Analysis Cycle Time Research Articles

Fast saliva analysis by GC-MS with Cold EI and Open Probe Fast GC-MS with Cold EI for the detection of cannabis usage.

Saliva is a body fluid that is much easier to collect and analyze than blood. Thus, saliva analysis for the detection of delta 9-tetrahydrocannabinol (delta 9-THC) can serve as a tool for law enforcement agents to detect cannabis consumption by drivers. Fast saliva analysis for the presence of delta 9-THC and/or cannabidiol (CBD) is described with both gas chromatography-mass spectrometry (GC-MS) with Cold electron ionization (EI) with good separation and in 10min and/or with Open Probe Fast GC-MS with Cold EI in under 1 min full analysis cycle time. Saliva was taken directly from donors' tongues on a thin glass rod that was used "as is" for analysis. The saliva was thermally desorbed with a modified ChromatoProbe device inside the gas chromatograph (GC) injector and in an Open Probe (Agilent name QuickProbe) for its sub-1-min analysis. Cold EI is based on coupling of the GC and mass spectrometer (MS) with a supersonic molecular beam and on EI of vibrationally cold sample molecules during their flight through a contact-free ion source (thereby named Cold EI). A revised type of Open Probe Fast GC-MS on the bench is also described. Our saliva analysis was characterized by: Saliva can be collected in the field and transported to the lab for analyses "as is" without any sample preparation. Easy detection of cannabis consumption from cigarettes and/or other cannabis products. Distinction between the isomers delta 9-THC and CBD. Ultra-fast analysis in under 1 min using Open Probe Fast GC-MS with Cold EI.

Automated sample preparation of protein solid dosage forms: Novel application for the tablet processing workstation.

Biological macromolecule solid dosage forms represent the frontier of orally administered pharmaceuticals. Analysis of these drug products poses new challenges compared to traditional small molecule tablets. In this study we demonstrate the first, to our knowledge, automated Tablet Processing Workstation (TPW) sample preparation of large molecule tablets. Tablets containing a modified version of human insulin were tested for content uniformity and the automated method was successfully validated for recovery, carryover and displayed manual method equivalency in repeatability and in-process stability. Per TPW's ability to process one sample sequentially, the total analysis cycle time is, in fact, increased. In lieu, a net gain in scientist productivity is realized by enabling continuous operation reducing analytical scientist labor time by 71% compared to manually conducted sample preparation.

Capillary Sodium Dodecyl Sulfate Agarose Gel Electrophoresis of Proteins

Capillary sodium dodecyl sulfate gel electrophoresis has long been used for the analysis of proteins, mostly either with entangled polymer networks or translationally cross-linked gels. In this paper capillary agarose gel electrophoresis is introduced for the separation of low molecular weight immunoglobulin subunits. The light (LC~24 kDa) and heavy (HC~50 kDa) chain fragments of a monoclonal antibody therapeutic drug were used to optimize the sieving matrix composition of the agarose/Tris-borate-EDTA (TBE) systems. The agarose and boric acid contents were systematically varied between 0.2–1.0% and 320–640 mM, respectively. The influence of several physical parameters such as viscosity and electroosmotic flow were also investigated, the latter to shed light on its effect on the electrokinetic injection bias. Three dimensional Ferguson plots were utilized to better understand the sieving performance of the various agarose/TBE ratio gels, especially relying on their slope (retardation coefficient, KR) value differences. The best resolution between the LC and non-glycosylated HC IgG subunits was obtained by utilizing the molecular sieving effect of the 1% agarose/320 mM boric acid composition (ΔKR = 0.035). On the other hand, the 0.8% agarose/640 mM boric acid gel showed the highest separation power between the similar molecular weight, but different surface charge density non-glycosylated HC and HC fragments (ΔKR = 0.005). It is important to note that the agarose-based gel-buffer systems did not require any capillary regeneration steps between runs other than simple replenishment of the sieving matrix, significantly speeding up analysis cycle time.

A novel method for the quantification of anlotinib in human plasma using two-dimensional liquid chromatography.

