Optimization and method validation of determining polyphenolic compounds by UFLC-DAD system using two biphenyl and pentafluorophenylpropyl columns

The development of efficient analytical methods for the identification and quantification of polyphenols in natural products is needed. The present study was aimed to optimize the polyphenols by UFLC using two different Ascentis express columns. Chromatographic separation was performed using UFLC-DAD connected with Ascentis Express Biphenyl column (system I) and Ascentis Express F5 pentafluorophenylpropyl (PFP) column (system II). Multistep gradient elution program was used in both analytical instruments to detect 34 compounds of different classes (phenolic acids, flavonoids, isoflavones, flavanols, flavonols and flavanones). The detection wavelength, linear calibration range, theoretical plate, tailing factor, limits of detection and of quantification were determined. The results were validated and the optimized method was proven to be precise, revealing good separation in the UFLC system II using Ascentis Express F5 PFP column. In conclusion, this study shows analytical preciseness and is useful in determining polyphenols in natural products.     

The ales laboratory automation system

A systenm for the automation of the analytical activities of a chemical laboratory is described. The system dieals with investigations of physical and chemical characteristics of telecommunication materials for prototype development and quality control. The architecture, software development and interfacing of different analytical instruments (mainly for calorimetry, spectroscopy, electrochemistry, chromatography and mechanics) for the ALES (Automatzione del Laboratorio Elettrochimico SIP) system are discussed in terms of analytical and management performance.     

User-oriented software for determination of the precision of signal-integrating analytical methods

A software package for the determination of the uncertainty in signal-integrating analytical techniques is presented. The program can be used to calculate detection limits and to determine baseline drift properties, the influence of drift-correcting procedures and integration time, etc. Complete analytical procedures, e.g., chromatography or atomic emission spectrometry, or parts of a system can be tested. No profound theoretical knowledge is required by the user; confidence intervals are calculated where necessary. The theoretical basis and the structure of the program are evaluated.     

Portable automated separation system for routine purification and/or pre-concentration of radionuclides based on column chromatography

In recent years the purification and/or pre-concentration of radionuclides before the measurement has grown increasing interest in analytical chemistry. In this study, a new compact and portable stand-alone equipment permitting automatisation of various separation tasks is developed. The new system allows performing quick and reliable automated separation of the selected radionuclide. Since there is no need for permanent manual control of the separation procedures (automatic loading of the sample, washing and stripping solution on the column are controlled via a computer program) the system can be operated overnight. The new system posses the possibility of more variable control for the separation process via new developed user-friendly software, is shielded against the chemical vapors and could be universally equipped with any available chromatographic column. For the automated separation of U, Pu and Am isotopes (achieved recoveries were in the range of 65–95 %, depending on the element separated. The data, presented, show that the application of the module should be also straightforward for other elements: simply by changing the chromatographic columns with the resin having high chemical selectivity for the target ion. The developed separation column module, software and hardware can be readily adapted in any laboratory to meet defined analytical requirements.     

Characterizing nonconstant instrumental variance in emerging miniaturized analytical techniques

Measurement variance is a crucial aspect of quantitative chemical analysis. Variance directly affects important analytical figures of merit, including detection limit, quantitation limit, and confidence intervals. Most reported analyses for emerging analytical techniques implicitly assume constant variance (homoskedasticity) by using unweighted regression calibrations. Despite the assumption of constant variance, it is known that most instruments exhibit heteroskedasticity, where variance changes with signal intensity. Ignoring nonconstant variance results in suboptimal calibrations, invalid uncertainty estimates, and incorrect detection limits. Three techniques where homoskedasticity is often assumed were covered in this work to evaluate if heteroskedasticity had a significant quantitative impact—naked-eye, distance-based detection using paper-based analytical devices (PADs), cathodic stripping voltammetry (CSV) with disposable carbon-ink electrode devices, and microchip electrophoresis (MCE) with conductivity detection. Despite these techniques representing a wide range of chemistries and precision, heteroskedastic behavior was confirmed for each. The general variance forms were analyzed, and recommendations for accounting for nonconstant variance discussed. Monte Carlo simulations of instrument responses were performed to quantify the benefits of weighted regression, and the sensitivity to uncertainty in the variance function was tested. Results show that heteroskedasticity should be considered during development of new techniques; even moderate uncertainty (30%) in the variance function still results in weighted regression outperforming unweighted regressions. We recommend utilizing the power model of variance because it is easy to apply, requires little additional experimentation, and produces higher-precision results and more reliable uncertainty estimates than assuming homoskedasticity.     

