[Simultaneous determination of pentostatin and 2'-amino-2'-deoxyadenosine in fermentation broth by high performance liquid chromatography-tandem mass spectrometry].

An analytical method was established for the simultaneously determination the pentostatin and 2'-amino-2'-deoxyadenosine contents in fermentation broth by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). After high-speed centrifugation, aqueous solution dilution, vortex shock, and microfiltration, the fermentation broth samples were analyzed by HPLC-MS/MS. The samples were separated on a Waters Atlantis® T3 column (100 mm×2.1 mm, 5 µm) using a gradient elution program with 10 mmol/L ammonium formate (containing 0.1% formic acid) and methanol (containing 0.02% formic acid) as the mobile phases. Moreover, a chromatographic protection column (5 mm×2.1 mm, 5 µm) was added to preserve the column efficiency. The flow rate, column temperature, and injection volume were set at 0.3 mL/min, 25 ℃, and 10 µL, respectively. Qualitative and quantitative analyses of the target compounds were performed using an ESI+ source. MS parameters such as the collision energies and tube lens offsets of pentostatin and 2'-amino-2'-deoxyadenosine were optimized. The quantitative ion pairs of pentostatin and 2'-amino-2'-deoxyadenosine were m/z 269.17>153.20 and m/z 267.00>136.10, respectively; the corresponding collision energies were 11 V and 18 V. The external standard method was used for quantitative analysis. The established method was verified rigorously in terms of the linear range, limit of detection, limit of quantification, recovery rate, and precision. Pentostatin and 2'-amino-2'-deoxyadenosine showed good linear relationships in the range of 1.0-250 µg/L. The correlation coefficients ranged from 0.9969 to 0.9996, and the relative standard deviations (RSDs) ranged from 6.51% to 8.35% (n=8). This result indicated good accuracy and exactitude in the detection of the pentostatin and 2'-amino-2'-deoxyadenosine. The recoveries (n=6) at three spiked levels (1.0, 5.0, and 25 µg/L) were in the ranges of 97.94%-104.46% and 89.96%-107.21% for the pentostatin and 2'-amino-2'-deoxyadenosine, respectively; the corresponding RSDs were in the ranges of 3.74%-4.88% and 4.81%-13.29%. The limits of detection (LODs, S/N≥3) and limits of quantification (LOQs, S/N≥10) of the 2'-amino-2'-deoxyadenosine and pentostatin in the fermentation broth were 0.003-0.060 µg/L and 0.010-0.200 µg/L, respectively. The validated experimental method was used for the detection of actual samples, viz. the stored multiple pentostatin-producing mutagenic strains in our laboratory. The HPLC-MS/MS method for the determination of the pentostatin and 2'-amino-2'-deoxyadenosine in fermentation broth offered the advantages of small sampling volume, strong maneuverability, good stability, and high sensitivity. Compared with previously published methods, this systematically established and optimized method significantly reduced the detection time, and matrix effects were well suppressed. Moreover, the peak shape and stability of the target compounds were greatly improved. This method provides a methodological basis and meaningful reference for the detection of the pentostatin and 2'-amino-2'-deoxyadenosine in fermentation broth.

Determination of tiadinil and its metabolite in flue-cured tobacco.

A novel and sensitive method was developed for the determination of residues of tiadinil and its metabolite, 4-methyl-l,2,3-thiadiazole-5-carboxylic acid, in flue-cured tobacco. The pesticides were extracted with acetone and purified by gel permeation chromatography and solid-phase extraction. Analysis was performed by ultra-performance liquid chromatography-tandem mass spectrometry in negative ionization mode. Two precursor-product ion transitions were monitored for both compounds in the multiple reaction monitoring mode. Quantification was conducted by using matrix-matched standard calibration. Recovery values of the proposed method for tiadinil ranged from 72.5 to 98.2%, with relative standard deviations ranging from 3.8 to 9.5%; recovery values for 4-methyl-l,2,3-thiadiazole-5-carboxylic acid ranged from 75.4 to 103.3% with RSDs ranging from 3.7 to 9.3%. The limit of quantification for both compounds was 0.01 mg/kg. This method is valuable for residual analysis, quality control and monitoring of tiadinil and its metabolite, 4-methyl-l,2,3-thiadiazole-5-carboxylic acid, in tobacco.

Simultaneous determination of 44 pesticides in tobacco by UPLC/MS/MS and a modified QuEChERS procedure.

A novel, simple, and rapid method for determination of the residues of 44 commonly used pesticides in tobacco was developed based on modified Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) procedures. The pesticides were extracted with acetonitrile and purified on a mixed sorbents column of primary secondary amine, C18, and graphitized carbon black. Quantitative determination of all pesticides was performed by ultra-performance LC/MS/MS in the positive or negative ionization mode by a single run due to the fast polarity switching capability of the mass spectrometer. Two precursor-product ion transitions were monitored for each compound in the multiple reaction monitoring mode. Quantification was carried out using matrix-matched standard calibration. The method has been validated by various tobacco cultivars, such as Burley, Oriental, and Virginia from different countries. Recoveries of the proposed method for spiked samples ranged from 71.1 to 109.8%, and RSD values were below 10%. The LOQ values were are all below the guidance residue levels proposed by the Agrochemical Advisory Committee of Cooperation Center for Scientific Research Relative to Tobacco. This method is valuable for measurement of pesticide residues in tobacco for QC and monitoring.

