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The aim of this study was to determine the content of rutin in Hemidesmus indicus and to optimize the high-performance thin-layer chromatography method. The method was validated in compliance with the International Council for Harmonisation guidelines Q2 (R1) for parameters such as linearity, accuracy, precision, robustness, limit of detection, and limit of quantitation. A Box-Behnken design and response surface methodology has been used to investigate the impact of independent variables on the response. Three independent variables, mobile phase composition (% v/v), mobile phase volume (mL), and duration of saturation (min), were studied. Rutin was verified, and its content was determined using a validated high-performance thin-layer chromatography method with good linearity within the range of 200-1000 ng spot -1 with r 2  = 0.9998 and correlation coefficient with calibration curve equation y = 0.0297x + 0.0001. The average percentage recovery values varied from 99.03 to 101.15 and 98.88 to 100.12%, respectively, for in-house and marketed mother tincture). The peak area determination at three different concentration levels shows low values of percentage relative standard deviation (<2%) for inter-day (0.04-0.06) and intra-day (0.04-0.05) precision of rutin. The average content of rutin in extract and marketed mother tincture was 229 ± 0.57 and 210 ± 0.57 μg g -1 . The proposed method was simple, precise, and accurate for the determination of rutin with frequent quality control assessment of H. indicus., (© 2021 John Wiley & Sons, Ltd.)

The Commelina erecta L. (C. erecta) also known as erva-de-santa-luzia is reported by local population to have medical properties against some pathological conditions. In this study, two extracts of C. erecta leaves (aqueous and ethanolic) were phytochemically analysed and evaluated for their in-vitro antioxidant activities by DPPH, TBARS, NO assays and cell viability assays. The ultra-high performance liquid chromatography followed by tandem mass spectrometry analysis showed the presence of rutin and caffeic acid in aqueous and ethanolic extract. The total polyphenols in aqueous and ethanolic extracts found were 142.7 ± 3.0 and 123.1 ± 5.8 μg/mL of GAE, respectively. The ethanolic extract (5 mg/mL) inhibits TBARS by 33.8%, and the aqueous extract (5 mg/mL) exhibited scavenger property against nitric oxide derivatives to an extent of 77.8%. In cell culture, both extracts improved cell survivability under H2O2 induced oxidative stress. Thus, C. erecta extract is a good candidate to become a phytotherapic medicine.

Alternanthera sessilis (red) (ASR) is an edible herbal plant with many beneficial health effects. This study aimed to investigate the antioxidant components and antioxidant activities of the edible leaves and stems of ASR extracted using solvent of varying polarities namely water, ethanol, ethyl acetate and hexane. ASR leaf extracts showed higher in both antioxidant components and activities than the stem extracts. Among the antioxidant components, the ethanol leaf extract showed higher phenolic (77.29 ± 1.02 mg GAE/g extract) content while the ethyl acetate leaf extract was rich in flavonoids (157.44 ± 10.19 mg RE/g extract), carotenoids (782.97 ± 10.78 mg BE/g extract) and betalains (betanin: 67.08 ± 0.49 mg/g extract; amaranthin: 93.94 ± 0.68 mg/g extract and betaxanthin: 53.92 ± 0.88 mg/g extract). Nevertheless, the ethanol leaf extract showed the highest DPPH radical scavenging activity and ABTS radical cation scavenging activity. It also exhibited highest ferric reducing activity among all the extracts. Four polyphenolic compounds from ASR leaf, namely ferulic acid, rutin, quercetin and apigenin, were identified and quantified using ultra high performance liquid chromatography. The existence of these compounds was further verified using tandem mass spectrometry. These current results indicate that ASR leaf particularly the ethanol extract has the potential to be exploited as a source of natural antioxidants., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Aim of this work was to develop a validated high performance liquid chromatography method for the analysis of extracts and final products of St. John's wort, according to international guidelines for bioanalytical method validation. Chromatographic separation was performed on a C18 column with a combination of gradient and isocratic steps; the mobile phase composed of ammonium acetate solution (pH 4.5; 10 mM), acetonitrile and methanol. Quantification and method validation was performed using extract spiked with external reference standards of chlorogenic acid, rutin, hyperoside, isoquercitrin, quercetin and hypericin. Validation study revealed that trans-chlorogenic acid is partially transformed into its cis-isomer during analysis. The method showed good linearity, precision and accuracy. Hyperforin was completely unstable. All other ingredients were stable at -18°C and after three freeze-thaw cycles, while stability of most ingredients was limited at room temperature and 4 - 8°C; quercetin was the most unstable one. The major ingredients of methanolic extracts, infusions and final products of Hypericum perforatum were completely resolved and quantified. Beyond its potential usefulness in the analysis of St. John's wort products, this study addresses the issue of validation from the perspective of the field of bioanalysis and reveals the wealth of critical information which can be derived., (© The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