A simple, efficient, and stable detection method of two-dimensional liquid chromatography (2D-LC) was established and validated to determine anlotinib in the human plasma. The 2D-LC system comprises a first-dimensional column (LC1), an intermediate transfer column, and a second-dimensional column (LC2). With simple protein precipitation treatment, the samples were processed directly for detection. The analysis cycle time was completed within 9.50 min. For the anlotinib concentrations, the calibration curve was linear over the 5.00-320.00 ng/mL range. The intra-day and inter-day precision ranges were 0.77-6.22% and 1.92-4.26%, respectively, for anlotinib concentrations. The recoveries were in the range of 97.85-102.50%. A total of 135 plasma samples from 94 patients were analyzed by our method. The plasma concentrations of patients were in the range of 5.17-106.38 ng/mL, in which the female had a higher plasma concentration (6.44-106.38 ng/mL). The simultaneous application of dexamethasone can increase the anlotinib concentration in the plasma. In our clinical application, we found that the factors that affect the plasma concentration include the time and dose of the medication, gender, and drug interactions. The method appears to be sensitive, precise, selective, and suitable for determining the concentration of anlotinib in the plasma sample.

Microfluidic Devices for Magnetic Separation of Biological Particles: A Review

Abstract Separation of microparticles and cells serves a critical step in many applications such as in biological analyses, food production, chemical processing, and medical diagnostics. Sorting on the microscale exhibits certain advantages in comparison with that on the macroscale as it requires minuscule sample or reagents volume and thus reduced analysis cycle time, smaller size of devices, and lower fabrication costs. Progresses have been made over time to improve the efficiency of these microscale particle manipulation techniques. Many different techniques have been used to attain accurate particle sorting and separation in a continuous manner on the microscale level, which can be categorized as either passive or active methods. Passive techniques achieve accurate manipulation of particles through their interaction with surrounding flow by carefully designed channel structures, without using external fields. As an alternative, active techniques utilize external fields (e.g., acoustic, electronic, optical, and magnetic field, etc.) to realize desired pattern of motion for particles with specific properties. Among numerous active methods for microfluidic particle sorting, the magnetic field has been widely used in biomedical and chemical applications to achieve mixing, focusing, and separating of reagents and bioparticles. This paper aims to provide a thorough review on the classic and most up-to-date magnetic sorting and separation techniques to manipulate microparticles including the discussions on the basic concept, working principle, experimental details, and device performance.

Highly sensitive retrospective determination of organophosphorous nerve agent biomarkers in human urine implemented in vivo in rabbit.

Highly toxic organophosphorous nerve agents (OPAs) have been used in several armed conflicts and terror attacks in the last few decades. A new method for retrospective determination of alkyl methylphosphonic acid (AMPA) metabolites in urine after exposure to VX, GB and GF nerve agents was developed. This method enables a rapid, sensitive and selective determination of trace levels of the nerve agent biomarkers ethyl methylphosphonic acid (EMPA), isopropyl methylphosphonic acid (IMPA) and cyclohexyl methylphosphonic acid (CMPA) in urine. The new technique involves a unique combination of two solid phase extraction (SPE) cartridges: a Ba/Ag/H cartridge for urine interference removal, and a ZrO2 cartridge for selective reconstitution and enrichment of the AMPAs. Extraction of AMPAs from the ZrO2 cartridge was accomplished with a 1% ammonium hydroxide (NH4OH) solution and was followed by analysis via liquid chromatography-mass spectrometry (LC-MS). The limits of quantitation (LOQs) were in the range of 10-100pg/mL with recoveries of 64-71% (± 5-19%) after fast sample preparation and a total LC-MS analysis cycle time of 15min and 13min, respectively. This method was successfully applied in vivo in a rabbit that was exposed to 0.5 LD50 (7.5µg/kg, i.v.) sarin for retrospective monitoring of the IMPA metabolite in urine. For the first time, IMPA was determined in rabbit urine samples for 15days post-exposure, which is longer than any reported post-exposure method for AMPAs. To the best of our knowledge, this new method is the most sensitive and rapid for AMPA determination in urine by LC-MS/MS analysis.

High-performance method for identification of super enhancers from ChIP-Seq data with configurable cloud virtual machines

A universal method for rapid identifying super-enhancers which are large domains of multiple closely-spaced enhancers is proposed. The method applies configurable cloud virtual machines (cVMs) and the rank-ordering of super-enhancers (ROSE) algorithm. To identify super-enhancers a сVM-based analysis of the ChIP-seq binding patterns of the active enhancer-associated mark is employed. The use of the proposed method is described step-by-step: configuration of cVM; ChIP-seq data alignment; peak calling; ROSE algorithm; interpretation of the results on a client machine. The method was validated for the search of super-enhancers using the H3K27ac mark in the sample datasets of a cell line (human MCF-7), mouse tissue (heart), and human tissue (adrenal gland). The total analysis cycle time of raw ChIP-seq data ranges from 15 to 48 min, depending on the number of initial short reads. Depending on the data processing step and availability of multi-threading, a cVM can be scaled up to a multi-CPU configuration with large amount of RAM. An important feature of the method is that it can run on a client machine that has low-performance with virtually any OS. The proposed method allows for simultaneous and independent processing of different sample datasets on multiple clones of a single cVM.•Cloud VMs were used for rapid processing of ChIP-seq data to identify super-enhancers.•The method can use a low-performance computer with virtually any OS on it.•It can be scaled up for parallel processing of individual sample datasets on their own VMs for rapid high-throughput processing.

