High Performance Liquid Chromatography Determination of flavonoids in semipinnata
Author: Gong Xian-ling, Zhi-Hong Chen, Nian-Ci
【Abstract】 Objective To determine semipinnata of luteolin and apigenin content. Methods ODS C18 analytical column (25
mm × 4.6 mm, 5 μm); mobile phase was methanol -0.4%
phosphoric acid solution (1:1); detection wavelength of 345 nm, flow rate 1 ml * min-1, column temperature at 25 ?.
Results of luteolin linear range 0.02 ~ 0.4 μg, r = 0.999 9,
the average recovery was 96.39% (RSD = 1.95%); apigenin linear range of 0.02 ~ 0.4 μg, r = 0.999 6, the average recovery was 98.10 % (RSD = 0.96%). Conclusion This method is simple, accurate, high sensitivity, good reproducibility can be used
as half of the flag of the determination of two flavonoids.
Key words semipinnata; flavonoids; high-performance liquid
Analysis and Determination of Flavonoids in Pteris semipinnata L. by HPLC
Abstract: ObjectiveTo analyze and determine the content of luteolin and apigenin in Pteris semipinnata L. by HPLC. MethodsThe HPLC system consisted of ODS-C18 column (25 mm ×
4.6 mm, 5 μm), methanol: 0.4% sodium mixture as a mobile phase , detection wavelength at 345nm, 1 ml * min-1 of flow
rate and column temperature at 25 ?. ResultsThe linear range
of luteolin and apigenin were 0.02 ~ 0.4 μg, r = 0.999 9 and
0.02 ~ 0.4 μg, r = 0.999 6 respectively , the recovery were
96.39% (RSD = 1.95%, n = 5) and 98.10% (RSD = 0.96%, n = 5) respectively.ConclusionThe method is simple, accurate, sensitive and reproducible. This method can be used for the quantitative determination of flavonoids in Pteris semipinnata L.
Key words: Pteris semipinnata L.; Flavonoids; HPLC
Pteris Pteris semipinnata L. flag for Pteridaceae Pteris species, widely distributed in Sichuan and the Yangtze River south of the provinces and autonomous regions, bitter, Xin, sexual cool, with evacuation cold, dampness swelling, heat and so the effectiveness of detoxification [ 1]. Semipinnata
entire grass-containing flavonoids [1,2], Fu Yuping et al  examined semipinnata content of total flavonoids, flavonoid compounds with anti-tumor, anti-inflammatory, antibacterial,
antiviral and other effects . In order to further clarify
the semipinnata in flavonoids, this paper analyzed using high performance liquid chromatography determination of semipinnata of luteolin and apigenin content.
An instrument and reagent
1.1 Instrument the U.S. Hewlett-Packard HP 1100 high-
performance liquid chromatography; Mettler's AE 240
electronic analytical balance.
1.2 Standard test drug flavonoids: luteolin (luteolin; 5,7,3 ', 4' TETRA hydroxyflavone), apigenin (apigenin; 5,7,4 ' trihydroxyflavone), provided by Tianjin peak
(content greater than 98%), methanol as a pure
chromatographic, and other reagents were of analytical grade; semipinnata 2002 05 samples collected from the suburbs of Guangzhou, Guangdong Ocean University, Associate Professor Yang Yanjun identification.
2 Methods and Results
2.1 The chromatographic conditions were ODS C18 column for the analytical column (25 mm × 4.6 mm, 5 μm); mobile
phase was methanol -0.4% of the phosphoric acid solution
(1:1); DAD detector, detection wavelength 345nm, flow rate 1
ml * min-1, the column temperature was 25 ?.
2.2 Preparation of solution for the test materials semipinnata crushed herbs, over 40 mesh sieve, saying that the powder obtained is about 2.0 g, plus 20 ml methanol, 80 ?
water bath heated to return the next 2 times, 2 h / times. Filtration, combined filtrate, moved into 25 ml flask, the constant volume, take a small amount of the solution using 0.45 μm microporous membrane filter, as a solution for the test materials, spare.
2.3 Preparation of reference substance solution
Weigh luteolin reference substance 2.50 mg, plus constant volume of methanol to 25 ml, as a reserve liquid. Weigh Apigenin reference substance 2.50 mg, plus constant volume of methanol to 25 ml, as a reserve liquid. Take two kinds of
stock solution of 15ml, plus methanol 45 ml, made of luteolin, apigenin concentrations were 0.02 μg * μl -1 mixture of the
reference substance, as a reference substance solution. Reposted elsewhere in the paper for free download http://
2.4 linear relationship between the study
Precision drawing reference substance solution 1,2.5,5,10,20 μl followed by injection, in accordance with the above-mentioned chromatographic conditions were measured peak area, peak area of luteolin, respectively
68.30,168.95,331.70,698.26,1415.26; apigenin peak area, respectively 74.91,161.53,346.67,743.72,1509.54. To peak area (Y) and quality (X) regression. Obtain the regression equation in Table 1. Table 1 Regression equations and linear range