A simple, rapid, sensitive and selective high performance liquid chromatographic (HPLC) method with ultraviolet detection has been developed for simultaneous determination of ascorbic acid and rutin in pure forms and pharmaceutical dosage form. HPLC separation was performed on Phenomenex C18 analytical column with 0.1% v/v acetic acid in water and acetonitrile (75:25, v/v), as mobile phase. The separation was done at ambient temperature with flow rate of 1mL·min(-1) in isocratic mode. HPLC measurements were carried out using ultraviolet detection wavelength at 257nm. The average retention times were 2.72 and 7.00min for ascorbic acid and rutin, respectively. The calibration plots were constructed over the concentration range of 5.0-30.0 for ascorbic acid and 10.0-60.0μg·mL(-1) for rutin. The limits of detection were 1.06 and 1.89μg·mL(-1) and limits of quantification were 3.54 and 6.31μg·mL(-1) for ascorbic acid and rutin, respectively. The proposed HPLC-UV method was successfully applied for determination of ascorbic acid in its tablets and for simultaneous determination of the studied drugs in their laboratory prepared mixtures and in pharmaceutical formulation. Statistical comparisons of the results with the reference method show an excellent agreement and indicate no significant difference in respect to accuracy and precision., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Malus domestica is the most widely cultivated fruit tree and is well known for its therapeutic value. Apple leaves are known to contain phenolic compounds but the nature of these has not been explored to the same extent as in apple fruit. A simple, rapid and sensitive ultra-performance liquid chromatography-diode array detection (UPLC-DAD) quantification method has been developed. Total polyphenol and flavonoid contents, as well as the antioxidant activity of golden and royal apple leaves were evaluated., Results: Four compounds, namely rutin, 3-hydroxyphloridzin, phloridzin and quercetin-3-O-arabinoside were identified by UPLC. The separation was achieved in less than 7 min. Total polyphenol and flavonoid contents were found to be slightly higher in apple golden variety than royal variety. The IC50 values determined by the DPPH assay were 49.94 µg mL(-1) for golden apple leaves and 43.89 µg mL(-1) for royal apple leaves. IC50 values determined by the ABTS assay were 47.10 and 66.53 µg mL(-1) for golden and royal apple leaves, respectively. Antioxidant activity was determined as 24.45 and 21.15 mg ascorbic acid g(-1) for golden and royal apple leaves, respectively, by using the FRAP assay., Conclusion: This study showed that apple leaves (both varieties) contain considerable amounts of polyphenols and flavonoids and are also a promising source of phloridzin., (© 2015 Society of Chemical Industry.)

Introduction: Peroxynitrite is involved in the pathogenesis of a number of significant diseases. Peroxynitrite scavengers thus have potential application in understanding and treating these diseases. It is, therefore, important to establish screening methods able to rapidly identify peroxynitrite scavengers from herbal plants., Objective: To develop effective and easily operable screening methods for identifying peroxynitrite scavengers in complex matrices, including Chinese herbal medicines., Methods: Two simple and efficient screening methods have been developed for the identification of natural peroxynitrite scavengers in Flos Lonicerae Japonicae (FLJ). Method I used HPLC-DAD-(luminol-peroxynitrite)-CL techniques combined with Q-TOF MS/MS analysis, while Method II used pre-column reaction of the sample with peroxynitrite, followed by HPLC separation and Q-TOF MS/MS analysis., Results: Five peroxynitrite scavengers (neochlorogenic acid, chlorogenic acid, 3,4-O-dicaffeoyl quinic acid, 3,5-O-dicaffeoyl quinic acid and 4,5-O-dicaffeoyl quinic acid) were identified in FLJ using Method I. Besides the compounds identified using Method I, three additional peroxynitrite scavengers (rutin, isoquercitrin and luteoloside) were identified using Method II., Conclusion: The two new methods proved to be complementary and the use of these methods should allow rapid detection of peroxynitrite-scavenging natural products from FLJ and other complex matrices., (Copyright © 2015 John Wiley & Sons, Ltd.)

To control the quality of Tibetan medicine Saussureae obvalltae, the quality control method and standard were established in this study. The water content, total ash, acid-insoluble ash and ethanol-soluble extractives of Saussureae obvalltae were determined according to the methods recorded in appendix of Chinese Pharmacopeia(2010 edition). Microscopical identification was performed for the plant and powder of the leaves. The TLC method was established, with chlorogenic acid and rutin as control substances, and a mixture of acetate-btuanone-formic acid-water (10∶6∶1∶2) as developing solvent and silica gel G as thin layer plate. Agilent ZORBAX Eclipse XDB-C₁₈ (4.6 mm×250 mm, 5 μm) column was adopted and eluted with the mobile phase of acetonitrile-0.1% phosphoric acid (17∶83) in a gradient mode at a flow rate of 1.0 mL•min⁻¹. The column temperature was 40 ℃ and the detection wavelength was 350 nm. As a result, the plant and leaves of Saussureae obvalltae of different origins showed constent microscopic features. Chlorogenic acid, rutin and the other constituents were well separated on TLC detected under the sun light. According to the results of the methodological study, chlorogenic acid and rutin were in good linear correlation in the ranges of 0.119 2-0.715 4 μg(r=0.999 9) and 0.160 7-0.964 4 μg(r=1.000), and the average recoveries were 105.4% (RSD 1.4%) and 99.50% (RSD 0. 91%), respectively. The content of ethanol-soluble extractives, water content, total ash and acid-insoluble ash were 26.01%-31.59%, 7.16%-8.04%, 8.46%-11.14%,0.50%-1.87%, respectively. According to the study, the established method was specific and accurate, which could be used for the quality control of this drug., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Hibisci mutabilis Folium (HMF), the dried leaf of Hibiscus mutabilis (Malvaceae), is commonly used in traditional Chinese medicine. This article aimed to establish a high-performance liquid chromatography method for the determination of rutin and isoquercitrin contents in the HMF, and to compare the content variation in all the samples, including nearly withered yellow leaf, in different collection periods, so as to provide research basis for the quality evaluation and determination of the optimal collection period. A reversed-phase HPLC separation method was employed, with a BDS Hypersil C18 column (4.6 m × 250 mm, 5 μm), under the following conditions: acetonitrile-0.3% phosphoric acid (15:85, v/v) solution as the mobile phase, flow rate 1.0 mL/min at 30°C and detection wavelength 254 nm. The calibration curves for rutin and isoquercitrin were linear over the range of 1.5-48 and 0.25-8 μg/mL, and the average recoveries were 99.92 and 100.45% (RSD: 2.39% and 2.11%, respectively)]. Based on the analysis results, it was found that contents of rutin and isoquercitrin in HMF (mature green leaf) harvested in different periods had significant difference, and reached the highest in mid-December. It was also found that the contents of the two components in the mature green leaf were much higher than those in the nearly withered leaf from the same collection period. In conclusion, the results indicated that the HPLC method was easy-to-operate and precise, and could be applied for the determination of rutin and isoquercitrin contents in the HMF. The experimental data also showed that early winter should be the most suitable collection period for HMF., (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

