Monolithic silica spin column extraction （MonoSpin-SPE） was developed as a simple, sensitive, and eco-friendly pretreatment method which combined with ultra-fast liquid chromatography-mass spectrometry （UFLC-MS） t...Monolithic silica spin column extraction （MonoSpin-SPE） was developed as a simple, sensitive, and eco-friendly pretreatment method which combined with ultra-fast liquid chromatography-mass spectrometry （UFLC-MS） to determine the levels of six phthalate esters, dimethyl- （DMP）, diethyl- （DEP）, dipropyl- [ DPrP], butyl-benzyl- （BBP）, dicyclohexyl- （DcHP）, and di-n-octyl-（DOP） phthalate in physiological saline samples. Under optimized experimental conditions, the method was linear in the following ranges： 0.2- 50 μg/L for DMP, DEP, DPrP, DcHP and DOP; 5- 100 μg/L for BBP. The correlation coefficients （R2 ） were in the range of 0. 9951 - 0. 9995 for all the analytes and the limits of detection （LODs） and limits of quantification （LOQs） were in the ranges of 0.02 - 0.9 μg/L and 0.08 - 2.7μg/L, respectively. The pretreatment process showed good reproducibility with inter-day and intra-day relative standard deviations （RSDs） below 8.5% and 11.2%, respectively. This method was used to determine the levels of six phthalate esters in physiological saline samples and the recoveries ranged from 71.2% to 107.3%. DMP and DEP were found in actual physical saline samples （brand A and brand B）.展开更多
Background:Nails are one of the objects that are more durable than other witness objects.This study was interested in whether the Spin column-based nucleic acid purification technique could extract DNA from nails to g...Background:Nails are one of the objects that are more durable than other witness objects.This study was interested in whether the Spin column-based nucleic acid purification technique could extract DNA from nails to give satisfactory results.Aims and Objectives:This study aimed to evaluate the yield and quality of DNA extracted from the nail.Materials and Methods:Nail samples from 15 males and 15 females over 18 were extracted using GeneAll®Exgene™Cell SV Mini Kit.The DNA concentration and purity were measured using a NanoDrop Spectrophotometer,and the quality of DNAwas assessed by polymerase chain reaction(PCR)using primers specific for human growth hormone(HGH).Results:The average yield of DNA was 0.816µg(0.141–2.706µg)obtained from the average nail weight of 30.2 mg(22.2–40.0 mg).The average DNA concentration was 27.2 ng/µL(4.7–90.2 ng/µL),corresponding to 30µL in volume.Almost all DNA samples(96.7%)had high purity(A260/A280≥1.80)and gave a band of PCR product of the HGH gene on agarose gel electrophoresis.Conclusion:Spin column-based nucleic acid purification is recommended for nail DNA extraction due to its simplicity and high quality.展开更多
A rapid, simple and selective method based on molecularly imprinted, spin column extraction coupled with fluorescence detection was successfully established for the determination of 2,4-dinitrophenol in serum samples....A rapid, simple and selective method based on molecularly imprinted, spin column extraction coupled with fluorescence detection was successfully established for the determination of 2,4-dinitrophenol in serum samples. The 2,4-dinitrophenol imprinted polymers exhibited highly selective recognition for the template molecule and the maximum adsorption capacity was 138.9 mg/g. The results