Development of a HPLC-MS/MS Method to Determine the 13 Elements of Semen Cuscutae and Application to a Pharmacokinetic Study in Rats
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Abstract:
This study developed a method for simultaneous determination of 13 elements of Semen Cuscutae (quercitrin, quercetin, hyperoside, caffeic acid, chlorogenic acid, luteolin, apigenin, kaempferol, isoquercitrin, cryptochlorogenic acid, isorhamnetin-3-O-glucoside, astragalin, and rutin) in rat plasma using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in the negative MRM mode. The analytes were analyzed with CORTECS®C18 column (4.6 × 150 mm, 2.7 μm) with mobile phases consisting of 0.1% formic acid in water (A) and acetonitrile (B). The intra- and interday precision of the target compounds were expressed as relative standard deviation (RSD) in the range of 0.5%-10.4%, and the accuracy of the target compounds was expressed as relative error (RE) not exceeding ±14.5% for all analytes. In the meantime, the extraction recovery of the target compounds in plasma samples ranged from 87.4% to 106.2% and matrix effect from 81.0% to 115.5%. The established method was successfully accomplished for the pharmacokinetic study of the analytes in rat plasma samples following oral administration of Semen Cuscutae extract, and the pharmacokinetic parameters of seven compounds were obtained.Keywords:
Hyperoside
Chlorogenic Acid
Astragalin
Isorhamnetin
Quercitrin
Ten flavonoids and chlorogenic acid were isolated from the leaves and stems of Cryptocarya alba (Lauraceae). The flavonoids were identified as isorhamnetin. kaempferol, quercetin and their glycosides isorhamnetin-3-O-rhamnoside, isorhamnetin-3-O-galactoside, isorhamnetin-3-O-glucoside, kaempferol-3-O-galactoside, quercetin-3-O-rhamnoside (quercitrin), quercetin-3-O- galactoside (hyperoside), and quercetin-3-O-glucoside (isoquercitrin).
Isorhamnetin
Quercitrin
Hyperoside
Flavonols
Galactosides
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To simultaneously determine the contents of six flavonoids(rutin,hyperoside,astragalin,quercetin,kaempferol and isorhamnetin)in lotus leaves and related tea products,eight lotus leaf samples from different companies were detected by HPLC.The prepared samples were separated by Agilent TC-C18(2)(150 mm × 4.6 mm,5μm)at 25℃ with a step linear gradient using 0.5%formic acid-water(A)and 0.1%formic acid-acetonitrile(B)at a flow rate of 1.0 mL/min.The detection wavelength was 360 nm.Results showed that the concentration ranges of rutin,hyperoside,astragalin,quercetin,kaempferide and isorhamnetin were 1.6-160,1.8- 180,2.16-216,1.4-140,2.12-212 and 1.6-160μg/mL,respectively.Between these ranges,the linearity of the six flavonoids were well-presented(R 2 0.9992)in a certain concentration range.The RSD values for the stability,repeatability,intra-day and inter-day precisions and recovery rates were all less than 2%.Eight lotus leaf samples from different companies were detected by the above method.The content of quercetin was the highest among the six flavonoids,and was highest in Batch No.130802 from Company G.The method used in this study accurately detected the contents of six flavonoids in lotus leaves and tea products,and provided an effective way in which to determine the functional components and quality of lotus leaf products.