Less than one minute low-pressure gas chromatography - mass spectrometry

Conventional gas chromatography - mass spectrometry (GC-MS) takes 20–40 min per sample, which is undesirably slow in any application if speed can be increased while still meeting analytical needs. In this study, we achieved reasonably good separations with full analysis cycle times of less than 1 min by combining for the first time low-pressure (LP) GC-MS with low thermal mass (LTM) resistive-heating for rapid temperature ramping and cooling of the capillary column. The analytical column is threaded into the LTM thin-walled metal tubing in an instrumental device known as “LTM Fast GC” that is mounted at the top of the gas chromatograph in a detector port. The column inlet and outlet are connected to the GC injector and MS transfer line as usual. For LPGC-MS, a 40 cm, 0.1 mm. i.d. uncoated flow restrictor capillary connected at the injector is coupled with a 2.6 m, 0.25 mm i.d., 0.25 µm film thickness analytical column leading to the MS. Thus, the inlet operates at normal GC pressures, but the analytical column is under vacuum, which increases the optimal helium carrier gas flow velocity thereby increasing speed of full range separations while maintaining acceptable quality of chromatography. This column configuration in LTM-LPGC-MS trades a 64-fold gain in speed of analysis vs. standard GC-MS for a 4-fold loss in chromatographic peak capacity, thereby converting analysis time from minutes into seconds in common applications. For example, jet fuel containing fatty acid methyl esters (akin to biofuel) was separated in 25 s with <1 min full analysis cycle time. An EPA Method 8270 mixture of 76 analytes was also analyzed in <1 min full cycle time by LTM-LPGC-MS. Other examples include very fast analysis of heroin in a street drug powder and elucidation of a new organic synthetic compound. In this report, we describe and discuss the several advantageous and practical features of LTM-LPGC-MS, as well as its trade-offs.

A novel and nonderivatization method for the determination of valproic acid in human serum by two-dimensional liquid chromatography.

A novel and robust two-dimensional liquid chromatography with ultraviolet detection method (2D-LC-UV) was developed and validated for high-throughput determination of the concentrations of valproic acid (VPA) in human plasma. This 2D-LC system was composed of a first-dimensional LC column, a second-dimensional LC column and an intermediate transfer column. The sample was directly injected into the 2D-LC system after an easy protein precipitation treatment. After online preconcentration and primary separation by the first-dimensional column, the target was captured by an intermediate column and then transferred to second-dimensional column for analysis. The system transferred the target through "central cutting" mode whereby the drug peak was not subject to interference from the matrix. The analysis cycle time was completed within 7.0 min. Compared with other methods that have been developed, the analysis time was reduced and the operation was much easier without any derivatization. The calibration curve was linear over the 5.90-188.94 μg/ml range for the VPA concentrations. The intra-day and inter-day precisions were <5.6%. The recoveries were in the range from 95.2 to 98.0%. This method appears to be sensitive, precise, rapid and low-cost for the quantification of VPA in serum sample.

Multicapillary Gas Chromatography-Temperature Modulated Metal Oxide Semiconductor Sensors Array Detector for Monitoring of Volatile Organic Compounds in Closed Atmosphere Using Gaussian Apodization Factor Analysis.

A unique metal oxide semiconductor sensor (MOS) array detector with eight sensors was designed and fabricated in a PTFE chamber as an interface for coupling with multicapillary gas chromatography. This design consists of eight transfer lines with equal length between the multicapillary columns (MCC) and sensors. The deactivated capillary columns were passed through each transfer line and homemade flow splitter to distribute the same gas flow on each sensor. Using the eight ports flow splitter design helps us to equal the length of carrier gas path and flow for each sensor, minimizing the dead volume of the sensor's chamber and increasing chromatographic resolution. In addition to coupling of MCC to MOS array detector and other considerations in hardware design, modulation of MOS temperature was used to increase sensitivity and selectivity, and data analysis was enhanced with adapted Gaussian apodization factor analysis (GAFA) as a multivariate curve resolution algorithm. Continues air sampling and injecting system (CASI) design provides a fast and easily applied method for continues injection of air sample with no additional sample preparation. The analysis cycle time required for each run is less than 300 s. The high sample load and sharp injection with the fast separation by MCC decrease the peak widths and improve detection limits. This homemade customized instrument is an alternative to other time-consuming and expensive technologies for continuous monitoring of outgassing in air samples.

