Ultra-fast Liquid Chromatography Research Articles

Identification of the functional food potency of çalkama: A traditional recipe with edible Mediterranean wild greens from Turkish cuisine

BACKGROUND: The phytochemical contents of traditional foods are necessary to further elucidate the impacts of the Mediterranean diet on health. Çalkama is traditional food prepared by using wild green plants including chard (Beta vulgaris var.cicl), wild fennel (Foeniculum vulgare spp), common mellow (Malva sylvestris L), common poppy (Papaver rhoeas L), dock (Rumex spp), common nettle (Urtica dioica L), sow-thistle (Sonchus asper (L) Hill), common stork’s bill (Erodium circutarium (L)L Hér) and wild leek (Allium ampeloprasum L). OBJECTIVE: In this research, the antioxidant activity and the flavonoid and phenolic acid composition were separately analyzed for each plant and çalkama. METHODS: Total phenolic content, total flavonoid content and total antioxidant capacity of each plant and çalkama were measured spectrophotometric assays. Ultra-fast liquid chromatography (UFLC) was performed to detect specific flavonoid groups. RESULTS: It was detected that one portion of çalkama (100 g) contained approximately 250 mg of flavonoids. Particularly, quercetin and apigenin contributed to the main flavonoid source and chlorogenic acid was the major phenolic acid in çalkama. CONCLUSIONS: According to our findings, this food can be considered as a good phenolic and flavonoid source which protects its high antioxidant capacity through preparation and cooking processes.

A Reliable and Cost-Effective Method for Determination of Endocrine-Disrupting Compounds in Coastal Waters, Suspended Particulate Matter, and Sediments by Ultrafast Liquid Chromatography Coupled to Photodiode Array and Fluorescence Detectors

Analytical methods for determining 14 endocrine-disrupting compounds (EDCs) in coastal waters, suspended particles, and sediment samples were successfully performed by ultrafast liquid chromatography with photodiode array and fluorescence detections (UFLC-PDA-FLD). Solid‑phase extraction (SPE) and ultrasound-assisted extraction (USE) were used for sample preparation. Two chromatographic methods have been developed. An isocratic separation method was used to separate bisphenol A (BPA) and steroids and another gradient elution method to separate phthalates and alkylphenols. The detection by fluorescence was used for alkylphenols, BPA, and steroids and photodiode array (PDA) for phthalates. Limits of detection (LOD) ranged from 0.41 (4-tert-octylphenol (4tOP)) to 63 ng L-1 (dibutylphthalate (DBP)), 0.41 (4tOP) to 63.2 ng g-1 dried weight (dw) coastal waters, and solid samples (suspended particulate matter (SPM) and sediment samples), respectively. Recoveries ranged from 52 (diethylphthalate (DEP)) to 116% (DBP) for water, from 54 (DEP) to 108% (estrone (E1)) for SPM, and from 62 (4-n-nonylphenol (4nNP)) to 117% (4-n-octylphenol (4nOP)) for sediment samples. Finally, with the minimization of reagents and energy, the proposed methods were applied to samples collected from Todos os Santos Bay (BTS), Bahia, Northeastern Brazil.

Adsorptive stripping voltammetric determination of chloramphenicol residues in milk samples using reduced graphene oxide sensor.

In this paper, the electrochemical response of chloramphenicol (CHL) was investigated on a bare glassy carbon electrode (GCE) and after modification with reduced graphene oxide (GCE/rGO). Preliminary studies by cyclic voltammetry demonstrated an adsorption-controlled mass transport regime of CHL species and a pH-dependent behavior on both electrode surfaces. An adsorptive stripping differential pulse voltammetry (AdSDPV) method was proposed and under optimized instrumental conditions, a comparison of the analytical characteristics of both sensors was performed. The GCE/rGO sensor showed an increase in sensitivity (10-fold), and an anticipation of the reduction potential (200 mV), compared to the bare electrode, due to the adsorptive character (pre-concentration of the CHL species) and the electrocatalytic effect of the nanomaterial. The method was applied to skimmed and whole milk samples, which were simply diluted (50-fold) in supporting electrolyte. The results by AdSDPV using GCE/rGO showed adequate detectability (0.22 μmol L-1), good precision with a 6% relative standard deviation (RSD) and satisfactory recovery ranging from 93 to 108%. The obtained results were statistically similar (95% confidence level) with those performed through ultra-fast liquid chromatography (UFLC). Furthermore, the sensor showed an improvement in the analytical performance for CHL detection, when compared to other sensors reported in the literature. Therefore, the developed method is reliable and promising for implementation in monitoring CHL residues in milk samples.

