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Adrenaline Determination by High Performance Liquid Chromatography

How to detect epinephrine by HPLC?

Adrenaline is a hormone and neurotransmitter released by the adrenal glands. Adrenaline will increase the contractility of the heart, dilate the blood vessels of the heart, liver, muscles and bones, and constrict the blood vessels of the skin and mucous membranes. It is a necessary product to save dying people or animals. Its biosynthesis is mainly to first form norepinephrine in medullary chromium cells, and then further through the action of phenylethanolamine N-methyl transferase (PNMT) to make norepinephrine methyl The formation of adrenaline.

1,Analytical equipment required for adrenaline testing

High performance liquid chromatograph: LC3100;
Binary high pressure gradient pump: P3100B;
Autosampler: AS3100;
Column thermostat: CT3100;
Detector: ECD3100 (ampere detector)

LC3100

2,Chromatographic conditions

Column Welch Ultimate XB-C18 4.6x150mm,5μm
Mobile phase A: 5% glacial acetic acid buffer solution (pH value is 3.42, add EDTA disodium salt 0.15mmol/L, sodium heptane sulfonate 2.7mmol/L) B: methanol (A:B=95:5 premix)
Flow rate 1.0mL/min
Column temperature 30℃ Conductivity cell temperature 30℃
ECD mode DC Ampere Voltage 0.65V
Sampling Rate 10SPS Injection volume 20μl

Reference substance

Liquid chromatograph adrenaline reference substance
Compound name Keep time Peak area Peak height Symmetry Theoretical plate number concentration(mg/L)
Adrenaline 5.52 671.164 76.573 1.01 9762 12.300mg/L

Standard curve

Standard curve series concentration(mg/L)
Serial number 1 2 3 4 5
Concentration 0.246 2.460 4.100 6.150 12.300
Overlapping chromatograms at each point of the standard curve
Standard curve linear graph

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