Quantitative determination of some food dyes using digital processing of images obtained by thin-layer chromatography

A high-performance thin-layer chromatographic method combined with image processing of scanned chromatograms was developed for the determination of some food dyes (tartrazine, azorubine and Sunset Yellow) in different products. Porous silica gel with 3-aminopropyl functional groups attached to the matrix was used as stationary phase and a mixture of isopropanol, diethyl ether and ammonia (2:2:1, v/v/v) formed the mobile phase. Quantitative evaluation was performed using special-purpose software. The linearity of the analytical procedure was sustained by the numerical parameters such as correlation coefficient (0.9952-0.9980) and standard error of determination (0.03-0.20). The limits of detection were found to be within the range of 5.21-9.34 ng/spot, and the limits of quantification between 10.21 and 18.09 ng/spot. Recovery studies performed on two levels of concentration gave values between 96.39 and 102.76%. These results show that the regression approach provides rigorous and realistic detection and quantification limits and as a consequence can be routinely applied to other analytical systems. This method does not require expensive analytical instruments compared with classical densitometry and provides a reliable quantitative evaluation with minimum of time.     

A Smartphone-assisted Paper-based Analytical Device for Fluorescence Assay of Hg2+

Rapid, efficiency and portable detection systems in low-resource settings with limited laboratory equipment and technical expertise are urgently needed. Herein, an integrated platform composed of a paper-based analytical device and a smartphone detection system for Hg²⁺ onsite testing was developed. Nitrogen-doped carbon dots(N-CDs) were synthesized by a simple hydrothermal method using citric acid as the carbon source and ethanediamine as the nitrogen source, which gave out bright blue fluorescence under the excitation at 350 nm UV light and the absolute fluorescence quantum yield was 17.1%. The fluorescence of the prepared N-CDs can be effectively quenched by Hg²⁺. In addition, an external attachment of smartphone for illumination and external light interference was designed to trace the fluorescence signals, and a software application of Android system with simple operation program was developed to perform snapshot and image processing. The smartphone-assisted detection system was combined with the N-CDs decorated paper chip to achieve the sensitive detection of Hg²⁺ in water samples. This integrated method for reliable sensing of Hg²⁺ shows a good linear detection range of 10-800 μmol/L(R²=0.9595) with detection limit of 1.07×10^-8 mol/L.     

Analytical Method Development for 19 Alkyl Halides as Potential Genotoxic Impurities by Analytical Quality by Design

Major issues in the pharmaceutical industry involve efficient risk management and control strategies of potential genotoxic impurities (PGIs). As a result, the development of an appropriate method to control these impurities is required. An optimally sensitive and simultaneous analytical method using gas chromatography with a mass spectrometry detector (GC–MS) was developed for 19 alkyl halides determined to be PGIs. These 19 alkyl halides were selected from 144 alkyl halides through an in silico study utilizing quantitative structure–activity relationship (Q-SAR) approaches via expert knowledge rule-based software and statistical-based software. The analytical quality by design (QbD) approach was adopted for the development of a sensitive and robust analytical method for PGIs. A limited number of literature studies have reviewed the analytical QbD approach in the PGI method development using GC–MS as the analytical instrument. A GC equipped with a single quadrupole mass spectrometry detector (MSD) and VF-624 ms capillary column was used. The developed method was validated in terms of specificity, the limit of detection, quantitation, linearity, accuracy, and precision, according to the ICH Q2 guideline.     

New tools: Expert systems for uncertainty budgets

An important part of quality assurance in any analytical laboratory is the production of comprehensive results integrating uncertainty measurements. Many testing laboratories face the problem that the expenditure required to evaluate small uncertainties (high precision and high accuracy) is often uneconomic. In most cases an uncertainty of high reliability has to be calculated from only a few data (one calibration, few replications, etc.). This problem can be solved by an expert system. To achieve this the analytical procedure has to be structured into a dialouge and divided into parts. The uncertainty has to be calculated for each part of the procedure. Addition of the individual uncertainties results in the combined and expanded uncertainty. During the dialouge the system should advise the analyst how to get an efficient and effective calculation of uncertainty. All calculations, mathematical and statistical procedures have to be surveyable but running the system should not be too time consuming for economic reasons. Within the scope of the EURECA-project initiated by the Eidgenössische Materialprüfungs- und Forschungsanstalt (EMPA), St. Gallen, Switzerland, expert system software is being developed in cooperation with other research institutes and manufacturers of analytical instruments. Using this software it will be possible to calculate the uncertainty for analytical procedures such as titration, atomic emission spectrometry (ICP-OES), atomic absorption spectrometry (AAS) and gas- and liquid chromatography (GC, HPLC).     