Evaluation of the Effect of Brazilian Savanna (Cerrado) Seasons in Flavonoids and Alkaloids Accumulation: The Case of Duguetia furfuracea

Environmental factors can influence secondary plant metabolism, inducing seasonal variation in chemical composition. Few works report how the seasons of Brazilian savanna (Cerrado), rainy and dry, impacts plant metabolism. We investigated the seasonal effect on the production of secondary metabolites in Duguetia furfuracea. Leaves of three individuals were collected monthly for one year and analyzed by ultra-fast liquid chromatography coupled to diode array detector and mass spectrometry (UFLC-DAD-MS), partial least squares-discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA), volcano plot, and Pearson correlation. The main chemical classes found were benzyltetrahydroisoquinoline derivatives and flavonoids. Alkaloids are virtually perennial, with qualitative variation during the year, and flavonoids accumulate during the rainy season. Dew point temperature seemed to be the most representative weather indicator in the metabolite change. These results showed for the first time the effect of the Cerrado environment on the levels of flavonoids and alkaloids in Duguetia furfuracea. They could be useful for exploring this species since it is used in popular medicine and accumulates valuable secondary metabolites.

Analysis of Orphenadrine Citrate in Various Chiral Stationary Phases: A Comparative Study

Background: Polysaccharide based chiral stationary phases (CSPs) were used to perform enantiomeric separation of Orphenadrine Citrate by Ultra-Fast Liquid Chromatography (UFLC) technique. Trials were conducted using the polar mode, reverse phase mode and normal phase mode. Amylose and Cellulose-based CSPs were used for the same. Materials and Methods: Eight Amylose-based CSPs and four Cellulose-based CSPs were used in the reverse phase mode. Five Amylose-based CSPs and two Cellulose-based CSPs were used in polar mode. The only Cellulose-based CSP used in the normal phase mode could effectively separate Orphenadrine Citrate enantiomers with a good resolution. Results and Discussion: Successful enantioseparation was obtained using Chiralcel OD-H containing Cellulose tris (3, 5-dimethylphenylcarbamate) as a chiral selector and n-hexane: Ethanol: Diethylamine (95: 05: 0.1, v/v/v) as the mobile phase. The developed method was validated in accordance with ICH guidelines (Q2R1). Conclusion: The proposed objectives were successfully accomplished as the developed method could effectively resolve Orphenadrine Citrate enantiomers.

Comparison of content-toxicity-activity of six ingenane-type diterpenoids between Euphorbia kansui before and after stir-fried with vinegar by using UFLC-MS/MS, zebrafish embryos and HT-29 cells

