Ultra Performance Liquid Chromatography-diode Array Research Articles

A comprehensive quality evaluation for Huangqi Guizhi Wuwu decoction by integrating UPLC-DAD/MS chemical profile and pharmacodynamics combined with chemometric analysis

Ethnopharmacological relevanceHuangqi Guizhi Wuwu Decoction (HGWD), a classical Chinese formula originally recorded in Jin Kui Yao Lue, was used for the treatment of human “blood impediment” (a type of “Bi” syndrome). In clinical practice, HGWD has been applied to treat rheumatoid arthritis (RA). Aim of the studyThe characterization of chemical markers reflecting both efficacy and chemical characteristics is of great significance for TCM quality control. With the anti-RA effects of HGWD as an example, the aim of this study was to develop a comprehensive strategy combining the overall chemical profile and biological activity data to identify chemical markers. Materials and methodsFirst, an ultra-performance liquid chromatography-diode array detector (UPLC-DAD) fingerprint was established and validated to evaluate the holistic quality of HGWD of different origins. Characteristic markers associated with HGWD from different geographical origins were screened by a combination of UPLC-DAD fingerprint and chemometrics methods. Second, the chemical profiles of the 15 batches of HGWD samples were characterized by UPLC coupled tohybrid linear ion trap-Orbitrap mass spectrometry (UPLC-HRMS). The in vitro anti-RA activities of the 15 HGWD samples were then evaluated. Third, spectrum-effect relationship analysis was performed to identify bioactive compounds that could potentially be used as quality markers. Finally, a UPLC-triple quadrupole tandem mass spectrometry approach was optimized and established for quantitative analysis of the characteristic and quality markers in 15 batches of HGWD. ResultsIn total, 30 common peaks were assigned in the UPLC-DAD fingerprint. Nine peaks were recognized and considered characteristic markers: protocatechuic acid, coumarin, cinnamic acid, oxypaeoniflorin, paeoniflorin, calycosin, formononetin, catechin, and albiflorin. Furthermore, ninety-five common compounds were identified in the UPLC-HRMS chemical profile. The pharmacological analysis indicated that the anti-RA activities of the 15 HGWD samples were vastly different. The spectrum-effect relationship analysis revealed 30 potential bioactive constituents positively correlated with anti-RA activity. Among them, five compounds with relative amounts >1%, paeoniflorin, astragaloside IV, hexahydrocurcumin, formononetin and calycosin-7-glucoside, were selected as quality markers, and their activity was verified in LPS-induced RAW264.7 macrophages. Finally, the above 12 representative components were simultaneously quantified in the 15 batches of HGWD samples. ConclusionCombining a holistic chemical profile with representative component evaluation, this systematic strategy could be a reliable and effective method to improve quality evaluations of HGWD.

Cytotoxic and Pro-Apoptotic Effects of Leaves Extract of Antiaris africana Engler (Moraceae).

Antiaris africana Engler leaves have been used in Senegalese folk medicine to treat breast cancer. The present study aimed to investigate the anticancer potential of Antiaris africana Engler leaves using several human cancer cell lines. The leaves of Antiaris africana Engler were extracted in parallel with water or 70% ethanol and each extract divided into three parts by successive liquid-liquid extraction with ethyl acetate and butanol. The phytochemical components of the active extract were investigated using ultra-performance liquid chromatography-diode array detector-quadrupole time-of-flight tandem mass spectrometry (UPLC-DAD-QTOF-MS/MS). The cytotoxic and cytostatic effects of each extract, as well as their fractions, were evaluated in vitro via flow and image cytometry on different human cancer phenotypes, such as breast (MCF-7), pancreas (AsPC-1), colon (SW-620) and acute monocytic leukemia (THP-1). Both hydro-alcoholic and aqueous extracts induced strong apoptosis in MCF-7 cells. The water fraction of the hydro-alcoholic extract was found to be the most active, suppressing the cell growth of MCF-7 in a dose-dependent manner. The half maximum effective concentration (EC50) of this fraction was 64.6 ± 13.7 μg/mL for MCF-7, with equivalent values for all tested phenotypes. In parallel, the apoptotic induction by this fraction resulted in a EC50 of 63.5 ± 1.8 μg/mL for MCF-7, with again equivalent values for all other cellular tested phenotypes. Analysis of this fraction by UPLC-DAD-QTOF-MS/MS led to the identification of hydroxycinnamates as major components, one rutin isomer, and three cardiac glycosides previously isolated from seeds and bark of Antiaris africana Engler and described as cytotoxic in human cancer models. These results provide supportive data for the use of Antiaris africana Engler leaves in Senegal.

