In this study, we established a comprehensive method for simultaneous identification and quantification of tetrodotoxin （TTX） in fresh pufferfish tissues and pufferfish-based products using liquid chromatography/qua...In this study, we established a comprehensive method for simultaneous identification and quantification of tetrodotoxin （TTX） in fresh pufferfish tissues and pufferfish-based products using liquid chromatography/quadrupole-linear ion trap mass spectrometry （LC-QqLIT-MS）. TTX was extracted by 1% acetic acid-methanol, and most of the lipids were then removed by freezing lipid precipitation, followed by purification and concentration using immunoaffinity columns （IACs）. Matrix effects were substantially reduced due to the high specificity of the IACs, and thus, background interference was avoided. Quantitation analysis was therefore performed using an external calibration curve with standards prepared in mobile phase. The method was evaluated by fortifying samples at 1, 10, and 100 ng/g, respectively, and the recoveries ranged from 75.8%--107%, with a relative standard deviation of less than 15%. The TTX calibration curves were linear over the range of 1-1 000 ~tg/L, with a detection limit of 0.3 ng/g and a quantification limit of 1 ng/g. Using this method, samples can be further analyzed using an information- dependent acquisition （IDA） experiment, in the positive mode, from a single liquid chromatography-tandem mass spectrometry injection, which can provide an extra level of confirmation by matching the full product ion spectra acquired for a standard sample with those from an enhanced product ion （EPI） library. The scheduled multiple reaction monitoring method enabled TTX to be screened for, and TTX was positively identified using the IDA and EPI spectra. This method was successfully applied to analyze a total of 206 samples of fresh pufferfish tissues and pufferfish-based products. The results from this study show that the proposed method can be used to quantify and identify TTX in a single run with excellent sensitivity and reproducibility, and is suitable for the analysis of complex matrix pufferfish samples.展开更多
A rapid, simple and accurate method using an immunoaffinity column (IAC) and capillary electrophoresis (CE) for the analysis of the major alkaloids in opium is developed. The IAC was synthesized by coupling specific m...A rapid, simple and accurate method using an immunoaffinity column (IAC) and capillary electrophoresis (CE) for the analysis of the major alkaloids in opium is developed. The IAC was synthesized by coupling specific morphine polyclonal antibodies to CNBr-actived Sepharose 4B. The IAC showed high selectivity and obvious enrichment to morphine, codeine, dionin and thebaine. The extraction solution was analyzed by CE with β-cyclodextrin as an additive. Recoveries of the four alkaloids from PBS were between 93%-105% with RSD value less than 5.0%. The result showed that this method was practical for the determination of morphine analogs in opium.展开更多
提出了题示方法测定衢枳壳样品中黄曲霉毒素B1、B2、G1、G2和赭曲霉毒素A的含量。样品经过体积比79∶20∶1的乙腈-水-乙酸混合溶液超声提取,离心,上清液用p H 7.0磷酸盐缓冲液稀释,用复合免疫亲和柱净化,过滤,滤液中的目标物在XDB C18...提出了题示方法测定衢枳壳样品中黄曲霉毒素B1、B2、G1、G2和赭曲霉毒素A的含量。样品经过体积比79∶20∶1的乙腈-水-乙酸混合溶液超声提取,离心,上清液用p H 7.0磷酸盐缓冲液稀释,用复合免疫亲和柱净化,过滤,滤液中的目标物在XDB C18色谱柱上用不同体积比的体积比1∶9的甲醇-5 mmol·L^(-1)乙酸铵混合溶液和体积比9∶1的甲醇-5 mmol·L^(-1)乙酸铵混合溶液的混合溶液分离,电喷雾离子源正离子(ESI+)和多反应监测(MRM)模式检测,同位素内标法定量。结果显示:5种真菌毒素与其同位素内标质量浓度比和对应的峰面积比在一定范围内呈线性关系,检出限为0.05~0.20μg·kg^(-1)。3个加标浓度水平下阴性样品中5种真菌毒素的回收率为82.7%~97.3%,测定值的相对标准偏差(n=6)为5.2%~12%。展开更多