The dried roots of Euphorbia kansui (EK) are especially beneficial for the treatment of edema, but the severe toxicity limits their clinical applications. Euphorbia kansui stir-fried with vinegar (VEK) is traditionally employed to reduce the toxicity of EK. However, the material basis for the toxicity reduction with effectivity conservation is still unclear. Therefore, in this study, a rapid, sensitive, and reliable ultra-fast liquid chromatography tandem mass spectrometry (UFLC-MS/MS) method was firstly established to simultaneously determine six ingenane-type diterpenoids, i.e. kansuiphorin C (1), 5-O-benzoyl-20-deoxyingenol (2), 20-deoxyingenol (3), 3-O-(2’E,4’E-decadienoyl)-20-O-acetylingenol (4), 20-O-(2’E,4’Z-decadienoyl)ingenol (5), and ingenol (6), in EK and VEK based on the processing conversion. Then, the toxicity evaluation on zebrafish embryos and modulation of the expression of aquaporin-3 (AQP3) proteins in HT-29 cells were employed to investigate the toxicity-activity of six compounds. Chromatographic separation was obtained on Waters BEH RP18 column (2.1 mm × 100 mm, 2.5 μm) with the mobile phase composed of 0.1 % formic acid in acetonitrile and water, respectively. The column temperature was 35 ℃ at a flow rate of 0.4 mL min−1. Multiple reaction monitoring was conducted in both positive and negative modes for quantitative analysis. The method was then successfully used for the determination of six compounds in EK and VEK. In addition, 1, 2, 4, and 5 had evident cardiotoxicity, intestinal irritation and nutrient absorption disorders on zebrafish larvae, while no in-vivo toxicity was seen for groups given 3 and 6 (LC50 > 200 μM). Meanwhile, 1, 2, 4, 5, and 6 significantly increased the expression of AQP3 protein (p < 0.05) to promote the excretion of water in the colon. This study demonstrated that toxic ingenane-type diterpenoids converted into the less toxic compounds with the same core structure through the breakage of multiple ester bonds in the side chain. At the same time, the laxative effect was retained, providing useful information for the optimization of the process of EK and quality evaluation of other similar toxic Chinese herbal medicines.

Bioguided study of the Antarctic alga Himantothallus grandifolius (A. Geep &amp; E.S.Geep) indicates 13E-Docosenamide as potential antileishmanial agent

Constant efforts to discover new antileishmanial agents are important because there are only a few available drugs for the treatment of leishmaniasis, which present several drawbacks including high toxicity and difficult route of administration. In this scenario, different sources of natural products have been explored regarding their potential to treat infectious diseases. Following this initiative, our research team has been analyzing the bioactive potential of Himantothallus grandifolius, an endemic macroalga of the Antarctic region, with the hypothesis that the hostile environment imposed on these organisms has shaped its arsenal of chemical constituents, thus consequently bringing new opportunities to discover bioactive compounds that might be useful against leishmaniasis. Herein, we report the antileishmanial property of the fatty acid 13E-docosenamide, identified during the fractionation of the hexanic extract of H. grandifolius after semipreparative high performance liquid chromatography separation and ultrafast liquid chromatography coupled with mass spectrometry analyses to track its antileishmanial constituents. 13E-docosenamide was found in HSG11and HSG12 fractions, presented promising antileishmanial activity (IC50 = 9.6 μg ml−1) and is 10 times more selective to the parasite rather than to the host cells (SI > 10.4).

Determination of Coumarins from Pterocaulon balansae by an Ultra-Fast Liquid Chromatography Method in Topical Applications

Determination of Coumarins from Pterocaulon balansae by an Ultra-Fast Liquid Chromatography Method in Topical Applications

Qualitative and quantitative analysis of the major constituents in Spatholobi Caulis by UFLC-Triple TOF-MS/MS and UFLC-QTRAP-MS/MS

There have been few comprehensive studies on the holistic chemical composition of Spatholobi Caulis (SC) and consequently, the information is lacking for the in-depth study of the major constituents. SC is a kind of widely used traditional Chinese medicine with its xylem and phloem alternately arranged in 3–10 rings, but the relationship of phloem ring number and the quality remains unclear. In this study, the characterization of the major constituents in SC was analyzed by ultra-fast liquid chromatography coupled with triple quadrupole-time of flight tandem mass spectrometry (UFLC-Triple TOF-MS/MS), and the content of 19 flavonoids in SC with different phloem ring numbers was simultaneously determined by ultra-fast liquid chromatography coupled with triple quadrupole-linear ion trap tandem mass spectrometry (UFLC-QTRAP-MS/MS). Correlation analysis was performed to evaluate the quality of SC with different phloem ring numbers according to the content of 19 flavonoids. Results showed that 50 constituents in SC were identified and the fragmentation pathways of different types of compounds were preliminarily deduced by the fragmentation behavior of the 50 constituents. In addition, the content of flavonoids increased with phloem ring number, which demonstrated that the content of flavonoids in SC was positively correlated with the number of phloem rings. Our research will contribute to the variety identification and quality evaluation of SC, and provide a scientific basis for evaluating the quality of medicinal materials based on its appearance and characteristics.