Screening and identification of tyrosinase inhibitors in edible plant materials by on-line UPLC-enzyme reactor coupled with UHPLC-FTMS

Tyrosinase plays a primary role in melanin biosynthesis and enzymatic browning of freshly cut fruits and vegetables. Herein, an on-line ultraperformance liquid chromatography diode array detector biochemical detection (UPLC-DAD-BCD) method was established to identify trace amount potent tyrosinase inhibitors and antibrowning agents in complex mixtures. The tyrosinase inhibition activities of some representative compounds were evaluated by using the established method and their chromatography-activity relationships were obtained. Then the proposed UPLC-DAD-BCD method was applied to screen tyrosinase inhibitors in edible herbal extracts and identified two tyrosinase inhibitors in green tea and three in cinnamon. The above active ingredients were determined by ultra-high-performance liquid chromatography linear ion trap/orbitrap high resolution mass spectrometry (UHPLC-FTMS). The on-line UPLC-DAD-BCD in combination with UHPLC-FTMS was confirmed to be a powerful technique to screen and elucidate the active ingredients in complex matrixes and could be applied to evaluate the integrated effects of multiple ingredients against corresponding targets.

Comprehensive characterization of flavonoid derivatives in young leaves of core-collected soybean (Glycine max L.) cultivars based on high-resolution mass spectrometry

Most previous studies have been focused on isoflavone profile with biological activities from soybean seed and its related products. However, in the present study, eighty-three flavonoid derivatives (55 flavonols, 9 flavones and 19 isoflavones) were comprehensively identified and quantified from young leaves of 21 core-collected soybean cultivars based on ultra-performance liquid chromatography-diode array detector with quadrupole time of flight/mass spectrometry (UPLC-DAD-QToF/MS). Among total flavonoids from soybean leaves (SLs), the abundant flavonols (83.6%) were primarily composed of di- and tri- glycosides combined to the aglycones (K, kaempferol; Q, quercetin; I, isorhamnetin). Particularly, K-rich SLs (yellow coated seed), Nongrim 51 (breeding line) and YJ208-1 (landrace) contained mainly kaempferol 3-O-(2″-O-glucosyl-6″-O-rhamnosyl)galactoside and 3-O-(2″,6″-di-O-rhamnosyl)galactoside, and were expected to be superior cultivars by their higher flavonoids. Besides, the new tri-I-glycosides (soyanins I–V) were presented as predominant components in Junyeorikong (landrace, black). Thus, this study suggest that the SLs can be considered as valuable edible resources due to their rich flavonoids. Also, these detailed profiles will support breeding of superior varieties with excellent biological activities as well as relationship with seed anthocyanins production, and contribute to perform metabolomics approach to investigate the changes of SLs flavonols during the leaf growth and fermentation in further research.

Anti-Inflammatory Effects of Jakyakgamcho-Tang in IL-4- and TNF-α-Stimulated Lung Epithelial Cells and Lipopolysaccharide-Stimulated Macrophages