indicated that when water is used as the loading solution, only 2,4-dinitrophenol could be adsorbed on the spin column without the remaining structural analogs（2-nitrophenol, 4-nitrophenol and phenol）. After eluting with acetonitrile/acetic acid（9/1, v/v）, 2,4-dinitrophenol in serum samples could be determined by using the fluorescence spectrometer, based on the fluorescence enhancement of fluorescein by the template molecule. Under the optimal conditions, the spiked recovery ranged from 95.8% to 103.4% and the detection limit was 1 nmol/L. The results confirmed the reliability and practicality of the protocol and revealed a good perspective of this method for biological sample analysis.展开更多
目的对两步磁珠法提取血浆游离DNA的方法进行评价,初步探讨血浆游离DNA定量对于肺癌患者实验诊断的意义。方法在健康志愿者血浆中掺入DNA标准品127 bp,208 bp和Quick-load~@100 bp DNA ladder(由100-1 500 bp DNA片段组成),采用两步磁...目的对两步磁珠法提取血浆游离DNA的方法进行评价,初步探讨血浆游离DNA定量对于肺癌患者实验诊断的意义。方法在健康志愿者血浆中掺入DNA标准品127 bp,208 bp和Quick-load~@100 bp DNA ladder(由100-1 500 bp DNA片段组成),采用两步磁珠法和分离柱提取法提取血浆中掺入的DNA标准品;用Qubit测定DNA浓度,安捷伦2100生物芯片分析仪和琼脂糖凝胶电泳检测分析DNA片段大小分布。用两步磁珠法和分离柱提取法分别提取15例肺癌患者的血浆cfDNA,利用安捷伦2100生物芯片分析仪和琼脂糖凝胶电泳比较两种方法提取的cf-DNA片段大小分布,分析cf-DNA浓度与肺癌分期的相关性。结果两步磁珠法第二步分离的DNA浓度与掺入标准品DNA(127 bp,208 bp)片段浓度呈正相关(r^2=0. 985 7,P <0. 001);两步磁珠法提取的Quick-load~@100 bp DNA ladder第一步分离的DNA片段≥300 bp,第二步分离的DNA片段≤300 bp,分离柱提取法重提取的Quick-load~@DNA ladder≤1 500 bp。两步磁珠法第二步分离的肺癌患者血浆cf-DNA浓度与肺癌分期呈正相关(r^2=0. 866 4,P=0. 004 9)。分离柱提取法提取的cf-DNA浓度与肺癌分期无显著相关性(r^2=0. 500 9,P=0. 214 5)。结论两步磁珠法可分别提取不同大小片段的DNA标准品,同时有效提取肺癌患者血浆中小片段游离DNA,且其浓度与肺癌分期呈正相关。展开更多
氢同位素核自旋异构体正-仲态比例影响氢同位素的低温物性,有必要对其比例进行测定。本文利用活性三氧化二铝多孔层开管(Porous Layer Open Tubular,PLOT)柱实现了正-仲氢同位素(氕、氘)的基线分离,发展了一种可在液氮温度下测定同核分...氢同位素核自旋异构体正-仲态比例影响氢同位素的低温物性,有必要对其比例进行测定。本文利用活性三氧化二铝多孔层开管(Porous Layer Open Tubular,PLOT)柱实现了正-仲氢同位素(氕、氘)的基线分离,发展了一种可在液氮温度下测定同核分子正-仲态比例的色谱分析技术。研究结果表明,与传统三氧化二铝填充柱相比,高效PLOT柱实现了正、仲氕(o-H_2、p-H_2)以及正、仲氘(o-D2、p-D2)的基线分离(分离度R_s大于1.5),当流量为5 m L·min-1时,分离度R_s(p-H_2,o-H_2)=6.9,R_s(o-D2,p-D2)=1.8。正仲态分离度与进样量、流量均有关系。根据峰面积的积分结果,常温(298 K)下正、仲氕比例为2.77:1,正、仲氘比例为1.78:1,与理论测算值基本符合。HD与o-H_2实现了部分分离,R_s(o-H_2,HD)=0.5,根据理论预测,实现HD与o-H_2的基线分离(R_s达到1.5),理论塔板数需要达到3.9×105。展开更多
基金supported by the National Natural Science Foundation of China(Nos.90813015,20935002)
文摘Monolithic silica spin column extraction （MonoSpin-SPE） was developed as a simple, sensitive, and eco-friendly pretreatment method which combined with ultra-fast liquid chromatography-mass spectrometry （UFLC-MS） to determine the levels of six phthalate esters, dimethyl- （DMP）, diethyl- （DEP）, dipropyl- [ DPrP], butyl-benzyl- （BBP）, dicyclohexyl- （DcHP）, and di-n-octyl-（DOP） phthalate in physiological saline samples. Under optimized experimental conditions, the method was linear in the following ranges： 0.2- 50 μg/L for DMP, DEP, DPrP, DcHP and DOP; 5- 100 μg/L for BBP. The correlation coefficients （R2 ） were in the range of 0. 9951 - 0. 9995 for all the analytes and the limits of detection （LODs） and limits of quantification （LOQs） were in the ranges of 0.02 - 0.9 μg/L and 0.08 - 2.7μg/L, respectively. The pretreatment process showed good reproducibility with inter-day and intra-day relative standard deviations （RSDs） below 8.5% and 11.2%, respectively. This method was used to determine the levels of six phthalate esters in physiological saline samples and the recoveries ranged from 71.2% to 107.3%. DMP and DEP were found in actual physical saline samples （brand A and brand B）.