Therapeutic Effects of Morinda citrifolia Linn. (Noni) Aqueous Fruit Extract on the Glucose and Lipid Metabolism in High-Fat/High-Fructose-Fed Swiss Mice.

The aim of this study was to evaluate the therapeutic effects of two different doses (250 and 500 mg/kg) of Morinda citrifolia fruit aqueous extract (AE) in high-fat/high-fructose-fed Swiss mice. The food intake, body weight, serum biochemical, oral glucose tolerance test (OGTT), and enzyme-linked immunosorbent assay (ELISA), as well as histological analyses of the liver, pancreatic, and epididymal adipose tissue, were used to determine the biochemical and histological parameters. The chemical profile of the extract was determined by ultra-fast liquid chromatography–diode array detector–tandem mass spectrometry (UFLC–DAD–MS), and quantitative real-time PCR (qRT-PCR) was used to evaluate the gene expressions involved in the lipid and glucose metabolism, such as peroxisome proliferative-activated receptors-γ (PPAR-γ), -α (PPAR-α), fatty acid synthase (FAS), glucose-6-phosphatase (G6P), sterol regulatory binding protein-1c (SREBP-1c), carbohydrate-responsive element-binding protein (ChREBP), and fetuin-A. Seventeen compounds were tentatively identified, including iridoids, noniosides, and the flavonoid rutin. The higher dose of AE (AE 500 mg/kg) was demonstrated to improve the glucose tolerance; however, both doses did not have effects on the other metabolic and histological parameters. AE at 500 mg/kg downregulated the PPAR-γ, SREBP-1c, and fetuin-A mRNA in the liver and upregulated the PPAR-α mRNA in white adipose tissue, suggesting that the hypoglycemic effects could be associated with the expression of genes involved in de novo lipogenesis.

Simultaneous determination of nine constituents by validated UFLC-MS/MS in the plasma of cough variant asthma rats and its application to pharmacokinetic study after oral administration of Huanglong cough oral liquid

Huanglong cough oral liquid (HL), an important traditional Chinese medicine prescription for treating pediatric cough variant asthma (CVA) in Nanjing hospital of traditional Chinese medicine for many years. In this study, a selective, accurate and sensitive ultra fast liquid chromatography extreme resolution coupled with mass spectrometer (UFLC-MS/MS) method was established and validated for the simultaneous determination of nine constituents including morusin, ephedrine, praeruptorin A, praeruptorin B, luteolin, rosmarinic acid, quercetin, amygdalin, caffeic acid in CVA rat plasma sensitized and challenged with ovalbumin and cinnamaldehyde. Plasma samples were prepared by protein precipitation with four-fold amount of methanol. UFLC separation was performed on a Thermo Scientific AcclaimTM RSLC 120 C18 column (2.1 mm × 100 mm, 2.2 μm) with mobile phase containing methanol and 0.1% formic acid-water by gradient elution in 8.1 min at total flow of 0.3 mL/min. The determination of target compounds in plasma was operated by multiple reaction monitoring (MRM) mode with positive and negative electrospray ionization (ESI) source. The correlation coefficients (r) of all compounds were from 0.9930 to 0.9994 in the linear range. Lower limit of quantification (LLOQ, ng/mL) was 0.81, 2.01, 2.11, 1.17, 1.04, 0.89, 0.67, 1.45 and 0.59 for morusin, ephedrine, praeruptorin A, praeruptorin B, luteolin, rosmarinic acid, quercetin, amygdalin and caffeic acid, respectively. Intra- and inter-day accuracy and precision, extraction recovery, matrix effect, carryover effect, dilution integrity, and stability were within the limits specified. The established method was effectively applied to a pharmacokinetic study of the nine compounds in CVA rat plasma following oral administration HL exact (7.5, 15, 30 g/kg).

