A new sensitive and specific HPLC–MS/MS method for the determination of α-fluoro-β-alanine (FBAL), the main metabolite of the antineoplastic drug 5-fluorouracil (5-FU), in urine for the biological ...monitoring survey of health care workers exposed to 5-FU is described. This procedure is characterized by a pre-column FBAL derivatization by 2,4-dinitrofluorobenzene followed by solid phase extraction sample clean-up. The chromatographic separation was achieved by hydrophilic interaction chromatography (HILIC) on a ZIC HILIC column (Sequant) and the quantification was performed by tandem mass spectrometry. The method offers high sensitivity with a quantification limit of 1
μg/l, which is an improvement on those previously reported. The within- and between-day precisions were less than 13% and 15% respectively at the LOQ and no significant relative matrix effect was observed for FBAL. The validated method was applied to the biological monitoring of occupational exposure to 5-FU in a French hospital. Pre- and post-shift urine samples were collected from 19 workers in a hospital pharmacy and an oncology ward over a period of 5 days. On a total of 121 analysed samples, measurable amounts of FBAL were detected in up to 29%, the concentrations range from LOQ to 22.7
μg/l, yielding evidence of occupational exposure to 5-FU. Such data are scarce and represent a step forward in assessing the occupational health risks associated with handling antineoplastic drugs.
The tea shrub is grown in long-standing orchards, an environment that is suitable for persistent weed growth, which is increasingly controlled by herbicides. Therefore, there is increasing concern ...that tea consumers may be exposed to herbicide residues. In this study, the levels of glufosinate-ammonium (GLU), glyphosate
-(phosphonomethyl) glycine; PMG, and its metabolite aminomethyl phosphoric acid (AMPA) were determined in tea samples by HPLC-MS/MS using several current purification methods and a new method that we developed herein. The matrix effect of our proposed method was between -27.3 and 27.7%, which was lower than that in other methods, indicating that this method effectively reduced the interference of tea matrix in the mass spectrometry process. This method was used to determine the levels of PMG, GLU, and AMPA in 780 samples, including six traditional Chinese teas (green tea, black tea, oolong tea, dark tea, white tea, and yellow tea) and a floral tea, from 14 provinces of China. Probability estimates showed that the 95
percentile risk entropy values of the three pesticide residues were far below the acceptable risk level. The risk assessment results showed that exposure to PMG, GLU, and AMPA caused by drinking tea beverages poses no significant risk to human health.
•The molecular recognition of cordycepin arabinoside in Cordyceps militaris by MS2 fragmentation rule.•A quantitative analysis method of cordycepin and its arabinoside was established by ...HPLC-MS/MS.•The cordycepin and its arabinoside were determined in fruiting body and pupa during growth process.•Cordyceps militaris as the optimum raw material of functional foods during 60th to 70th days.
Cordyceps militaris is an edible fungus that is widely used as a functional food in many countries. In order to objectively evaluate its nutritional value, free and glycosidic cordycepins need to be analyzed. The cordycepin arabinoside molecule was recognized by the MS2 fragmentation rule, and both cordycepin and its arabinoside were quantitatively analyzed in the fruiting body and pupa of Cordyceps militaris by high-performance liquid chromatography with tandem mass spectrometric (HPLC-MS/MS). The method had good linear regression (R2 = 0.9999), with a detection limit of 0.021 ng/mL. The recovery range was 94.32–103.09% in the fruiting body and pupa. The content of cordycepin and its arabinoside showed an upward trend with growth, and the total contents reached the highest level at the mature stage (60-70th day) without mildew. This study provides a useful reference for the evaluation and application of Cordyceps militaris as a functional food resource.
•Development of LC-MS/MS method based on IDA approach for CAs in M. Koenigii.•Application of 2 survey scans (MRM+PIS) and an EPI as a dependent scan.•Effective extraction and clean-up developed and ...validated with FDA guidelines.•Build-in-house database by means of in silico MS/MS spectra prediction.
