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Abstract

Toddalia asiatica (Linn) Lam (T. asiatica) as a traditional Miao medicine was investigated to find rational alternative medicinal parts for T. asiatica root bark and its antitumor chemical constituents by quantitative pharmacognostic microscopy, high performance liquid chromatography (HPLC) fingerprint and multivariate statistical analysis. A bivariate correlation analysis method based on microscopic characteristics and content of chemical constituents was established for the first time, there were some regular discoveries between powder microscopic characteristics and common chromatographic peaks of T. asiatica through quantitative pharmacognostic microscopy, cork cells, calcium oxalate square crystal, brown clump, starch granule and phloem fiber, as powder microscopic characteristics may be placed where the main chemical constitutes were enriched. Scores plot of principal component analysis (PCA) and dendrogram of hierarchical clustering analysis (HCA) showed that 18 T. asiatica samples were distinguished correctly, clustered clearly into two main groups as follows: S01∼S03 (root bark) and S07∼S09 (stem bark) in cluster 1, S04∼S06 and S10∼S18 in cluster 2. Nineteen common peaks were obtained in HPLC fingerprint of T. asiatica, loadings plot of PCA indicated seven compounds played important roles in different part of samples (P10 > P08 > P07 > P14 > P16 > P17 > P19), peaks 04, 06, 07, 08, 10 were identified as hesperidin, 4-methoxycinnamic acid, toddalolactone, isopimpinlline and pimpinellin. MTT assay was used to determine the inhibitory activity of different medicinal parts of T. asiatica on human breast cancer MCF-7 cells, all parts of T. asiatica had different inhibitory effects on MCF-7 cell lines, root and stem barks of T. asiatica showed the best inhibitory activity. The relationship between chemical constituents and the inhibitions on MCF-7 cell had been established, significant antitumor constituents of T. asiatica were identified by correlation analysis, the order of the antitumor effect of the main compounds was P07 (toddalolactone) > P16 > P06 (4-methoxycinnamic acid), P11 > P18 > P10 (pimpinellin) > P08 (isopimpinellin) > P01 > P19 > P14 > P04 (hesperidin) > P17, which were antitumor chemical constituents of T. asiatica root bark. T. asiatica stem bark was the most rational alternative medicinal part for T. asiatica root bark.

Open access
Authors: Ramia Z. Al Bakain, Yahya S. Al-Degs, James V. Cizdziel and Mahmoud A. Elsohly

Abstract

In this research, cannabis varieties represent 23 USA States were assayed by GC-FID to generate their complex chemical profiles informative for plants clustering. Results showed that 45 cannabinoids and terpenoids were quantified in all plant samples, where 8 cannabinoids and 18 terpenoids were identified. Among organics, Δ9-THC, CBN (cannabinoids) and Fenchol (terpenoid) not only showed the highest levels overall contents, but also were the most important compounds for cannabis clustering. Among States, Washington, Oregon, California and Hawaii have the highest cannabis content. GC-FID data were subjected to PCA and HCA to find (1) the variations among cannabis chemical profiles as a result of growing environment, (2) to reveal the compounds that were responsible for grouping cultivars between clusters and (3) finally, to facilitate the future profile prediction and States clustering of unknown cannabis based on the chemical profile. The 23 cannabis USA States were grouped into three clusters based on only Δ9-THC, CBN, C1 and Fenchol content. Cannabis classification based on GC-profile will meet the practical needs of cannabis applications in clinical research, industrial production, patients' self-production, and contribute to the standardization of commercially-available cannabis cultivars in USA.

Open access

Abstract

Fifty four domestically produced cannabis samples obtained from different USA states were quantitatively assayed by GC–FID to detect 22 active components: 15 terpenoids and 7 cannabinoids. The profiles of the selected compounds were used as inputs for samples grouping to their geographical origins and for building a geographical prediction model using Linear Discriminant Analysis. The proposed sample extraction and chromatographic separation was satisfactory to select 22 active ingredients with a wide analytical range between 5.0 and 1,000 µg/mL. Analysis of GC-profiles by Principle Component Analysis retained three significant variables for grouping job (Δ9-THC, CBN, and CBC) and the modest discrimination of samples based on their geographical origin was reported. PCA was able to separate many samples of Oregon and Vermont while a mixed classification was observed for the rest of samples. By using LDA as a supervised classification method, excellent separation of cannabis samples was attained leading to a classification of new samples not being included in the model. Using two principal components and LDA with GC–FID profiles correctly predict the geographical of 100% Washington cannabis, 86% of both Oregon and Vermont samples, and finally, 71% of Ohio samples.

Open access

Abstract

Sodium polystyrene sulfonate (SPS) powder is in use for over 50 years for the treatment of hyperkalemia. SPS powder is official in United States Pharmacopoeia, British Pharmacopoeia and European Pharmacopoeia. However, till date, no study has been published on the assessment of organic impurities for this drug. The organic impurities in bulk drug and finished product are associated with their safety, efficacy and stability. A simple, rapid, specific, precise and an accurate HPLC method has been developed for the estimation of toxic organic impurities like styrene, naphthalene, divinyl benzene (DVB) and ethylvinyl benzene (EVB) from SPS bulk drug and finished product. The developed method was validated for specificity, accuracy, precision, linearity, limit of detection (LOD), limit of quantitation (LOQ), solution stability, ruggedness and robustness. The influence of acid, alkali, oxidative stress, photolytic stress, thermal stress and humidity stress conditions on SPS bulk powder and finished product has been studied and reported. The proposed method can be successfully employed for the impurity testing of commercial batches of the bulk drug and finished products of both sodium salt and calcium salt of polystyrene sulfonate.

Open access

Abstract

Mono- and bis-pyridinium quaternary aldoximes (K-oximes) have long been employed as cholinesterase reactivator components of antidotes against lethal cholinesterase-inhibiting organophosphorous chemicals. Their positive charge poses difficulties in their chromatographic analysis, resulting in the publication of different approaches for each K-oxime. A multiplexed method is presented for the rapid quantitation of 10 K-oximes in blood with its utility demonstrated in vivo. Liquid chromatography with absorbance detection was employed. Reversed-phase separation was achieved on a highly nonpolar stationary phase. Method validation was based on the respective guideline of the European Medicines Agency. Times to peak concentrations and 120-min areas under the time–concentration curves were determined in rats following intraperitoneal administration. Adequate retention and separation of K-oximes with acceptable peak shapes in short isocratic runs was achieved by adjusting ionic strength, organic content and the concentration of the ion-pairing agent of the mobile phase. Chromatographic properties were governed by optimizing the concentration of dissolved ions. Accurate adjustment of the organic content was indispensable for avoiding peak drifting and splitting. Dose-adjusted exposure to K-347 and K-868 was exceptionally low, while exposure to K-48 was the highest. The method is suitable for screening systemic exposure to various K-oximes and can be extended.

Open access

Abstract

This study presents the optimization and validation of methods for the analysis of retinol, thiamine, niacin, pyridoxine, folic acid, cyanocobalamin, zinc, and iron in fortified kernels (coated and extruded) and in fortified rice. The analyses were performed by HPLC-UV/FLD/MS and ICP-OES. The optimized methods showed good resolution of the analyte peaks, excellent recovery (87–108%), reproducibility with relative standard deviation (SD) of analyte content between 1.8 and 11% and high correlation coefficient of the calibration curves (R2 > 0.997). Limit of detection was from 2.8 E-4 mg/kg for pyridoxine to 1.26 mg/kg for zinc and limit of quantification was from 9.2 E-4 mg/kg for pyridoxine to 4.21 mg/kg for zinc. Thereby the optimized methods demonstrated reliability and sensitivity in the detection and quantification of these micronutrients and that they are suitable for routine analysis of fortified kernels (coated and extruded) and fortified rice.

Open access
Authors: Fatema Moni, Suriya Sharmin, Satyajit Roy Rony, Farhana Afroz, Shammi Akhter and Md. Hossain Sohrab

Abstract

This study describes the development and validation of a simple, specific, accurate, and precise method for quantitative determination of Esomeprazole in human serum using Pantoprazole as internal standard (IS). After the addition of internal standard, Esomeprazole from serum samples was extracted simply by protein precipitation method followed by centrifugation and the supernatants were directly injected into the high performance liquid chromatography (HPLC). The chromatographic separation of the compounds was obtained on Hitachi Lachrom C8 column (5 µm, 250 × 4.6 mm) with a mobile phase consisting of 5 mM potassium dihydrogen phosphate pH 7.4 and acetonitrile in a ratio of 70:30 with UV detection at 302 nm with a flow rate of 1 mL/min. The method was sensitive and specific, and validated over a concentration range of 0.06–6.0 µg/mL. The limit of detection (LOD) and lower limit of quantification (LOQ) was 0.03 µg/mL and 0.06 µg/mL, respectively. The precision and accuracy expressed as relative standard deviation were less than 15%. The average recovery of Esomeprazole from serum was 97.08%.