A new ultra-performance liquid chromatography (UPLC) using conventional and chemometric calibrations was improved for the simultaneous estimation of chlorogenic acid (CA) and rutin (RUT) in leaves of Ribes L. species (R. rubrum, R. biebersteinii, R. nigrum, R. uva-crispa, R. alpinum, R. orientale, R. multiflorum and R. anatolica). The UPLC separation for CA and RUT in samples were performed on a Waters UPLC BEH phenyl column (100 mm × 1.0 mm i.d., 1.7 μm) and mobile phase consisting of acetonitrile and 0.1 M formic acid buffer, pH = 3.77 (15:85, v/v) containing 1.0 mL triethylamine in 1,000 mL mobile phase. Multi-wavelength UPLC chromatograms of CA and RUT in calibration and plant samples were obtained by the photodiode array (PDA) detection at the wavelength set from 290 to 360 nm with the interval of Δλ = 10 nm. Conventional UPLC-single and chemometric calibrations were subjected to the analysis of the related compounds. By using UPLC data, conventional and chemometric calibrations in the linear concentration range between 2.5 and 40.0 μg/mL for all compounds were applied for the simultaneous quantitative analysis of CA and RUT in plant samples of seven different Ribes species., (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Introduction: Flavonoids are polyphenolic compounds found ubiquitously in foods of plant origin. They are commonly extracted from plant materials with ethanol, methanol, water, their combination or even with acidified extracting solutions. The disadvantages of these methods are the use of high quantity of organic solvent, the possible loss of analytes in the different steps and the laborious process of the techniques. In addition, the complexity of the phenolic mixtures present in plant materials requires a preliminary clean-up and fractionation of the crude extracts., Objective: To develop a hollow fibre liquid phase micro-extraction (HF-LPME) method for a one step clean-up and pre-concentration of flavonoids., Methodology: Two flavonoids (catechin and rutin) has been extracted by HF-LPME and analysed by HPLC. The related driving force for the liquid membrane has been studied by means of facilitated and non-facilitated transport. Different ionic and non-ionic water insoluble compounds [trioctylamine (TOA), tributyl phosphate (TBP), trioctylphosphine oxide (TOPO) and methyltrioctylammonium chloride (aliquat 336)] were used as carriers. The liquid membrane was constituted by a solution of n-decanol in the presence or absence of carriers., Results: Maximum enrichment factors were obtained with n-decanol/aliquat 336 (20%) as organic liquid membrane, sodium hydroxide (NaOH) (0.1 M) as donor solution, sodium chloride (NaCl) (2 M) as acceptor solution and 3 h as extraction time. Under these conditions, good results for validation parameters were obtained [for linearity, limit of detection (LOD), limit of quantitation (LOQ) and repeatability]., Conclusions: The developed method is simple, effective and has been successfully applied to determine catechin and rutin in ethanolic extracts of faba beans., (Copyright © 2015 John Wiley & Sons, Ltd.)

The present study is to determine the flavonoid glycosides, terpene lactones, biflavones, gingko acid and procyanidins of ginkgo pollen. UPLC-TQ-MS technology was used for the determination of 24 kinds of resource chemical composition in ginkgo pollen qualitatively and quantitatively. The results shows that the contents of rutin, quercetion 3-O-[4-O-(α-L-rhamnosyl )-β-D-glucoside] and kaempferolis were 120.9, 114.0, 222.1 μg x g(-1). In this paper, the contents of 24 kinds of chemical components of ginkgo pollen were determinated by UPLC-TQ-MS for the first time. This method is simple and quick, which will be benefit for recycling utilization of ginkgo pollen.

Objective: To set up the quality standard of Chimonanthus nitens leaf., Methods: Five compounds were identified by double-development thin layer chromatography, the plate was colorized with 1% vanillin-H2 SO4 solution and warmed up in 110 degrees C; The content of four compounds was determined by HPLC, the chromatography separation was performed on the Elite hypersil ODS2 (250 mm x 4.6 mm, 5 microm) column with gradient elution. The mixture of acetonitrile and water as the mobile phase at a flow rate of 1.0 mL/min was used, the detection wavelength was 344 nm and the column temperature was set at 40 degrees C., Results: The TLC identification was highly specific and spots were clear. In HPLC method, the calibration curve was linear in the range of 5.665 - 56.65 microg/mL for scopoletin (r = 0.9999), the average recovery was 97.06% and RSD was 1.27%; The calibration curve was linear in the range of 2.4312 - 24.312 microg/mL for isofraxidin (r = 0.9999), the average recovery was 102.86% and RSD% was 0.93%; The calibration curve presented a linear range from 2.444 - 24.44 microg/mL for scoparone (r = 0.9999), the average recovery was 102.18% and RSD was 1.81%. The standard curve presented a linear range from 9.012 - 90.12 microg/mL for rutin (r = 0.9992), the average recovery was 104.44% and RSD was 4.2%., Conclusion: These methods are reproducible, sensitive and simple and can be used to control the quality of Chimonanthus nitens leaf.