Hyperoside
Astragalin
Isorhamnetin
Quercitrin
Lotus effect
Repeatability
Tincture (heraldry)
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Results of seasonal changes of phenolic compounds are presented in the article for leaves of Dasiphora mandshurica grown in culture of the south of the Amur Region. The phenolic compounds were analyzed by the method of a high-performance liquid chromatography. Six glycosides of flavonol (hyperoside, isoquercitrin, rutin, avicularin, quercitrin, astragalin), two aglycones (quercetin and rhamnetin) and tannins (ellagic acid and its glycoside) were found. It was found that phenolic composition of D. mandshurica is constant, but the changes of qualitative composition occur at the expense of minor compounds. The largest number of phenolic components (25) was established in stage the period of bud swelling of the leaf buds and periods of beginning and the mass of a blossoming. A higher total content of phenolic compounds in leaves of D. mandshurica was established in the stages the period of full isolation of leaves (35.3 mg/g), of total aglycones – in the period of ending of a blossoming (0.48 mg/g), and of total flavonols – in the period of mass blossoming (22.2 mg/g). Quercetin, kaempferol, rhamnetin glycosides were found in all stages of development. The largest glycosides of flavonol were found in the phases of the vegetation, budding and blossoming, and aglycones (quercetin and rhamnetin) at the beginning of vegetation and the ending of a blossoming. A fact of contrariety of the dynamics of accumulation of glycosides and their aglycones was revealed. A higher content of most individual phenolic compounds was found as in young leaves in the vegetation and budding, so in mature leaves in the blossoming phase. Avicularin and hyperoside are the predominant glycosides during the growing season. A higher content of the tannins was established in young leaves, ellagic acid dominated in the phase of the vegetation whereas ellagic acid glycoside was the predominant in the phase of the budding.
Hyperoside
Quercitrin
Astragalin
Flavonols
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以上述べた茶葉フラボンに関する成果を要約すれば次の通りである. 1. 2次元ぺーパークロマトグラフによる分析の結果,茶葉中には約23種類のフラボン類が存在し,其の中9種類はkaempferolを,残りはquercetinをアグリコンとする. 2. カラムクロマトグラフ及びクロマトパイルを用いる事により次の11種類のフラボン類を分離決定した,即ち, i. kaempferol, ii. quercetin, iii. quercitrin, iv. astragalin, v. isoquercitirin, vi. kaempferol-3-rhamnoglucaside, vii. rutin. viii. kaempferol-3-rhamnodiglucoside. ix. quercetin-3-rhamnodiglucoside, x. kaempferol-triglucoside, xi. quercetin-triglucoside 3. 以上フラボン類中iv以下の配糖体は水溶性に富み,緑茶及び紅茶の水色の主因をなすものであり,これらの全量は定性的には ix>vii>viii>x>v>xi>vi>iii>iv>ii>i であり,特にix及びxの含量が極めて大きい.
Astragalin
Quercitrin
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OBJECTIVE To study the estimate method of C. chinensis and C. australia. METHOD HPLC was used to determine the contents of four kinds of flavones of C. chinensis and C. australia growing on different hosts. RESULT C. chinensis and C. australia growing on different hosts both had hyperoside, quercetin, kaempferol and isorhamnetin. The content range of hyperoside was 2.790-6.502 mg/g and was higher than other flavones. The content ranges of quercetin, kaempferol and isorhamnetin were 0.025-0.176 mg/g, 0.001-0.213 mg/g and 0.001-0.077 mg/g, respectively. CONCLUSION The contents of hyperoside and quercetin are higher in C. chineasis than in C. australia. The contents of kaempferol and isorhamnetin are lower in C. chinensis than in C. australia. The hosts influence flavones content of C. chinensis and C. australia.
Hyperoside
Isorhamnetin
Flavones
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Isorhamnetin
Hyperoside
Quercitrin
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The production techniques of Houttuynia cordata Thunb steeping liquor had been optimized as follows by use of HPLC and UV with flavonoids(kaempferol-3-O-glucoside,hyperoside,quercitrin,rutin,quercetin,kaempferol and isorhamnetin) and phenolics content as assaying indexes to investigate the effects of alcohol concentration,liquid-solid ratio and steeping time: the best alcohol concentration was 55 %vol,liquid-solid ratio was 10∶1(mL/g),and the best steeping time was 40 d.After 40 d storage,the content of kaempferol-3-O-glucoside,hyperoside,quercitrin,rutin,quercetin,kaempferol,isorhamnetin and phenolics in steeping liquor could theoretically reach up to 333.6 mg/L,157.4 mg/L,67.3 mg/L,137.3 mg/L,44.8 mg/L,161.7 mg/L,54.6 mg/L and 1921.6 mg/L,respectively.