Open access

Abstract

A rapid and sensitive High-Performance Liquid Chromatography-tandem Mass Spectrometry (HPLC/MS/MS) method for determining apremilast in beagle dog plasma and urine samples was developed and validated using clopidogrel as the internal standard (IS). Apremilast was extracted from the plasma and urine samples by liquid–liquid extraction using methyl tert-butyl ether. Chromatographic separation was performed using a C8 column with gradient elution and a mobile phase containing methanol and 0.1% formic acid. Quantification was achieved in multiple reaction monitoring (MRM) mode with a transition of m/z 461.3→178.2 for apremilast and m/z 322.2→184.1 for clopidogrel (IS). This method was validated regarding its specificity, linearity, precision, accuracy, and stability. The lower limit of quantification (LLOQ) for this method was 5 ng/mL, and the calibration curve was linear over 5–1,000 ng/mL. The intra- and inter-run coefficients of variance (CV) of aprelimast in plasma samples were less than 12.92% and 10.64%, respectively, while in urine samples, the CV were less than 11.84% and 10.20%, respectively. The samples were stable under the tested conditions. This method was successfully applied to a pharmacokinetic study in beagle dogs following oral administration of 10 mg of apremilast.

Open access

Abstract

Calycanthine is an important class of alkaloids extracted and isolated from the roots, leaves, flowers and fruits of Chimonanthus praecox. In this work, the UPLC-MS/MS method was used for determination of calycanthine in rat plasma, and the pharmacokinetics in rats were investigated. Midazolam was used as an internal standard (IS), and methanol precipitation method was used to pretreatment the rat plasma samples. Chromatographic separation was achieved on a UPLC BEH C18 (50 × 2.1 mm, 1.7 μm) column with the mobile phase of methanol- 0.1% formic acid aqueous solution with gradient elution. Multiple reaction monitoring (MRM) mode with positive ionization was applied for quantitative analysis, m/z 347.3 → 246.7 and 326.2 → 291.4 for calycanthine and IS, respectively. The results indicated that within the range of 1–200 ng/mL, linearity of calycanthine in rat plasma was good (r > 0.995), and the lower limit of quantification (LLOQ) was 1 ng/mL. Accuracy range was between 90.6 and 109.4%, precision (RSD) of calycanthine was less than 14%. The matrix effect was between 97.9% and 105.4%, the recovery was better than 85.6%. The developed UPLC-MS/MS method was successfully applied in the pharmacokinetics of calycanthine in rats after oral and intravenous administration. The absolute bioavailability of the calycanthine was 37.5% in rats.

Open access
Authors: Jinzhao Yang, Huamin Liu, Yuan Cai, Yazhen Wu, Xiaoxin Xu, Xianqin Wang and Chongliang Lin

Abstract

Twelve Sprague-Dawley rats were randomly divided into two groups: Citrus suavissima Hort. ex Tanaka group and control group (n = 6). The rats in Citrus suavissima Hort. ex Tanaka group were given Citrus suavissima Hort. ex Tanaka juices (1 mL/100 g) by oral administration each day, continued for 14 days; the rats in control group were given Stroke-physiological saline solution (1 mL/100 g) by oral administration each day, continued for 14 days. The rats of these two groups were given a single oral administration of erlotinib (20 mg/kg) on the 15th day. After blood sampling at different time points and processing, the concentrations of erlotinib in rat plasma were determined by the established ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method. Chromatographic separation was achieved using a UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with erlotinib-d6 as an internal standard (IS). The initial mobile phase consisted of acetonitrile and water (containing 0.1% formic acid) with gradient elution. Multiple reaction monitoring (MRM) modes were utilized to conduct quantitative analysis. The sensitive, rapid and selective UPLC-MS/MS method was successfully applied to analyse the effect of Citrus suavissima Hort. ex Tanaka on pharmacokinetics of erlotinib in rat plasma. There were no significant differences in AUC(0−t), t1/2, Tmax, CL, Cmax between the two groups (P > 0.05). While MRT(0−t) was decreased (P < 0.05) in Citrus suavissima Hort. ex Tanaka group, compared to the control group. It showed that Citrus suavissima Hort. ex Tanaka could not affect the metabolism of erlotinib.

Open access

Abstract

A simple, inexpensive and sensitive method was developed for the simultaneous determination of three pesticide residues (carbendazim, thiophanate-methyl, and imidacloprid) in fruit and vegetable samples using high performance liquid chromatography (HPLC) based on a combined pretreatment of ultrasound-assisted deep eutectic solvent extraction (UA-DES-E) and liquid-liquid extraction (LLE). In this study, various types of deep eutectic solvents (DESs) were synthesized and the extraction efficiency was compared as extraction solvents. Results showed that glycerol-proline = 9:4 (GP-5) obtained the highest extraction efficiency among different types of DESs. Experiment conditions, including DES volume, extraction time and pH, were systematically optimized using single-factor experiment. Under the optimum conditions, the limits of detection (LODs) and quantification (LOQs) were in the ranges of 0.05–0.2 μg·mL−1 and 0.1–0.5 μg·mL−1, respectively. The relative recoveries of the three pesticides in the fruit and vegetable samples ranged from 85.7 to 113.0% at two spiked levels. Meanwhile, the method achieved excellent linearity with determination coefficients (r) greater than 0.999. Furthermore, the method was successfully applied to the analysis of the pesticides in real fruit and vegetable samples (apple, tomato, and grape).

Open access

Abstract

A simple, accurate and sensitive method of high performance liquid chromatography (HPLC) with diode array detector was established to identify Xinfeng capsules and systematically evaluated its quality, based on chromatographic fingerprint integrated with the similarity analysis, hierarchical cluster analysis and the quantitative analysis of multi-components by single marker (QAMS). In this study, 18 peaks were selected as the common peaks to evaluate the similarities among different batches (S1–S10) of Xinfeng capsules samples, which were manufactured in the First Affiliated Hospital of Anhui University of Chinese Medicine with a three-year span. Compared to control fingerprint, the similarities values for 10 batches of samples were more than 0.90. Moreover, by analyzing the reference of astragalus, the chromatogram of astragalus was developed, and 10 common peaks of astragalus were identified. More importantly, simultaneous quantification of three markers in Xinfeng capsule, including Calycosin-7-glucoside, calycosin and Formononetinaldehyde was performed, the three constituents showed good regression (R > 0.999) within linear ranges, and their recoveries were within the range of 97.6–101.5%. The validation results showed that the developed method was specific, accurate, precise and robust. This study demonstrated that the developed method offers an efficient, reliable and practical approach for systematic quality evaluation of Xinfeng capsule.

Open access

Abstract

A simple capillary electrophoresis (CE) method with ultraviolet (UV) detection was developed for the determination of hexachlorophene (HCP) in cosmetics. Separation conditions were obtained in 20 mM Na2B4O7, 10% MeOH (pH 9.20), with 25 kV applied voltage and UV detection at 208 nm. Under the selected conditions, electrophoretic analysis was completed in about 4 min, with limit of detection (LOD) of 0.06 µg·mL−1 for HCP. The method was successfully applied to determine HCP in three kinds of cosmetics with relative standard deviations (RSD) of 0.52–3.02% and recoveries from 90.0 to 96.4% for the spiked samples. The results indicated that the proposed method was reliable. Comparative experiments were also carried out with high-performance liquid chromatography (HPLC)-UV method described in National Standards of People's Republic of China. The validation results of the two methods are comparable, but the proposed CE method is simple, rapid, which makes separation and analyte quantification in shorter time with much less reagent consumption.

Open access

Abstract

The major processes for introducing polycyclic aromatic hydrocarbons (PAHs) in food are smoking and grilling of different products. But in addition, PAHs can permeate in the food chain due to their high lipophilicity and ability to be accumulated in specific tissue, through contaminated animal feed. Further, when some parts of these animals are marketed as food, the accumulated PAHs can go to the human organism. Some of them are classified as highly toxic, carcinogenic and mutagenic for animal and human organisms so they are under consideration of International and European legislation. This work reports development and validation of simple and fast GC/MS method for 16 PAHs determination. Comparison of two methods for sample preparation in pork meat matrix standard extraction/saponification procedure and modified QuEChERS method is also done. In addition, this paper report the calibration step of instrument and a recovery study for 16 PAHs in model pork meat, using modified QuEChERS procedure for sample pretreatment. The calibration step with accessible and suitable for use in real laboratory conditions internal standard (chrysene D12) is done in the range 10–100 ppb using toluene as solvent. The obtained results show very good linearity (R2 = 0.99 to 1.00). For the recovery study six model samples were spiked with 16 PAHs and they all are subjected to QuEChERS procedure. The recovery is calculated and the obtained data (71–120%) is in a good correlation with requirements of international legislation. Finally, LOD values for all 16 investigated compounds of modified GC/MS method and for the instrument were determined.