An optimized microwave-assisted extraction (MAE) method and RP-HPLC method were developed for the simultaneous extraction and determination of rutin, forsythiaside A, and phillyrin in the fruits of Forsythia suspensa. The key parameters of the open-vessel MAE process were optimized. A mixed solvent of methanol and water (70:30, v/v) was most suitable for the simultaneous extraction of the three components. The sample was soaked for 10 min before extraction. The optimized conditions were: microwave power 400 W, temperature 70°C, solvent-to-material ratio 30 mL/g, and extraction time 1 min. Compared to conventional extraction methods, the proposed method can simultaneously extract the three components in high yields and was proved to be a more rapid method with a lower solvent consumption. The optimized HPLC-photodiode array detection analysis was validated to have good linearity, precision, accuracy, and sensitivity. The developed MAE followed by RP-HPLC is a fast and appropriate method for the simultaneous extraction and determination of rutin, forsythiaside A, and phillyrin in the fruits of F. suspensa., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Heartsease, also known as wild pansy (Viola tricolor L.), contains considerable amounts of polyphenols: 109.32 ± 1.29 mg of Gallic acid equivalent/g of extract. This study investigates their phytoconstituents and antioxidant capacity and validates a method for the quantification of rutin in the crude extract of the flowers of V. tricolor and in the extract incorporated in gel. Much better antioxidant capacity was found for the extract [inhibition concentration (IC50) of 16.00 ± 0.78 µg/mL] than the standard ascorbic acid (IC50 of 16.57 ± 0.95 µg/mL); these excellent results may be attributable to the amounts of polyphenols, flavonoids and condensed tannins. The high-performance liquid chromatography method for the quantification of rutin in the extract and gel was linear, sensitive, precise, specific, accurate and robust. This validated method can be used to control the quality of the extract and the gel.

Objective: To Set up a method for determining the contents of the five flavonoids simultaneously in the HuangKui capsule and analyze their specific chromatograms., Methods: HPLC method was used. The analytical column was Thermo scientific Hypersil GOLD (250 mm x 4.6 mm, 5 microm). The mobile phase was composed of acetonitrile (A) and 0.2% orthophosphoric acid (B) with gradient elution. The flow rate was 0.8 mL/min and detection wavelength was 360 nm. The column temperature was maintained at 30 degrees C., Results: Contents of the five flavonoids (Rutin, Hyperoside, Isoquercitrin, Myricetin, Quercitrin) had good resolution with the correlation coefficients exceed 0.9999 and the average percent recovery lied in 98.46% to 100.33%. The chromatograms of the HuangKui capsule shared 15 common peaks in which 5 of them were recognized by the reference standard. Chromatograms of 10 lots of HuangKui capsule were analyzed with the similarities over 0.95., Conclusion: The proposed method of contents determination and chromatogram analysis has strong characteristic and specificity. This method is fast, easy and reliable, and can be applied for quality control of the preparation.

Introduction: Dianthus versicolor is used in traditional Mongolian medicine against liver impairment. Fractions enriched in flavone-di- and triglycosides were shown to enhance bile secretion. Therefore, reliable and accurate analytical methods are needed for the determination of these flavonoids in the crude drug and extracts thereof., Objective: To provide a validated HPLC-DAD (diode array detector) method especially developed for the separation of polar flavonoids and to compare the data obtained with those evaluated by UV spectrophotometry., Methodology: Separations were carried out on an Aquasil® C₁₈-column (4.6 mm × 250.0 mm, 5 µm) with a linear gradient of acetonitrile and water (adjusted to pH 2.8 with trifluoroacetic acid) as mobile phase. Rutoside was employed as internal standard with linear behavior in a concentration range of 0.007-3.5 mg/mL. Accuracy was determined by spiking the crude drug with saponarin resulting in recoveries between 92% and 102%., Results: The method allows the quantification of highly polar flavonoid glycosides and the determination of their total content. For saponarin a linear response was evaluated within the range 0.007-3.5 mg/mL (R²  > 0.9999). It was proven that threefold sonication represents a time-saving, effective and cheap method for the extraction of the polar flavonoid glycosides. The contents determined by HPLC were shown to be in agreement with those obtained employing UV spectrophotometry., Conclusion: The study has indicated that the newly developed HPLC method represents a powerful technique for the quality control of D. versicolor. Ultraviolet spectrophotometry may be used alternatively provided that the less polar flavonoids are removed by purification., (Copyright © 2011 John Wiley & Sons, Ltd.)

Tobacco consists of at least 3,800 chemical constituents. Among them, rutin is an important polyphenolic secondary metabolite in tobacco, which has positive actions such as antiallergic, anti-inflammatory and vasoactive, antitumor, antibacterial, antiviral and anti-protozoal properties. A high performance liquid chromatography method was used to analyze rutin in tobacco and filters, mainstream smoke, and burned ash of ten varieties of cigarettes made in China. The chromatographic analysis was performed on a Hypersil ODS2 column with a gradient elution of acetonitrile and water at a flow rate of 1.0 mL/min. Detection was carried out at 350 nm using a photodiode array detector. The calibration curves for the determination of analytes showed good linearity over the investigated ranges (R2 > 0.9998). Precision and reproducibility were evaluated by six replicated analyses, and the R.S.D. values were less than 0.59% and 1.53%. The recoveries were between 98.47 and 100.84%. Under the optimized conditions, namely 45 mL/g of solvent to solid ratio, 30 min of extraction time and 200 W of ultrasound power, the concentrations of rutin in tobacco and filter, mainstream smoke, burned ash of different brands cigarettes were 10.20-63.98, 0.10-0.32, 0.06-0.16 and 0 μg/per cigarette, respectively.

