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Agrokémia és Talajtan
Authors:
Gabriella Szabóné Kele
and
Péter Szabó
Restricted access

Abstract

Rationale

The bark of Eucommia ulmoides and the roots of Achyranthes bidentata are commonly used in traditional Chinese medicine, and their pairing appears in many traditional Chinese medicine formulas as a recognized compatible unit. However, the changes and interactions of the main components of these two formulas when paired remain unclear, and there is currently no standard or method for their quality control and assessment of pharmacological effects.

Methods

An optimized ultra-high-performance liquid chromatography triple-quadrupole mass spectrometry (UHPLC-MS/MS) method was established for the simultaneous identification of 10 components in E. ulmoides and A. bidentata using in vitro and in vivo models. Tributyltin methacrylate was the internal standard solution, and the blood samples were treated by an organic solvent precipitation method. Gradient elution was conducted on a C18 column at 25 °C with 0.1% formic acid water:acetonitrile as the mobile phase at a flow rate of 0.5 mL min−1. Dynamic multiple response monitoring was performed in negative-ion mode using an Agilent Jet Stream electrospray ionization ion source.

Results

In negative-ion detection mode, eucommiol exhibited a good response, and the isomers ginsenoside Ro and achyranthoside C could also be well separated. The developed method accurately detected the five components with a low blood content. Compared to controls, the levels of ginsenoside Ro, chikusetsusaponin Ⅳa, and achyranthoside C increased; the contents of geniposidic acid and pinoresinol diglucoside were unchanged; and the levels of eucommiol, geniposide, β-ecdysterone, genipin, and achyranthoside D decreased in vitro. In vivo, the contents of geniposidic acid, geniposide, pinoresinol diglucoside, and β-ecdysterone were reduced; the contents of eucommiol and ginsenoside Ro were unchanged; and those of achyranthoside D, chikusetsusaponin Ⅳa, and achyranthoside C increased compared to the corresponding levels in the internal control.

Conclusions

A method for the quality control of the E. ulmoides-A. bidentata drug pair was established for the first time and the main components in 10 drug pairs could be determined simultaneously in vitro and in vivo. These findings show that the E. ulmoides and A. bidentata drug pair cause a compositional change, providing new ideas for the development of this combination to improve clinical efficacy.

Open access

Abstract

We developed and validated a sensitive, heart-cutting, two-dimensional liquid chromatography–tandem mass spectrometry (2D-LC‒MS/MS) method to determine the concentration of mometasone furoate in human plasma after nasal spray administration. Isotopically labeled mometasone furoate-13C,d6 was used as an internal standard (IS). Plasma samples were prepared using a solid-phase extraction (SPE) method. With this 2D-LC strategy, the analytes were trapped in the first dimension (1D) column, and only judiciously selected portions of the 1D effluent were transferred to the second dimension (2D) column for further separation to obtain high-resolution information. MS/MS quantification was performed in positive ionization mode via multiple-reaction monitoring (MRM). This analytical method was fully validated according to related regulatory guidance, and the results showed that the method is robust and sensitive enough for pharmacokinetic investigation of mometasone furoate with satisfactory linearity from 0.25 to 30 pg mL−1. This method was successfully applied to a bioequivalence (BE) study of mometasone furoate aqueous nasal sprays in healthy volunteers.

Open access

Abstract

A simple, rapid, sensitive and eco-friendly liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of free cordycepin (3′-deoxyadenosine) and isocordycepin (2′-deoxyadenosine) in 10 kinds of Cordyceps samples. The samples were prepared by ultrasonic extraction at 75 °C for 30 min with boiling water as the extraction solvent. The LC separation was performed on an Agilent poroshell 120 SB-Aq C18 column (3.0 × 50 mm, 2.7 μm) in isocratic mode with an eco-friendly mobile phase (2% ethanol containing 0.2% acetic acid) at a flow rate of 0.6 mL min−1, and detected by MS/MS in positive mode with multiple reaction monitoring (MRM). The developed method showed good linearity (r > 0.9990), sensitivity (LODs = 0.04 pg, LOQ = 0.1 pg), precision (RSD ≤ 3.8%) and stability (RSD ≤ 3.6%). The recoveries of developed method were 94.4–109.5% (RSD ≤ 5.5%). Compared with reported methods, the current method was rapid (less than 35% analytical time), sensitive (more than 5 folds), and eco-friendly (less than 10 μL harmful organic solvent). 10 different kinds of Cordyceps samples (40 batches) were tested by the developed method. Codycepin was only found in Cordyceps millitaris and C. millitaris fruiting body, and isocordycepin was detected in Cordyceps sinensis and other 6 Cordyceps samples. The developed method would be an improved method for the quality evaluation of Cordyceps samples.

Open access

Abstract

The study was aimed to validate and optimize high performance liquid chromatographic (HPLC) method for the determination of coumarin-3-carboxylic acid (C3A) in the heart and liver issue of Sprague-Dawley (SD) rats after intragastric administration of extractive of leaves of Ficus virens var sublanceolata. And simple ADME and target prediction analyses were performed for C3A. Ethyl acetate was employed to precipitate protein with appropriate sensitivity and acceptable matrix effects. The satisfactory separation was developed on an ODS2 column (4.6 mm × 250 mm, 5 μm) by gradient elution with a methanol-acetic acid solution (pH = 3.0) as the mobile phase. The flow rate was 1.0 mL min−1, the column temperature was maintained at 30 ± 2 °C, the injection volume was 20 μL, and the detection wavelength was set as 309 nm. The method was fully validated in terms of selectivity, linearity, accuracy, precision, extraction recovery and stability. The results of the ADME analysis found that C3A has excellent characteristics of drug-likeness, consistent with good bio-absorption. And the predicted 12 target protein belongs to the amine oxidoreductase and carbonic anhydrase target class. This method is simple, rapid, sensitive, and accurate for the determination of coumarin-3-carboxylic acid in the heart and liver tissue of SD rats.