Open access

Abstract

In the last few years, the use of surfactants as mobile phase additives in reversed phase liquid chromatography (RPLC) has been steadily developing and improving. Surfactants modify the polarity of the stationary phase which in turn decreases the amount of organic solvent required for elution of the analytes rendering the methodologies linked to them greener and more eco-friendly. Brij-35 is a fatty alcohol ethoxylates non ionic surfactant, which is less widely used as mobile phase additive. Brij-35 can decrease stationary phase polarity while remaining neutral. In this research, Brij-35 was studied in the separation and determination of marketed antihypertensive combination therapy composed of triamterene (TRM) and xipamide (XIP). TRM and XIP are diuretics used for treatment of essential hypertension and associated edema conditions. Chromatographic separation was achieved on RP-C18 column (Kinetix®, 5 µm, 15 cm × 4.6 mm) at flow rate 1  mL  min−1 and UV-detection at 254 nm. Isocratic elution was performed using mobile phase composed of 0.1 M Brij-35: methanol (MeOH) (60:40, v/v). The analytes were well separated and quantified within linearity ranges of 5–50 µg mL−1 for both drugs in short retention time (2.6 and 5.3 min. for TRM and XIP, respectively). Since claiming greenness is not enough, Green Analytical Procedure Index (GAPI) was used to demonstrate the superiority of the proposed method over the previously reported methods. GAPI is a new metric for evaluation of the ecological impact of analytical procedures. The proposed method was validated according to ICH guidelines and applied successfully for simultaneous determination of the drugs in their co-formulated tablets.

Open access

Abstract

A simple HPLC technique has been utilized for rapid and sensitive quantitative analysis of two mixtures of drugs that are used during pregnancy and lactation. Drugs of the first mixture are used to manage gastrointestinal tract illness that are common during early stages of pregnancy, while pharmaceutical agents of the second mixture are administered over the counter as galactagogues or to overcome postpartum depression. Mixture I includes famotidine (FMT), ranitidine (RNT), nizatidine (NZT), and pantoprazole (PNT), which were separated on a C18 column using a mobile phase composed of methanol: 0.02 M sodium dihydrogen phosphate (60:40, v/v) of pH 6.9, adopting UV detection at 240 nm at a flow rate of 1 mL/min. Mixture II on the other hand, consists of domperidone (DOM), metoclopramide (MET), and sulpiride (SUL). These drugs were eluted using the same column and flow rate as those in mixture I, using a mobile phase consisting of acetonitrile: 0.075 M sodium dihydrogen phosphate (30:70, v/v) of pH 6 adopting a detection wavelength 270 nm. Two optimization protocols were utilized to optimize the chromatographic separation conditions, namely one factor at a time (OFAT) and design of experiments (DOE) where face centered cube response surface experimental design was chosen for this investigation. Comparison of the results obtained from both protocols reveals the accordance between them.

Full validation procedure under guidance of United States Pharmacopoeia (USP) was applied to the proposed methods which enabled their application to separate the drugs of both mixtures in spiked rat whole blood samples and in vivo analysis of rat heart blood.

Open access
Authors: Jianbo Li, Yujie Hu, Yajin Wu, Tiantian Feng, Congcong Wen and Xiajuan Jiang

Abstract

Palmatine is a compound with good water solubility extracted from Coptis chinensis, Fibraurea recisa Pierre, Cortex Phellodendri Chinensis. Palmatine has good antibacterial activity and mainly used for the treatment of bacterial dysentery, gynecological inflammation, surgical infection, and conjunctivitis. It has anti-diabetic, anti-oxidant, and cognitive-enhancing activities. In this study, we used UPLC-MS/MS to determinate palmatine in rat plasma, and investigated its pharmacokinetics. Coptisine was utilized as an internal standard (IS), and acetonitrile precipitation method was used to process the plasma samples. Chromatographic separation was achieved using a UPLC BEH C18 column using mobile phase of acetonitrile- 0.1% formic acid with gradient elution. Electrospray ionization (ESI) tandem mass spectrometry in multiple reaction monitoring (MRM) mode with positive ionization was applied. The results indicated that within the range of 1–500 ng/mL, linearity of palmatine in rat plasma was acceptable (r > 0.995), and the lower limit of quantification (LLOQ) was 1 ng/mL. Intra-day and inter-day precision RSD of palmatine in rat plasma were less than 14%. Accuracy range was between 93.7 and 107.1%, and matrix effect was between 101.6 and 109.4%. The method was successfully applied in the pharmacokinetics of palmatine in rats after oral and intravenous administration. The absolute bioavailability of the palmatine was 15.5% in rats.

Open access

Vicia faba, also known as “bakla” in Turkey, is a species of Fabaceae family that is widely grown in Africa and Asia. It is rich in levodopa, a medicinal substance used to treat Parkinson's disease. Levodopa produced by chemical synthesis is expensive and causes various side effects. Therefore, it is recommended to use natural levodopa sources to prevent possible side effects. A Central Composite Design technique has been used in this study to optimize levodopa extraction from Vicia faba. First, a single factor analysis examined 3 variables such as extraction temperature, extraction time, and concentration of acetic acid. The purpose of this study was to assess the effects of variables chosen on levodopa's extraction performance. By using variance and regression analyses, a second-order regression equation was determined as a predicted model. The value of R2 is 0.9882, which shows that the equation fits well. The best conditions are as follows: a temperature of 59.85 °C, an extraction time of 18.74 min, and an acetic acid content of 0.28%. Under optimum conditions, the maximum levodopa yield calculated from the predicted module was 4.53%. Extraction efficiency was determined as 4.54% experimentally under optimum conditions. A good relationship has been found between the experimental result and the predicted value.

Open access

Cortisol and cortisone are 2 important glucocorticoids produced in the human hypothalamus–pituitary–adrenal (HPA) axis that respond to stress. An analytical method to determinate cortisol and cortisone in serum and saliva using high-performance liquid chromatography–tandem mass spectrometry following a supported liquid extraction (SLE) was developed. Serum and saliva samples of 0.2 mL were extracted by SLE three times using 0.4 mL of methyl tert-butyl ether each time. The chromatographic separation was obtained on an Agilent Poroshell column using a 0.01% formic acid buffer and acetonitrile (60:40, v/v) as the solvent with a flow rate of 0.3 mL/min. Optimized quantitative mass transitions for cortisol, cortisone, and cortisone d-4 were 363.2/121.0 (m/z), 361.2/163.1 (m/z), and 367.1/270.7 (m/z), respectively. The method validation was achieved according to regulatory guidance. The lower limit of quantification (LLOQ) in serum were 2 ng/mL for cortisol and 1 ng/mL for cortisone, and the LLOQ in saliva were 0.1 ng/mL for cortisol and 0.2 ng/mL for cortisone. The developed method showed convenient and efficient extraction, a lower LLOQ, and a short running time. Modest correlations between serum and saliva cortisol and cortisone concentrations were found. The method was successfully applied in assessing the HPA condition of patients with depressive disorders.

Open access
Authors: Jovana Tomić, Branka Ivković, Slavica Oljačić, Katarina Nikolić, Nevena Maljurić, Ana Protić and Danica Agbaba

The aim of this study was to develop a novel reversed-phase high-performance liquid chromatography (RP-HPLC) method for efficient separation of ivabradine and its 11 impurities. Similar polarity of impurities in the sample mixture made method optimization challenging and accomplishable only when different chemometric tools, such as principal component analysis (PCA), Box–Behnken design (BBD), and desirability function as a multicriteria approach, were employed. The presence of 3 positional isomers (impurities III, V, and VI), keto–enol tautomerism of impurity VII, and diastereoisomers of impurity X made separation of this complex mixture even more challenging. Chromatographic retention parameters obtained with the mobile phase consisting of 30 mM phosphate buffer and acetonitrile (80:20, v/v) on four different RP-HPLC columns at varying pH values (3.0, 4.0, and 5.0) were subjected to the PCA analysis to select the column with the most appropriate selectivity. Then the column temperature, pH of the aqueous component of mobile phase, phosphate buffer molarity and the organic solvent content in the mobile phase were estimated employing BBD. Valid and reliable mathematical models towards resolution of twelve critical peak pairs were obtained. After determination of the desirability making criteria for all responses, desirability functions were established and used in optimization. The proposed optimal chromatographic conditions included the Zorbax Eclipse Plus C18 chromatographic column (100 × 4.6 mm, 3.5 μm), the column temperature of 34 °C, the mobile phase flow rate of 1.6 mL min−1 and the UV detection at 220 nm. The mobile phase consisted of the 28 mM phosphate buffer at pH 6.0 and acetonitrile (85:15, v/v). Separation of one pair of positional isomers was not achieved, so methanol was added to the organic part of mobile phase in small increments with the optimal ratio of methanol to acetonitrile 59:41, v/v. The overall organic component of the mobile phase also increased to 18%, accelerating the chromatographic analysis.

Open access

We developed and validated an assay for determination of glyphosate (GLYP) and glufosinate (GLUF) in human serum. Serum samples were extracted by using a MonoSpin® TiO column and analyzed by liquid chromatography–tandem mass spectrometry (LC–MS/MS). MonoSpin® TiO tends to specifically bind to phosphate groups. The assay was linear over a concentration range of 1–250 μg/mL. The recoveries for the 2 compounds were 1.6%–2.3%. The intra- and inter-day variations were <15%. Precision and accuracy were 5.6%–12.7% and 97.0%–103.9%, respectively. The validated method was applied to quantify the GLYP and GLUF content in the serum of GLYP and GLUF-poisoned patients. In conclusion, the method was successfully applied for accurate determination of GLYP and GLUF in serum obtained from patients with GLYP and GLUF poisoning.