Asthma is a chronic respiratory disease mediated by airway inflammation. Jakyakgamcho-tang (JGT), a traditional medicine, is widely subscribed to common diseases such as muscle pain and cramps in East Asian countries. Although the efficacy of JGT on peripheral neuropathy, gouty arthritis, and colitis has been reported, the effect of JGT on airway inflammation related to asthma is not clearly investigated. In this study, we aimed to evaluate the effects of JGT water extract (JGTW) on factors related to airway inflammation using the human bronchial epithelial BEAS-2B and the mouse monocyte-macrophage RAW264.7 cell lines. Furthermore, the constituents in JGTW were quantitatively and qualitatively studied for future reference of JGTW standardization. JGTW reduced the generation of several airway inflammation mediators such as eotaxins, regulated on activation normal T-cell expressed and secreted (RANTES), and matrix metalloproteinase-9, and expressions of adhesion molecules (ICAM-1 and VCAM-1), which attracts leukocytes to the site of inflammation in interleukin-4 + tumor necrosis factor-α (IT)-stimulated BEAS-2B cells. In lipopolysaccharide-stimulated RAW264.7 cells, JGTW effectively suppressed inducible nitric oxide synthase (iNOS) induction by inhibiting the MAPK and NF-κB signaling. In addition, JGTW treatment showed decreased inflammatory cells and Th2 cytokines in bronchoalveolar lavage fluid and decreased IgE levels in plasma in the OVA-induced asthmatic mice model. In the ultra-performance liquid chromatography-diode array detector-tandem mass spectrometry analysis, 24 phytochemicals were identified in JGTW, and paeoniflorin (63.971 mg/g) and glycyrrhizin (11.853 mg/g) were found to be the most abundant. These findings suggest that JGTW has anti-inflammatory effects on airway inflammation by regulating inflammatory response-related factors, possibly through MAPK and NF-κB in pulmonary epithelial cells and macrophages.

Inhibitory Effects of Gyeji-Tang on MMP-9 Activity and the Expression of Adhesion Molecules in IL-4- and TNF-α-Stimulated BEAS-2B Cells.

Gyeji-tang (GJT), a traditional herbal formula composed of five herbal medicines, is commonly used to treat the common cold, exogenous febrile disease, fever and headaches in Korea, China and Japan. Although various pharmacological activities of GJT have been reported in several studies, the effect of GJT water extract (GJTWE) on airway inflammation has not yet been investigated. This study aimed to evaluate the effects of GJTWE on airway inflammation-related factors using human bronchial epithelial BEAS-2B cells, and to identify the phytochemicals in GJTWE by ultra-performance liquid chromatography-diode array detector-tandem mass spectrometry (UPLC-DAD-MS/MS) analysis. GJTWE significantly decreased the production of chemokines, including eotaxin-3, eotaxin-1, regulated on activation normal T-cell expressed and secreted (RANTES), and matrix metalloproteinase-9, and the expression of the adhesion molecules, intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, in interleukin-4 + tumor necrosis factor-α (IT)-stimulated BEAS-2B cells. In the UPLC-DAD-MS/MS analysis, 21 phytochemicals, including six flavonoids, two chalcones, five terpenoids, six phenolics, one phenylpropanoid and one coumarin, were identified in GJTWE. The findings suggested that GJTWE might exhibit anti-inflammatory effects on airway inflammation by regulating the expression of inflammatory response-related factors in IT-stimulated BEAS-2B cells; further studies are required to determine the bioactive compounds involved in the inhibition of airway inflammation.

Determination of Nitrofuran Metabolites in Fish by Ultraperformance Liquid Chromatography-Photodiode Array Detection with Thermostatic Ultrasound-Assisted Derivatization.

Nitrofuran (NF) is a class of broad-spectrum antibiotics that are used illegally in animal feeding. NF and its metabolites have proven to pose potential risk to human health. To address the current analytical needs to quantify low levels of NF metabolites in animal foods, a sensitive method was developed for simultaneous detection of four NF metabolites in fish products by an ultraperformance liquid chromatography-diode array detector (UPLC-DAD). With 2-nitrobenzaldehyde (2-NBA) as the derivatizing reagent, the metabolites were hydrolyzed and derivatized under the assistance of thermostatic ultrasound. Compared with the current detection methods, the time of the derivatization reaction has been shortened from 16 to 2 h. The relative coefficient of four NF metabolite derivatives reached more than 0.998, with excellent linear relationship. The limits of detection (LODs) and limits of quantification (LOQs) of six repeated determinations reached 0.25–0.33 and 0.80–1.10 μg/kg, respectively. For all four NF metabolites, the limit of detection of the method was below the minimum required performance limit (MRPL) of 1.0 μg/kg, which makes it compatible with the EU requirements. The recoveries ranging from 89.8 to 101.9% with relative standard deviation below 6.5% were obtained for all of the NF metabolites. What’s more, this method was successfully applied for the determination of four NF metabolites in the fish products. As a promising approach, this method could also be extended for the quantitation of NF metabolites in aquaculture and poultry products.