Open access

Húsz éves avarmanipulációs kísérlet hatásai barna erdőtalaj szén tartalmára és vízkapacitására: Síkfőkút DIRT Project

The effects of a twenty-year litter manipulation experiment on the carbon content and water retention capacity of the examined Luvisols: Síkfőkút DIRT Project

Agrokémia és Talajtan
Authors:
István Fekete
,
Áron Béni
,
Katalin Juhos
, and
Zsolt Kotroczó

A talajok a szárazföldi széntároló rendszerek egyik legjelentősebb tagját jelentik, melyek szénelnyelése, illetve szénkibocsátása jelentős mértékben hat a klímára, ugyanakkor a klímaváltozás is befolyásolja a talajok szénraktározó képességét. Az avar produkció mennyiségi és minőségi változásai jelentősen befolyásolják ezeket a folyamatokat, azonban ezek mértéke, sőt időnként iránya sem ismert pontosan.

A klímaváltozás mellett a területhasználat változások is befolyásolják a talajba kerülő szerves anyagok mennyiségét és ezen keresztül számos egyéb talajfizikai, kémiai és biológiai paramétert. Ezeknek a hatásoknak a rendszerszintű vizsgálatát segítik a nemzetközi avarmanipulációs projektek, melyek azonos kezeléseket alkalmazva, de eltérő klímaviszonyok mellett vizsgálják a mesterségesen átalakított avar inputok hatását a talajrendszerekre. A Síkfőkút project területén, mely 2000-ben csatlakozott a nemzetközi DIRT projecthez, vizsgáltuk az avar input növekedésének és csökkenésének hatásait egy cseres tölgyes erdőben a talajok szén körforgalmára, illetve a vízháztartására. Ezeken a kutatásokon belül vizsgáltuk a kezelések talajaiban a talajnedvesség tartalmat, vízkapacitást és térfogatsűrűséget, valamint CNS analizátorral a talajok szerves szén tartalmát.

Eredményeink azt mutatták, hogy az avar produkció mennyiségi változása, éghajlati viszonyoktól függően, eltérően hat a talajok SOC tartalmának változásaira. A kezelésekkel modellezett avar produkció változások nemcsak közvetlen úton hatnak a talajok szerves anyag tartalmára, de közvetett módon a megváltozott mikroklimatikus viszonyok révén is befolyásolják a talajok szén és vízforgalmi viszonyait. A nagyobb avar produkció a szárazabb síkfőkúti erdőben növelte a talajok szén tartalmát (szemben a nedvesebb amerikai területeken tapasztalt visszaeséssel, vagy stagnálással) és magasabb szerves anyag tartalom társulva a vastagabb avartakaróval magasabb átlagos talajnedvességet és vízmegtartó képességet eredményezett az avar elvonásos kezelésekkel szemben. Ezek a hatások összefüggésben lehetnek azzal is, hogy az avarelvonásos kezeléseknél szignifikánsan magasabb térfogattömeg értékeket mértünk, ami a pórustérfogat csökkenését jelentheti ebben az esetben, csökkentve ezzel a talajban tárolható víz mennyiségét.

Az általunk végzett avarmanipulációs kísérletek nemzetközi kontextusában közelebb juthattunk a biogeokémiai ciklusok, ezáltal a mineralizáció és a humifikáció közötti összefüggések megértéséhez különböző erdőtípusokban és különböző klimatikus feltételek között.

Open access

Abstract

In the present study, an LC-MS/MS method allowing to quantify pretomanid and pyrazinamide simultaneously in rat plasma was developed. Chromatographic separation was achieved on an Agilent Eclipse plus C18 column (100 mm × 2.1 mm, 3.5 μm; Agilent, USA) and maintained at 30 °C. Multiple reaction monitoring (MRM) using positive-ion ESI mode to monitor ion transitions of m/z 360.1 → m/z 175.1 for pretomanid, m/z 124.1 → m/z 81.0 for pyrazinamide, m/z 172.1 → m/z 128.1 for metronidazole (IS). The calibration curves showed good linear relationships over the concentration range of 50–7,500 ng mL−1 for pretomanid and 500–75,000 ng mL−1 for pyrazinamide. The precision and accuracy were below 15% and within ±15% of the nominal concentrations, respectively. The selectivity, recovery and matrix effect of this method were all within acceptable limits of bioanalytics. The method was applied to the analysis of plasma samples from pharmacokinetic studies in rats. The results show that the main pharmacokinetic parameters of pyrazinamide, namely, T max, t 1/2, and AUC(0–t), decreased in the combined group than in the alone group.

Open access

Abstract

Muscle relaxants and pain killers with their different types are widely used as combination approach for treatment of pain associated with several muscle spasm conditions. A sensitive and simple HPLC-UV detection method was developed in this work for simultaneous assay of Dantrolene (DNT) and co-administrated: Ibuprofen (IBU) and Diclofenac (DIC). After simple protein precipitation, separation was achieved using C18 column (150 × 4.6 mm) with a mobile phase of acidified water with orthophosphoric acid (pH = 3.5) and acetonitrile using gradient elution with a flow rate of 1 mL/min. The DAD was adjusted at 380, 219, 280 and 240 nm to measure DNT, IBU, DIC, and dexamethasone (internal standard), respectively. Linearity was demonstrated over the range from 0.1 to 3 μg/mL, 1 to 40 μg/mL, and 0.1 to 2 μg/mL for DNT, IBU, and DIC, respectively. The validated method was applied successfully to compare the effect of co-administration of IBU or DIC on the pharmacokinetic profile of DNT.

Open access