Amatoxins and phallotoxins are toxic cyclopeptides found in the genus Amanita and are among the predominant causes of fatal food poisoning in China. In the treatment of Amanita mushroom poisoning, an early and definite diagnosis is necessary for a successful outcome, which has prompted the development of protocols for the fast and confirmatory determination of amatoxins and phallotoxins in human biological fluids. For this purpose, a simple, rapid and sensitive multiresidue UPLC-MS/MS method for the simultaneous determination of α-amanitin, β-amanitin, γ-amanitin, phalloidin (PHD) and phallacidin (PCD) in human plasma, serum and urine was developed and validated. The diluted plasma, serum and urine samples were directly purified with a novel PRiME technique on a 96-well μElution plate platform, which allowed high-throughput sample processing and low reagent consumption. After purification, a UPLC-MS/MS analysis was performed using positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode. This method fulfilled the requirements of a validation test, with good results for the limit of detection (LOD), lower limit of quantification (LLOQ), accuracy, intra- and inter-assay precision, recovery and matrix effects. All of the analytes were confirmed and quantified in authentic plasma, serum and urine samples obtained from cases of poisoning using this method. Using the PRiME μElution technique for quantification reduces labor and time costs and represents a suitable method for routine toxicological and clinical emergency analysis.

A risk assessment of deoxynivalenol (DON) was recently conducted for the residents in Henan province, China, where wheat as the staple food are highly consumed. A high-throughput sensitive UPLC-MS/MS method following 96-well μElution solid-phase extraction (SPE) were developed and validated for the determination of DON biomarkers in human urine. Isotope labelled internal standard, ¹³C-DON, was used for accurate quantification. Urinary samples collected from 151 healthy Chinese aged 2–78 years were processed with and without enzyme hydrolysis to determine total and free biomarkers, respectively. DON, and de-epoxy-deoxynivalenol (DOM-1) to a lesser extent, can be frequently detected in these samples both with and without enzyme hydrolysis. Free DOM-1 was detected at low level in human urine for the first time. Total DON was detected in all samples with a mean concentration at 47.6 ng mL⁻¹. The mean and median probable daily intakes (PDI) for the whole participants, estimated to be 1.61 μg/kg bw and 1.10 μg/kg bw, both exceeded the PMTDI (1 μg/kg bw/day), indicating a potential risk for the residents in this area, especially for children and adolescents.

Zearalenone is a widespread mycotoxin with high estrogenic activity. This study aimed to characterize the exposure of ZEN in a Chinese population during harvest season in 2016. Exposure to ZEN was measured using both duplicate diet method and human biomonitoring approaches. Duplicate diet samples from 199 individuals (4-80 years old) and their following morning urine samples were collected and analyzed using LC-MS/MS methods sensitive for ZEN, ZAN, α/β-ZEL and α/β-ZAL. ZEN was detected in 59.8% of the food samples at a mean level of 1.21 ± 2.15 μg/kg. The estimated daily intake (EDI) of ZEN was calculated from food contamination and consumption data at a mean level of 25.6 ± 38.6 ng/kg bw/day, representing 10.2% of the tolerable daily intake (TDI) set by EFSA and 5.1% of the provisional maximum tolerable daily intake (PMTDI) set by JECFA, respectively. Wheat appears to be the main diet source of ZEN exposure, contributing over 80% of the mean EDI. Children had the highest EDI at 37.5 ± 56.3 ng/kg bw/day (p < 0.05). Urine samples were analyzed both before and after enzymatic hydrolysis to determine the free and total amounts of ZEN biomarkers. The majority of ZEN was excreted as conjugates with the mean fZEN/tBM ratio of 25.4%. Adolescents had the highest excretion of ZEN biomarkers among all age groups (p < 0.05). Probable daily intake (PDI) was calculated from ZEN biomarkers and an excretion rate of 36.8%, giving a mean value of 41.6 ± 65.5 ng/kg bw/day. Significant correlation between internal and external exposure measurement was evidenced in this study (r = 0.344, p < 0.01). Although the mean PDI was approximately 1.6 times the mean EDI, these two approaches resulted in similar calculated degrees of ZEN exposure, both markedly below the health-based guidance value. This study is the first to compare ZEN exposure in a same population based on both diet study and human biomonitoring approaches. Significant differences of PDI/EDI ratios were found in different age groups (p < 0.05), possibly indicative of diversified excretion capabilities and metabolism patterns within the population.

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.

Liposomes have made remarkable achievements as drug delivery vehicles in the clinic. Liposomal products mostly benefited from remote drug loading techniques that succeeded in amphipathic and/or ionizable drugs, but seemed impracticable for nonionizable and poorly water-soluble therapeutic agents, thereby impeding extensive promising drugs to hitchhike liposomal vehicles for disease therapy. In this study, a series of weak acid drug derivatives were designed by a simplistic one step synthesis, which could be remotely loaded into liposomes by pH gradient method. Cabazitaxel (CTX) weak acid derivatives were selected to evaluate regarding its safety profiles, pharmacodynamics, and pharmacokinetics. CTX weak acid derivative liposomes were superior to Jevtana® in terms of safety profiles, including systemic toxicity, hematological toxicity, and potential central nerve toxicity. Specifically, it was demonstrated that liposomes had capacity to weaken potential toxicity of CTX on cortex and hippocampus neurons. Significant advantages of CTX weak acid derivative-loaded liposomes were achieved in prostate cancer and metastatic cancer therapy resulting from higher safety and elevated tolerated doses.