In Vivo Kinetics and Biotransformation of Aflatoxin B1 in Dairy Cows Based on the Establishment of a Reliable UHPLC-MS/MS Method

The in vivo kinetics of aflatoxin B1 (AFB1) and its carry-over as aflatoxin M1 (AFM1) in milk as well as the toxin loads in the tissue of dairy cows were assessed through a repetitive feeding trial of an AFB1-contaminated diet of 4 μg kg−1 body weight (b.w.) for 13 days. This was followed by a clearance period that ended with a single dose trial of an AFB1-contaminated diet of 40 μg kg−1 b.w. An ultra-high performance liquid chromatography tandem mass spectrometry method was developed and successfully validated by the determination of linearity (R2 ≥ 0.990), sensitivity (lower limit of quantification, 0.1–0.2 ng ml−1), recovery (79.5–111.2%), and precision relative standard deviation (RSD) ≤14.7%) in plasma, milk, and various tissues. The repetitive ingestion of AFB1 indicated that the biotransformation of AFB1 to AFM1 occurred within 48 h, and the clearance period of AFM1 in milk was not more than 2 days. The carry-over rate of AFM1 in milk during the continuous ingestion experiment was in the range of 1.15–2.30% at a steady state. The in vivo kinetic results indicated that AFB1 reached a maximum concentration of 3.8 ± 0.9 ng ml−1 within 35.0 ± 10.2 min and was slowly eliminated from the plasma, with a half-life time (T1/2) of 931.1 ± 30.8 min. Meanwhile, AFM1 reached a plateau in plasma (0.5 ± 0.1 ng ml−1) at 4 h after the ingestion. AFB1 was found in the heart, spleen, lungs, and kidneys at concentrations of 1.6 ± 0.3, 4.1 ± 1.2, 3.3 ± 0.9 and 5.6 ± 1.4 μg kg−1, respectively. AFM1 was observed in the spleen and kidneys at concentrations of only 0.7 ± 0.2 and 0.8 ± 0.1 μg kg−1, respectively. In conclusion, the in vivo kinetics and biotransformation of AFB1 in dairy cows were determined using the developed UHPLC-MS/MS method, and the present findings could be helpful in assessing the health risks to consumers.

Download Full-text

Analysis of the Carry-Over of Ochratoxin A from Feed to Milk, Blood, Urine, and Different Tissues of Dairy Cows Based on the Establishment of a Reliable LC-MS/MS Method

Molecules ◽

10.3390/molecules24152823 ◽

2019 ◽

Vol 24 (15) ◽

pp. 2823 ◽

Cited By ~ 5

Author(s):

Zhiqi Zhang ◽

Zhichen Fan ◽

Dongxia Nie ◽

Zhihui Zhao ◽

Zheng Han

Keyword(s):

Dairy Cows ◽

Ochratoxin A ◽

Limit Of Quantification ◽

Fresh Milk ◽

Liquid Chromatography Tandem Mass ◽

Ochratoxin Α ◽

Carry Over ◽

First Time ◽

Different Tissues

A rapid and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of ochratoxin A (OTA) and its metabolite ochratoxin α (OTα), for the first time, in dairy cow plasma, milk, urine, heart, liver, spleen, lung, and kidney. The established method was extensively validated by determining the linearity (R2 ≥ 0.990), sensitivity (lower limit of quantification, 0.1–0.2 ng mL−1), recovery (75.3–114.1%), precision (RSD ≤ 13.6%), and stability (≥83.0%). Based on the methodological advances, the carry-over of OTA was subsequently studied after oral administration of 30 μg/kg body weight OTA to dairy cows. As revealed, OTA and OTα were detected in urine, with maximal concentrations of 1.8 ng mL−1 and 324.6 ng mL−1, respectively, but not in milk, plasma, or different tissues, verifying the protection effects of rumen flora against OTA exposure for dairy cows. Moreover, 100 fresh milk samples randomly collected from different supermarkets in Shanghai were also analyzed, and no positive samples were found, further proving the correctness of the in vivo biotransformation results. Thus, from the currently available data, regarding OTA contamination issues on dairy cows, no significant health risks were related to OTA exposure due to the consumption of these products.

