ChP HPLC Analysis of Uridine, Guanosine and Adenosine in Ban-Lan-Gen Granules on Purospher® STAR RP-18e
Materials
related product
Purospher® STAR RP-18 endcapped (5 µm)
L × I.D. 250 mm x 4.6 mm LiChroCART™ HPLC cartridge, manuCART™ holder required for useused together
Milli-Q® IQ 7000 Ultrapure Water Purification System
output: type 1 water (18.2 MΩ·cm), the most advanced Milli-Q® ultrapure (Type 1) water purification system that is intelligent, intuitive, and green.CONDITIONS
column
Purospher® STAR RP-18e 250x4.6 mm, 5 µm
mobile phase
[A] Water; [B] methanol.
gradient
97%A for 3 min, to 90%A in 17min, to 30%A in 20 min, 30%A for 10 min, 97%A for 10 min
flow rate
0.8 mL/min
pressure
1140 - 1920 psi
column temp.
30 oC
detector
UV @254 nm (DAD)
injection
10 µL
sample/matrix
Standard: 10 µg/mL each compound mixed standard solution in 5% aqueous methanol
Granule Sample: Banlan Gen granules ground, 1.4 g dissolved in 10 mL 5% methanol solution (ultrasonication for 5 min, cool down to room temperature, filtered trough a PVDF filter, fill-up to mark).
Description
General description
Ban-Lan- Gen (Banlangen or Ban Lan Gen) Granules, a famous traditional Chinese herbal medicine, is composed of isatis root. It is used to treat sore throat and dry oropharynx caused by excessive heat in the lungs and stomach. Uridine, Guanosine and Adenosine are the main active ingredients of Banlan Gen granules.
Acceptance Criteria for Standard Solution in the Chinese Pharmacopoeia (ChP) monograph: Theoretical plate number calculated by peak of Uridine: NLT 10000
Analysis Note
The theoretical plate count for Uridine was 16105 (>10000) with a tailing factor of 1.16.
The method′s linear range was determined from 0.1 to 50 µg/mL with R2 between 0.9994-0.9998 for the 3 compounds. The LOD and LOQ of uridine are 0.05 µg/mL and 0.15 µg/mL respectively. The LOD and LOQ of guanosine are 0.09 µg/mL and 0.28 µg/mL respectively. LOD and LOQ of adenosine is 0.04 µg/mL and 0.13 µg/mL separately.
Reproducibilities for a 10 mg/mL standard solution were between 0.83-0.87% RSD for the compounds of interest.
Other Notes
APP_532
Legal Information
Purospher is a registered trademark of Merck KGaA, Darmstadt, Germany