Hyperoside
Quercitrin
Isorhamnetin
Steeping
Houttuynia cordata
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Daily changes in the composition and content of phenolic compounds in the aerial parts Potentilla fruticosa L., growing in culture of the Novosibirsk Region, using the method of high performance liquid chromatography (HPLC) was investigated. Six glycosides of flavonol (hyperoside, isoquercitrin, rutin, avicularin, quercitrin and astragalin), two aglycones (quercetin and kaempferol) and tannins (ellagic acid and its glycoside) were identified. Established that phenolic composition of P. fruticosa is the time of day remains constant. The highest total content of phenolic compounds in the leaves (28–29 mg/g of absolutely dry solid matter) and stems (4–4,5 mg/g) of P. fruticosa found in the evening and at night. In the flowers, in contrast, the maximum content of phenolic compounds was observed in the middle of the day (42 mg/g). The level of variability of the total content of phenolic compounds was evaluated as the lowest for leaves and flowers (СV = 10–11%). It was found a threefold increase in free aglycones - quercetin and kaempferol in the leaves with 3 to 6 o'clock in the morning on the background of reduction of the relevant glycosides in 1,5–2,5 times. Maximum accumulation of glycosides of quercetin and minimum content of free quercetin in flowers was observed at 12 o'clock. It is shown predominant accumulation of ellagic compounds quercitrin, component 10, hyperoside, isoquercitrin and rutin in the leaves in the evening and at night, in the flowers – in the middle of the day. The level of variability content of most of the individual components in the above-ground organs of P. fruticosa during the day was estimated as high (СV ≥ 21%).
Hyperoside
Quercitrin
Astragalin
Potentilla
Ellagic Acid
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This paper describes a validated high-performance liquid chromatographic (HPLC) - photodiode array (PDA) detection method to quantitate five flavonol components as markers; rutin, quercitrin, quercetin, kaempferol and isorhamnetin for use in the quality control of Ginkgo biloba dosage forms.Separation was achieved using a minibore Phenomenex Luna 5microm C(18) (2) column with dimensions 250 x 2.00mm at 45 degrees C with a one step linear gradient using acetonitrile:formic acid (0.3%) at a flow rate of 0.4ml/min.The limits of quantitation for the flavonols were 2.76, 0.77, 1.11, 1.55 and 1.03microg/ml for rutin, quercitrin, quercetin, kaempferol and isorhamnetin, respectively. This method is linear over concentration ranges of 3-26microg/ml for all flavonols. Recoveries for rutin, quercitrin and kaempferol were above 94% while quercetin and isorhamnetin had average recoveries of 83% and 76%, respectively. Intraday precision did not exceed 6% and with-in day precision was better than 12% for all compounds.A suitable method was developed to identify and quantitate five relevant flavonol marker compounds and was successfully used to assay some commercially available solid oral dosage forms of Ginkgo biloba .
Quercitrin
Isorhamnetin
Flavonols
Hyperoside
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To establish a quick and easy method of qualitative and quantitative analysis for determining the content of hyperoside,quercetin,kaempferol and isorhamnetin in lotus leaves by HPLC.Results:Under conditions of Gemini C18 for the chromatographic column and 40℃ for column temperature,with methanol-0.1% phosphoric aicd(55∶45,v∶v)of the mobile phase and 1mL/min of flow velocity,360 nm for the detection wavelength,the linear ranges of hyperoside,quercetin,kaempferol and isorhamnetin were 1.5~48.0μg/mL,1.0~32.0μg/mL,1.0~32.0μg/mL,0.7~24.0μg/mL,respectively,the recovery was 90%~103%and the precision was 3.5%~4.2%,the contents of four kinds of flavonoids in lotus leaves were respectively 1.94%,0.092%,0.025% and 0.021%.The proposed method was suitable for the determination of hyperoside,quercetin,kaempferol and isorhamnetin in lotus leaves.
Hyperoside
Isorhamnetin
Lotus effect
Phosphoric acid
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