scholarly journals Determination of Urinary Mycotoxin Biomarkers Using a Sensitive Online Solid Phase Extraction-UHPLC-MS/MS Method

Toxins ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 418
Author(s):  
Jessica Schmidt ◽  
Benedikt Cramer ◽  
Paul C. Turner ◽  
Rebecca J. Stoltzfus ◽  
Jean H. Humphrey ◽  
...  
Keyword(s):  
Solid Phase Extraction ◽  
Solid Phase ◽  
Fumonisin B1 ◽  
Human Biomonitoring ◽  
Aflatoxin M1 ◽  
Phase Extraction ◽  
Alternariol Monomethyl Ether ◽  
Method Performance ◽  
Online Spe ◽  
Online Solid Phase Extraction

In the course of assessing the human exposure to mycotoxins, biomarker-based approaches have proven to be important tools. Low concentration levels, complex matrix compositions, structurally diverse analytes, and the large size of sample cohorts are the main challenges of analytical procedures. For that reason, an online solid phase extraction-ultra high-performance liquid chromatography-tandem mass spectrometry (online SPE-UHPLC-MS/MS) method was developed, allowing for the sensitive, robust, and rapid analysis of 11 relevant mycotoxins and mycotoxin metabolites in human urine. The included spectrum of analytes comprises aflatoxin M1 (AFM1), altenuene (ALT), alternariol monomethyl ether (AME), alternariol (AOH), citrinin (CIT) and its metabolite dihydrocitrinone (DH-CIT), fumonisin B1 (FB1), ochratoxin A (OTA), and zearalenone (ZEN) as well as α- and β-zearalenol (α- and β-ZEL). Reliable quantitation was achieved by means of stable isotope dilution, except for ALT, AME and AOH using matrix calibrations. The evaluation of method performance displayed low limits of detection in the range of pg/mL urine, satisfactory apparent recovery rates as well as high accuracy and precision during intra- and interday repeatability. Within the analysis of Zimbabwean urine samples (n = 50), the applicability of the newly developed method was shown. In addition to FB1 being quantifiable in all analyzed samples, six other mycotoxin biomarkers were detected. Compared to the occurrence rates obtained after analyzing the same sample set using an established dilute and shoot (DaS) approach, a considerably higher number of positive samples was observed when applying the online SPE method. Owing to the increased sensitivity, less need of sample handling, and low time effort, the herein presented online SPE approach provides a valuable contribution to human biomonitoring of mycotoxin exposure.

Simultaneous Determination of Five Hormones in Milk by Automated Online Solid-Phase Extraction Coupled to High-Performance Liquid Chromatography

2020 ◽  
Vol 103 (1) ◽  
pp. 265-271 ◽  
Author(s):  
Kun Liu ◽  
Kai Kang ◽  
Nan Li ◽  
Jing An ◽  
Kaoqi Lian ◽  
...  
Keyword(s):  
Solid Phase Extraction ◽  
Solid Phase ◽  
Growth Promotion ◽  
Operation Time ◽  
Hplc Method ◽  
Cow Milk ◽  
Contamination Level ◽  
Phase Extraction ◽  
Online Spe ◽  
Online Solid Phase Extraction

Abstract Background: Many hormones show the effects of protein assimilation and growth promotion, and they are frequently used as veterinary drugs in livestock, which has harmful effects on human health. It is necessary to determine their contamination level in animal-derived food, especially in milk. Objective: In this study, a detailed procedure is described for an automated online solid-phase extraction (SPE)–HPLC method capable of detecting five hormones (i.e., estriol, prednisone acetate, hydrocortisone, diethylstilbestrol, and estrone) in cow milk. Methods: The corresponding milk samples were precipitated by addition of acetonitrile and then purified as well as enriched by a polar-enhanced polymer (PEP) online SPE column. The supernatants were directly injected into the online SPE–HPLC system using methanol–water as the mobile phase mixture. Results: The linearity range of the method was 0.1–25 μg/mL for prednisone acetate, hydrocortisone, and diethylstilbestrol, 0.2–25 μg/mL for estriol, and 0.5–25 μg/mL for estrone, with correlation coefficients (r) ranging from 0.9994 to 0.9996. The recovery rates determined at three concentration levels for the five compounds were in the range of 70.82–112.90%. LODs of estriol, prednisone acetate, hydrocortisone, diethylstilbestrol, and estrone were 0.023, 0.005, 0.006, 0.004, and 0.054 μg/mL, respectively. Conclusions: This automated online SPE–HPLC method was both effective and reliable in the simultaneous measurement of five hormones, and the method was successfully applied to the detection of five hormone species in milk. Highlights: An automated online SPE–HPLC method has been developed for the analysis of five hormones in cow milk. Online SPE proved to be a powerful technique for determining five hormones simultaneously. This method ensured simple sample pretreatment and less operation time. The established method was successfully applied to the analysis of five hormone species in milk.