Download Full-text

High-Performance Liquid Chromatography-Tandem Mass Spectrometry for Buprenorphine Evaluation in Plasma—Application to Pharmacokinetic Studies in Rabbits

Molecules ◽

10.3390/molecules26020437 ◽

2021 ◽

Vol 26 (2) ◽

pp. 437

Author(s):

Marta Tikhomirov ◽

Błażej Poźniak ◽

Tomasz Śniegocki

Keyword(s):

High Performance ◽

Pharmacokinetic Profile ◽

Pharmacokinetic Studies ◽

Limit Of Quantification ◽

Reliable Determination ◽

Main Challenge ◽

Analytical Protocol ◽

Liquid Chromatography Tandem Mass ◽

Cost Efficient

The precise and reliable determination of buprenorphine concentration is fundamental in certain medical or research applications, particularly in pharmacokinetic studies of this opioid. The main challenge is, however, the development of an analytical method that is sensitive enough, as the detected in vivo concentrations often fall in very low ranges. Thus, in this study we aimed at developing a sensitive, repeatable, cost-efficient, and easy HPLC analytical protocol for buprenorphine in rabbit plasma. In order to obtain this, the HPLC-MS2 system was used to elaborate and validate the method for samples purified with liquid-liquid extraction. Fragment ions 468.6→396.2 and 468.6→414.2 were monitored, and the method resulted in a high repeatability and reproducibility and a limit of quantification of 0.25 µg/L with a recovery of 98.7–109.0%. The method was linear in a range of 0.25–2000 µg/L. The suitability of the analytical procedure was tested in rabbits in a pilot pharmacokinetic study, and it was revealed that the method was suitable for comprehensively describing the pharmacokinetic profile after buprenorphine intravenous administration at a dose of 300 µg/kg. Thus, the method suitability for pharmacokinetic application was confirmed by both the good validation results of the method and successful in vivo tests in rabbits.

Download Full-text

Determination of myrislignan levels in BALB/c mouse plasma by LC-MS/MS and a comparison of its pharmacokinetics after oral and intraperitoneal administration

BMC Veterinary Research ◽

10.1186/s12917-021-02990-y ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Jili Zhang ◽

Hongfei Si ◽

Jichao Sun ◽

Kun Lv ◽

Biqing Yan ◽

...

Keyword(s):

Formic Acid ◽

Intraperitoneal Administration ◽

Internal Standard ◽

Pharmacokinetic Parameters ◽

Clinical Effects ◽

Limit Of Quantification ◽

Mouse Plasma ◽

Relative Standard ◽

Liquid Chromatography Tandem Mass

Abstract Background Myrislignan is a natural product from Myristica sp. with diverse pharmacological activities. Recently, the anti-Toxoplasma gondii (T. gondii) activity of myrislignan has been proposed, and in vivo studies of its pharmacokinetics in BALB/c mice are necessary to further evaluate the clinical effects of myrislignan. Results In this study, a sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to quantify myrislignan levels in mouse plasma using dehydrodiisoeugenol as an internal standard (IS) in positive ion mode. Chromatographic separation of the analytes was achieved using an ACE Ultracore Super C18 analytical column (2.5 μm, 2.1 × 50 mm) at 30 °C. A gradient mobile phase consisting of water (0.1 % formic acid) and acetonitrile (0.1 % formic acid) was delivered at a flow rate of 0.4 mL/min. Myrislignan and the IS eluted at 1.42 and 1.71 min, respectively. A good excellent linear response across the concentration range of 1-1000 ng/mL was achieved (r2 = 0.9973). The lower limit of quantification (LLOQ) was 1 ng/mL, and the inter- and intra-day accuracy and precision of the method showed relative standard deviations (RSDs) less than 10 %. The method was applied to examine the pharmacokinetics of myrislignan in mouse plasma following a single oral administration of 200 mg/kg or intraperitoneal administration of 50 mg/kg myrislignan, and the bioavailability (F) of orally administered myrislignan was only 1.97 % of the bioavailability of intraperitoneally administered myrislignan. Conclusions A rapid and sensitive LC-MS/MS method has been was developed, validated and successfully used to determine myrislignan levels in mice after oral or intraperitoneal administration. This study is the first to report the pharmacokinetic parameters of myrislignan in mice and to compare its pharmacokinetics after oral and intraperitoneal administration, which will be useful for further research on the administration of myrislignan in animals and humans.

Download Full-text

Determination of Methylene Blue and Its Metabolite Residues in Aquatic Products by High-Performance Liquid Chromatography–Tandem Mass Spectrometry

Molecules ◽

10.3390/molecules26164975 ◽

2021 ◽

Vol 26 (16) ◽

pp. 4975

Author(s):

Xuan Zhang ◽

Yunhua Hui ◽

Changling Fang ◽

Yuan Wang ◽

Feng Han ◽

...