Comparison of endometrial prostanoid profiles in three infertile subgroups: the missing part of receptivity?

To study the prostanoid profile of the endometria of patients with recurrent implantation failure (RIF), unexplained infertility (UIF), and recurrent miscarriages (RM), and to compare them with the endometria of healthy fertile controls. Prospective cohort study. University hospital. Fifteen patients with RIF, 18 patients with UIF, 16 patients with RM, and 23 fertile controls were recruited. Endometrial samples were taken during the window of implantation. After tissue homogenization and extraction, analysis with ultra-performance liquid chromatography diode array detector electrospray ionisation tandem mass spectrometry was performed. Concentrations of prostaglandin (PG) D1, PGE1, PGF1α, 6-ketoPGF1α, PGD2, PGE2, PGF2α, 15-deoxy-Δ12,14-PGJ2, PGD3, PGE3, PGF3α, thromboxane B2, 13,14-dihydro-PGE1, 13,14-dihydro-PGF1α, 13,14-dihydro-PGF2α, 13,14-dihydro-15-keto-PGE1, 13,14-dihydro-15-keto-PGE2, and 13,14-dihydro-15-keto-PGF2α were assessed. Comparison of the endometria of patients with UIF and the controls showed no statistically significant differences. When the endometria of patients with RIF were compared with the controls, thromboxane B2 (TXB2) was found significantly higher (843.1 pg/mg vs. 133.5 pg/mg). When the endometria of patients with RM were compared with controls, 13,14-dihydro-15-keto PGF2α and TXB2 were found significantly higher (3907.30 pg/mg vs. 17.80 pg/mg and 858.7 pg/mg vs. 133.5 pg/mg respectively). We identified increased endometrial presence of TXB2 in patients with RM and RIF, and 13,14-dihydro-15-keto PGF2α in patients with RM. Although common ground is observed for RM and RIF, prostanoids, on the other hand, might make their own contribution to endometrial receptivity as important as genes and proteins. Attempts to normalize the prostaglandin profile of the endometrium via enzymatic activity can open new therapeutic options.

Simultaneous quantification and identification of Amaryllidaceae alkaloids in Narcissus tazetta by ultra performance liquid chromatography-diode array detector-electrospray ionisation tandem mass spectrometry

Narcissus tazetta is used traditionally for treatment of sores, wounds, skin diseases, cancer in different parts of world. Present study focus on the analysis of amaryllidaceae alkaloids in this plant using an ultra-performance liquid chromatography-diode array detection method. The method was developed for simultaneous quantification of eight Amaryllidaceae alkaloids i.e. pseudolycorine (1), lycorine (2), galanthamine (3), 8-O-demethylhomolycorine (4), N-methylhaemanthidine chloride (5), homolycorine (6), narciclasine (7) and zefbetaine (8) in Narcissus tazetta. The method was validated using a BEH C18 column with linear gradient. Standard calibration curve for the analytes showed good linearity ( r2≥0.999). The method was validated for intra-day (RSDs<0.91%) and inter-day (RSDs<0.65%) precisions and accuracy (recovery 92.2–112.5%). The developed method was successively applied for studying the variation of alkaloids in different parts of Narcissus tazetta, i.e. bulbs, roots, flowers, flower stalks and leaves. The study showed a significant variation of these alkaloids in different parts of the plant. Among the alkaloids under investigation, pseudolycorine had highest content in all the parts. Furthermore, application of the developed method to the identification of phytocomponents allowed the identification of sixteen alkaloids.

