Simultaneous determination of six herbal components in intestinal perfusate by high-performance liquid chromatography.
An effective, accurate and reliable HPLC with UV detection method was developed and validated for quantitation of six components: baicalin, berberine hydrochloride, quercetin, kaempferol, isorhamnetin and baicalein in intestinal perfusate using rotundin as an internal standard. The chromatographic separation was performed on a Welchrom-C(18) column (250 x 4.6 mm i.d. with 5.0 microm particle size) with a mobile phase consisting of acetonitrile, water, phosphoric acid and triethylamine (30:70:0.2:0.1,v/v) at a flow rate of 1.0 mL/min and a UV detection at 270 nm. The method had a chromatographic run time of 30 min and excellent linear behavior over the investigated concentration ranges observed with the values of r higher than 0.99 for all the analytes. The lower limit of quantification of the analytical method was 0.09 microg/mL for berberine hydrochloride, quercetin, kaempferol and baicalein and 0.18 microg/mL for baicalin and isorhamnetin. The intra- and inter-day precisions measured at three concentration levels were all less than 10% for all analytes. The bias ranged from -6.91 to 4.33%. The validated method has been successfully applied to investigate the rat intestine absorption profiles of baicalin, berberine hydrochloride, quercetin, kaempferol, isorhamnetin and baicalein.
Wang
Zhanguo
Z
West China School of Pharmacy, Sichuan University, Chengdu 610041, People's Republic of China.
Du
Qingqing
Q
Qiu
Xuelan
X
Liu
Fang
F
Tan
Fei
F
Lan
Ke
K
Jiang
Xuehua
X
Jiang
Qian
Q
D000818:Animals; D001599:Berberine / Q000032:analysis / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D044950:Flavanones / Q000032:analysis / Q000493:pharmacokinetics; D005419:Flavonoids / Q000032:analysis* / Q000493:pharmacokinetics*; D044948:Flavonols / Q000032:analysis / Q000493:pharmacokinetics; D007408:Intestinal Absorption; D044949:Kaempferols / Q000032:analysis / Q000493:pharmacokinetics; D008297:Male; D011794:Quercetin / Q000032:analysis / Q000493:pharmacokinetics / Q000031:analogs & derivatives; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity
D044950:Flavanones; D005419:Flavonoids; D044948:Flavonols; D044949:Kaempferols; C047368:3-methylquercetin; D001599:Berberine; C038044:baicalin; C006680:baicalein; C006552:kaempferol; D011794:Quercetin