目的基于HPLC研究柏子仁中4种黄曲霉毒素的污染情况,并进行风险分析,评估柏子仁的用药安全。方法采用Agilent Eclipse Plus C 18(4.6×250 mm 5μm)为色谱柱,柱后光化学衍生法检测,以甲醇∶乙腈∶水(35∶10∶55)为流动相,流速1.0 mL...目的基于HPLC研究柏子仁中4种黄曲霉毒素的污染情况,并进行风险分析,评估柏子仁的用药安全。方法采用Agilent Eclipse Plus C 18(4.6×250 mm 5μm)为色谱柱,柱后光化学衍生法检测,以甲醇∶乙腈∶水(35∶10∶55)为流动相,流速1.0 mL·min^(-1);柱温:40℃;荧光检测器检测。结果黄曲霉毒素B 1、B 2、G 1、G 2分别在0.35~20.8μg·L^(-1)、0.13~7.6μg·L^(-1)、0.36~21.6μg·L^(-1)、0.13~7.6μg·L^(-1)范围内呈良好的线性关系,黄曲霉毒素B 1、B 2、G 1、G 2平均回收率分别为90.6%、83.46%、87.84%、86.58%,相对偏差分别为2.9%、3.6%、3.6%、4.7%。23批柏子仁中有14批检出黄曲霉毒素B 1和总量,残留量分别在1~4μg·kg^(-1)和1~5μg·kg^(-1)之间,均符合规定,但检出率高达61%,存在安全隐患。结论该方法简单、准确、方便,能有效评价柏子仁中黄曲霉毒素的污染情况。警示柏子仁易受黄曲霉毒素的污染,需加强柏子仁监管力度和不定期的专项抽检,降低安全风险,以确保临床用药安全。展开更多
基金Supported by the National Natural Science Foundation of China(No.41106109)the China National Food Safety Standards Development Project(No.ZHENGHE-2015-356)
文摘In this study, we established a comprehensive method for simultaneous identification and quantification of tetrodotoxin （TTX） in fresh pufferfish tissues and pufferfish-based products using liquid chromatography/quadrupole-linear ion trap mass spectrometry （LC-QqLIT-MS）. TTX was extracted by 1% acetic acid-methanol, and most of the lipids were then removed by freezing lipid precipitation, followed by purification and concentration using immunoaffinity columns （IACs）. Matrix effects were substantially reduced due to the high specificity of the IACs, and thus, background interference was avoided. Quantitation analysis was therefore performed using an external calibration curve with standards prepared in mobile phase. The method was evaluated by fortifying samples at 1, 10, and 100 ng/g, respectively, and the recoveries ranged from 75.8%--107%, with a relative standard deviation of less than 15%. The TTX calibration curves were linear over the range of 1-1 000 ~tg/L, with a detection limit of 0.3 ng/g and a quantification limit of 1 ng/g. Using this method, samples can be further analyzed using an information- dependent acquisition （IDA） experiment, in the positive mode, from a single liquid chromatography-tandem mass spectrometry injection, which can provide an extra level of confirmation by matching the full product ion spectra acquired for a standard sample with those from an enhanced product ion （EPI） library. The scheduled multiple reaction monitoring method enabled TTX to be screened for, and TTX was positively identified using the IDA and EPI spectra. This method was successfully applied to analyze a total of 206 samples of fresh pufferfish tissues and pufferfish-based products. The results from this study show that the proposed method can be used to quantify and identify TTX in a single run with excellent sensitivity and reproducibility, and is suitable for the analysis of complex matrix pufferfish samples.
文摘A rapid, simple and accurate method using an immunoaffinity column (IAC) and capillary electrophoresis (CE) for the analysis of the major alkaloids in opium is developed. The IAC was synthesized by coupling specific morphine polyclonal antibodies to CNBr-actived Sepharose 4B. The IAC showed high selectivity and obvious enrichment to morphine, codeine, dionin and thebaine. The extraction solution was analyzed by CE with β-cyclodextrin as an additive. Recoveries of the four alkaloids from PBS were between 93%-105% with RSD value less than 5.0%. The result showed that this method was practical for the determination of morphine analogs in opium.