Simultaneous detection and quantification of 57 compounds in Spatholobi Caulis applying ultra-fast liquid chromatography with tandem mass spectrometry.

A method of ultra-fast liquid chromatography in series with tandem mass spectrometry for the rapid and sensitive detection of 57 compounds in Spatholobi Caulis (the vine stem of Spatholobus suberectus Dunn) within 35min was established. This assay can simultaneously determine a variety of compounds without matrix interference in multiple reaction monitoring mode including evaluating the quality of different batches of Spatholobi Caulis from several areas and further identifying the characteristic compounds efficiently. After comprehensive validation, this method can be used to determinate samples rapidly, precisely, accurately, repeatably, and sensitivity. There were significant content differences in 12 batches of Spatholobi Caulis, which were further classified and systematically differentiated applying multivariate statistical analysis. Furthermore, orthogonal partial least squares discrimination analysis results indicated that (-)-gallocatechin (10), (-)-epiafzelechin (20), 4,7,2'-trihydroxy-4'-methoxyisoflavanol (51), and biochanin A (53) characterize compounds to discernment internal quality of Spatholobi Caulis, and recommended as quality control indicators. Hence, presented work provides a method for further study on pharmaceutic preparation, metabolism, as well as for the design, production optimization process, and clinical application.

Fumonisin B1 Production by Fusarium Species and Mycotoxigenic Effect on Larval Zebrafish.

Fumonisin B1 (FB1) is a common mycotoxin produced by Fusarium species particularly F. proliferatum and F. verticillioides. The toxin produced can cause adverse effects on humans and animals. The objectives of this study were to detect the production of FB1 based on the amplification of FUM1 gene, to quantify FB1 produced by the isolates using Ultra-fast Liquid Chromatography (UFLC) analysis, to examine the embryotoxicity effect of FB1 and to determine EC50 toward the larvae of zebrafish (Danio rerio). Fifty isolates of Fusarium species were isolated from different hosts throughout Malaysia. Successful amplification of the FUM1 gene showed the presence of this gene (800 bp) in the genome of 48 out of 50 isolates. The highest level of FB1 produced by F. proliferatum isolate B2433 was 6677.32 ppm meanwhile F. verticillioides isolate J1363 was 954.01 ppm. From the assessment of embryotoxicity test of FB1 on larvae of zebrafish, five concentrations of FB1 (0.43 ppm, 0.58 ppm, 0.72 ppm, 0.87 ppm and 1.00 ppm) were tested. Morphological changes of the FB1 exposed-larvae were observed at 24 to 168 hpf. The mortality rate and abnormality of zebrafish larvae were significantly increased at 144 hpf exposure. Meanwhile, the spontaneous tail coiling showed a significant difference. There were no significant differences in the heartbeat rate. As a conclusion, the presence of FUM1 in every isolate can be detected by FUM1 gene analysis and both of the species produced different concentrations of FB1. This is the first report of FB1 produced by Fusarium species gave a significant effect on zebrafish development.

