The Application of IP-RPHPLC in Determination of ATP,ADP and AMP in Liver of the Rats Treated by Electric Acupuncture
10.14148/j.issn.1672-0482.2016.0371
- VernacularTitle:I P-RPHPLC 法检测电针后肥胖大鼠肝脏组织中 ATP、ADP 和 AMP 含量
- Author:
Xiao-Lin BI
1
;
Xiao-Qian ZHU
;
Mu XUE
;
Mei-Rong GONG
Author Information
1. 南京中医药大学药学院
- Keywords:
IP-RPHPLC;
ATP;
ADP;
AMP;
the quantitative determination
- From:
Journal of Nanjing University of Traditional Chinese Medicine
2016;32(4):371-374
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To establish an ion-pair reversed-phase high performance liquid chromatography(HPLC) method and determine ATP,ADP and AMP in liver of the rats treated by electric acupuncture.METHODS Separation of ATP,ADP and AMP in the liver tissue samples of rats was performed on a Megres-C1 8 water resistant column(5 μm,250 mm×4.6 mm) by Shimadzu SIL-20AT.A mobile phase was 0.18 mol/L phosphate dihydrogen containing 5% methanol (pH =6.25).The detection wavelength was set at 254 nm and the flow rate was 0.8 mL/min.The injection volume was 20 μL and the column temperature was set at 25 ℃.RESULTS The linear ranges of ATP,ADP and AMP were 1.1~212.2 μg/mL(r =0.999 9), 0.9~180 μg/mL(r =0.999 9) and 0.9 ~ 181.8 μg/mL (r =0.999 7).The average recovery rates of ATP,ADP and AMP were 96.4%,98.3% and 100.8%;and RSD were 2.50%,2.88% and 4.14%,respectively.The RSD of intra-day and inter-day were 0.06%~0.40% and 0.06%~0.69%.The detection limits for ATP,ADP and AMP were 0.1 1,0.09 μg/mL and 0.045μg/mL,respectively.CONCLUSION The established method for the determination of ATP,ADP and AMP in the liver of rats is simple,quick,accurate and reliable,which will lay a foundation for the determination of small molecular bio-active substances in the animal tissues treated by acupuncture intervention.