RETRACTED: Organometallic cerium oxide nanostructures acted as nanofertilizer during callogenesis and organogenesis of recalcitrant plant Berberis lycium Royle

Abstract We report organometallic cerium oxide nanostructures (OM-CeO2-NS) mediated in vitro callus induction and direct shoot regeneration of medicinally important and recalcitrant plant (Berberis lycium Royle). To achieve greater biocompatibility, Hypericum scabrum flower extract was used during the green synthesis process. Ultra-performance Liquid Chromatography-Diode Array Detector (UPLC-DAD) coupled with Mass spectrometry (MS) was employed to detect the potential secondary metabolites in H. scabrum flower extract which acted as reducing as well as the surface stabilizing agents for tailoring the morphology of OM-CeO2-NS. High resolution Transmission Electron Microscopy (Hr-TEM) and Atomic Force Microscopy (AFM) examination of the prepared OM-CeO2-NS revealed homogenous morphology with an average size of 38 ± 5 nm. Moreover, Fourier Transform Infrared (FT-IR) spectroscopy examination and X-ray Diffraction (XRD) technique also confirm organometallic chemistry of prepared CeO2-NS. Interestingly, nutritive behaviour of OM-CeO2-NS was observed in B. lycium tissue culturing primarily due to the extremely small size and organometallic nature. The results reveal that callus induction is enhanced to 90% with low concentration of OM-CeO2-NS (4 mg/L) and direct shoot regeneration is improved to 79% with increased concentration of 20 mg/L. The current study concludes that OM-CeO2-NS can be used as nanofertilizer in tissue culturing of the recalcitrant plant species.

Geographical Discrimination in Curcuminoids Content of Turmeric Assessed by Rapid UPLC-DAD Validated Analytical Method.

A fast and reliable ultra-performance liquid chromatography-diode array detection method was developed and validated for the quantitative assessment of turmeric extracts from different geographical locations. Acclaim RSLC PolarAdvantage II column (2.2 μm, 2.1 × 100 mm) was used to analyze individual curcuminoids (curcumin, demethoxycurcumin and bisdemethoxycurcumin) from turmeric samples. The detection was done on ultraviolet absorbance at 425 nm and the column temperature was maintained at 45 °C. A mobile phase consisting of acetonitrile and water was found to be suitable for separation, at a flow rate of 1 mL/min with linear gradient elution. Linearity, specificity, precision, recovery and robustness were measured to validate the method and instrument. Under the described conditions, curcuminoids were collected within one minute. The calibration curve of each curcuminoid showed good linearity (correlation coefficient > 0.999). The relative standard deviations (RSD) of intra-day, inter-day precision and repeatability were less than 0.73%, 2.47% and 2.47%, respectively. In the recovery test, the accuracy ranged from 98.54%-103.91% with RSD values of less than 2.79%. The developed method was used for quantification of individual curcuminoids of turmeric samples. Analysis of turmeric samples from Nepal and South Korea revealed that curcuminoid content was related to geographical location. Turmeric cultivated in warmer climates were found to have higher curcumionoid content than turmeric samples from cooler climates, the southern part of Nepal was found to have two times higher content of curcuminoids than turmeric from the north.

Qualitative identification of impurities and principal component quantitative analysis of zearalanone on purity certified reference material

A liquid chromatography method was established for the determination of zearalanone (ZAN) raw material. The qualitative analysis of ZAN and its trace impurities was performed by ultra performance liquid chromatography-diode array detector (UPLC-DAD) and ultra performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS), and the response factors of each impurity were calculated. The three main organic impurities in the ZAN raw material were identified as β -zearalanol, α -zearalanol and a dehydration product of zearalanol with relative response factors of 0.5352, 0.8594 and 0.6973, respectively. The main component of the ZAN raw material was determined by the calibration factor normalization method. The purity of zearalanone was determined to be 99.6% with a standard deviation of 0.01%. This method can provide a technical support for the development of ZAN standard materials.