Introduction of measurement uncertainty estimation into analytical instrument software: mission impossible?

In this paper we argue that introduction of ISO GUM based uncertainty estimation into analytical equipment software is a “mission possible” and is wholly realistic at this stage of development of the art. A possible general scheme of implementation of uncertainty estimation into analytical instrument software is presented based on the example of high-performance liquid chromatography (HPLC) but is also applicable to most other analytical instruments. This implementation would be very beneficial for the analysts/practitioners as the uncertainty would be handled within their everyday software environment.     

基于氧化还原循环信号放大技术的单分子电化学研究进展

现代分析科学的整体发展对分析方法的灵敏度、选择性以及快速响应等有了更高的要求。在单分子水平上实现对目标分子的检测及控制是化学家们长期以来梦寐以求的一项富有挑战性的前沿领域,也是近年来分析科学很重要的前沿发展方向。用电化学方法直接检测单分子面临的一项挑战是单个分子在氧化还原过程中得失电子产生的电流变化太小,现代仪器无法对如此小的电流进行识别。使电极表面氧化还原过程中的电子交换实现多次循环可以放大产生的电流,从而实现单分子水平的直接电化学分析。本文对近期通过循环电子交换过程放大电流信号的技术和装置进行了综述,将各类方法进行对比,并对单分子电化学未来的发展方向进行了展望。     

Operation of an academic open access mass spectrometry facility with particular reference to the analysis of synthetic compounds

Open access mass spectrometry now provides the opportunity to move this spectroscopic method to the beginning of the analytical chain, a place formerly the exclusive province of NMR and TLC. To date this transition has been occurring in industrial settings but there has been less change in the academic environment. This paper provides one blueprint for setting up such a facility, primarily in support of organic synthesis but also for the use of biological scientists. The open access format used at UCI utilizes four instruments: an ESI-TOFMS system used in the flow injection mode, two GC/MS systems (one in EI and one in CI) and a MALDI-TOFMS system. The first three instruments have autosamplers and open access software whereas the MALDI system has a fully automated plate handling interface. This level of automation allows access to the instruments by a user community of more than 100 users, day or night. The decisions made in setting up these instruments were based on a 'keep it simple' philosophy, given the fact that the primary type of data of interest is the molecular mass of the analyte and that data are required for a very wide range of structures.     

Introduction of measurement uncertainty estimation into analytical instrument software: mission impossible?

In this paper we argue that introduction of ISO GUM based uncertainty estimation into analytical equipment software is a “mission possible” and is wholly realistic at this stage of development of the art. A possible general scheme of implementation of uncertainty estimation into analytical instrument software is presented based on the example of high-performance liquid chromatography (HPLC) but is also applicable to most other analytical instruments. This implementation would be very beneficial for the analysts/practitioners as the uncertainty would be handled within their everyday software environment.

     

Autonomous microfluidic system for phosphate detection

Miniaturization of analytical devices through the advent of microfluidics and micro total analysis systems is an important step forward for applications such as medical diagnostics and environmental monitoring. The development of field-deployable instruments requires that the entire system, including all necessary peripheral components, be miniaturized and packaged in a portable device. A sensor for long-term monitoring of phosphate levels has been developed that incorporates sampling, reagent and waste storage, detection, and wireless communication into a complete, miniaturized system. The device employs a low-power detection and communication system, so the entire instrument can operate autonomously for 7 days on a single rechargeable, 12 V battery. In addition, integration of a wireless communication device allows the instrument to be controlled and results to be downloaded remotely. This autonomous system has a limit of detection of 0.3 mg/L and a linear dynamic range between 0 and 20 mg/L.     

Teaching of process analytical chemistry

In the teaching of analytical chemistry for chemical engineering students it is essential today to teach the chemical analysis of dynamic systems, not only in the process control of the modern technological systems, where the control of composition or structure of different material streams is necessary, but also in all other instances where analysis, decision and intervention follow each other, forming a closed cycle. Teaching can be made effective if students already have a knowledge of the basic disciplines (including analytical chemistry). The schedule of the teaching programme should include the mathematical statistical treatment of process signals, quality of the signals, signal-improvement methods, characteristics of instruments, calibration and an introduction to sensors, analysers suitable for continuous or periodical measurements and local area networks. As practical exercizes an apparatus for investigation of the dynamic properties of a thermoanalytical detector system, a computer program for simulating process variables and the control loop including the measuring system are presented.     