文摘Background:Nails are one of the objects that are more durable than other witness objects.This study was interested in whether the Spin column-based nucleic acid purification technique could extract DNA from nails to give satisfactory results.Aims and Objectives:This study aimed to evaluate the yield and quality of DNA extracted from the nail.Materials and Methods:Nail samples from 15 males and 15 females over 18 were extracted using GeneAll®Exgene™Cell SV Mini Kit.The DNA concentration and purity were measured using a NanoDrop Spectrophotometer,and the quality of DNAwas assessed by polymerase chain reaction(PCR)using primers specific for human growth hormone(HGH).Results:The average yield of DNA was 0.816µg(0.141–2.706µg)obtained from the average nail weight of 30.2 mg(22.2–40.0 mg).The average DNA concentration was 27.2 ng/µL(4.7–90.2 ng/µL),corresponding to 30µL in volume.Almost all DNA samples(96.7%)had high purity(A260/A280≥1.80)and gave a band of PCR product of the HGH gene on agarose gel electrophoresis.Conclusion:Spin column-based nucleic acid purification is recommended for nail DNA extraction due to its simplicity and high quality.
基金supported by National Key Technology R&D Program in the 11th Five-Year Plan of China(No.2009BADB9B02)
文摘A rapid, simple and selective method based on molecularly imprinted, spin column extraction coupled with fluorescence detection was successfully established for the determination of 2,4-dinitrophenol in serum samples. The 2,4-dinitrophenol imprinted polymers exhibited highly selective recognition for the template molecule and the maximum adsorption capacity was 138.9 mg/g. The results indicated that when water is used as the loading solution, only 2,4-dinitrophenol could be adsorbed on the spin column without the remaining structural analogs（2-nitrophenol, 4-nitrophenol and phenol）. After eluting with acetonitrile/acetic acid（9/1, v/v）, 2,4-dinitrophenol in serum samples could be determined by using the fluorescence spectrometer, based on the fluorescence enhancement of fluorescein by the template molecule. Under the optimal conditions, the spiked recovery ranged from 95.8% to 103.4% and the detection limit was 1 nmol/L. The results confirmed the reliability and practicality of the protocol and revealed a good perspective of this method for biological sample analysis.
文摘氢同位素核自旋异构体正-仲态比例影响氢同位素的低温物性,有必要对其比例进行测定。本文利用活性三氧化二铝多孔层开管(Porous Layer Open Tubular,PLOT)柱实现了正-仲氢同位素(氕、氘)的基线分离,发展了一种可在液氮温度下测定同核分子正-仲态比例的色谱分析技术。研究结果表明,与传统三氧化二铝填充柱相比,高效PLOT柱实现了正、仲氕(o-H_2、p-H_2)以及正、仲氘(o-D2、p-D2)的基线分离(分离度R_s大于1.5),当流量为5 m L·min-1时,分离度R_s(p-H_2,o-H_2)=6.9,R_s(o-D2,p-D2)=1.8。正仲态分离度与进样量、流量均有关系。根据峰面积的积分结果,常温(298 K)下正、仲氕比例为2.77:1,正、仲氘比例为1.78:1,与理论测算值基本符合。HD与o-H_2实现了部分分离,R_s(o-H_2,HD)=0.5,根据理论预测,实现HD与o-H_2的基线分离(R_s达到1.5),理论塔板数需要达到3.9×105。
