[Rapid determination of 15 N-nitrosamines in air-dried yak meat using one-step QuEChERS-gas chromatography-triple quadrupole mass spectrometry].

A method based on gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) coupled with one-step QuEChERS technique was developed for the simultaneous determination of 15 N-nitrosamines in air-dried yak meat. The hydration volume, extraction solvent, extracting salt, and cleaning material were optimized according to the characteristics of the N-nitrosamines and sample matrix. The optimized conditions were as follows: 10 mL of purified water for sample hydration, acetonitrile as the extraction solvent for the sample after hydration, 4.0 g of anhydrous MgSO4 and 1.0 g of NaCl as extracting salts, 500 mg of MgSO4+25 mg of C18+50 mg of PSA as cleaning materials. Favorable recoveries of the 15 N-nitrosamines were obtained when the extraction solution was incompletely dried. Thus, the final extract was dried to below 0.5 mL under a mild nitrogen stream and then redissolved to 0.5 mL with acetonitrile. After filtration, 200 µL of the sample was transferred to an autosampler vial for GC-MS/MS analysis. The 15 N-nitrosamines were determined using GC-MS/MS on a DB-HeavyWAX column (30 m×0.25 mm×0.25 µm) with an electron impact ion source in multiple-reaction monitoring (MRM) mode, and quantified using an external standard method. Under the optimized experimental conditions, the results showed that the calibration curves exhibited good linearities for the 15 N-nitrosamines, with correlation coefficients (r2) greater than 0.9990. The limits of detection (LODs) and the limits of quantification (LOQs) ranged from 0.05 to 0.20 µg/kg and from 0.10 to 0.50 µg/kg, respectively. At spiked levels of 1LOQ, 2LOQ, and 10LOQ, the average recoveries were 79.4%-102.1%, 80.6%-109.5%, and 83.0%-110.6%, respectively, and the relative standard deviations were in the range of 0.8%-16.0%. The low matrix effects of the 15 N-nitrosamines indicated the high sensitivity of the proposed method. The method was applied to detect representative commercial air-dried yak meat samples obtained using different processing techniques. Seven N-nitrosamines, including N-nitrosodimethylamine, N-nitrosodiisobutylamine, N-nitrosodibutylamine, N-methyl-N-phenylnitrous amide, N-ethyl-N-nitrosoaniline, N-nitrosopyrrolidine, and N-nitrosodiphenylamine were detected in all samples. The average contents of the seven N-nitrosamines was 0.08-20.18 µg/kg. The detection rates and average contents of the N-nitrosamines in cooked air-dried yak meat samples were higher than those in traditional raw air-dried yak meat samples. Compared with the manual QuEChERS method, the one-step QuEChERS method developed integrated the extraction and clean-up procedures into one single run, and the detection efficiency was considerably improved. The developed method is simple, rapid, highly sensitive, and insusceptible to human errors. Thus, it is useful for the determination of N-nitrosamines in air-dried yak meat and can be extended to the qualitative and quantitative analysis of N-nitrosamines in other meat products. It also provides method support and a data reference for the general determination of N-nitrosamines, which is of great significance for food safety.

Association of radiotherapy with thoracic vertebral fractures in esophageal squamous cell carcinoma: A retrospective cohort study.

To investigate the association between radiotherapy (RT) and thoracic vertebral fractures in esophageal squamous cell carcinoma (ESCC) and explore the risk factors of thoracic vertebral fracture in ESCC who underwent RT. This retrospective cohort study including 602 consecutive ESCC patients examined the association between RT and thoracic vertebral fractures using multivariable Cox proportional hazard models and relevant risk factors of thoracic vertebral fractures based on clinical and RT parameters in patients with ESCC. Followed for a median follow-up of 24 months, 54 patients had thoracic vertebral fractures. The multivariable analysis revealed RT as an independent risk factor after adjusting for clinical risk factors. Univariable analyses associated a 5-Gy increase in vertebral dose to single vertebrae and a 1-time increase in RT fraction with higher risk of vertebral fracture. Adding RT factors (vertebral dose and fraction) and mean vertebral hounsfield unit to the Cox models containing conventional clinical risk factors significantly improved the χ2 value for predicting vertebral fractures (all P < .001). This study revealed RT, as well as increased vertebral dose and RT fractions, as a significant, consistent, and strong vertebral fracture predictor in ESCC. Combined vertebral dose, RT fractions, and vertebral hounsfield unit provided optimal risk stratification for ESCC patients.