Keyword(s):

Methylene Blue ◽

High Performance ◽

Correlation Coefficients ◽

Limit Of Quantification ◽

Aquatic Products ◽

Azure B ◽

Chromatography Tandem Mass Spectrometry ◽

Relative Standard ◽

Liquid Chromatography Tandem Mass

A sensitive and reliable method was developed to determine methylene blue (MB) and its metabolite residues, including azure A (AZA), azure B (AZB), and azure C (AZC) in aquatic products by HPLC–MS/MS. The samples were extracted by acetonitrile and cleaned up by alumina-neutral (ALN) cartridges. The analytes were separated on a Sunfire C18 column (150 mm × 2.1 mm, 5 µm). The method was validated according to the European criteria of Commission Decision 2002/657/CE. Good linearity between 1–500 µg/L was obtained with correlation coefficients (R2) greater than 0.99. The limit of quantification (LOQ) was 1.0 µg/kg. The average recoveries at three levels of each compound (1, 5, and 10 µg/kg) were demonstrated to be in the range of 71.8–97.5%, with relative standard deviations (RSDs) from 1.05% to 8.63%. This method was suitable for the detection of methylene blue and its metabolite residues in aquatic products.

Download Full-text

Calcium/Copper alginate framework doped with CuO nano-particles as a novel adsorbent for micro-extraction of benzodiazepines from human serum

Current Pharmaceutical Analysis ◽

10.2174/1573412916666200210150914 ◽

2020 ◽

Vol 16 ◽

Author(s):

Nadereh Rahbar ◽

Fatemeh Ahmadi ◽

Zahra Ramezani ◽

Masoumeh Nourani

Keyword(s):

Human Serum ◽

High Performance ◽

Limit Of Detection ◽

Solid Phase ◽

Nano Particles ◽

Limit Of Quantification ◽

Analytical Methodology ◽

Fiber Fabrication ◽

Relative Standard ◽

Green Procedure

Background: Sample preparation is one of the most challenging phases in pharmaceutical analysis, especially in biological matrices, affecting the whole analytical methodology. Objective: In this study, a new Ca(II)/Cu(II)/alginate/CuO nanoparticles hydrogel fiber (CCACHF) was synthesized through a simple, green procedure and applied for fiber micro solid phase extraction (FMSPE) of diazepam (DIZ) and oxazepam (OXZ) as model drugs prior to high-performance liquid chromatography-UV detection (HPLC-UV). Methods: Composition and morphology of the prepared fiber were characterized and the effect of main parameters on the fiber fabrication and extraction efficiency have been studied and optimized. Results: In optimal conditions, calibration curves were linear ranging between 0.1–500 µg L−1 with regression coefficients of 0.9938 and 0.9968. Limit of detection (LOD) (S/N=3) and limit of quantification (LOQ) (S/N=10) of the technique for DIZ and OXZ were 0.03 to 0.1 µg L−1. Within-day and between-day relative standard deviations (RSDs) for DIZ and OXZ were 6.0–12.5% and 3.3–9.4%, respectively. Conclusion: The fabricated adsorbent has been substantially employed to extraction of selected benzo-diazepines (BZDs) from human serum real specimens and the obtained recoveries were also satisfactory (82.1-109.7%).

Download Full-text

Determination of 77 Multiclass Pesticides and Their Metabolitesin Capsicum and Tomato Using GC-MS/MS and LC-MS/MS

Molecules ◽

10.3390/molecules26071837 ◽

2021 ◽

Vol 26 (7) ◽

pp. 1837

Author(s):

Harischandra Naik Rathod ◽

Bheemanna Mallappa ◽

Pallavi Malenahalli Sidramappa ◽

Chandra Sekhara Reddy Vennapusa ◽

Pavankumar Kamin ◽

...