Murraya koenigii (M. koenigii) is recognized as one of the most significant Indian medicinal plants, due to its therapeutic properties. In fact, various biological activities, including anti-inflammatory and antifungal, are attributed to carbazole alkaloids (CAs) and their metabolites, but it is still difficult to achieve a full classification of these compounds because of their heterogeneous structures. For this reason, the development of new strategies for their identification is necessary. In this work, a reliable method, including the simultaneous quantification of three main CAs of Murraya (Mahanimbine, Koenimbine and Koenigicine) and a putative identification for other compounds belonging to the huge family of CAs, was developed by means of HPLC-MS/MS. Also, an efficient extraction procedure followed by a suitable clean-up step was presented, in order to obtain reliable recoveries (resulted from 60 to 85% for all the analytes). The analyses were performed by using predictive multi experiment approach based on information-dependent acquisition (IDA), coupling multiple reaction monitoring (MRM) and precursor ion (PI) as survey scans, and enhanced product ion scan (EPI) as dependent scan. Competitive Fragmentation Modeling-ID (CFM-ID) was used to predict MS/MS spectra from the chemical structures of the compounds in order to create a suitable MRM inclusion list. The obtained results showed that this method can simultaneously provide quantitative information for the target analytes (Mahanimbine (7.22–5.62 mg/kg), Koenimbine (1.26–1.62 mg/kg) and Koenigicine (0.44–1.77 mg/kg)) and a putative identification for several compounds belonging to different classes of CAs which are not included in the target list, thanks to PI survey scan.
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A high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method and an ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determining eight ...coccidiostat (halofuginone, lasalocid, maduramicin, monensin, narasin, nigericin, robenidine and salinomycin) residues in beef were developed and compared. Samples were extracted with a mixture of acetic acid, acetonitrile and ethyl acetate and were then purified on a C18 solid-phase extraction (SPE) column. The purified samples were analyzed by HPLC-MS/MS and UPLC-MS/MS, using 0.1% formic acid-water solution (A) and pure methanol (B) as the mobile phase. The samples were fractionated on a C18 column using different gradient elution procedures, followed by qualitative analysis using a mass spectrometer operated in multiple reaction monitoring (MRM) mode with positive electrospray ionization; the external standard method was used for quantitation. At spiked levels that ranged from the limit of quantification (LOQ) to 100 μg/kg, the average recoveries were 71.96%–100.32% and 71.24%–89.24%, with relative standard deviations (RSDs) of 2.65%–12.38% and 2.98%–14.86% for UPLC-MS/MS and HPLC-MS/MS, respectively. The limits of detection (LODs) and LOQs of the eight coccidiostats were 0.14–0.32 μg/kg and 0.43–1.21 μg/kg, respectively, for UPLC-MS/MS analysis and 0.16–0.58 μg/kg and 0.53–1.92 μg/kg, respectively, for HPLC-MS/MS analysis. Both methods had good accuracy and precision, but UPLC-MS/MS had higher sensitivity than HPLC-MS/MS.
•Two methods were established for the determination of eight coccidiostat residues in beef.•HPLC-MS/MS and UPLC-MS/MS methods were compared to determine eight coccidiostats in beef.•These two methods provide reliable accuracy for the determination of the eight coccidiostats.•The two methods are novel, fast and highly sensitive.
The influence of ultrasonic pretreatment on the release and antioxidant activity of potential antioxidant peptides after in-vitro simulated gastrointestinal digestion of β-lactoglobulin (BLG) were ...measured by HPLC-MS/MS, chemical and cellular-based assays. The gastrointestinal digest was fractionated into four fractions by Sephadex G-25 gel filtration column, and fractions showed a considerable ABTS·
scavenging ability. The fraction with the strongest antioxidant activity was produced by ultrasonicated BLG after gastrointestinal digestion, which relies on ultrasonic-promoted proteolysis to produce many small-molecule antioxidant peptides. The best active fraction has better cellular antioxidant activity and protection of H
O
-induced oxidative HepG2 cell model, which significantly increases the activities of antioxidant enzyme, and is concentration-dependent. HPLC-MS/MS analysis showed that there were more potential antioxidant peptides in the best active fraction. This research will provide a basis for the further application of ultrasonic in dairy products, which can promote the release of more potential antioxidant peptides-derived from gastrointestinal digestion.