Physiological responses of Microcystis aeruginosa PCC7806 to nonanoic acid stress.

A recent study has shown that nonanoic acid (NA) is one of the strongest allelochemicals to a cyanobacterium Microcystis aeruginosa, but the physiological responses of M. aeruginosa to NA stress remain unknown. In this study, physiological characters such as the growth rate, photosynthetic processes, phosphorus and nitrogen uptake kinetics, and the contents of intracellular microcystin of M. aeruginosa PCC7806 were studied under the NA stress. The results showed that the growth rates of M. aeruginosa PCC 7806 were significantly inhibited in all NA stress treatments during first 3 days after exposure, and the growth rate was recovered after 5-day exposure. After 2-day exposure, the contents of both phycocyanin and allophycocyanin per cell decreased at NA concentration of 4 mg L(-1), and oxygen evolution was inhibited even at the concentration of 0.5 mg L(-1), but carotenoid content per cell was slightly boosted in NA stress. Physiological recovery of M. aeruginosa PCC7806 was observed after 7-day exposure to NA. It was shown that NA stress had no effect on uptake of nitrogen, but could stimulate the uptake of phosphorus. The contents of intracellular microcystin have not been affected in all NA treatments in contrast with the control.

[Biodegradation of LAS in the anaerobic water of a representative inlet of Lake Dianchi].

Biodegradation and degradation kinetics of anion-surfactant (LAS) in the anaerobic water of a representative inlet (Haihe River) of Lake Dianchi under different incubation conditions were studied by the 'river die-away' test method. The influences of temperature, pH, initial concentration of LAS, aeration condition and added nutrients (NH4Cl or NaH2PO4) on the biodegradation of LAS in the water were investigated. The results demonstrate that LAS can be biodegraded by microorganisms in the water and that the percentage of degradation of LAS was more than 95% after 26 d. The biodegradation of LAS fit the second kinetic model. Incubation temperature, initial concentration of LAS, aeration and added nutrients (NH4Cl or NaH2PO4) can all affect the biodegradation of LAS. When the incubation temperature increased from 10 degrees C to 25 degrees C, the biodegradation rate (p) of LAS increased from 0.21 d(-1) to 0.90 d(-1). The LAS degradation rate increased from 0.72 d(-1) under anaerobic condition to 1.97 d(-1) under continuous aeration condition. The increased initial concentrations of LAS lead to decrease of the biodegradation rate. NaH2PO4 accelerated the degradation of LAS but added NH4Cl instead inhibited degradation. In our experiment, pH value (7.05-9.44) had little influence on the biodegradation of LAS.

[Preparative purification and characterization of [Dha7] microcystin RR].

One variant of microcystins was isolated and purified with cyanobacteria natural bloom as the starting material, which was collected in Dianchi Lake, China. The separation protocol involved extraction of cyanobacterial cells by 75% aqueous methanol, isolation by reversed-phase flash chromatography, and purification by reversed-phase semipreparative HPLC. The structure and purity of purified microcystin was identified with electrospray ionization mass spectrometry, UV spectrophotometer, and analytical HPLC. The purified microcystin was assigned as [Dha7]MCRR (purity > 95%), which was a demethylated variant of MCRR. The structure of purified microcystin was identified as cyclo-(Ala-Arg-MeAsp-Arg-Adda-Glu-Dha) with molecular weight of 1023. There was a maximum absorbance at 239 nm in its UV spectrum (200-300 nm). This variant of microcystins occurred frequently, and sometimes could become the main variant in waterbloom from Dianchi Lake.

[Subchronic toxic effects of fish muscle-bound MCLR].

Subchronic oral gavage toxicity of MCLR in water and in fish muscle was examined in male Balb/C mice for 13 weeks to assess the safety of aquatic products. The results showed that the liver coefficient (p < 0.05), the activities of ALT and AST (p < 0.01) increased significantly and distinct centrilobular to midzonal hepatucellular occurred after oral gavage of dissolved MCLR at a dose of 68.75 microg/kg (body weight), but neither influence on the activities of BUN and Cr nor histological changes on kidney were observed at any time point. In contrast, the administration of fish muscle-bound MCLR at the same dose resulted in no obvious subchronic toxicity in mice, except that the increase of liver coefficient (p < 0.05) and the activity of ALT (p < 0.01) can be observed only at the first week. It was concluded that the toxicity of fish muscle-bound MCLR was much lower than that of dissolved MCLR.