A high performance liquid chromatographic (HPLC) method was developed to determine the contents of seven compounds in Lonicera japonica and Folium Lonicerae with maximum wavelength conversion program, and analyze the content differences between them. The separation was performed on an Agilent Eclipse Plus C18 column (250 mm x 4.6 mm, 5 microm) operated at normal temperature with the gradient elution by two mobile phases of water (including 0.3% (v/v) formic acid) (A) and acetonitrile (B) at a flow rate of 1 mL/min. The maximum detection wavelength was set at 330 nm and 350 nm by conversion. The contents of chlorogenic acid and caffeic acid in Lonicera new leaves were 2.572% and 1.498 per thousand respectively, both higher than those cited in Chinese Pharmacopoeia, indicating that Lonicera new leaves have the necessity to be further studied and developed. This method is simple, rapid and highly sensitive. It is suitable for the simultaneous determination of the seven compounds and the quality control of Lonicera japonica.

Gynostemma pentaphyllum (Thunb.) Makino, also known as jiaogulan, has been shown to have antioxidant, antiproliferative, and anti-inflammatory activities. Flavonoid is considered a major contributor for these beneficial effects. To obtain chemical patterns of flavonoids in G. pentaphyllum of different genotypes (di- versus tetraploids) and different parts (leaf versus whole plant) of plants, the extraction condition was optimized and a fingerprinting approach was established by means of high-performance liquid chromatography coupled with diode array detection and mass spectrometry (HPLC-DAD-MS). Eight flavonoids were identified, among which rutin and quercetin were quantified. The highest levels of rutin and quercetin were 23.03 and 12.10 mg/g, respectively, observed in the diploid leaf sample 2L3 and 2L2, while the lowest levels of rutin and quercetin were 1.92 and 0.25 mg/g in the tetraploid whole-plant sample 4W3. The chemical patterns were further analyzed by similarity calculation and principal component analysis (PCA). Seven common characteristic peaks were found in all of the tested samples. Flavonoid patterns of tetraploids were significantly different from those of diploids, whereas different parts of plants showed less difference. The flavonoid pattern of the diploid leaf sample was most similar to that of the reference botanical G. pentaphyllum. The combination of chromatographic fingerprint and quantification analysis could be used for quality assessment of G. pentaphyllum and its derived nutraceutical products.

Asparagus (Asparagus officinalis L.) spears are rich in bioactive compounds such as protodioscin, a saponin, and rutin, a flavonoid. Protodioscin and rutin are routinely quantified separately, and an approach permitting simultaneous measurement would significantly improve speed of analysis. We have optimized an extraction procedure and modified a method of high-performance liquid chromatography by coupling to an ultraviolet detector to simultaneously analyze protodioscin and rutin in asparagus extracts. An acidic ethanol solvent was more efficient than methanol, acetonitrile, or water in coextraction of protodioscin and rutin. Protodioscin and rutin were detected at 210 nm, with retention times of 12.6 min and 7.9 min, respectively. The method was validated by high linear correlations between 3.13 and 1000.0 μg/mL for protodioscin (r(2)= 0.9999), and between 0.3 and 1087.5 μg/mL for rutin (r(2)= 0.9997). The limit(s) of detection and quantification for protodioscin were 1.6 μg/mL and 3.13 μg/mL, respectively, and for rutin 0.2 μg/mL and 0.3 μg/mL, respectively. White asparagus spears and the crown of the plants were revealed to be rich sources of protodioscin and contained 2.59 to 10.4 mg/g dry weight. Green asparagus spears, particularly the upper portion, were rich in rutin and contained between 1.51 and 7.29 mg/g dry weight., (© 2010 Institute of Food Technologists®)

Objective: To develop an HPLC method for simultaneous determination of rutin, isoquercitrin and chlorogenic acid in Farfarae Flos., Method: The analysis was carried out on a Phenomenex Synergi POLAR-RP 80A column (4.6 mm x 250 mm, 4 microm) with gradient elution using methanol-acetonitrile-water (adjusted to pH 2.5 with formic acid) as mobile phase. The flow rate was 1.2 mL x min(-1) and the detection wavelength was at 255 nm., Result: The calibration curves were linear over the range of 0.2-2 000 microg x L(-1) for rutin and isoquercitrin, 10-2 000 microg x L(-1) for chlorogenic acid, respectively. The average recoveries were 99.5% for rutin, 100.1% for isoquercitrin and 99.4% for chlorogenic acid, respectively, with RSD not more than 3.0%., Conclusion: The described method is reliable and could be used for the quality control of Farfarae Flos.