Keyword(s):

Limit Of Detection ◽

Solid Phase ◽

Graphitized Carbon Black ◽

Limit Of Quantification ◽

Gas And Liquid Chromatography ◽

Relative Standard ◽

The Matrix ◽

Liquid Chromatography Tandem Mass ◽

Primary Secondary Amine

A quick, sensitive, and reproducible analytical method for the determination of 77 multiclass pesticides and their metabolites in Capsicum and tomato by gas and liquid chromatography tandem mass spectrometry was standardized and validated. The limit of detection of 0.19 to 10.91 and limit of quantification of 0.63 to 36.34 µg·kg−1 for Capsicum and 0.10 to 9.55 µg·kg−1 (LOD) and 0.35 to 33.43 µg·kg−1 (LOQ) for tomato. The method involves extraction of sample with acetonitrile, purification by dispersive solid phase extraction using primary secondary amine and graphitized carbon black. The recoveries of all pesticides were in the range of 75 to 110% with a relative standard deviation of less than 20%. Similarly, the method precision was evaluated interms of repeatability (RSDr) and reproducibility (RSDwR) by spiking of mixed pesticides standards at 100 µg·kg−1 recorded anRSD of less than 20%. The matrix effect was acceptable and no significant variation was observed in both the matrices except for few pesticides. The estimated measurement uncertainty found acceptable for all the pesticides. This method found suitable for analysis of vegetable samples drawn from market and farm gates.

Download Full-text

Dissipation, Processing, Leaching, and Safety Evaluation of Flonicamid and Its Metabolites in Tea

Journal of AOAC International ◽

10.1093/jaoacint/qsaa052 ◽

2020 ◽

Vol 103 (6) ◽

pp. 1441-1450

Author(s):

Ya Yang ◽

Xiangwu Liu ◽

Qingtao Zhang ◽

Ya Chen ◽

Sumei Zhang ◽

...

Keyword(s):

Green Tea ◽

High Performance ◽

Alcoholic Beverage ◽

Safety Evaluation ◽

Tea Infusions ◽

Relative Standard ◽

Liquid Chromatography Tandem Mass ◽

Chinese Consumer ◽

Transfer Behavior ◽

Single Factor Analysis

Abstract Background Tea is a popular traditional non-alcoholic beverage worldwide. Flonicamid is a selective systemic pyridine carboxamide insecticide that is widely used for controlling tea leafhopper in tea. Objective The leaching rates, dissipation dynamics, and residue levels of flonicamid and its metabolites in tea leaves during processing and transferring were investigated to validate the safe risk in tea and transfer behavior using high performance liquid chromatography–tandem mass spectrometry with a convenient pretreatment method. Method The extracting method and immersion rate experiments were optimized by single factor analysis and orthogonal tests. The acetonitrile extracting solvent with 0.5% formic acid was used and optimal leaching conditions were obtained with a regime of 15 min immersion time, 100°C temperature, three immersions and a tea-to-water ratio of 1:50. Results Average recoveries in processed green tea and infusions were 80.85–98.75% with relative standard deviations <5.87%. LODs and LOQs of flonicamid, 4-trifluoromethylnicotinic acid (TFNA), N-(4-trifluoromethylnicotinoyl) glycine (TFNG), and 4-trifluoromethylnicotinamide (TFNA-AM) were 0.0013–0.350 and 0.004–1 μg/g, respectively. The processing factor of flonicamid was 0.36–5.52 during green tea manufacture. The leaching rates were 22.9-97.4% from processed tea to infusion. Conclusions The risk of long-term and short-term dietary intake of flonicamid was safe in tea infusions with the risk quotient (RQ) values <1 for the Chinese consumer. This work may provide guidance for safe and reasonable consumption of flonicamid in tea in China. Highlights The suitable leaching factors of flonicamid and its metabolites in tea infusions were optimized by orthogonal experimentation for the first time.

Download Full-text

Validation of developed method by RP-HPLC for estimation of Prasugrelin human plasma and studying the stability of the drugs in plasma

Kathmandu University Journal of Science Engineering and Technology ◽

10.3126/kuset.v13i1.21263 ◽

2018 ◽

Vol 13 (1) ◽

pp. 65-75

Author(s):

Kumar S. Ashutosh ◽

Debnath Manidipa ◽

Rao J.V.L.N. Seshagiri ◽

Sankar D. Gowri

Keyword(s):