UPLC-DAD quantification of chemical constituents of Potentilla atrosanguinea roots and their antioxidant activity.

Potentilla atrosanguinea, native to Himalayan region, is well known for its curative effects in traditional medicinal system. An ultra performance liquid chromatography-diode array detection method for the quantification of constituents of root part of P. atrosanguinea has been developed along with antioxidant activity evaluation. A simple and sensitive quantification method developed for seven compounds however only four compounds; p-coumaric acid (4), rutin (7), tiliroside (14) and kaempferol (16) were quantified as others were in lesser amount. Syringic acid and quercetin were found in trace amount whereas chlorogenic acid was absent in the ethanol extract of roots of P. atrosanguinea. Total polyphenolic and flavonoid contents were determined to be 21.75mg of gallic acid equivalent and 8.57mg of quercetin equivalent per gram of dry plant material, respectively. Antioxidant activity of extract was assessed using three assays; 2,2'-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and ferric reducing antioxidant power (FRAP). The IC50 values; 35.75μg/ml and 30.35μg/ml by DPPH and ABTS assays for ethanolic extract showed excellent free radical scavenging potential of its root part. The ferric reducing ability (FRAP) value, 26.67mg of ascorbic acid per gram also indicated its higher antioxidant potential.

Stability Study and Identification of Degradation Products of Caffeoylgluconic Acid Derivatives from Fructus Euodiae.

Caffeoylgluconic acid derivatives are characteristic constituents isolated from the aqueous extract of Fructus Euodiae. In this research focusing on caffeoylgluconic acid derivatives, trans-caffeoyl-6-O-d-gluconic acid (CGA), trans-caffeoyl-6-O-d-gluconic acid methyl ester (CGA-ME), and trans-caffeoyl-6-O-d-glucono-γ-lactone (CGA-LT), a systematic study of stability was performed under different temperatures and pH levels by ultra performance liquid chromatography-diode array detector (UPLC-DAD) and ultra performance liquid chromatography-diode array detector/electrospray ionization-quadrupole-time of flight mass spectrometry (UPLC-DAD/ESI-Q-TOF MS). From the concentration–time curves and sensitivity index (SeI), it was found that compared to CGA, which is inert to the variation of temperature and pH in the tested range, CGA-ME and CGA-LT were more sensitive, with stabilities more likely to be influenced by temperature. Considering the stability index (StI), the integrated stability of CGA was the best, and that of CGA-ME was the worst. In terms of the quasi-molecular and fragment ions of the tested compounds, the degradation products were identified or tentatively characterized, which could shed light on the degradation pathways. CGA-ME and CGA-LT were easily converted to CGA by hydrolytic reaction, all of which were susceptible to the formation of isomers. This study elucidated the degradation mechanism of caffeoylgluconic acid derivatives, contributing to better guidance on manufacturing and controlling the quality of drugs.

Identification Markers of Adulteration in Korean Red Ginseng (Panax Ginseng) Products Using High-Performance Liquid Chromatography (HPLC) and Liquid Chromatography–Mass Spectrometry (LC-MS)

ABSTRACTThe commercial products of Panax ginseng have increasing market demand and high prices due to the pharmacological activities. To obtain high profits, P. ginseng may be adulterated with lower priced morphologically similar species such as Platycodon grandiflorum, Codonopsis lanceolata, and Pueraria lobata. This study was designed to validate accurate methods for the analysis of adulteration in P. ginseng products. High-performance liquid chromatography (HPLC) and ultraperformance liquid chromatography–diode array detector–electrospray ionization–ion trap–time of flight–mass spectrometry (UPLC–DAD–ESI-IT-TOF-MS) were validated to analyze the raw plant materials, self-prepared formulations, and commercial products of P. ginseng, C. lanceolata, P. grandiflorum, and P. lobata. The developed analytical methods were confirmed by quality assurance parameters such as linearity, sensitivity, precision, and accuracy. Lobetyolin and ononin were identified as marker compounds by HPLC and confirmed by accurate mass measurement with ESI-IT-TOF-MS. HPLC analysis of self-prepared formulations indicated that by increasing the ratio of C. lanceolata, P. grandiflorum, and P. lobata in P. ginseng extracts, the peak area is increased at the same retention time. The limits of detection and quantification for lobetyolin and ononin were 0.098 and 0.171, and 0.108 and 0.726 mg/kg, respectively. Furthermore, the intraday precision (<1.0%) measurements confirmed that the developed analytical methods fulfill the required criteria for characterization of these products. The results demonstrated that the developed liquid chromatographic and mass spectrometric methods accurately characterized adulteration in P. ginseng commercial products.