Stability Indicating Ultra-Fast Liquid Chromatography Coupled with Photodiode Array Method for Simultaneous Estimation of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Formulations

Stability Indicating Ultra-Fast Liquid Chromatography Coupled with Photodiode Array Method for Simultaneous Estimation of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Formulations

Development and Validation of Spectrophotometric and Liquid Chromatographic Methods for Estimation of Tigecycline in Injections

Three new analytical methods were optimized and validated for the estimation of tigecycline (TGN) in its injection formulation. A difference UV spectroscopic, an area under the curve (AUC), and an ultrafast liquid chromatographic (UFLC) method were optimized for this purpose. The difference spectrophotometric method relied on the measurement of amplitude when equal concentration solutions of TGN in HCl are scanned against TGN in NaOH as reference. The measurements were done at 340 nm (maxima) and 410nm (minima). Further, the AUC under both the maxima and minima were measured at 335-345nm and 405-415nm, respectively. The liquid chromatographic method utilized a reversed-phase column (150mm×4.6mm, 5µm) with a mobile phase of methanol: 0.01M KH2PO4 buffer pH 3.5 (using orthophosphoric acid) in the ratio 80:20 %, v/v. The flow rate was 1.0ml/min, and diode array detection was done at 349nm. TGN eluted at 1.656min. All the methods were validated for linearity, precision, accuracy, stability, and robustness. The developed methods produced validation results within the satisfactory limits of ICH guidance. Further, these methods were applied to estimate the amount of TGN present in commercial lyophilized injection formulations, and the results were compared using the One-Way ANOVA test. Overall, the methods are rapid, simple, and reliable for routine quality control of TGN in the bulk and pharmaceutical dosage form.

Back Cover: Enantiomeric separation and estimation of stiripentol by liquid chromatography mass spectrometer in human plasma

DOI: 10.1002/sscp.202000034 The cover picture shows achiral Stiripentol that was to be extracted from the human plasma for a bioequivalence study for a test and a reference product of 500 mg dose. The enantiomers, R and S Stiripentol behave differently in the human body, hence it was necessary to first extract them from human plasma and then separate them using ultra-fast liquid chromatography (UFLC) and chiral chromatography. High sensitivity for the method was achieved by using triple quad mass spectrometry in unit resolution mode. All these ideas put together resulted in a fantastic enantiomeric resolution with a complete base to base separation for Stiripentol with a resolution factor of more than 4.

Anti-Inflammatory and Chemopreventive Effects of Bryophyllum pinnatum (Lamarck) Leaf Extract in Experimental Colitis Models in Rodents.

Inflammatory bowel diseases, mainly ulcerative colitis and Crohn’s disease are characterized by chronic inflammation in the intestine. Currently several therapeutic strategies available to treat inflammatory bowel diseases. Though, most treatments can be associated with serious adverse effects what justifies the search for new treatments. In this sense, we highlight the interest in herbal products rich in bioactive compounds which immunomodulatory and antioxidant properties as is the case of Bryophyllum pinnatum (Crassulaceae). This plant is used in traditional medicine in Brazil for treating inflammatory diseases. We hypothesized that hydroethanolic B. pinnatum leaf extract has intestinal anti-inflammatory effects on two experimental colitis models: 2.4-dinitrobenzene sulfonic acid (DNBS) in rats, and dextran sulfate sodium (DSS) in mice. Ultra-fast liquid chromatography method used for the quantification of the main compounds indicated good linearity, specificity, selectivity, precision, robustness and accuracy. The major flavonoids (mg/g of the extract) quantified were: quercetin 3-O-α-L-arabinopyranosyl-(1→2)-α-L-rhamnopyranoside (35.56 ± 0.086 mg/g), kaempferol 3-O-α-L-arabinopyranosyl-(1→2)-α-L-rhamnopyranoside (4.66 ± 0.076 mg/g) and quercetin-3-O-rhamnopyranoside (4.56 ± 0.026 mg/g). The results obtained in the DNBS and DSS models indicate that extract has both chemopreventive and anti-inflammatory effects, observing a significant reduction in the disease activity index score, and less macroscopic and microscopic damage. The extract promoted downregulation of Toll-like receptor and kappa B p65 nuclear factor gene expression, leading to a reduction in pro-inflammatory and oxidative mediators, chemokines, and cell adhesion molecules. This immunomodulatory property was proposed that one of the possible action mechanisms of extract. An improvement in intestinal damage was also associated with a reduction in oxidative stress and infiltration of leukocytes, as evidenced by the reduction in malonaldialdehyde and myeloperoxidase activity and increase in total glutathione in the colonic tissue. Moreover, the extract improved the cytoarchitecture of the colonic tissue and the integrity of the intestinal epithelial barrier by restoring the expression of the proteins associated with mucosa protection. In view of the beneficial effects showed by the B. pinnatum leaf extract in preclinical rodent models of colitis there is the potential to conduct some future clinical studies to ensure safe and effective development of a phytotherapeutic treatment for human inflammatory bowel diseases.