Open access
Authors: Lenche Velkoska-Markovska, Mirjana S. Jankulovska, Biljana Petanovska-Ilievska and Kristijan Hristovski

Coffee is one of the most widely consumed beverages in the world. It contains many bioactive compounds, including chlorogenic acid which possesses various biological properties. In this study, in order to determine concentration of chlorogenic acid in green coffee, a reverse-phase rapid resolution liquid chromatography (RP-RRLC) method with diode-array detection (DAD) was developed. Successful separation was achieved on a Poroshell 120 EC-C18 (50 mm × 3 mm; 2.7 μm) column using acetonitrile–water with 1% phosphoric acid (10:90, v/v) as a mobile phase, at a flow rate of 1 mL/min, and with UV detection at 325 nm. The identification was made with comparison of the retention time of pure analytical standard with the retention time of chlorogenic acid in the analyzed samples. The developed method was validated using the following parameters: linearity, sensitivity, selectivity, precision, and accuracy. Excellent linearity over the range 12.33–143.50 μg/mL was achieved with R 2 values greater than 0.99. The intra-day precision was validated with the %RSD values, which confirmed that the method for determination of chlorogenic acid was repeatable. The mean recovery rate of the method ranged between 97.87% and 106.67% with %RSD values lower than 1%. The limit of detection and limit of quantification values under the used chromatographic conditions were 0.29 and 0.96 pg, respectively. This method was successfully employed for quantitative determination of chlorogenic acid in green coffee samples.

Open access

The advent of disposable micro-columns will be a hope of workers of chromatography-related laboratories. A very critical and important requirement is the formation of affordable inlet frits. Welding a metal screen to a column inlet is not recommended because of the risk of damage to stationary phase. In this study, the Tollens probe (silver mirror reaction) was adopted to make affordable frits. Silver is reduced on the particle surface and in an empty space among the particles, forming a solid silver network structure at the column inlet area by injecting the reaction solution into the packed column at a depth of one third (10 cm) of the packed bed (0.5 mm × 300 mm). The silver cement structure was successfully formed, and the silver cement frit endured mobile phase flow well when C18 modified ground silica monolith particles were used to make the packed bed. The formation of the silver cement frit was not successful when the stationary phase based on conventional spherical silica particles was used. Negligible reduction of chromatographic performance by the silver cemented frit was observed. This study serves as the first step toward realization of disposable micro-columns.

Open access

This study aimed to develop a chromatographic method to quantitatively determine phenol in fish tissues. This method involves solvent extraction of acidified samples, followed by derivatization to phenyl acetate and analysis with gas chromatography coupled with mass spectrometry (GC–MS). Phenol in a representative tissue sample (belly, gill, or renal tubules), which was homogenized with 2 N sulfuric acid, was extracted with ethyl acetate and derivatized to phenyl acetate using acetic anhydride and K2CO3 in water. An n-butyl acetate extract was injected into the GC–MS. The linearity (r 2) of the calibration curve was greater than 0.996. The analytical repeatability, which is expressed as the relative standard deviation, was less than 6.14%, and the recovery was greater than 96.3%. The method detection limit and the limit of quantitation were 8.0 μg/kg and 26 μg/kg, respectively. The proposed method is also applicable to the analysis of other biological tissues for phenol and its analogs, such as pentachlorophenol.

Open access
Authors: Xi Bao, Bingge Huang, Yiting Mao, Zhiguang Zhang, Yunfang Zhou, Congcong Wen and Quan Zhou

Byakangelicol is one of coumarins from Baizhi and has been shown to inhibit the release of PGE2 from human lung epithelial A549 cells in a dose-dependent manner. A sensitive ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed and full validated for the quantification of byakangelicol in rat plasma. The pharmacokinetics of byakangelicol after both intravenous (5 mg/kg) and oral (15 mg/kg) administrations were studied. Chromatographic separation was performed on an ultra-performance liquid chromatography ethylene bridged hybrid (UPLC BEH) C18 column with acetonitrile and 0.1% formic acid as the mobile phase at a flow rate of 0.4 mL/min; fargesin was used as the internal standard (IS). The following quantitative analysis of byakangelicol was utilized in the multiple reaction monitoring mode. The samples were extracted from rat plasma via protein precipitation using acetonitrile. In the concentration range of 1–2000 ng/mL, the method correlated linearity (r > 0.995) with a lower limit of quantitation (LLOQ) of 1 ng/mL. Intra-day precision was less than 11%, and inter-day precision was less than 12%. The accuracy was between 92.0% and 108.7%, the recovery was better than 89.6%, and the matrix effect was between 85.9% and 98.6%. The method was successfully applied to a pharmacokinetic study of byakangelicol after intravenous and oral administration, and the absolute bioavailability was 3.6%.

Open access
Authors: Lianguo Chen, Qingwei Zhang, Yijing Lin, Xiaojie Lu, Zuoquan Zhong, Jianshe Ma, Congcong Wen and Cheng Ding

An ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was established to determine the hapepunine in mouse blood, and the pharmacokinetics of hapepunine after intravenous (1.0 mg/kg) and intragastric (2.5, 5, and 10 mg/kg) administrations was studied. Delavinone was used as an internal standard. The UPLC ethylene bridged hybrid (BEH) C18 column was used for chromatographic separation. The mobile phase consisted of acetonitrile and 0.1% formic acid with a gradient elution flow rate of 0.4 mL/min. Multiple reaction monitoring (MRM) mode was used for quantitative analysis of hapepunine in electrospray ionization (ESI) positive interface. Proteins from mouse blood were removed by acetonitrile precipitation. The verification method was established in accordance with the US Food and Drug Administration (FDA) bioanalytical method validation guidelines. In the concentration range of 1–1000 ng/mL, the hapepunine in the mouse blood was linear (r 2 > 0.995), and the lower limit of quantification was 1.0 ng/mL. In the mouse blood, the intra-day precision coefficient of variation (CV) was less than 12%, the inter-day precision CV was less than 14%. The accuracy ranged from 91.7% to 109.3%. The average recovery was higher than 76.7%, and the matrix effect was between 86.0% and 106.4%. The UPLC–MS/MS method was sensitive, rapid, and selective and was successfully applied to the pharmacokinetic study of hapepunine in mice. The absolute bioavailability of hapepunine was 22.0%.

Open access
Authors: Yanqin Zhu, Ping Du, Shaojun Huang, Qinhong Yin and Yaling Yang

A fingerprint analysis method was established for the quality control of Moringa seed shells by high-performance liquid chromatography with diode array detection (HPLC–DAD). The HPLC–DAD separation was performed on a Thermo Hypersil Gold C18 (4.6 mm × 250 mm, 5 μm) column by gradient elution with acetonitrile–water as mobile phase. The fingerprint of Moringa seed shells was established with good precision, reproducibility, and stability obtaining within 60 min, and 13 common peaks in the fingerprint were designed. Similarity analysis, principal component analysis (PCA), and hierarchical clustering analysis (HCA) were carried out to analyze the obtained fingerprints. The similarity among 11 batches of samples in addition to No. 5 and 6 was no less than 0.92. Eleven samples could be classified into 2 clusters. The HPLC fingerprint technology and application of chemical pattern recognition can provide a more comprehensive reference for the quality control of medicinal plants.

Open access

An isocratic reversed-phase high-performance liquid chromatography (RP-HPLC) method has been developed for rapid and simultaneous separation and estimation of 3 antidiabetic drugs, namely, metformin, pioglitazone, and glimepiride, in human plasma within 3 min. Separation was carried out on a MAGELLEN 5U C18 (5 μm, 150 mm × 4.60 mm) using a mobile phase of MeOH–0.025 M KH2PO4 adjusted to pH 3.20 using ortho-phosphoric acid (85:15, v/v) at ambient temperature. The flow rate was 1 mL/min, and the maximum absorption was measured at 235 nm. The retention time of metformin, pioglitazone, and glimepiride was noted to be 1.24, 2.32, and 2.77 min, respectively, indicating a very short analysis time compared to that of other reported methods. Also, limits of detection were reported to be 0.05, 0.26, and 0.10 μg/mL for metformin, pioglitazone, and glimepiride, respectively, showing a high degree of method sensitivity. The method was then validated according to the FDA guidelines for the determination of the three drugs clinically in human plasma, in particular, regarding pharmacokinetic and bioequivalence simulation studies.

Open access
Authors: Stefano Dugheri, Nicola Mucci, Alessandro Bonari, Giorgio Marrubini, Giovanni Cappelli, Daniela Ubiali, Marcello Campagna, Manfredi Montalti and Giulio Arcangeli

In the last decade, the development and adoption of greener and sustainable microextraction techniques have been proved to be an effective alternative to classical sample preparation procedures. In this review, 10 commercially available solid-phase microextraction systems are presented, with special attention to the appraisal of their analytical, bioanalytical, and environmental engineering. This review provides an overview of the challenges and achievements in the application of fully automated miniaturized sample preparation methods in analytical laboratories. Both theoretical and practical aspects of these environment-friendly preparation approaches are discussed. The application of chemometrics in method development is also discussed. We are convinced that green analytical chemistry will be really useful in the years ahead. The application of cheap, fast, automated, “clever”, and environmentally safe procedures to environmental, clinical, and food analysis will improve significantly the quality of the analytical data.