This work was designed to develop a simple, effective, and reliable LC system to identify a chemical marker and compare Sambucus nigra L. and Sambucus australis Cham. et Schltdl. flower extracts (American and European elder). Rutin was the main constituent of both species. The developed method showed a linear response in the range of 10 to 45 microg x mL(-1) for rutin and 1.75 to 3.25 microg x mL(-1) for samples of the Sambucus species. Precision was determined and the relative standard deviations were 1.75 % for HSN and 1.28 % for HSA for intraday precision and 1.28 % and 1.51 % for inter-day precision, respectively, while accuracy was 97.9 % for HSN and 99.41 % for HSA. Quantification and detection limits as well as robustness were determined, presenting adequate results. The LC method showed an adequate performance for the separation of flavonoid glycosides in S. nigra and S. australis extracts, since the presence of interference had been previously evaluated. The analysis of thirty different samples of S. NIGRA and S. australis of different origins did not show significant variability among them. An accelerated stability study revealed a significant decrease in the first 30 days reaching 57 % in 90 days for S. australis samples and a total decrease of 25 % in 90 days for S. nigra samples, considering rutin as the chemical marker. These results will contribute to quality control analysis routines of these raw materials in pharmaceutical production facilities., (Georg Thieme Verlag KG Stuttgart.New York.)

RP HPLC-UV-MS was optimized for simultaneous determination of several prominent flavonoids (quercetin, naringenin, naringin, myricetin, rutin and kaempferol) using a commercially available monolithic column and isocratic elution. The concentration of organic modifier (methanol and ACN) was tested in order to obtain the best resolution. The LODs for a UV detector at 254 nm were comprised between 0.1 microg/mL for myricetin and quercetin and 0.5 microg/mL for rutin, naringin and kaempferol with the precision in the range of 1.5-3.0% RSD. MS detection with multiply reaction monitoring (MRM) offers a considerable gain in selectivity as well as sensitivity LODs for flavonoids were in the range of 1-20 ng/mL. The developed method was used for monitoring the content of some flavonoids in tomatoes during their maturation process.

Phyllanthus niruri L., commonly known in Brazil as 'quebra-pedra', has long been used in the treatment of diverse diseases and especially urolithiasis. The therapeutic effects of P. niruri are attributed to various compounds present in the plant, including the hydrolysable tannin corilagin. In the present study, high-performance liquid chromatography (HPLC-/PAD) profiles of leaves and commercial extracts of P. niruri were examined and three compounds, found to be present in all of the samples studied, were isolated by open column chromatography over C18)silica gel followed by preparative HPLC. These compounds were identified by nuclear magnetic resonance as corilagin, rutin and ethyl 3,4,5-trihydroxybenzoate. Corilagin, which has been proposed as a phytochemical marker for P. niruri, was employed as an external standard in the development and validation of a rapid and efficient qualitative and quantitative HPLC assay for the analyte. The method may be applied in the standardization of herbs and phytomedicines commercialized in Brazil as quebra-pedra.

Neurodegenerative disorders (NDD) have become the common global health burden over the last several decades. According to World Health Organization (WHO), a staggering 30 million people will be affected by Alzheimer's disease in Europe and the USA by 2050. Effective therapies in this complex field considering the multitude of symptoms associated with NDD indications, have not been found yet. Based on the results of NDD related studies, prevention appears to be the promise alternative. Antioxidative and anti-inflammatory properties are hypothesized for natural phenolics, a group of plant secondary products that may positively impact neurodegenerative diseases. In these studies, phenolic-rich extracts from less common fruit species: Blue honeysuckle (Lonicera edulis, Turcz. ex. Freyn), Saskatoon berry (Amelanchier alnifolia Nutt.), and Chinese hawthorn (Crateagus pinnatifida Bunge) were obtained and analyzed to detect neuroprotective substances content and establish a potential therapeutic value. High performance liquid chromatography with electrochemical detection was optimized and further applied on analysis of the extracts of less common fruit species. It was observed that Chinese hawthorn and Blue honeysuckle extracts are potent source of neuroprotective phenolic antioxidants. In accordance the results, it appears that the fruit or formulated products may have the potential for the prevention of neurodegenerative diseases.

A simple high-performance liquid chromatography method using a diode array detector (DAD) is developed for the simultaneous analysis of five major catechins: (+)-catechin (C), (-)-epicatechin (EC), (-)-gallocatechin (GCT), (-)-epigallocatechin (EGC), (-)-epigallocatechin gallate (EGCG), and the phenolic plant metabolites gallic acid (GA) and rutin (RT) in lyophilized extracts of Cistus species. The optimal analytical conditions are investigated to obtain the best resolution and the highest UV sensitivity for the quantitative detection of catechins. The optimized conditions (acetonitrile-phosphate buffer 50 mM, pH 2.5, gradient elution system on a C18 reversed-phase column with a flow rate of 1 mL/min and UV absorbance at 210 nm) allowed a specific and repeatable separation of the studied analytes to be achieved. All compounds are successfully separated within 32 min. Calibration curves are linear in the 2-50 microg/mL range for GCT, C, and EGCG and in the 5-50 microg/mL range for GA, EGC, EC, and RT. The limit of detection values ranged from 0.24 to 0.74 microg/mL. The limit of quantitation limit values ranged from 0.77 to 1.94 microg/mL. The validated method is applied to the determination of the specific phytochemical markers GA, GCT, C, and RT in Cistus incanus and Cistus monspeliensis lyophilised extracts. The recovery values ranged between 78.7% and 98.2%. The described HPLC method appears suitable for the differentiation and determination of the most common catechins together with the glycoside rutin and the phenolic compound gallic acid and can be considered an effective and alternative procedure for the analyses of this important class of natural compounds.

Objective: To study the influence of steaming processing on components in Chinese medicine., Methods: RP-HPLC was used to separate and detect every, components and all the same components' peak area were compared between crude-sun-cured and stcam-sun-cured Chinese medicine., Results: Steaming processing showed some advantatages to reserve most but not every compo-nent., Conclusion: Steaming processing can be applied as a superior method.