Antioxidant activity and phenolic compounds of sumac extracts were investigated. Sumac was extracted in methanol and subjected to solvent–solvent partitioning to yield two fractions as ethyl acetate ...and aqueous. Methanol extract was further fractioned over Sephadex LH-20 column. Antioxidant activity of extracts and fractions were screened using ferric thiocyanate and DPPH radical scavenging methods. Phenolic composition of active fraction(s) was determined by HPLC–MS systems. Those fractions which exhibited strong antioxidant activity were rich in anthocyanins and hydrolysable tannins. While gallic acid was the main phenolic acid in the extracts, anthocyanin fraction contained cyanidin, peonidin, pelargonidin, petunidin, and delphinidin glucosides and coumarates. Pentagalloyl glucose was abundant in the hydrolysable tannin fraction. Effective scavenging concentration (EC
50) on DPPH radical was 0.70
μg/mL both in ethyl acetate and tannin fractions, and 5.33
μg/mL in anthocyanin rich fraction. Same extracts and fractions showed moderate lipid peroxidation inhibition effect compared with the synthetic antioxidants. The findings demonstrate that sumac can be used as a natural antioxidant.
The current approach involves the development of dual drug loaded nanoparticles of Paclitaxel (Pac) and Erlotinib (Erl) with subsequent thorough optimization in order to achieve sustained and ...simultaneous delivery of Pac and Erl. The formulation is developed to produce maximum efficacy with least side effects. The dual loaded paclitaxel and erlotinib nanoparticles (PE-NPs) were prepared by modified single emulsion method with appropriate optimization. The formulation was optimized for size, polydispersity index (PDI), zeta potential, surface morphology, encapsulation efficiency and loading capacity. The XRD and DSC results of a physical mixture and PE-NPs together confirmed that there was not any physical or chemical interaction among the drugs and polymer (PLGA). Further, the XRD and DSC analyses revealed that the crystalline forms of both drugs in PE-NPs were changed into amorphous form. The optimized PE-NPs were observed to have the particle size of 145.5±3.2nm, zeta potential of −9.3±1.4mV and PDI of 0.135 which indicated the particles to be of uniform size with optimum stability. The entrapment efficiency of paclitaxel was found to be 90.36% and that of erlotinib was found to be 62.23% from PE-NPs. PE-NPs further showed the sustained release of drugs from the particles with total release of 65.25% for paclitaxel and 74.83% for erlotinib in 120h. Accelerated and intermediate stability studies also indicated that there was a minimal variation in size and entrapment efficiency of PE-NPs.
•Exploration of PLGA for simultaneous oral delivery of Paclitaxel and Erlotinib•Optimum mean size of 145nm and low PDI 0.135nm for dual drug loaded PLGA nanosystem•High encapsulation efficiency of 90.36% for Paclitaxel and 62.23% for Erlotinib•Augmented gut permeation of approximately 57% for Paclitaxel and 35% for Erlotinib•Sustained drug release of Paclitaxel and Erlotinib followed the Higuchi model.
Detection of vitamin A, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and vitamin E is important in neurological diseases such as epilepsy. Traditional HPLC-based methods require labor intensive and ...time-consuming sample extraction procedures, which limits the applications of these methods. In this work, a high-throughput and robust HPLC-MS/MS method with simple sample preparation for the simultaneous quantification of vitamin A, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and vitamin E in rat plasma was developed. These four fat-soluble vitamins were separated on a Kinetex C18 column (50 × 3.0 mm, 2.6 μm) using gradient elution. The LLOQs of vitamin A, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and vitamin E were 50 ng/ml, 1 ng/ml, 5 ng/ml and 1 μg/ml respectively. The intra-day precision and accuracy were in the range of 1.31–13.0% and 85.8–114% respectively, and the inter-day precision and accuracy were 2.47–12.8% and 86.1–115% respectively. After fully validated, the HPLC-MS/MS method was applied to our pentylenetetrazole (PTZ)-induced kindling rat model.
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•A fast and robust HPLC-MS method for the simultaneous quantification of vitamin A, D and E in rat plasma.•One-step sample pretreatment procedure using a Protein Precipitation & Phospholipid Removal Plate.•Simultaneous quantification of vitamins A, D and E in our PTZ-induced kindling rat model.