Open access
Authors: Abdul Shakoor, Mahmood Ahmed, Rabia Ikram, Sajad Hussain, Arifa Tahir, Badrul Mohamed Jan and Ahmad Adnan

The present work aimed to develop and validate a simple, rapid, sensitive, accurate, and precise method for simultaneous determination of metformin hydrochloride and vildagliptin in tablet and biological samples. Isocratic elution of both the analytes was performed at 35 °C by injecting 20 μL into Thermo Hypersil ODS C18 column (5 μm, 4.6 mm× 250 mm), while the flow rate was set to 0.8 mL/min. The mobile phase comprised of methanol, acetonitrile, and phosphate buffer (5:30:65, v/v, pH 3.5), and wavelength was selected at 212 nm. The overall run time per sample was 7.0 min with a retention time of 3.36 and 5.41 min for metformin hydrochloride and vildagliptin, respectively. The calibration curve was linear from 10–140 μg/mL for metformin and 1–14 μg/mL for vildagliptin with a coefficient of determination (R 2) ≤ 0.9919, while repeatability and reproducibility (expressed as relative standard deviation) were lower than 1.13 and 0.97%, respectively. Force degradation studies indicated a complete separation of the analytes in the presence of their degradation products providing a high degree of method specificity. The proposed reversed-phase high-performance liquid chromatography (RP-HPLC) method was demonstrated to be simple and rapid for the determination of metformin hydrochloride and vildagliptin in commercially available tablet and biological samples providing recoveries ranged between 100.13–100.29%.

Open access
Authors: Leonel Vinicius Constantino, Douglas Mariani Zeffa, Alessandra Koltun, Mariana Ragassi Urbano, Alisson Wilson Santos Sanzovo and Suzana Lucy Nixdorf

An optimal condition for extraction of soluble sugars from green coffee using water and a validated chromatographic method for its separation and quantification were proposed in this research. An orbital incubator shaker (OIS) and microwave-assisted extraction (MAE) were the 2 techniques used to extract soluble sugars. In such experiments, the variables: sample amount (300, 400, and 500 mg), time (30, 60, and 90 min), and temperature (30, 45, and 60 °C) were tested. The separation of sugars was performed in a chromatographic system (high-performance liquid chromatography refractive index detector [HPLC-RID]), which presented the selectivity for the analytes, a limit of detection of 0.020 g/L, a limit of quantification of 0.0625 g/L, and recovery rates greater than 95%. The repeatability and inter-day precision had low dispersion, RSD < 2.0% and < 3.0%, respectively. Sucrose content ranged from 0.65 to 2.39 g/L using an OIS and from 1.19 to 2.72 g/L by MAE, while glucose and fructose concentration varied from 0.08 to 0.12 g/L using both methods. The OIS technique is preferably indicated for extraction of soluble sugars at the following conditions: 500 mg of grounded green coffee, 90 min, and 60 °C. The proposed method for soluble sugar extraction and quantification may be applied in research laboratories and food industries since it is a low-cost and environment-friendly technique.

Open access
Authors: Mohammad Al Bratty, Neelaveni Thangavel, Ramalingam Peraman, Vinod Kumar, Padmanabha Reddy, Krishna Veni Nagappan and Hassan Al Hazmi

A reversed-phased high-performance liquid chromatography–diode-array detection (HPLC–DAD) method has been developed for investigating the stress-dependent degradation of pantoprazole (PTZ) by a photolytic and oxidative mechanism. The developed method separated PTZ from its degradation products on a C18 column with a mobile phase consisted of methanol and water (60:40, v/v; pH 3.0) at a flow rate of 1 mL/min. The linear regression coefficient of 0.9995 was obtained for a concentration range from 5 to 25 μg/mL. The % relative standard deviation for repeatability and intermediate precision were below 0.5% and 1.5%, respectively, while the sensitivity of the method was demonstrated by a limit of detection value of 0.25 μg/mL. The stress sample analyses for PTZ results revealed the formation of a total of 18 degradation products, and out of them, 9 degradation products were common for both photolytic and oxidative degradations. Further, the oxidation by azobisisobutyronitrile produced the highest number of degradation products (11 impurities), 3 of which are more hydrophobic than PTZ. In photolytic degradation, 8 and 7 degradation products were observed with UV radiation and sunlight exposure, respectively. Furthermore, the degradation of pantoprazole sodium injection formulation was carried out under the same stress conditions, and it revealed the formation of 3 common impurities under both stress conditions, but other impurities were not detected in the formulations. Finally, 3 common impurities formed in formulations of PTZ injections, viz., sulfone, N-oxide, and N-oxide sulfone impurities, were identified by spike analyses.

Open access

Phyllostachys edulis (PES), the most important bamboo species in China, is widely distributed in East Asia. Flavonoids, which are important bioactive natural compounds, often have similar structures, making their structural elucidation difficult. The aim of this study was to represent valuable, reliable mass spectral data for the identification of flavonoids in plant leaves. Ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC–Q-TOF-MS/MS) method was established for characterization and identification of the major flavonoids in PES leaf extract. A total of 13 flavonoids were simultaneously characterized, and their proposed characteristic product ions and fragmentation pathways were investigated. Thirteen compounds were separated on an Agilent Zorbax RRHD SB-C18 column (150 mm × 2.1 mm, 1.8 μm). On the basis of comparing with the 4 reference standards and the literature data, the other 9 flavonoids were identified by tandem mass spectrometry (MS/MS). Eight compounds (compounds 1, 4, 5, 8, 9, 10, 11, and 12) were found in PES leaves for the first time. An efficient UPLC–QTOF-MS/MS method was successfully applied for the structural identification of flavonoids in PES leaves. These results have practical applications for the rapid identification and structural characterization of these compounds in crude bioactive extracts or mixtures.

Open access

Isocorynoxeine is one of the main alkaloids in Chinese medicinal herbs, and has pharmacological activities such as antihypertensive, sedative, anticonvulsant, and neuronal protection. It is an effective component of Uncaria for the treatment of hypertension. In this study, we used a fast and sensitive ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) to detect isocorynoxeine in rat plasma and investigated its pharmacokinetics in rats. Six rats were given isocorynoxeine (15 mg/kg) by intraperitoneal (i.p.) administration. Blood (100 μL) was withdrawn from the caudal vein at 5 and 30 min and 1, 2, 4, 6, 8, 12, and 24 h after administration. Chromatographic separation was achieved using a UPLC BEH C18 column using a mobile phase of acetonitrile–0.1% formic acid with gradient elution. Electrospray ionization (ESI) tandem mass spectrometry in the multiple reaction monitoring (MRM) mode with positive ionization was applied. Intra-day and inter-day precisions (relative standard deviation, %RSD) of isocorynoxeine in rat plasma were lower than 12%. The method was successfully applied in the pharmacokinetics of isocorynoxeine in rats after intraperitoneal administration. The t 1/2 of isocorynoxeine is 4.9 ± 2.1 h, which indicates quick elimination.

Open access

The authorities have identified an emerging trend where over-the-counter products, represented as dietary supplements, contain hidden active ingredients that could be harmful. Consumers may unknowingly take products laced with varying quantities of approved prescription drug ingredients, controlled substances, and untested and unstudied pharmaceutically active ingredients. Hidden ingredients are increasingly becoming a problem in products promoted for sexual enhancement, weight loss, or bodybuilding. The tests have revealed the presence of some undesired substances like sildenafil, tadalafil, vardenafil, and their analogues in tainted sexual enhancement products. The content of these substances is usually around the daily curative dose. A simple high-performance liquid chromatography (HPLC) method for simultaneously determination of sildenafil, vardenafil, tadalafil, dapoxetine, yohimbine, and sibutramine was developed and validated. InfinityLab Poroshell 120 EC-C18 (150 '4.6 mm '4 μm particles) was used, as well as a diode-array detector (DAD) at 230 nm, and a gradient flow with 0.030 М ammonium acetate buffer and acetonitrile. The method is linear in the following range: 2.5–37.5 μg/mL for yohimbine, 2.06–30.9 μg/mL for vardenafil, 2.0–30.0 μg/mL for sildenafil, 3.1–46.5 μg/mL for tadalafil, 1.98–29.7 μg/mL for dapoxetine, and 2.2–66.0 μg/mL for sibutramine. The linearity coefficient is R 2 = 1 for all substances. Model matrices were spiked, and the analytical recoveries for all substances are in the range 97.5%–99.5%. The method exhibited an upper hand compared with previously reported methods in terms of speed and simplicity. Additionally, the mobile phase (also used as extracting, column washing, and diluting solvent) was composed of only buffer and acetonitrile, which rendered the method much cheaper than others.

Open access
Authors: Jianbo Li, Zheng Yu, Cheng Han, Zhening Wang, Yujie Hu, Congcong Wen and Chongliang Lin

In this study, we used UPLC–MS/MS to determine diosmetin-7-o-β-d-glucoside in rat plasma and investigated its pharmacokinetics in rats. Six rats were given diosmetin-7-o-β-d-glucoside (5 mg/kg) by intravenous (i.v.) administration. The blood (150 μL) was withdrawn from the caudal vein after administration. Diazepam was used as an internal standard (IS), and a one-step acetonitrile precipitation method was used to process the plasma samples. Chromatographic separation was achieved using a UPLC BEH C18 column using a mobile phase of acetonitrile–0.1% formic acid with gradient elution. Electrospray ionization (ESI) tandem mass spectrometry in multiple reaction monitoring (MRM) mode with positive ionization was applied, 463.1 → 301.0 for diosmetin-7-o-β-d-glucoside, m/z 285.1 → 193.0 for diazepam (IS). Intra-day and inter-day precision of diosmetin-7-o-β-d-glucoside in rat plasma were less than 14%. The method was successfully applied in the pharmacokinetics of diosmetin-7-o-β-d-glucoside in rats after intravenous administration. The t 1/2 of diosmetin-7-o-β-d-glucoside is 1.4 ± 0.4 h, which indicates the quick elimination.