Can an instrument learn from experiments done by itself?

The hardware and software requirements for intelligent analytical instruments are discussed. Existing technology in both fields allows the production of a new generation of instruments that can learn from their own operation and even transfer the acquired knowledge to machines of a later generation. The hardware needed is a processor with the power of at least the Intel 80386 processor, 4–5 Mbyte RAM, storage equivalent to a 1.6-Gbyte hard disk and a special encode/decode processor for compressing complex instrumental information. This hardware can support the transmission, transformation, storage and handling of about 100 000 complex measurements (up to 3 kbyte of numbers per measurement) and a knowledge base of about the same size. In this context, the space for software can be neglected or, in the case of neural networks, the “software” can be hardwired. At preent, there are two different approaches to handling the required amount of data, extracting vital information, and acquiring (in close cooperation with experts) new knowledge. The first is neural networks and the second is hierarchical clustering. It is argued that combination of both methods forms a very powerful basis for software development in intelligent instruments.     

Developing multiple high-throughput GLP methods for an investigational drug candidate in various matrices within a 96-well plate

In early pharmaceutical product development, an investigational drug candidate is typically dosed to various species for toxicological and pharmacokinetic studies. Most of these studies require multiple analytical methods that have to be validated with good laboratory practice (GLP) prior to the application in regulated studies. Usually, these analytical methods are developed in either a serial or parallel approach. For either approach, the development of multiple analytical methods takes tremendous work from scientists and instruments, and thus is not cost-effective. In this respect, a new strategy has been developed for simultaneous GLP method development using liquid chromatographic separation and tandem mass spectrometric detection. This high-throughput approach allows system suitability, carryover, calibration curve, accuracy, precision, matrix effect and selectivity to be evaluated in one 96-well plate. The strategy has been successfully implemented for multiple investigational drug candidates at Abbott Laboratories. The methods developed with this strategy are accurate, precise, selective, robust and matrix-independent. As an example, ABT-279 was used to demonstrate the feasibility of this strategy.     

Compressed Air-Driven Continuous-Flow Thermocycled Digital PCR for HBV Diagnosis in Clinical-Level Serum Sample Based on Single Hot Plate

We report a novel compressed air-driven continuous-flow digital PCR (dPCR) system based on a 3D microfluidic chip and self-developed software system to realize real-time monitoring. The system can ensure the steady transmission of droplets in long tubing without an external power source and generate stable droplets of suitable size for dPCR by two needles and a narrowed Teflon tube. The stable thermal cycle required by dPCR can be achieved by using only one constant temperature heater. In addition, our system has realized the real-time detection of droplet fluorescence in each thermal cycle, which makes up for the drawbacks of the end-point detection method used in traditional continuous-flow dPCR. This continuous-flow digital PCR by the compressed air-driven method can meet the requirements of droplet thermal cycle and diagnosis in a clinical-level serum sample. Comparing the detection results of clinical samples (hepatitis B virus serum) with commercial instruments (CFX Connect; Bio Rad, Hercules, CA, USA), the linear correlation reached 0.9995. Because the system greatly simplified the traditional dPCR process, this system is stable and user-friendly.     

LSI-based amperometric sensor for bio-imaging and multi-point biosensing

We have developed an LSI-based amperometric sensor called "Bio-LSI" with 400 measurement points as a platform for electrochemical bio-imaging and multi-point biosensing. The system is comprised of a 10.4 mm × 10.4 mm CMOS sensor chip with 20 × 20 unit cells, an external circuit box, a control unit for data acquisition, and a DC power box. Each unit cell of the chip contains an operational amplifier with a switched-capacitor type I-V converter for in-pixel signal amplification. We successfully realized a wide dynamic range from ±1 pA to ±100 nA with a well-organized circuit design and operating software. In particular, in-pixel signal amplification and an original program to control the signal read-out contribute to the lower detection limit and wide detection range of Bio-LSI. The spacial resolution is 250 μm and the temporal resolution is 18-125 ms/400 points, which depends on the desired current detection range. The coefficient of variance of the current for 400 points is within 5%. We also demonstrated the real-time imaging of a biological molecule using Bio-LSI. The LSI coated with an Os-HRP film was successfully applied to the monitoring of the changes of hydrogen peroxide concentration in a flow. The Os-HRP-coated LSI was spotted with glucose oxidase and used for bioelectrochemical imaging of the glucose oxidase (GOx)-catalyzed oxidation of glucose. Bio-LSI is a promising platform for a wide range of analytical fields, including diagnostics, environmental measurements and basic biochemistry.