Identification and characterization of matrix components in spinach during QuEChERS sample preparation for pesticide residue analysis by LC-ESI-MS/MS, GC-MS and UPLC-DAD.

In this article matrix components in spinach were investigated in detail. The samples were prepared using two QuEChERS (quick, easy, cheap, effective, rugged and safe) methods, AOAC and CEN. Liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), gas chromatography-mass spectrometry (GC-MS) and ultra performance liquid chromatography-diode array detector (UPLC-DAD), were applied for component identification. The strategies of identification by LC-ESI-MS/MS include accurate mass spectra of the parent ion, comparison with previous literature data and investigation of the mass spectral decomposition pattern. Overall, fourteen components were identified by LC-ESI-MS/MS in each methods of AOAC and CEN, which were phytosteroids, flavonoids, fatty acids and fatty acid amides. Fifty components using AOAC method and fifty-seven components using CEN method were identified in GC-MS by comparing mass data with the National Institute of Standards and Technology (NIST, U.S.) database. The results indicate that the major components of the matrix are terpenoids, fatty acids and fatty acid esters. Moreover, three pigments (neoxanthin, violaxanthin and lutein) in the AOAC method and eight pigments (neoxanthin, violaxanthin, zeaxanthin, lutein, chlorophyll a, chlorophyll b, pheophytin a and beta-carotene) in the CEN method that gave a characteristics peak at 440nm were identified by the UPLC-DAD. According to the sample preparation condition using different amounts of graphitized carbon black (GCB) in this study, the AOAC method had higher matrix component removal efficiency than the CEN method. A better understanding of matrix components would increase the current knowledge for improvement of existing QuEChERS methodology, as well as contribute to new method developments.

Effects of thermal treatments on 10 major phenolics and their antioxidant contributions in Acer truncatum leaves and flowers.

This study aimed to investigate effects of thermal treatments on major phenolics and their antioxidant contributions in Acer truncatum leaves and flowers (ATL and ATF, respectively). With ultra performance liquid chromatography-diode array detector-quadrupole time-of-flight-mass spectrometer/mass spectrometer, phenolic compositions of ATF were first characterized and compared with those of ATL. An optimized high performance liquid chromatography fingerprint was then established, and 10 major phenolics existing in both ATL and ATF were quantified. Gallic acid derivatives and flavonol-3-O-glycosides were found to be their dominant phenolic constituents, with the former being key constituents which was affected by thermal treatments and further influencing the variations of total phenols. Moreover, the mechanism underlining the changes of phenolics in ATL and ATF by the treatments was characterized as a thermolhydrolysis process. During thermal treatments, polymerized gallotannins were hydrolysed to 1,2,3,4,6-pentakis-O-galloyl-β-d-glucose, ethyl gallate and gallic acid, resulting in more than fivefold and twofold increase of their contents in ATL and ATF, respectively. By contrast, contents and antioxidant contributions of flavonol-3-O-glycosides gradually decreased during the process.\\absbreak Overall, this is, to our knowledge, the first report on the effects of thermal treatments on phenolics and their antioxidant contributions in ATL and ATF, and the three gallic acid derivatives with potentially higher bioactivity could be efficiently achieved by thermal treatments.

Preventive Effects of an UPLC-DAD-MS/MS Fingerprinted Hydroalcoholic Extract of Citrus aurantium in a Mouse Model of Ulcerative Colitis.