An effective UFLC\u2013MS/MS method used to study pharmacokinetics of major constituents of Fukeqianjin formula in rat plasma

BackgroundFukeqianjin formula (FKQJF) is a Chinese medicine prescription, which has been widely used individually or in combination with other western medicine for the treatment of various gynecological inflammatory diseases, including chronic cervicitis, chronic pelvic inflammatory disease and endometritis, so on and so force.MethodsThe ultra-fast liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UFLC–MS/MS), a quick and efficient method was established and applied to quantify the major constituents of Fukeqianjin formula in rat plasma, and its pharmacokinetics of oral absorption was studied. Nineteen components in Fukeqianjin formula were detected and identified as the major compounds absorbed into the blood according to their chromatographic behavior, molecular weight, ion fragments and other information of these compounds. Furthermore, the plasma drug concentration–time curves were established and the related kinetic parameters were analyzed.ResultsThe results showed that all the 19 compounds could be rapidly absorbed by the gastrointestinal tract, the plasma drug concentration of most compounds could reach a peak at around 1–2 h, and the double-peaks on behalf of the enterohepatic circulation were found in most drug concentration–time curves. The method used in this experiment was validated comprehensively including specificity, linearity, precision, accuracy, stability, matrix effect, and recovery.ConclusionsThese results showed that the developed method was suitable for pharmacokinetic analysis of the main components of Fukeqianjin formula in rat plasma, and may provide useful information for the subsequent distribution studies in vivo.

Development and Validation of a Sensitive Method for Analysis of Ellagic Acid in Dietary Supplements from Punica granatum

Ellagic acid is a popular antioxidant dietary supplement. It is a natural phenolic compound common to multiple botanical sources. The ellagic acid based dietary supplements are manufactured by hydrolysis of ellagitannins from Punica granatum. However, use of ellagic acid from sources other than P. granatum is not uncommon. Currently, there is no robust analytical methodologies for quantification and confirmation of the enriched ellagic acid (›40%) source in commercial products. Therefore, we have developed and validated ultra-fast liquid chromatography - photodiode array detector and ultra-fast liquid chromatography - tandem mass spectrometry methods for quantification and source identification of ellagic acid in commercial products. The results of the study confirm punicalin A-B, punicalagin A and B as positive markers and catechin and chebulinic acid as adulteration markers.

Evolution of liquid chromatography: Technologies and applications

Liquid chromatographic offers efficient analyte separation employing high pressure pumps. The reversed phase high performance liquid chromatography (RP-HPLC) is widely utilized in the purity testing and quantitative determination of pharmaceuticals and neutraceuticals. The limitations of traditional liquid chromatography such as particle size, resolution and selectivity demanded for the developments and Waters Corporation developed ultraperformance liquid chromatography (UPLC). Ultrafast liquid chromatography (UFLC) is another milestone, which offers faster and efficient separation. Multidimensional UHPLC provides separation of complex molecules. The particle size decrease enhances the resolution of LC separation. Ethylene bridged hybrid (BEH), Charged surface hybrid (CSH) and Peptide separation technology (PST) offer better performance in. The amalgamation of chromatographic and spectroscopic detectors namely fluorescence detector (FD) and mass spectrometry (MS) provides efficient separation. Liquid chromatography (LC) offers the analysis of pharmaceuticals, biological, food materials, and natural products. This review covers technologies and recent pharmaceutical and biomedical applications of liquid chromatography technologies