Open access

In reversed-phase liquid chromatography, in case of the absence of additives, cationic basic compounds give rise to asymmetrical and broad peaks as a result of interactions of analyte cations with residual free silanols on silica-based stationary phases. Ionic liquids, added to the mobile phase, have been suggested as alternatives to amines to block the activity of free silanols. The different parameters affecting the retention behavior, symmetry of peak, system efficiency, and separation selectivity of selected psychotropic drugs, especially the effect of concentration of ionic liquid, kind and concentration of organic modifiers of mobile phases, and kind of stationary phases were investigated. The most selective and efficient systems are used for separations of psychotropic drug standards' mixture and for determination of selected psychotropic drugs in human serum.

Open access

Abstract

The existing effective domestic regional development framework requires analyses for increasingly wider areas (micro, meso and even macro regions) before operational – short-term – local developments to be prepared and implemented.

Such comprehensive complex studies or larger-term programmes may demonstrate the profitability of the given project and can complement it with combined utilization technologies; in the case of Himesháza several locally known renewable energy sources could facilitate geothermal heat, later electricity supply, e.g. local biomass (biogas-based) recovery technology (organic waste of the local pig farm) and, for example, the construction of a low-power “dwarf” hydroelectric power plant chain based on rich watercourses of the region (the “southern dwarves” in Hungary) and the connection of existing solar utility facilities to a modern “smart grid” system in the longer term.

Himesháza, located in southern Hungary in Baranya county, is developing; it has a detailed feasibility study of a thermal energy supply network and an energy supply development plan.

Based on the geothermal characteristics of Baranya county it would be reasonable to encourage the development of smaller-scale, decentralized heating systems for dynamic settlements. Several settlements in close proximity to Himesháza have already explored thermal wells. Power generation with a small scale, closed-loop system can be used in the project region for thermal water with an outflow temperature of 90 °C. The heating system may also be able to fulfill the needs of recreational, vacation-based or complex thermal spa facilities formerly planned in the region. Moreover, the system could also be capable of utilizing a larger spectrum of renewable energy through its combination with photovoltaic technology.

Due to the country's favorable agricultural characteristics, Hungary's biomass potential is higher than the European average. The utilization of organic waste from agricultural and farming sectors is highly recommended in Baranya county; biogas production seems to be the most suitable in the region of Himesháza too, broadening the utilization of renewable resources.

The realization of the current project could contribute to shifting the energy resource sector in a more modern, environmentally conscious direction.

The background for shorter-term plans and investment (carried out within the framework of operational programs) necessary for the optimal operation and maintenance of longer-term (25–50 years) energy development strategies is created by the analysis (at multiple scales) of complex regional characteristics and future potential, and the selection of optimal sites.

Open access

Összefoglalás

A klímaváltozásnak köszönhetően a következő évtizedekben a talajok defláció veszélyezettségének mértéke emelkedni fog hazánkban (CSORBA et al. 2012). A kutatásunk során arra kerestünk választ, hogy a talajtani alaptulajdonságok miként befolyásolják a kritikus indítósebességet és jelenleg mennyire defláció veszélyeztetettek a Dél-Alföld talajai.

Mintaterületként a Szeged környéki talajokat választottuk. A vizsgálataink során megállapítottuk, hogy az összes vizsgált talajparaméter közül az agronómiai szerkezet, azon belül is a rögfrakció az, ami leginkább befolyásolja a kritikus indítósebességet. Ez felhívja a figyelmet az ember szerepére, aki megfelelő agrotechnikával képes lenne a széleróziós kockázat csökkentésére (BODOLAY 1966; SHAHABINEJAD et al. 2019). A szélcsatornás kritikus indítósebesség vizsgálatok eredményeit összevetettük az időjárási adatokkal, és ezek eredményeit kivetítettük Csongrád megye területére. Kutatásunk során meghatároztuk a széleróziós események jellemző éves előfordulását (homok: 16,8 esemény; homokos vályog és vályog 1,6 esemény; agyagos vályog: 0,4 esemény), ezen események átlagos hosszát (homok: 3,0 óra; homokos vályog és vályog 1,4 óra; agyagos vályog: 1,0 óra) és a deflációnak kitett területetek aránya Csongrád-megyében (homok: 8,4%; homokos vályog és vályog 29,1%; agyagos vályog: 20,0%).

Kutatásunkkal képet kaptunk arról, hogy a defláció mekkora területet érint és mennyire jelentős talajvédelmi probléma Csongrád megyében. Korábbi kutatásaink bizonyítják, hogy egy párperces széleróziós esemény is súlyos veszteséget okozhat a talajok tápanyagtartalmából (FARSANG et al. 2011; FARSANG 2016), mely csupán csak egy aspektusa a szélerózió negatív hatásainak.

Open access
Authors: Qinghua Weng, Lianguo Chen, Luxin Ye, Xiaojie Lu, Zheng Yu, Congcong Wen, Yichuan Chen and Gang Huang

The aim of this study was to establish a rapid, sensitive, and selective ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method to quantify the concentrations of licochalcone A and applicate the technique to its pharmacokinetic study. Analytes were separated on an UPLC ethylene bridged hybrid (BEH) C18 column (2.1 mm × 50 mm, 1.7 μm). The mobile phase was consisted of acetontrile and 0.1% formic acid with a flow rate of 0.4 mL/min in a gradient elution mode. Multiple-reaction monitoring (MRM) was carried out in a negative mode for licochalcone A (m/z 337.2 → 119.7) and the internal standard (IS) (m/z 609.0 → 300.9). The linearity of licochalcone A was great from 0.53 to 530 ng/mL. The lower limit of quantification and the lower limit of detection were 0.53 ng/mL and 0.26 ng/mL, respectively. The intra-day precision was less than 14%, and the inter-day precision was no more than 11%. The accuracy was from 91.5% to 113.9%, the recovery was over 90.5%, and the matrix effect was between 84.5% and 89.7%. The results of stability were in an acceptable range. The bioavailability was only 3.3%, exhibiting poor absorption. The developed method was successfully applicable for determining the concentrations of licochalcone A and its pharmacokinetic study.

Open access

Metaldehyde is a molluscicide allowed for use in the control of slugs and snails in agriculture and horticulture in many countries. A simple, fast, and precise gas chromatography method was developed and single-laboratory validated for determination of metaldehyde in different formulations of plant protection products. The proposed method involves extraction of active substance from samples by sonication with acetone and analysis using gas chromatography–flame ionization detection (GC–FID). The suggested analytical procedure is accurate, precise, and repeatable. Moreover, it is environmentally friendly and useful for laboratories as it uses a no time- and no solvent-consuming reference chromatography technique for quality control of commercially available pesticide formulations. Advantages of the proposed method are consistent with the ideas of sustainable development, which are in accordance with the principles of Green Analytical Chemistry. Analysis of real samples of commercial pesticide formulations confirmed that the proposed method is fit for its purpose.

Open access

A new reversed-phase high-performance liquid chromatographic method with ultraviolet detection (RP-HPLC-UV) for simultaneous determination of phenytoin impurities, benzophenone and benzil, was developed and validated according to the International Council for Harmonization (ICH) guidelines. Chromatographic separation was performed on a C8 column using acetonitrile–1% acetic acid (60:40, v/v). The correlation coefficients of the calibration lines were greater than 0.999 with 95% confident interval of y-intercept over the origin. The analytical method showed good precision, intra-day precision ≤1.00 and inter-day precision ≤1.53. The standard solution of each compound exhibited good stability 99.18–99.70%, after storage at room temperature for 24 h. The limit of detection (LOD) and limit of quantification (LOQ) were 0.0015 and 0.005 μg/mL, respectively. The resolution of the impurities was 2.935 ± 0.009. The proposed analytical method was successfully applied to determine the amount of benzophenone and benzil in marketed products. The amount of benzophenone was found at 3.09–5.91 × 10−3%, while benzil was not detected in the samples.

Open access

The present study aimed to develop and validate an analytical method for determination of marbofloxacin (MAR) in veterinary chewable tablets. The isocratic reversed-phase chromatographic method was developed and validated using a Vertisep®, RP C18 column (150 mm × 4.6 mm, 5.0 μm). The mobile phase was composed of water–acetonitrile (55:45, v/v) with pH adjusted to 3.0 with ortho-phosphoric acid and a flow rate set at 0.4 mL/min. The proposed method was validated for linearity in a concentration range of 2.5 to 17.5 μg/mL with a correlation coefficient of 0.99991. The mean content of MAR found in chewable tablets was 104.40% with RSD below 2%. The accuracy expressed as average recovery of the proposed method was 98.74%, and the precision expressed as relative standard deviation among repeated analysis was 0.55%. The method has adequate sensitivity with detection and quantitation limits of 0.25 and 0.81 μg/mL, respectively. Based on the presented results and according to the ICH and AOAC guidelines on validation of analytical methods, the proposed method was considered precise, accurate with adequate sensitivity, and robust in the MAR quantitative analysis. Therefore, the method can be used in the quality control of chewable veterinary tablets containing MAR.