Although traditionally used to improve indigestion, diarrhea, dysentery, and constipation, the therapeutic effects of Citrus aurantium on intestinal inflammation remain unclear. The aim of this study was to evaluate the beneficial effects and to identify the active components of a hydroalcoholic extract of C. aurantium (HECA) on ulcerative colitis. HECA was prepared with 70% ethanol solution in water and extracted at 37 °C for 12 h in triplicate, filtered through a sieve, and lyophilized. Phytochemical identification of HECA was performed by ultra-performance liquid chromatography-diode array detector-tandem mass spectrometry (UPLC-DAD-MS/MS). Animals were randomly assigned to one of four groups based on the treatment conditions. Ulcerative colitis was induced by administration of 2.5% dextran sodium sulfate (DSS) in drinking water for 5 d. Body weight, clinical signs, colon length, pro-inflammatory cytokine expression levels, and histopathological findings were evaluated. In UPLC-DAD-MS/MS analysis, the identified phytochemical components of HECA included four alkaloids, seven coumarins, 18 flavonoids, two lignans, two phenolics, and 10 terpenoids. HECA markedly protected against body weight loss and colon shortening. In pathological examination, HECA alleviated DSS-related mucosal inflammatory lesions in the colon. Moreover, HECA markedly reduced the expression levels of interleukin-6, interferon-γ, tumor necrosis factor-α, and monocyte chemotactic protein-1 in colonic inflammation. Taken together, HECA has potential to relieve mucosal inflammation in the colon, suggesting that the putative active ingredients are responsible for the anti-ulcerative effects.

Extraction Optimization of Flavonoids from Hypericum formosanum and Matrix Metalloproteinase-1 Inhibitory Activity.

Hypericum formosanum is a valuable herb in Taiwan. In this study, response surface methodology was employed to optimize the ultrasound-assisted extraction of flavonoids from Hypericum formosanum. A central composite design with three variables (ethanol concentration, extraction time, and extraction temperature) was applied. Experimental results were fitted to the second order polynomial model and one-way analysis of variance was used to determine the goodness of fit of the model and the optimal conditions for responses. The optimal conditions for the maximum extraction yield of total flavonoid content (101.1 mg/g) using ultrasound-assisted extraction were ethanol concentration, 73.5%; extraction time, 38.3 min; and extraction temperature, 62.5 °C. The predicted result was consistent with the experimental result obtained under optimal extraction conditions. Hyperoside, astilbin, quercitrin, and quercetin from Hypericum formosanum extract (HFE) were identified by Ultra performance liquid chromatography-diode array detector-mass (UPLC-DAD-MS). HFE significantly reduced matrix metalloproteinase-1 protein expression in human skin keratinocyte cells, induced by advanced glycation end products.

Anti-oxidative and anti-inflammatory activities of devil's club (Oplopanax horridus) leaves.

This study aimed to investigate the anti-oxidative properties of the ethanolic extracts of the devil's club (Oplopanax horridus) leaves, stems, and roots. Furthermore, the anti-inflammatory activity of the leaf extract was analyzed. The leaf extract had higher total phenolic and flavonoid contents and anti-oxidative activity (radical scavenging, reducing power, and inhibition of lipid oxidation) than the root and stem extracts. The leaf extract also had anti-inflammatory effects. It significantly reduced lipopolysaccharide (LPS)-induced nitric oxide (NO; 71.0% at 50 μg/mL), tumor necrosis factor (TNF)-α (87.6% at 100 μg/mL), and interleukin (IL)-6 (36.2% at 100 μg/mL) production in murine RAW 264.7 macrophages. Furthermore, LPS-induced inducible nitric oxide synthase (iNOS) expression was decreased by the leaf extract (IC50=24.4 μg/mL). The ultra performance liquid chromatography-diode array detector (UPLC-DAD) analysis showed that the leaf extract contained gallic acid, protocatechuic acid, chlorogenic acid, and maltol. These findings suggest that the leaf extract could be utilized as a functional food material because of its anti-oxidative and anti-inflammatory activities.