographic conditions

Column: phenomenex Kinetex® XB-C18 5μm 4.6*250mm

Column temperature:35℃ Flow rate:1.0mL/min

Injection volume:10μL

Mobile phase A:0.1%TFA Aqueous solution mobile phase B:0.1%TFA Acetonitrile solution

Gradient conditions:

CAD Conditions:

Instrument model:INSTRUMAX CADetector a1

Drift tube temperature:50℃ Atomized air flow:3L/min

Corona current:1μA Corona airflow:1L/min

Gain:0.2 RF:2.0

2.2 Sample information

blank solvent 1 branch (blank),STD Sample:15µg/mL Standard sample 1 branch, sample 1 Branch (Sample)

Correspond STD sample 10%,20%,30%,50%,80%,120%,150%and 200%Concentration level samples, i.e.1.5µg/mL,3.0µg/mL,4.5µg/mL,7.5µg/mL,12.0µg/mL,18.0µg/mL,22.5µg/mL,30.0µg/mL standard sample, each 1 Branch

2.3 Experimental results

2.3.1 Exclusiveness

The blank solvent does not interfere with the determination of the target peak and has good specificity.

picture 1 Blank solvent (thinner)

2.3.2 Sensitivity

10%Concentration level samples (Sensitivity samples), using Agilent CDS,P2P The method calculates the signal-to-noise ratio, and the signal-to-noise ratio is 43.51, meeting the detection sensitivity requirements, indicating that Charged Aerosol Detector (CAD) Responds well to low concentration samples.

picture 2 10%Concentration level standards - signal-to-noise ratio plot

2.3.3 Linear

9 Each concentration level standard sample was injected separately.1 needle(concentration linear);STD Inject the standard sample into different injection volumes (injection volume is linear), and use concentration and peak area to make a linear equation. The results show that the concentration linear correlation coefficient R² is 0.9980, the linear correlation coefficient R² of the injection volume is 0.9995, indicating that the method is linear.

2.3.4 Repeatability

STD Continuous injection of standards 6 Needle, calculate retention time, peak area and peak height RSD%.

The results show: retention time RSD% is 0.184%, peak area and peak height RSD% are respectively 1.38%and 1.65%, all less than 2.0%, indicating that the method has good repeatability.

picture 3 STD Standard sample chart-repeatability chart

2.3.5 Sample test

Sample injection 10μL, target peak piperidine The detection was good, and the peak area was 23.542, use the concentration linear standard curve to calculate its content, which is 2.9890µg/mL. Other peaks in the sample have higher responses, as shown in the following spectrum:

picture 4 sample diagram - full scale

picture 4 sample picture-enlarged picture

1. Experimental discussion:
2. In the reference method, Waters T3 Chromatographic column, due to conditional restrictions, this test screening uses other C18 Column, final screening using phenomenex Kinetex XB-C18 Basically meets the experimental requirements, but the baseline at the target peak is slightly uneven.
3. This experiment examined several aspects such as specificity, sensitivity, linearity, and repeatability.CAD For the suitability of the sample, the results show that CAD The sensitivity, linearity, repeatability and other results are good and meet the experimental needs.