scholarly journals Development of a High-Throughput Online Solid-Phase Extraction/Tandem Mass Spectrometry Method for Cytochrome P450 Inhibition Screening

2010 ◽  
Vol 15 (4) ◽  
pp. 447-452 ◽  
Author(s):  
Kheng B. Lim ◽  
Can C. Özbal ◽  
Daniel B. Kassel
Keyword(s):  
Mass Spectrometry ◽  
Cytochrome P450 ◽  
Tandem Mass Spectrometry ◽  
Solid Phase Extraction ◽  
High Throughput ◽  
Solid Phase ◽  
Tandem Mass ◽  
Phase Extraction ◽  
Online Spe ◽  
Online Solid Phase Extraction

A high-throughput online solid-phase extraction/tandem mass spectrometry (online SPE/MS/MS) system has been developed to support rapid evaluation of drug discovery compounds for possible drug-drug interaction (DDI). Each compound is evaluated for its DDI potential by incubating over a range of 8 test concentrations and against a panel of 6 cytochrome P450 (CYP) enzymes, 1A2, 2C8, 2C9, 2C19, 2D6, and 3A4. Previously, a postassay pooling and a 2-min gradient LC/MS/MS method had been reported to increase sample throughput, allowing for a 96-well plate of samples to be analyzed in under 4 h. The development of a new online SPE/MS/MS system has reduced the analysis time to less than 15 min per 96-well plate, translating to a 15-fold time savings compared to the 2-min LC/MS/MS method. Sampling precision without internal standard correction ranged from 3.1% to 5.6% relative standard deviation, and the carryover was determined to be between 1.0% and 4.1%. One hundred twenty in-house compounds were assayed and pooled for analyses using both the online SPE/MS/MS and LC/MS/MS, and the correlation coefficients ranged from 0.89 to 1.13, when comparing the IC50 results obtained from the 2 approaches for each of the CYP enzymes.

scholarly journals Development of a Sensitive and Reliable UHPLC-MS/MS Method for the Determination of Multiple Urinary Biomarkers of Mycotoxin Exposure

Toxins ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 193 ◽  
Author(s):  
Zhezhe Liu ◽  
Xiaoxue Zhao ◽  
Libiao Wu ◽  
Shuang Zhou ◽  
Zhiyong Gong ◽  
...  
Keyword(s):  
Human Urine ◽  
Solid Phase ◽  
Accurate Determination ◽  
Fumonisin B1 ◽  
High Sensitivity ◽  
Human Biomonitoring ◽  
Urine Samples ◽  
Specific Method ◽  
Alternariol Monomethyl Ether

A variety of mycotoxins from different sources frequently contaminate farm products, presenting a potential toxicological concern for animals and human. Mycotoxin exposure has been the focus of attention for governments around the world. To date, biomarkers are used to monitor mycotoxin exposure and promote new understanding of their role in chronic diseases. The goal of this research was to develop and validate a sensitive UHPLC-MS/MS method using isotopically-labeled internal standards suitable for accurate determination of 18 mycotoxin biomarkers, including fumonisins, ochratoxins, Alternaria and emerging Fusarium mycotoxins (fumonisin B1, B2, and B3, hydrolyzed fumonisin B1 and B2, ochratoxin A, B, and alpha, alternariol, alternariol monomethyl ether, altenuene, tentoxin, tenuazonic acid, beauvericin, enniatin A, A1, B, and B1) in human urine. After enzymatic digestion with β-glucuronidase, human urine samples were cleaned up using HLB solid phase extraction cartridges prior to instrument analysis. The multi-mycotoxin and analyte-specific method was validated in-house, providing satisfactory results. The method provided good linearity in the tested concentration range (from LOQ up to 25–500 ng/mL for different analytes), with R2 from 0.997 to 0.999. The limits of quantitation varied from 0.0002 to 0.5 ng/mL for all analytes in urine. The recoveries for spiked samples were between 74.0% and 133%, with intra-day precision of 0.5%–8.7% and inter-day precision of 2.4%–13.4%. This method was applied to 60 urine samples collected from healthy volunteers in Beijing, and 10 biomarkers were found. At least one biomarker was found in all but one of the samples. The high sensitivity and accuracy of this method make it practical for human biomonitoring and mycotoxin exposure assessment.

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