Cultivation of high-performance liquid chromatography in the second Shegan irigenin content
【Abstract】 Objective high-performance liquid chromatography
in the second planting Shegan irigenin content, examining the inherent quality of its ingredients. Methods using high
performance liquid chromatography column was TSK ge1 ODS 80 TS250 mm × 4.6 mm; mobile phase was methanol phosphate
solution (pH = 3): 53%: 47%; a flow rate of 1 ml / min; detection wavelength was 266 nm; column temperature was 35 ?.
The results of sub-irigenin in the range of 0.0118 ~ 0.118 μg
chromatographic peak area within the linear relationship (r = 0.999 9); The average recovery was 99.84%, RSD of 2.059%, sub-
irigenin content of 0.1252%. Conclusion The method is simple,
accurate, sensitive, cultivated in the sub-Shegan irigenin the
content is more than 0.10%, in line with "Chinese Pharmacopoeia" requirement.
Key words high-performance liquid chromatography Shegan times irigenin
Abstract: ObjectiveTo determine irisflorentin in
cultivated Belamcanda chinensis (L.) DC by HPLC, and to inspect the quality of its ingredients.MethodsUsing HPLC with TSK ge1 ODS Column80 (250 mm × 4.6 mm), the mobile phase was
composed of a mixture of methanol-phosphoric aicd aqueous
soluton (pH = 3): 53%: 47%, the flow rate was 1 ml / min, the UV detection wavelength was set at 265 nm, and the temperature was at 35 ?. ResultsIrisflorentin showed linear relationship with peak areas in the range of 0.0118 ~ 0.118 μg (r = 0.999
9). The average recovery was 99.84%, RSD was 2.509%. ConclusionThe method is simple, rapid and accurate.The quantity of irisflorentin in Cultivated Belamcanda Chinensis (L.) DC exceeds 0.10% and meets the Chinese Pharmacopoeia.
Key words: HPLC; Belamcanda chinensis (L.) DC;
Shegan yes Iridaceae Belamcanda Shegan drying plant roots, for commonly used traditional Chinese medicine before they set out in the "Shen Nong's Herbal Classic", as beneath contempt. Bitter, cold, portal triad, lung. With Qingrejiedu,
Liyan expectorant efficacy, mainly for sore throats, and lungs carbuncle, sputum cough asthma embolism, for the treatment of sore throat Houbi to the drug . Shegan of isoflavones have anti-inflammatory, anti-virus, bacteria and eliminate free
radicals. Times irigenin for Shegan in one of the active ingredient. With the Shegan dwindling wild resources, in order to conserve resources, in recent years Shegan biology, introduction and cultivation of more and more research. In this paper, were measured by HPLC in the second cultivation Shegan irigenin content. According to measurements and to carry out quality evaluation.
An equipment and reagents
Diana P680 high-performance liquid chromatography,
UVD170U detector, quaternary gradient pump, HKZJZ-830171BAF;
Diana chameleon HPLC data processing system; Column: TSK ge1 ODS 80TS (250 mm × 4.6 mm); pre - treatment column: Allechc18
(United States); AP-9908S FILTER (Automatic Science Lastrument Co.Ltd); BP-211D type one hundred thousandth analytical
balance (West Germany Sartorius); methanol, phosphoric acid: analytically pure; re-distilled water.
Reference substance: sub-irigenin (Lot No. :1557-200101
20 mg / sticks) were purchased from Chinese medicine and biological products.
Herbal: 2005 11 12 corporations Fengxian Shegan
taken from GAP planting base of cultivation Shegan by Professor Liu Hegang hospital identification, the original plants Iridaceae plants Shegan Belamcanda chinensis (L.) DC.
2 Methods and Results
2.1 The conditions of chromatographic separation the mobile phase: methanol phosphate solution (pH = 3): 53%:
47%; flow rate: 1 ml / min; column temperature was 35 ?.
Detection wavelength: 266 nm; injection volume: 5 μl .
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2.2 Preparation of reference substance solution precision that take time irigenin reference substance 1.47 mg, buy 25 ml flask, add sufficient amount of methanol dissolved, will allow to scale, may reference substance concentration of 58.8 μg /
ml, precision drawn the above reference substance solution 1 ~ 5 ml flask, add methanol to constant volume scale, dubbed in 11.76 μg / ml of the solution, as a reference substance solution.
2.3 Preparation of solution for the test materials after drying herbs by crushing, over on the 4th screen, take samples of about 0.1 g, precision that set, and placing a plug conical bottle, precision adding methanol 25 ml, said that given the weight, heat reflux 1 h , put cold, and then said that given
the weight loss by using methanol to complement the weight, shake, porous membrane filtration, the filtrate continued to take for the test items as a solution .
2.4 A methodological survey
2.4.1 standard curve and linear in the range of precision drawing the above-mentioned reference substance solution
1,2,4,6,8,10 μl injection, according to the above
chromatographic conditions, to HPLC determination. With reference standards into the sample volume and peak area
mapping, linear relationship, shown in Figure 1. Regression equation was Y = 0.829 3X-0.060 4, r = 0.999 9. Linear range
between 0.011 8 ~ 0.118 μg.
2.4.2 Experimental precision precision drawing reference substance solution 5 μl sample to repeat 5 times,
respectively, record the peak area, calculated RSD of 1.3%, indicating good precision instruments. The results in Table 1. Table 1 precision experiment (omitted)
2.4.3 Stability test for the test products according to the preparation of sample solution was prepared by cultivation of Shegan a solution, respectively 0,2,4,6,8 h each sample 5 μl. The results in Table 2. Calculated RSD was 1.4%, indicating that the sample solution was stable for at least 8
h. Table 2 Stability Experiment (abbreviated)
2.4.4 Reproducibility experiment precision that take the same group of Shegan powder 5 copies, according to test materials for the preparation of sample solution prepared
sample solution, injection 5 μl, measured times irigenin
content. The results in Table 3. Calculated RSD of 1.1%, indicating good reproducibility of this method. Table 3 reproduce the experiment (omitted)
2.4.5 Experimental precision recoveries that were known
to take five samples of powder content in each Shegan 0.1 g, were added to sub-irigenin reference substance solution, 2 ml (concentration of 58.8 μg / ml), according to sample
preparation method Preparation of sample solution for the test
items to measure times irigenin were calculated recovery rate, the average recovery was 99.84%, RSD of 2.509%, proved that the method stable and reliable. The results in Table 4. Table 4 The average recovery experiment (omitted)
2.5 Determination of the sample for the test items
according to the above preparation method of sample solution and the chromatographic separation conditions, sample injection 5 μl, measured Phoenix Mission GAP planting base three different groups of batch cultivation in the sub-Shegan
irigenin of content. The results shown in Table 5 and Figure 2 ~ 3. Table 5 ingredients in the second irigenin content (abbreviated)
HPLC method used in this experiment shows the times measured Shegan irigenin content, good reproducibility and
high recovery rate, stable and reliable, this method can be used to determine the times Shegan irigenin content. According to measuring results in the second planting Shegan irigenin content of "0.10%, in line with" Chinese Pharmacopoeia
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