RP-HPLC with UV photodiode array detection (UV-DAD) was developed and validated for the simultaneous determination of vitexin-2"-O-glucoside, vitexin-2"-O-rhamnoside, rutin, and hyperoside in the extract of hawthorn (Crataegus pinnatifida Bge.) leaves. The analytes of interest were separated on a Diamonsil C18 column (250 x 4.6 mm id, 5 microm) with the mobile phase consisting of THF/ACN/methanol/ 0.05% phosphoric acid solution (pH 5.0) (18:1:1:80 v/vl/v). The flow rate was set at 1.0 mL/min and the eluent was detected at 340 nm for the four flavonoids. The method was linear over the studied range of 1.00-100 microg/mL for the four analytes of interest with the correlation coefficient for each analyte greater than 0.999. The LOD and LOQwere 0.03 and 0.10 microg/mL, 0.03 and 0.10 microg/mL, 0.05 and 0.15 pg/mL, 0.10 and 0.30 microg/mL for vitexin-2"-O-glucoside, vitexin-2"-0-rhamnoside, rutin, and hyperoside, respectively. The optimized method was successfully applied to the analysis of four important flavonoids in the extract of hawthorn leaves. The total amounts of the four flavonoids were 22.2, 62.3, 4.27, and 8.24 mg/g dry weight for vitexin-2"-O-glucoside, vitexin-2"-O-rhamnoside, rutin, and hyperoside in the extract of hawthorn leaves, respectively.

A simple and sensitive method has been developed for determining rutin, catechin, epicatechin, and epicatechin gallate in buckwheat (Fagopyrum esculentum Moench) flour and seeds by micro-high-performance liquid chromatography with electrochemical detection. Chromatography was performed using an octadecylsilica column, acetonitrile-water-formic acid (13:87:1, v/v/v) as a mobile phase, and an applied potential at +0.5 V vs Ag/AgCl. We found that Japanese buckwheat flour contains rutin (12.7 mg/100 g), catechin (3.30 mg/100 g), epicatechin (20.5 mg/100 g), and epicatechin gallate (1.27 mg/100 g). The relative standard deviations for rutin, catechin, epicatechin, and epicatechin gallate peak heights were less than 0.86% (n = 5). The detection limit of rutin was 0.86 ng/mL. Moreover, the present method was applied to the distribution analysis of these compounds in buckwheat seed. The embryo proper and cotyledons of a mature buckwheat seed contained rutin with the highest concentration as compared to other parts. This method is useful in determining rutin, catechin, epicatechin, and epicatechin gallate in buckwheat with a small amount of sample for quality control in the food industry.

Quantification of bioactive principles through modern analytical tools is essential for establishing the authenticity, credibility, prescription and usage of Ayurvedic medicines/herbal formulations. 'Chyavanprash' is one of the oldest and most popular Ayurvedic preparations, used widely as a health promotive and disease preventive 'Rasayana' drug in India and elsewhere. The rejuvenating and tonic properties of 'Chavanprash' are considered mainly due to their antioxidant principles, which in turn is due to the presence of phenolic compounds. A simple high-performance liquid chromatography (HPLC) method for the separation and quantitative determination of the major antioxidant compounds from 'Chyavanprash' has been developed. The use of Waters Symmetry column and an acidic mobile phase enabled the efficient separation of phenolic compounds (catechin, quercetin-3-O-rutinoside, syringic acid and gallic acid) within a 35 min analysis. Validation of the method was done with a view to demonstrate its selectivity, linearity, precision, accuracy and robustness. In addition optimization of the complete extraction of phenolic compounds were also studied.

A rapid and sensitive assay for quantitative determination of rutin in oral dosage forms based on isocratic reversed phase high performance liquid chromatography (RP-HPLC) was developed and validated. Using a C(18) reverse-phase analytical column, the following conditions were chosen as optimal: mobile phase methanol-water 1:1 (v/v), pH 2.8 (adjusted with phosphoric acid), flow rate=1 mL min(-1) and temperature T=40.0 degrees C. Linearity was observed in the concentration range 8-120 microg mL(-1) with a correlation coefficient of 0.99982 and the limit of detection (LOD)=2.6 microg mL(-1), and limit of quantification (LOQ)=8.0 microg mL(-1). Intra- and inter-day precision were within acceptable limits. Robustness test indicated that the mobile phase composition and pH influence mainly the separation. The proposed method allowed direct determination of rutin in pharmaceutical dosage forms in the presence of excipients, but is not suitable for preparations where compounds structurally/chemically related to rutin may be present.

Platycladus orientalis (L.) Franco (Cupressaceae), a traditional Chinese herb and food additive, has been used for treatments of gout, rheumatism, diarrhoea and chronic tracheitis. To evaluate the quality of P. orientalis (L.) Franco, a sensitive, simple and accurate reversed-phase high-performance liquid chromatographic (RP-HPLC) separation method with a photodiode array detector (DAD) was developed for the determination of four main bioactive flavonoids, rutin, quercitrin, quercetin and amentoflavone. Separation of the four compounds was achieved by the HPLC assay (Agilent Eclipse XDB-C 18 column with mobile phase, methanol-acetonitrile-18 mM sodium acetate buffer (pH 3.5) and recorded at UV 356 nm). This method showed good linear relation in the range of 0.8-80 microg/ml for rutin, 1.84-184 microg/ml for quercitrin, 0.72-72 microg/ml for quercetin and 0.72-72 microg/ml for amentoflavone. The correlation coefficients of the calibration curve for the analysis were all higher than 0.999. In addition, the contents of those four flavonoids in P. orientalis (L.) Franco growing in 12 different locations in China were compared to establish the effectiveness of the method.