Impact of instrument and column parameters on high-throughput liquid chromatography performance

The speed and separation performance of high-throughput liquid chromatography (HT LC) was investigated. We evaluated the contributions of various experimental parameters to the total analysis cycle time, including column length (column void time), gradient delay, flow rate, auto-sampler (A/S) speed, and software related delays. The best case injection-to-injection cycle time of 22s was achieved using 12s gradient time and 2.1×20mm columns packed with 1.7μm C18 particles. The total 22s analytical duty cycle consisted of 2.5s column void time, 1.8s gradient delay, 12s gradient time, and approximately 5.7s for software setup delay time that served as column re-equilibration time. The achieved peak capacity for an alkylphenone sample was approximately 35, giving a peak production rate of 95 peaks per minute. We estimated the impact of LC system dispersion on peak capacity and peak production rate (peak capacity per unit of time). For HT LC scenarios (5–50mm columns and 4.8–120s long gradients) we observed that even a minor amount of system dispersion (2μL2) reduces the achievable peak capacity and peak productivity rate significantly. HT LC–MS analysis using 2.1×5mm guard column with duty cycle of 22s was successfully demonstrated.

Open Probe fast GC-MS - combining ambient sampling ultra-fast separation and in-vacuum ionization for real-time analysis.

An Open Probe inlet was combined with a low thermal mass ultra-fast gas chromatograph (GC), in-vacuum electron ionization ion source and a mass spectrometer (MS) of GC-MS for obtaining real-time analysis with separation. The Open Probe enables ambient sampling via sample vaporization in an oven that is open to room air, and the ultra-fast GC provides ~30-s separation, while if no separation is required, it can act as a transfer line with 2 to 3-s sample transfer time. Sample analysis is as simple as touching the sample, pushing the sample holder into the Open Probe oven and obtaining the results in 30s. The Open Probe fast GC was mounted on a standard Agilent 7890 GC that was coupled with an Agilent 5977A MS. Open Probe fast GC-MS provides real-time analysis combined with GC separation and library identification, and it uses the low-cost MS of GC-MS. The operation of Open Probe fast GC-MS is demonstrated in the 30-s separation and 50-s full analysis cycle time of tetrahydrocannabinol and cannabinol in Cannabis flower, sub 1-min analysis of trace trinitrotoluene transferred from a finger onto a glass surface, vitamin E in canola oil, sterols in olive oil, polybrominated flame retardants in plastics, alprazolam in Xanax drug pill and free fatty acids and cholesterol in human blood. The extrapolated limit of detection for pyrene is <1fg, but the concentration is too high and the software noise calculation is untrustworthy. The broad range of compounds amenable for analysis is demonstrated in the analysis of reserpine. The possible use with alternate standard GC-MS and Open Probe fast GC-MS is demonstrated in the analysis of heroin in its street drug powder. The use of Open Probe with the fast GC acting as a transfer line is demonstrated in <10-s analysis without separation of ibuprofen and estradiol. Copyright © 2017 John Wiley & Sons, Ltd.

EVALUASI TINGKAT PELAYANAN JALAN DAN PENGARUHNYA TERHADAP ANGKA HENTI STOP

The number of students who coming from the outside of Yogyakarta is the main cause of the rapid growth. Hence, it’s impacted to transportation sector. In addition, one of the densenly traffic areas in Yogya phone’s Intersection. The purposes of this paper are to analize and evaluate the level of service provided by jogja phone’s intersection, and also identify the effect of saturation degree to number of stop. Based on traffic analysis cycle time, the service level to the traffic flow at the Jogja phone’s intersection at the present time is very low, that is F category. After the rearrangement process using MKJI 1997, it obtained a B level of service. In addition, the results of the analysis using SPSS 15, there is no significant effect between the degrees of saturation to the numbers of stop. Keywords: Degree of Saturation, Number of Stop, dan Regression

Static fault localization of subtle metallization defects using near infrared photon emission microscopy

