The present invention discloses a preparation method of a simple, low-cost, multi-channel microfluidic chemiluminescent paper 
chip and an on-site detection method by using the same. By using a full print mode, a plurality of multi-channel microfluidic chemiluminescent paper chips are printed out on a common A4-size 
filter paper in batch. The printing process includes: printing hydrophobic 
wax patterns in batch; melting 
wax for molding; printing chemiluminescent 
reagent ink patterns in batch; printing oxidase ink pattern in batch; 
cutting the microfluidic chemiluminescent paper chips into certain shape; and performing plastic 
package to the prepared microfluidic chemiluminescent paper chips. An on-site detection method by using the microfluidic chemiluminescent paper 
chip comprises the steps of: placing the plastic packaged microfluidic chemiluminescent paper 
chip into the 
cartridge of a handheld luminescent 
detector; dropwise adding a sample solution into the sample region; then covering the 
cartridge with a lid, and starting the test. The existence and contents of glucose, polyphenols, 
xanthine, 
cholesterol, 
uric acid, and 
hemoglobin are sequentially determined via the size of six 
chemiluminescence peaks sequentially appearing.