[Determination of 14 mycotoxins in Chinese herbs by liquid chromatography-tandem mass spectrometry with immunoaffinity purification].

A method was developed for the determination of 14 mycotoxins, aflatoxins, T-2, HT-2, fumonisins, ochratoxin A, zearalenone, etc. in Chinese herbs by liquid chromatography-tandem quadrupole mass spectrometry (LC-MS/MS). The sample was extracted with phosphate buffer solution (PBS) and methanol in turn, and then purified by a high selective multi-functional immunoaffinity column. The column was washed by PBS (containing 0.1% Twain) and water, and then eluted by methanol. The eluate was dried under nitrogen, dissolved in methanol-10 mmol/L NH4Ac (40 : 60, v/v) solution. The mycotoxins were separated on a Waters Xterra C18 MS column (100 mm x 2.1 mm, 3.5 microm) and detected by MS/MS. The limits of quantification (LOQs) of the 14 mycotoxins were from 1.0 to 5.0 microg/kg. The average recoveries of the 14 mycotoxins spiked in Chinese herbs (Ginseng, Campanulaceae, Radix and Ophiopogonis) ranged from 71.9% to 99.7% at the three spiked levels of 1.0, 5.0, 10.0 microg/kg, and the relative standard deviations (RSDs, n = 6) were between 4.8% and 15.8%. The method is rapid, sensitive and accurate, and suitable for the determination of the 14 mycotoxins in Chinese medicines. The quantification limits of aflatoxins can meet the domestic and foreign requirements.

[Analysis the pollution level of organotins in aquatic food and port wine].

OBJECTIVE: To determine the level of organotins in the aquatic food originated from Dalian, Shandong and Tianjin and port wine collected in Beijing market and accumulate data for risk analysis of organotins in food. METHODS: The samples were extracted by tetrahydrofuran- hydrobromic acid (20/1) and tropolone-hexane (0.03%), and purified by gel-permeation chromatograph. And the pentyl products (penty 1 methyltins, butyltins and phenyltins) were purified by Florisil and detected by gas chromatography pulsed fire photometric detector. RESULTS: The positive ratios of butyltins for shellfish, snail and fish were 86.4%, 33.3% and 38.9%, respectively. As for phenyltins, the ratios were 68.2%, 16.7% and 44.4%, respectively. The positive ratio of tributyltins, dibutyltins, and triphenyltin in shellfish were 81.8%, 59.1% and 68.2%, and the average grosses of butyltins and organotins were 4.5 microg/kg and 11.1 microg/kg. The positive ratios of tributyltin, dibutyltin and triphenyltin for fish were 38.9%, 22.2% and 50.0%, and the average grosses of butyltins and organotins were 1.3 microg/kg and 2.8 microg/kg. The positive ratio of dimethyltin, dibutyltin and monobutyltin in port wine were 76.4%, 58.8% and 41.2%, respectively. The levels of dimethyltin, dibutyltin and monobutyltin were from 0.4 to 5.5 microg/L, 0.1 to 1.7 microg/L and 0.2 to 0.8 microg/L respectively. The results of significant difference (P < 0.05) were gained in statistic analysis of shellfish from different area, and the difference between shellfish, fish and spiral shell also had statistic significance (P < 0.05). CONCLUSION: The positive ratios of organotins were relative high in aquatic food and port wine. The pollution of butyltins and phenyltins in aquatic food and dimethyltin in port wine were prevalent.

[Study on the contamination level and intake of organotins of Chinese dietary].

OBJECTIVE: To obtain the baseline data of organotins' pollution of Chinese meal in order to carry on primary danger analysis of the exposure. METHODS: The samples of the third Chinese total diet study were determined by gas-chromatography pulsed flame photometric detector to estimate dietary intake of organotins. The dietary intake of organotins was estimated according to the contamination level of organotins and food consumption. RESULTS: Only several kinds of organotin were founded in several foods and no organotins was found in fruit, sugar and alcoholic beverages. Dimethyltin (DMT) were detected in some samples from Southern 1 area, the content ranged from 1.5 microg/kg to 4.1 microg/kg. Butyltin compounds existed in seafoods from Southern 1 area, the contents of tributyltin (TBT), dibutyltin (DBT) and monobutyltin (MTB) being 0.9 microg/kg, 1.1 microg/kg, 1.4 microg/kg respectively. The lower limit and upper limit of exposure to tributyltin were from 0.003 microg x kg(-1) x d(-1) to 0.006 microg x kg(-1) x d(-1) and from 0.004 microg x kg(-1) x d(-1) to 0.019 microg x kg(-1) x d(-1) respectively. Comparing to ADI of tributyltin (WHO), the Chinese dietary intake of tributyltin only accounted for 2.5% and that of butyltin only accounted for 3.5%. To identify the contamination source of organotins in Southern 1 area, the individual samples of aquatic food from individual province were analyzed, revealing that Fujian province and Shanghai City were the main contributors of organotins pollution in this area. The belt fish and yellow croaker were typical pollution samples. Higher levels of DMT were detected in seafood samples from Shanghai. CONCLUSION: The exposure level of Chinese dietary was relative low, however the sources of organotin pollution needs further investigation.