Open access
Authors: Francielle Q. Soares, Bruna F. Alvarenga, Marçal A. Ruggiero, Monise C. Casanova, Eliana M. Lima, Denilson Rabelo and Andréa R. Chaves

A reliable method using disposable pipette extraction (DPX) based on composite of pernigraniline and styrene–divinylbenzene (Sty–DVB) copolymer was applied to the analysis of dexamethasone in synovial fluid using high-performance liquid chromatography and ultraviolet (UV) detector (HPLC–UV).

DPX variables, namely, number of draw/eject cycles for extraction or desorption, sample pH, volume, and desorption solvent, were optimized to establish the best sorption equilibrium and analysis time. The highest extraction efficiency value was obtained with 50 μL of synovial fluid mixed with 1950 μL of water, in five cycles of 300 μL of sampling, followed by liquid desorption of the drug with 300 μL of methanol in three cycles. The developed method demonstrated a linear response over the range from 10 to 100 ng/mL, with R 2 = 0.993. The limit of quantification (LOQ) was 10 ng/mL. Based on the validation results, the proposed method can be a useful tool to detect dexamethasone levels in synovial fluid.

Open access

Abstract

Hungary was one of the main countries in the world as regards the yields reached in maize production. The research was conducted to appraise the effect of NPK fertilizer on traits of different hybrid maize (Fao410, Fao340) at the University of Debrecen and our experiment was carried out in Centre for Agricultural Sciences, Institute of Crop Sciences at Látókép in 2018. NPK fertilizer was applied in six different combinations (0-0-0 control, 30-23-27 first dose, 60-46-54 second dose, 90-69-81 third dose, 120-92-108 fourth dose and 150-115-135 fifth dose kg · ha−1). The result of compound variance showed the level of fertilizer and interaction between fertilizer and genotypes were significant in one percent. Effect of genotypes was a variable level of fertilizer and providing a different yield in the level of fertilizer. The weight of seeds in ear and weight of ear were important traits in the average yield on Fao410 hybrid. Also, the fourth of the fertilizer level was the best level of fertilizer for yield on Fao410 and Fao340. the weight of fresh plant and weight of seeds in ear were highest relation with yield in H340 hybrid. The results of this research can successfully contribute to the science of maize cultivars, the given adapted hybrid to the discovery of their traits and to an application of fertilizers.

Open access

Összefoglalva

A hazai talajosztályozás diagnosztikus szemléletű megújítását minden erőmmel és képességemmel támogatom, ám a Vitaanyagban ismertetett módon történő leváltását jó szívvel nem tudom javasolni. Javaslom ugyanakkor az egyes részletkérdések (akár munkacsoportokban történő) megvitatását, a terepi és laboratóriumi módszertan fejlesztését, a módszerek konvertálhatóságának megteremtését, illetve a Vitaanyag módszertanának – mint 1. verziójú javaslatnak – a meglévő módszertannal párhuzamosan történő tesztelését szelvényfeltárások és a talajtérképezési munka során.

Open access
Authors: Hella Fodor, Ádám Csorba, Márta Fuchs, Károly Penksza and Erika Michéli
Open access

A novel method coupling spin column extraction with high-performance liquid chromatography–mass spectrometry was developed for simultaneous extraction of β-blockers and calcium channel blockers from human serum. Sample loading, washing, and elution were accomplished via centrifugation of the column, in which mixed-mode monolithic silica bonded to a C18 reversed phase, and a cation-exchange phase was packed in a spin column. The serum sample (0.2 mL) pH was adjusted to 3 and the analytes adsorbed onto the column were eluted with 0.1 mL MeOH containing 2% NH3. The recoveries of the tested drugs were 76–108%. A linear curve was observed up to a concentration of 500 ng/mL of the target drugs in serum (r 2 > 0.996). The intra-day relative standard deviations at three different concentrations were 0.6–9.6%. The limits of detection were 2 ng/mL. The proposed method was successfully applied to clinical and forensic cases.

Open access

Two pairs of homoisoflavonoid analogues, 6-aldehydo-isoophiopogonanone A (1) / 6-aldehydo-isoophiopogonanone B (2) and methylophiopogonanone A (3) / methylophiopogonanone B (4), were obtained from the fibrous roots of Ophiopogon japonicus (L. f.) Ker-Gawl. (FROJ) by silica gel column chromatography (SGCC) and recycling high-speed counter-current chromatography (rHSCCC). First, the ethyl acetate fraction from the 70% ethanol extract was pre-separated by SGCC with a petroleum ether–ethyl acetate gradient (50:1–2:1, v/v). Then, the two sub-fractions containing homoisoflavonoid analogues were further separated by rHSCCC with n-hexane–ethyl acetate–methanol–acetonitrile–water (3:2:3.5:1:0.5, v/v) and n-hexane–ethyl acetate–methanol–acetonitrile–water (3:2:2.5:1:1.5, v/v). Finally, 6-aldehydo-isoophiopogonone A (16 mg), 6-aldehydo-isoophiopogonanone B (26 mg), methylophiopogonanone A (46 mg), and methylophiopogonanone B (148 mg) were obtained with purities of 97.82%, 96.70%, 97.76%, and 94.62%. Their structures were identified by high-resolution quadrupole-time-of-flight mass spectrometry (HR-QTOF-MS), ultraviolet (UV), and nuclear magnetic resonance spectroscopy (1H-NMR and 13C-NMR). These results demonstrated that rHSCCC could be used for the large scale preparation of homoisoflavonoid analogues from FROJ, which provides scientific support for utilization of untraditional medicinal part of O. japonicus and also for reduction in waste of plant resources. Additionally, an online antioxidant activity assay was investigated with hyphenated HSCCC-DPPH (1-diphenyl-2-picrylhydrazyl) radical scavenging detection.

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A gyümölcstermesztésben a megfelelő termésbiztonság elérésében és fenntartásában kulcsszerepe van a rendszeres és szakszerű tápanyag-utánpótlásnak. Munkánk során nitrogén, foszfor, kálium és magnézium műtrágyakészítmények (Kontrol, NP, NPK, NPKMg) hatását vizsgáltuk a fák vegetatív és generatív teljesítményére intenzív almaültetvényben ’Golden Reinders’ fajtán hároméves kísérletben (2016-2018). Eredményeink alapján a vizsgált időszakban a legnagyobb mértékű (80%) törzsgyarapodást a NPKMg kezelésben rögzítettük. 2016-ban és 2018-ban a fák optimális termésmennyiséget produkáltak (0,83-1,25 kg cm−2, valamint 0,78-1,15 kg cm−2) míg 2017-ben a kisugárzási fagyok miatt alacsonyabb hozamokat kaptunk (0,21-0,60 kg cm−2). Valamennyi évben és kezelésben a gyümölcsök mérete elérte az étkezési piac által támasztott követelményeket (75 mm átmérő), de azok alakulásában tendenciózus kezeléshatást nem tapasztaltunk. A levelek méretében ugyanakkor 6-10% körüli gyarapodás jelentkezett és azok relatív klorofill-tartalma, azaz a SPAD értéke, 2-5% körüli mértékben növekedett a vizsgálatok 2. és 3. évében az alkalmazott műtrágya kezelésekben.

Open access
Authors: Anita Szabó, Klára Pokovai, Péter Ragályi, Márk Rékási, Renáta Sándor, Botond Bernhardt, József Koncz, Rita Kremper and Péter Csathó

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Dolgozatunkban a Kádár Imre által 1991 tavaszán meszes csernozjom talajon 13 potenciálisan toxikus mikro-/károselem (Al, As, Ba, Cd, Cr, Cu, Hg, Mo, Ni, Pb, Se, Sr és Zn) 0-90-270-810 kg ha−1 szintjeivel beállított szabadföldi tartamkísérlet 1-20. évi főtermés veszteség eredményeit értékeljük.

Első megközelítésben, a főtermések relatív termésben kifejezett százalékos terméscsökkenéseit az egyes elemek átlagos fitoxicitása sorrendjében mutattuk be. A 90-270-810 kg ha−1 kezelések átlagában az egyes elemek fitotoxicitása az alábbi sorrendben csökkent: Se > Cr > Cd > Al > Pb > As > Mo > Hg > Cu > Ni > Zn > Ba > Sr. A legkisebb – a kontroll %-ában kifejezett - relatív terméseket, azaz a legnagyobb fitotoxikus hatást a Se, a Cr és a Cd mutattak, míg legkevésbé fitotoxikusnak a Ni, a Zn, a Ba és a Sr bizonyultak.

A fitotoxikus évek számát illetően az elemek sorrendje az alábbiak szerint alakult: Se = As > Cr > Cd > Hg > Cu = Zn > Al = Mo > Ni = Pb > Ba = Sr. Legtöbb évben a Se, az As, a Cr és a Cd, míg legkevesebb évben a Ni, a Pb, a Ba és a Sr voltak fitotoxikusak.