In this paper, two electroluminescence phenomena, which enabled the static electrical fault localization of subtle back-end-of-line metallization defects using near-infrared photon emission microscopy in the logic circuitry and the memory array, are described. In the logic circuitry, through the study of the defect-induced hot carrier emissions from the combinational logic gates, distinctive differences in emission characteristic between open and short defects are identified. Using this defect induced emission characterization approach, together with layout trace and analysis, the type of defect can be predicted. The defect physical location, which yielded no detectable hotspot signal, can also be narrowed down along the long failure net. This allows for the selection of the most appropriate physical failure analysis approach for defect viewing and thus achieving significant reduction in failure analysis cycle time. In the memory array, the weak emission from partially turned-on pass gate transistor is leveraged to localize marginal opens and shorts on the wordline node of the pass-gate transistor. These approaches are applied with great success in the foundry environment to localize yield limiting defects that resulted in SCAN and memory build-in self-test failure, without memory bitmap, diagnostic support or measurable IDD leakage, on advanced technology nodes devices. A discussion on the factors that influence the success rate of this approach is also provided.

High-throughput determination of vancomycin in human plasma by a cost-effective system of two-dimensional liquid chromatography

Therapeutic drug monitoring (TDM) is one of the most important services of clinical laboratories. Two main techniques are commonly used: the immunoassay and chromatography method. We have developed a cost-effective system of two-dimensional liquid chromatography with ultraviolet detection (2D-LC-UV) for high-throughput determination of vancomycin in human plasma that combines the automation and low start-up costs of the immunoassay with the high selectivity and sensitivity of the liquid chromatography coupled with mass spectrometric detection without incurring their disadvantages, achieving high cost-effectiveness. This 2D-LC system offers a large volume injection to provide sufficient sensitivity and uses simulated gradient peak compression technology to control peak broadening and to improve peak shape. A middle column was added to reduce the analysis cycle time and make it suitable for high-throughput routine clinical assays. The analysis cycle time was 4min and the peak width was 0.8min. Compared with other chromatographic methods that have been developed, the analysis cycle time and peak width for vancomycin was reduced significantly. The lower limit of quantification was 0.20μg/mL for vancomycin, which is the same as certain LC–MS/MS methods that have been recently developed and validated. The method is rapid, automated, and low-cost and has high selectivity and sensitivity for the quantification of vancomycin in human plasma, thus making it well-suited for use in hospital clinical laboratories.

Fast Heroin and Cocaine Analysis by GC–MS with Cold EI: The Important Role of Flow Programming

A fast GC–MS method was developed based on the use of GC–MS with Cold EI. This new method was applied for the analysis of the street drugs heroin and cocaine and it enabled 2 min chromatography time and 3 min full analysis cycle time. GC–MS with cold EI provides mass spectra with enhanced molecular ions that are library compatible (with increased identification probabilities) and allows the use of short, 5 m 0.25 mm ID columns, which facilitates fast GC–MS. A central ingredient of our unique cold EI-based fast GC–MS analysis method is the use of column flow programming from 1 up to 32 ml min−1 column flow rate. Column flow programming can reduce the analysis time by about a factor of two and unlike temperature programs of GC ovens the carrier gas flow rate can be raised and lowered very quickly (in a few seconds). The fast GC–MS with Cold EI method is demonstrated by the analysis of heroin in its street drug powder and cocaine on paper money and it can be applied for other drugs of abuse as a general fast drugs analysis method.

Development of an Automated Headspace Gas Chromatography Instrument for the Determination of Residual Solvents in Pharmaceutical Compounds and Reaction Mixtures

The use of gas chromatography with headspace sampling is commonplace in analytical laboratories for the analysis of residual solvents. In this article, we discuss the use of an internal standard-based calibration, utilizing relative response factors, to enable the generation of accurate weight % solvent data on 25 common solvents in a single chromatographic run. The total cycle time for analysis is less than 30 min. To facilitate this technology into the process chemistry environment, bespoke open access software has been developed to simplify the sample submission process such that virtually no training is required to analyze the sample, process the data, and generate a report. Furthermore, an automated calibration check workflow has been implemented to validate the quality of the data on a daily basis and alert the system administrator in the event of a problem with the system.

Quantitative analysis of pasireotide (SOM230), a cyclic peptide, in monkey plasma using liquid chromatography in combination with tandem mass spectrometry.