Human Plasma ◽

High Performance ◽

Method Development ◽

Potassium Dihydrogen Phosphate ◽

Array Detector ◽

Dihydrogen Phosphate ◽

Limit Of Quantification ◽

Rp Hplc ◽

Relative Standard ◽

The Stability

This paper is concern with a reverse phase high performance liquid chromatography (RP-HPLC) bio-analytical method development and validation for Prasugrel in human plasma using photo diode array detector (PDA detector). The HPLC separation was carried out in an isocratic mode on an X-Terra C18 column (4.6 x 150 mm; 5 μm) with a mobile phase consisting of potassium dihydrogen phosphate [pH 3.0] and acetonitrile in the ratio of 30:70 v/v at a flow rate of 1.0 mL/min. The run time was maintained for 5 mins and the detection was monitored at 210 nm. The percentage recovery was found 99.61-100.06 in human plasma. This reveals that the method is quite accurate. The linearity was found 15-40 μg/mL in human plasma. The inter-day and intra-day precision in plasma was found within the limits. The lower limit of quantification (LLOQ) obtained by the proposed method was 0.05 μg/mL. The percentage relative standard deviation (%RSD) obtained for the drug spiked in plasma for stability studies were less than 2 %.Kathmandu University Journal of Science, Engineering and TechnologyVol. 13, No. 1, 2017, Page: 65-75

Download Full-text

Simultaneous Determination of Eight New Generation Amide Insecticide Residues in Complex Matrix Agri-food by Multi-walled Carbon Nanotube Cleanup and UPLC-MS/MS

10.21203/rs.3.rs-952153/v1 ◽

2021 ◽

Author(s):

Tao Lin ◽

Xinglian Chen ◽

Li Wang ◽

Haixian Fang ◽

Maoxuan Li ◽

...

Keyword(s):

Mass Spectrometry ◽

Simultaneous Determination ◽

High Performance ◽

Rapid Determination ◽

Complex Matrix ◽

Limit Of Quantification ◽

Relative Standard ◽

The Matrix ◽

Multi Walled Carbon Nanotubes

Abstract The simultaneous determination method of 8 amide pesticides by multi-walled carbon nanotubes (MWCNs) cleanup, combined with QuEChERS method and ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry has been developed and successfully applied in complex matrix such as green onions, celery, leeks, citrus, lychees, avocado. The matric effect of MWCNs is optimized and compared with QuEChERS materials. The results show that MWCNs can effectively reduce the matrix effect in sample extraction. The mass spectrometry is optimized, through their chemical structure skeletons, the ESI+ and ESI- mode are simultaneously scanned in the method. The coefficient (r) is greater than 0.9990, the limit of quantification ranges from 0.03 to 0.80 μg/kg, the recovery rate ranges from 71.2% to 120%, and the relative standard deviation (RSD) ranges from 3.8% to 9.4%. The method is fast, simple, sensitive, and has good purification effect. It is suitable for the rapid determination of amide pesticides in complex matrix agri-food.

Download Full-text

METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF THYMOL AND EUGENOL BY USING RP-HPLC IN PURE AND IN EMULGEL FORMULATION

INDIAN DRUGS ◽

10.53879/id.58.07.11895 ◽

2021 ◽

Vol 58 (07) ◽

pp. 59-65

Author(s):

Vinita C. Patole ◽

Shilpa P. Chaudhari ◽

Keyword(s):

High Performance ◽

Method Development ◽

Limit Of Detection ◽

Hplc Method ◽

Simultaneous Estimation ◽

Limit Of Quantification ◽

Relative Standard ◽

Peak Areas ◽

80 A 20 ◽

Development And Validation

An attempt was made to develop a simple, selective, rapid and precise high-performance liquid chromatography (HPLC) method for simultaneous estimation of thymol and eugenol. Analysis was performed on a C18 column with the mobile phase consisting of solvent %A (water) and solvent %B (acetonitrile) with the following gradient: 0–1 min, 80 % A, 20 % B; 1–7 min, 40 % A and 60 % B; 7–12 min, 10 % A and 90 % B; and 12–15min, 80 % A and 20 % B at a flow rate of 0.6 mL/min. The compounds were well separated on a Thermo Scientific Hypersil BDS RP C18 column (4.6 mm × 150 mm, dp = 5 µm) and ultraviolet detection at 280 nm. The retention times of eugenol and thymol were 10.5 min and 11.6 min, respectively. Validation of the proposed method was carried out according to the guidelines of the International Council on Harmonization (ICH). The linearity of the method is good for thymol and eugenol over the concentration range of 1–50 ppm, and the r 2 values were 0.9996 for both thymol and eugenol. The calculated limit of detection (LOD) value was 0.5ppm and the limit of quantification (LOQ) value was 1ppm for both the analytes. The intra and interday relative standard deviation (RSD) of the retention time and peak areas was less than 3 %.The established method was appropriate, and the two markers were well resolved, enabling efficient quantitative analysis of thymol and eugenol.

Download Full-text