油气回收装置检测标准器测量结果的不确定度评定

近年来,油站安装的油气回收装置,减少油站油气挥发造成的大气污染排放、改善空气环境质量起到了一定的作用。油气回收装置检测标准器的测量不确定度分析就显得格外重要,表征油站油气回收装置检测标准器的测量结果的准确程度亦备受重视。

Novel biosensor-based microarray assay for detecting rs8099917 and rs12979860 genotypes

AIM:To evaluate a novel biosensor-based microarray(BBM) assay for detecting rs12979860 and rs8099917 genotypes.METHODS:Four probes specific for rs8099917C/T or rs12979860G/T detection and three sets of quality control probes were designed,constructed and arrayed on an optical biosensor to develop a microarray assay.Two sets of primers were used in a one tube polymerase chain reaction(PCR) system to amplify two target fragments simultaneously.The biosensor microarray contained probes that had been sequenced to confirm that they included the rs8099917C/T or rs12979860G/T alleles of interest and could serve as the specific assay standards.In addition to rehybridization of four probes of known sequence,a total of 40 clinical samples collected from hepatitis C seropositive patients were also tested.The target fragments of all 40 samples were amplified in a 50 μL PCR system.Ten μL of each amplicon was tested by BBM assay,and another 40 μL was used for sequencing.The agreement of the results obtained by the two methods was tested statistically using the kappa coefficient.The sensitivity of the BBM assay was evaluated using serial dilutions of ten clinical blood samples containing 10 3-10 4 white cells/μL.RESULTS:As shown by polyacrylamide gel electrophoresis,two target segments of the interleukin 28Bassociated polymorphisms(SNPs) were successfully amplified in the one-tube PCR system.The lengths of the two amplified fragments were consistent with the known length of the target sequences,137 and 159 bps.After hybridization of the PCR amplicons with the probes located on the BBM array,the signals of each allele of both the rs8099917 SNPs and rs12979860 SNPs were observed simultaneously and were clearly visible by the unaided eye.The signals were distinct from each other,could be interpreted visually,and accurately recorded using an ordinary digital camera.To evaluate the specificity of the assay,both the plasmids and clinical samples were applied to the microarray.First,30 PCR amplicons of the various SNP alleles were hybridized on the BBM microarray.Full agreement between plasmids and the BBM assay was observed,with 30/30 correct matches(100%).The kappa value for the BBM assay with plasmids was 1.00(P < 0.05).For the 40 clinical blood samples,the BBM assay hybridization and direct sequencing results were compared for each amplicon.For patient blood samples,agreement was 28/28 for rs8099917T/T,9/11 for rs8099917T/G,1/1 for rs8099917G/G,24/24 for rs12979860C/C,11/14 for rs12979860C/T,and 2/2 for rs12979860T/T.Only five clinical samples of amplicon assay and direct sequencing results were discordant and heterozygotes:2/11 rs8099917T/G and 3/14 rs12979860C/T.The agreement of outcomes between BBM assay and direct sequencing for the detection of rs8099917 and rs12979860 was 95% and 92.5%,respectively;and the corresponding kappa values were 0.88 and 0.85(A kappa value > 0.75 was defined as substantial agreement).The BBM assay and sequencing had similar specificities for detection and identification of the two SNPs and their alleles.The sensitivity evaluation showed that the BBM assay could detect and identify SNP sequences present in blood samples containing as few as 10 2 white blood cells/μL.CONCLUSION:This biosensor microarray assay was highly specific,sensitive,rapid and easy to perform.It is compatible with clinical practice for detection of rs8099917 and rs12979860.

Low leakage 3×VDD-tolerant ESD detection circuit without deep N-well in a standard 90-nm low-voltage CMOS process

A new low leakage 3×VDD-tolerant electrostatic discharge(ESD)detection circuit using only low-voltage device without deep N-well is proposed in a standard 90-nm 1.2-V CMOS process.Stacked-transistors technique is adopted to sustain high-voltage stress and reduce leakage current.No NMOSFET operates in high voltage range and it is unnecessary to use any deep N-well.The proposed detection circuit can generate a 38 mA current to turn on the substrate triggered silicon-controlled rectifier(STSCR)under the ESD stress.Under normal operating conditions,all the devices are free from over-stress voltage threat.The leakage current is 88 nA under 3×VDD bias at 25°C.The simulation result shows the circuit can be successfully used for 3×VDD-tolerant I/O buffer.

Simultaneous quantification of lopinavir and ritonavir in human plasma by high performance liquid chromatography coupled with UV detection

High performance liquid chromatography was coupled with UV detection for simultaneous quantification of lopinavir （LPV） and ritonavir （RTV） in human plasma. This assay was sensitive, accurate and simple, and only used 200μL of plasma sample. Samples were liquid-liquid extracted, and diazepam was used as an internal standard. The chromatographic separation was achieved on a C18 reversed-phase analytic column with a mobile phase of acetonitrile-sodium dihydrogen phosphate buffer （10 mmol L-1, pH 4.80） （60：40, v/v）. UV detection was conducted at 205 nm and the column oven was set at 40℃. Calibration curves were constructed between 0,5-20 μg mL-1 for LPV and 0.05-5 μg mL-1 for RTV. The relative standard deviations were 2.16%-3.20% for LPV and 2.12%-2.60% for RTV for intra-day analysis, and 2.34%-4.04% for LPV and 0.31%-4.94% for RTV for inter-day analysis. The accuracy was within 100%＋10%. The mean extraction recoveries were 79.17%, 52.26% and 91.35% for RTV, LPV and diazepam, respectively. This method was successfully applied to human plasma samples from patients orally administered a salvage regimen of lopinavir-ritonavir tablets.

A new analytical method for quantifying of sterane and hopane biomarkers

An analytical method for quantifying biomarker compounds of the sterane and the hopane existing in saturated hydrocarbons has been established by using comprehensive two-dimensional gas chromatography-flame ionization detector(GC×GC-FID) with optimized operating parameters. The new method achieves the quantification by using a GC×GC-FID system which is able to completely separate steranes from hopanes. The data obtained by the new method are of good repeatability and reliability. Compared with the original data, the relative standard deviations(RSDs) of 12 reference compounds are less than 5%. The RSDs of the quantitative results of the biomarkers based on seven separate analyses are also less than 5%. Compared with the traditional method of gas chromatography-mass spectrometry(GC-MS), the new method has a number of advantages, such as common internal standards(ISs), high resolution, no co-eluting peak, and no interference caused by diagnostic ion peaks. The new method provides petroleum geologists with an effective and scientific means in future researches.