A new chromatographic procedure is proposed for rapid (35 min) complete fractionation of flavonoids in plant raw material; the method well discriminates between components with antioxidant activity (rutin, apigenin, quercetin, hesperidin). The method is based on reverse-phase high performance liquid chromatography using ODS-S5CN carrier and linear 10-60% pyridine elution gradient.

A reversed-phase high-performance liquid chromatography/electrospray ionisation mass spectrometry (RP-HPLC/ESI-MS) method was developed and validated for the simultaneous determination of ten major active components in Ginkgo biloba extract (bilobalide, ginkgolides A, B, C, quercetin, kaempferol, isorhamnetin, rutin hydrate, quercetin-3-beta-D-glucoside and quercitrin hydrate) which have not been previously reported to be quantified in a single analysis. The ten components exhibit baseline separation in 50 min by C18 chromatography using a water/1:1 (v/v) methanol/acetonitrile gradient. Quantitation was performed using negative ESI-MS in selected ion monitoring (SIM) mode. Good reproducibility and recovery were obtained by this method. The sensitivity of both UV and different mass spectrometry modes (full scan, selected ion monitoring (SIM), and selected reaction monitoring (SRM)) were compared and both quantitation with and without internal standard were evaluated. The analysis of Ginkgo biloba commercial products showed remarkable variations in the rutin and quercetin content as well as the terpene lactone contents although all the products satisfy the conventional quality control method., (Copyright 2006 John Wiley & Sons, Ltd.)

In order to continue our previous studies concerning Geranium robertianum L., herb Robert (Geraniaceae), we have realised a HPLC study of some polyphenols using an original method created by a group of young researchers from the University of Medicine and Pharmacy of Cluj-Napoca. We have identified and measured in the dried Geranii robertiani herba (harvested from Valea Runcului, district of Alba-Iulia) the following compounds: hyperoside (3.64 microg/100 mg), ellagic acid (7599.76 microg/100 mg), isoquercitrin (49.49 microg/100 mg), quercetrin (83.92 microg/100 mg), kaempferols (143.43 microg/100 mg), caftaric acid (166.92 microg/100 mg), rutoside (72.23 microg/100 mg). We have also analysed a hydrolysed sample of the same drug in which we have identified and measured: caffeic acid (6.62 microg/100 mg), ellagic acid (10550.65 microg/100 mg), quercetrin (203.44 microg/100 mg), kaempferols (231.80 microg/100 mg), caftaric acid (47.41 microg/100 mg). We have indirectly proved the presence of ellagic tannins (the amount of ellagic acid increases after acid hydrolise) and the existence of bi- or polycaffeoil derivatives (the caffeic acid is present only in the hydrolysed sample). The flavonoid aglycones exist in both forms: as free compounds and as part of the flavonoid molecules.

A comprehensive two-dimensional liquid chromatographic separation system based on the combination of an immobilized liposome chromatographic (ILC) column and an ODS column was developed for the separation of components in Ginkgo biloba, a traditional Chinese medicine. Two columns were coupled by a two-position, eight-port valve equipped with two storage loops, and the system was controlled by a computer. The effluent was detected both by a diode array detector and by an atmospheric pressure chemical ionization (APCI) mass spectrometer. Under the optimization separation conditions with the separation system, more than 41 components in the methanol extract of Ginkgo biloba were resolved. According to their UV and mass spectra, 13 of them were preliminarily identified as ginkgolide B, ginkgolide C, bilobalide, rutin, quercetin, quercetin-3-O-beta-D-glucosyl (1-2)-alpha-L-rhamnoside, quercetin-3-O-beta-D-glucoside, isorhamnetin, kaempferol-3-O-beta-D-glucosyl (1-2)-alpha-L-rhamnoside, isohamnetin-3-O-beta-D-rutinoside, kaempferol-3-O-beta-D-glucoside, kaempferol, kaempferol-3-O-beta-D-rutinoside.

Purpose: 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., Methods: 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., Results: 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., Conclusion: 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 .

A liquid chromatography/mass spectrometry (LC/MS) method with selected ion monitoring was developed and validated to analyze the contents of protodioscin and rutin in asparagus. The distribution of rutin and protodioscin within the shoots was found to vary by location, with the tissue closest to the rhizome found to be a rich source of protodioscin, at an average level of 0.025% tissue fresh weight in the three tested lines, while the upper youngest shoot tissue contained the highest amount of rutin at levels of 0.03-0.06% tissue fresh weight. The lower portions of the asparagus shoots that are discarded during grading and processing should instead be considered a promising source of a new value-added nutraceutical product.

Quantitative analysis of phenolic compounds from four apple varieties (Golden and Red Delicious, Granny Smith and Green Reineta) using high-performance liquid chromatography with diode-array detection was carried out. For each variety, both peel and pulp were analysed. The identification of phenolic compounds was made by comparing their retention times and UV spectra with those of standards. The results revealed differences between pulp and peel in all cases studied. The highest levels of phenolic compounds were found in the peel. High levels of catechins and flavonol glycosides, especially rutin, were found in apple peels. Chlorogenic acid was the major peak in the pulp for all apple varieties studied except for Granny Smith. Significant quantitative differences between the apple varieties were also found, the Golden Delicious variety showing the lowest content of phenolic compounds and Green Reineta variety the highest. The recovery of phenolic compounds from both peel and pulp was measured in all apple varieties. The values ranged between 95 and 105%, indicating close to quantitative recovery for the method used.