Átlagos terméscsökkentő hatásukat tekintve a Se, a Cr és a Cd már a 90 kg ha−1 adagtól, a Pb és a Hg a 270 kg ha−1 adagtól, míg az As és a Mo a legnagyobb, 810 kg ha−1 adagtól/adagnál mutatott 10%-nál nagyobb terméscsökkenést a fitotoxikus évek átlagában.

Mind a fitotoxikusság mértéke, mind a fitotoxikus évek száma tekintetében, általában, a legnagyobb növényi károsodások az anionos formában kijuttatott elemekhez voltak köthetők, ezen belül is, főleg a nem esszenciális mikroelemekhez.

Második megközelítésben, a fitotoxicitás mértékét a kísérleti növények, ill a kísérlet beállítása óta eltelt idő függvényében is nyomon követtük.

A kísérleti növények közül legnagyobb terméscsökkenéseket a napraforgó, a spenót és az őszi árpa, míg legkisebbeket a harmadik, a második és az első éves lucerna növényekben kaptunk.

Az idő múlásával egyre kisebb terméscsökkenéseket tapasztaltunk, a kísérlet 13. évétől kezdve az átlagos fitotoxicitás mértéke még a 10%-ot sem érte el.

Megkülönböztetett figyelemmel kell nyomon kísérni a kadmiumot, amely az idő múlásával egyre kevésbé volt fitotoxikus, viszont még a kísérlet 18. évében is igen nagy könnyen oldható elemtartalmakat mutatott a talaj szántott rétegében.

Open access
Authors: Anita Szabó, Klára Pokovai, Péter Ragályi, Márk Rékási, Renáta Sándor, Botond Bernhardt, József Koncz, Rita Kremper and Péter Csathó

Összefoglalás

Dolgozatunkban Kádár Imre 1991 tavaszán meszes csernozjom talajon 13 potenciálisan toxikus mikro- / károselem (Al, As, Ba, Cd, Cr, Cu, Hg, Mo, Ni, Pb, Se, Sr és Zn) 0-90-270-810 kg ha−1 szintjeivel beállított szabadföldi tartamkísérlete 1-20. évi talajvizsgálati-, illetve talaj visszanyerési százalék eredményeit értékeltük.

A LE-oldható (NH4-acetát+EDTA) és „összes” (cc.HNO3+cc.H2O2) elemtartalmak aboszlút értékeinek változásai mellett figyelemmel kísértük az elemek visszanyerési százalékainak időbeni változásait is.

A kísérlet 4. évében a legnagyobb talaj LE-oldható visszanyerési százalékokat a kationos formában kiadott elemek esetében kaptunk, míg az anionos formában kijuttatottak (Se, As, Mo, Cr) az elemsorrend második felében, illetve a végén helyezkedtek el.

A kísérlet 18. évére a legtöbb elem LE módszer szerint kimutatott oldhatósága, és így visszanyerési százaléka kisebb-nagyobb mértékben csökkent. Legnagyobb csökkenéseket a Sr, Pb, Zn, míg legkisebbeket a Mo, Cr és Cd elemek mutattak. A tápláléklánc szennyeződése szempontjából kedvezőtlen, hogy a kijuttatott karcinogén Cd jelentős hányada maradt a könnyen oldható frakcióban, és ez a frakció a 14 év alatt is csak minimálisan csökkent a meszes csernozjom talajon.

Az elemek átlagában, a LE-oldható elemtartalomban mért visszanyerési százalék 1994-ben 41%, míg 2008-ban 22% volt. A 14 év alatt tehát az átlagos, LE-oldható elemtartalomban mért visszanyerési százalék 19 abszolút %-kal csökkent, azaz gyakorlatilag megfeleződött.

Az egyes elemek „összes” elemtartalomban a kísérlet 4. évében mért visszanyerési százalékának sorrendje a LE-oldható tartalmaknál leírtakhoz hasonlóan alakult; a legkisebb visszanyerési százalékokat itt is az anionos formában kijuttatott elemeknél kaptunk. Míg a LE-oldható tartalmaknál a Hg, addig az „összes” elemtartalmaknál a Cd, mint nehézfém ékelődött be az anion formában kijuttatott elemek közé.

A kísérlet 18. évében az „összes” elemtartalomban mért visszanyerési százalékok esetében a legnagyobb csökkenéseket a Se, Sr, Pb, míg legkisebbeket a Cd, Mo és Cr elemek mutattak. Az utóbbi három elem közül humán egészségügyi szempontból a Cd tekinthető a legveszélyesebbnek.

Az elemek átlagában, az „összes” elemtartalomban mért visszanyerési százalék 1994-ben 81%, míg 2008-ban 49% volt. A 14 év alatt tehát az átlagos összes elemtartalomban mért visszanyerési százalék 32 abszolút %-kal Csökkent.

A meszes csernozjom, meszes homok, és savanyú, agyagos vályog textúrájú barna erdőtalajon beállított károselem-terheléses tartamkísérlet adatainak tanúsága szerint a LE-oldható elemtartalomban kifejezett, és az „összes” elemtartalomban kifejezett visszanyerési százalékok egymáshoz viszonyított aránya inkább elem-, mint talajtulajdonság-függő.

Úgy találtuk, hogy a három, egymástól jelentősen eltérő tulajdonságú talajon viszonylag azonos időben, azonos mennyiségben és azonos formában kijuttatott potenciálisan káros elemeknek a LE-oldható elemtartalomban mért visszanyerési százalékok és ”összes” (cc.HNO3+cc.H2O2-oldható) elemtartalomban mért visszanyerési százalékok arányainak sorrendje állandó, tehát a talajtulajdonságoktól független volt.

Ezen felismerés alapján fogalmaztuk meg a Szabó-Csathó-Kádár-féle potenciálisan toxikus talaj-mikroelem kiterjeszthetőségi szekvens megnevezést. Természetesen ezen hipotézisünket még számos más talajtípuson, hasonló feltételekkel beállított tartamkísérletben is szükséges lenne bizonyítani.

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Összefoglalás

Egy vegyület akkor tekinthető ösztrogén hatásúnak, ha alacsony affinitással is, de az ösztrogén receptorokhoz (ER) kötődni képes, az ösztrogén érzékeny sejtekben, szövetekben biológiai hatást képes kiváltani, melyek hasonlóak a petefészek eredetű ösztrogén hormonok által előidézett folyamatokhoz.

Irodalmi adatok alapján a hígtrágyával történő öntözés és intenzív állattartásból származó trágya termőföldre juttatása következtében a szteroid ösztrogének, beleértve az ösztron E1, ösztradiol E2, ösztriol E3, valamint a szintetikus ösztrogén EE2, mindenütt jelen vannak a termőtalajban. A növények felhalmozhatják az ösztrogéneket gyökereikben és hajtásaikban, bekerülve a táplálékláncba akár hatást fejthetnek ki a humán egészségre. A vegyületek ösztrogén potenciálját legtöbbször a 17β-ösztradiol referencia vegyülethez viszonyított relatív potenciálként fejezik ki. Meghatározó szerepe van a vegyületek minőségének, ugyanis az egyes ösztrogénhatású vegyületek különböző hatással lehetnek a receptorokra

Ahogy korábban említettük, hígtrágya csak talajtani szakvéleményre alapozott talajvédelmi hatósági engedély birtokában juttatható ki mezőgazdasági területre, melyben az is meghatározásra kerül, hogy a területre milyen mennyiségű hígtrágya helyezhető el. A különböző típusú hígtrágyák nemcsak tápanyag összetételben, hanem EDC tartalomban is különböznek egymástól. A természetes hormon kiválasztódás és a hormonhatású készítmények felhasználása tekintetében fontos az állatállomány ivararányát és korcsoportját figyelembe venni. A jövőre nézve fontos, hogy toxikológiai vizsgálatokat végezzünk, és nagyobb hangsúlyt fektessünk a mezőgazdasági melléktermékek által kijuttatott hormonhatású anyagok mennyiségére és lehetséges hatásaira.

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Atractylenolide III is one of the major bioactive compounds in Atractylodes japonica rhizome; it has been used clinically for the treatment of gastrointestinal disorders. In the present study, a simple, rapid, and selective analytical method was developed and validated for the quantification of atractylenolide III in rat plasma samples using ultra-performance liquid chromatography–ion trap mass spectrometry (UPLC–ion trap MS). Liquid–liquid extraction with ethyl acetate was used for plasma sample preparation. Bergapten was used as an internal standard (IS). The separation of compounds was carried out on a C18 column, with isocratic elution of 0.1% formic acid in water–acetonitrile (45:55, v/v) at 35 °C. Mass detection was performed in the positive ion mode, under optimized conditions for an electrospray ionization source at m/z 249.1 for atractylenolide III and m/z 217.0 for the IS. The methods of instrumental analysis and plasma sample extraction were validated in terms of precision, accuracy, matrix effect, and extraction recovery, with acceptable values. The present method was successfully applied to the pharmacokinetic study of atractylenolide III in rat plasma samples after oral administration of A. japonica rhizome extract.

Open access

This paper presents a new, simple, precise, and accurate rapid resolution liquid chromatography (RRLC)