A novel liquid chromatographic method with tandem mass spectrometric detection (LC-MS/MS) for the determination of Pasireotide (SOM230) was developed and validated with a dynamic range of 0.5-250ng/ml using 50μl of monkey plasma. SOM230 and the internal standard, [M+6]SOM230, were extracted from monkey plasma via μElution SPE. The acidified sample matrix was loaded onto the preconditioned Waters SPE plate for further processing. The analyte was eluted from the SPE plate using freshly prepared elution solvent, which was followed by dilution and LC-MS/MS analysis. By eliminating a step of evaporation of the SPE eluent, instead, injecting the eluent after a simple dilution, compound loss due to non-specific binding to the 96-well materials was prevented. Furthermore, freshly prepared elution solution was found a key to optimal extraction recovery of the analyte from monkey plasma. The optimal chromatographic separation was achieved on an Atlantis dC18 (50×2.1mm, 5μm particle size) column using gradient elution with a total analysis cycle time approximately 4min per injection. The mobile phases were water containing 0.5% acetic acid and 0.05% trifluoroacetic acid (TFA) (mobile phase A) and acetonitrile containing 0.5% acetic acid and 0.05% TFA (mobile phase B). The incorporation of TFA (0.05%, v/v) and acetic acid (0.5%, v/v) into the mobile phases was accompanied by the improved chromatography and minimized carryover due to the HPLC column. The current method was validated for specificity, sensitivity, matrix effect, recovery, linearity, accuracy and precision, dilution integrity, batch size and stability. The accuracy and precision for the LLOQs (0.5ng/ml) were within ±5.6% bias and ≤7.8% CV, respectively. From the intra-day and inter-day evaluations, the precision of the other QC samples (1.5, 7.5, 75 and 190ng/ml) ranged from 2.7 to 4.9% CV and the accuracy (% bias) from -1.3 to 7.3%, respectively. Additional assessment of incurred sample reanalysis (ISR) was conducted to demonstrate the ruggedness and robustness of the assay method. The validated method was successfully implemented to support a toxicity study in monkeys administered with 5 and 30mg of SOM230 in a single intramuscular injection of a long acting release (LAR) formulation.

Open Probe fast GC–MS—Real time analysis with separation

An Open Probe inlet was combined with a low thermal mass (LTM) ultra fast gas chromatograph (GC) and mass spectrometer (MS) of GC–MS for obtaining real time analysis with separation. The Open Probe is based on a vaporization oven that is open to room air while having helium purge flow protection to eliminate air leakage into the oven and MS ion source. Sample introduction into the Open Probe is as simple as touch the sample, push the sample holder into the Open Probe oven and have the results in 20–30s. The Open Probe is mounted onto the LTM fast GC that is coupled with the MS of either GC–MS with Cold EI or standard GC–MS. Open Probe fast GC–MS provides real time analysis in combination with GC separation, library identification, absence of ion suppression effects and uniform electron ionization response for improved quantitation, and it uses the low cost MS of GC–MS. The operation of the Open Probe fast GC–MS is demonstrated in the 20s separation and 40s full analysis cycle time of heroin in its street drug powder, cockroach repeller liquid residue on tomato in 30s and trace TNT on human hand in 40s.

Measurement of mercury species in human blood using triple spike isotope dilution with SPME-GC-ICP-DRC-MS.

The measurement of different mercury compounds in human blood can provide valuable information about the type of mercury exposure. To this end, our laboratory developed a biomonitoring method for the quantification of inorganic (iHg), methyl (MeHg), and ethyl (EtHg) mercury in whole blood using a triple-spike isotope dilution (TSID) quantification method employing capillary gas chromatography (GC) and inductively coupled dynamic reaction cell mass spectrometry (ICP-DRC-MS). We used a robotic CombiPAL(®) sample handling station featuring twin fiber-based solid-phase microextraction (SPME) injector heads. The use of two SPME fibers significantly reduces sample analysis cycle times making this method very suitable for high sample throughput, which is a requirement for large public health biomonitoring studies. Our sample preparation procedure involved solubilization of blood samples with tetramethylammonium hydroxide (TMAH) followed by the derivatization with sodium tetra(n-propyl)borate (NaBPr(4)) to promote volatility of mercury species. We thoroughly investigated mercury species stability in the blood matrix during the course of sample treatment and analysis. The method accuracy for quantifying iHg, MeHg, and EtHg was validated using NIST standard reference materials (SRM 955c level 3) and the Centre de Toxicologie du Québec (CTQ) proficiency testing (PT) samples. The limit of detection (LOD) for iHg, MeHg, and EtHg in human blood was determined to be 0.27, 0.12, and 0.16 μg/L, respectively.