Internal standard tracked dilution to overcome challenges in dried blood spots and robotic sample preparation for liquid chromatography/tandem mass spectrometry assays

Dried blood spot (DBS) technology is an emerging alternative for sample collection in bioanalysis. Dilution for DBS samples is a challenge due to its solid sample format. Currently, DBS samples requiring dilution were first extracted as regular samples and then diluted with extracted blank samples containing internal standard (IS). Since the dilution step is a volume-critical step, extra care has to be taken to achieve accurate dilution when dealing with limited volume extracted samples. Here, we introduce an alternative sample dilution for liquid chromatography/tandem mass spectrometry (LC/MS/MS) assays using IS to track the dilution step. Dilution factor-adjusted IS working solution was first added to the sample requiring dilution before sample processing; subsequently, the processed sample was approximately diluted into the assay linear response range before LC/MS/MS analysis. We define this approach as "IS-tracked dilution". The advantage of this approach is that the diluting step is tracked by the IS and is no longer a volume-critical step. Another recognized challenge related to sample dilution is automatic sample dilution using a liquid handler. This "IS-tracked dilution" may also help address some of the challenges for automatic sample dilution of liquid samples. This new dilution approach was proven to be effective and convenient in both plasma assays and DBS assays using omeprazole as a probe compound.     

A serial dilution microfluidic device using a ladder network generating logarithmic or linear concentrations

In this paper, we propose a serial dilution microfluidic chip which is able to generate logarithmic or linear step-wise concentrations. These concentrations were generated via adjustments in the flow rate of two converging fluids at the channel junctions of the ladder network. The desired dilution ratios are almost independent of the flow rate or diffusion length of molecules, as the dilution device is influenced only by the ratio of volumetric flow rates. Given a set of necessary dilution ratios, whether linear or logarithmic, a serial dilution chip can be constructed via the modification of a microfluidic resistance network. The design principle was suggested and both the logarithmic and linear dilution chips were fabricated in order to verify their performance in accordance with the fluorescence intensity. The diluted concentrations of a fluorescein solution in the microfluidic device evidenced relatively high linearity, and the cytotoxicity test of MCF-7 breast cancer cells via the logarithmic dilution chip was generally consistent with the results generated with manual dilution.     

Cascade system for rapid on-line dilutions in flow-injection analysis

A system for on-line dilutions in flow-injection analysis (FIA) is described. The system requires two flow-through pumps (peristaltic) and a conventional rotary injection valve. No precision timing or computer-controlled valves are required. The "cascade" dilution technique is demonstrated by the determination of 0-1.7M chloride. Viscosity effects are studied for samples with viscosities of 1-150 relative to water. Sample-zone volume reduction by splitting streams is combined with dilution by merging streams to achieve effective dilution factors of nearly 500. The reciprocal mole fraction is proposed as a variable for the characterization of open flow-injection systems and compared with the dispersion coefficient. An equation is included to predict this factor as a function only of the flow-rates used in the system. The cascade dilution system yields reproducible dilution factors with relative standard deviations of less than 3%, with sampling rates up to 100/hr.     

电渗泵—流动注射在线稀释法在火焰原子吸收光谱分析高浓度样品中？…

采用电渗泵流动注射－火焰原子吸收法（ＦＩ－ＦＡＡＳ）,研究了高浓度镁样品的在线稀释和分析。电渗泵在低流量时有很好的稳定性。利用控制采样时间的办法,可以准确采集微量样品,在线稀释。同时使用Ｔ型客对雾化量进行补偿,在线稀释倍数可达１０００以上,相对标准偏差〈１．５％（ｎ＝６）。通过改变采样电压（速度）,载流速度和采样时间可以方便地得到不同的稀释倍数,测定速度可达１００个／ｈ样品以上。     

Study on the capacities,heats of dilution and properties of concentrated potassium carnallite solutions at 298.15 K

The heats of dilution of the infinitesimally dilute potassium carnallite solutions at 298.15 K have been studied by continuous titration from 1.8942 to 0.01044 mol·kg^?1, and an equation for the curve of heats of dilution has been fitted. It was shown that the enthalpy of dilution for the same concentration of the carnallite solution is equal to the sum of those of KCl and MgCl₂ solutions. The equation for the curve of enthalpy of dilution corresponds to that of natural carnallite.     

Enthalpies of dilution of mono-, di- and poly-alcohols in dilute aqueous solutions at 298.15 K

The enthalpies of dilution of aqueous solutions of methanol, ethanol, l-propanol, 2-propanol, 1-butanol, l-pentanol, 1-hexanol, cyclohexanol, 1,3-butanediol, 1,4-butanediol, 2,3-butanediol and poly-alcohol(cyclohexaamylose) have been determined at high dilution as a function of the mole fraction of alcohol at 298.15 K, by a rocking twin-microcalorimeter of the heat-conduction type. A smoothing equation of the enthalpies of dilution against the mole fractions of alcohols are given. The graphical comparison of experimental results with their smoothed values or literature ones, taking into account the dependence of the mole fractions, are also presented. It has been found for the aqueous solutions of shorter n-alcohols than hexanol that at very high dilution, exothermic values of molar enthalpies of dilution from a definite mole fraction of alcohols to infinite dilution with the change of mole fraction is proportional to carbon number of n-alcohols. The molar enthalpies of infinite dilution of aqueous butanediol isomers and 1-hexanol were very large. Molar enthalpies of infinite dilution of aqueous poly-alcohol (cyclohexaamylose) were endothermic. This revised version was published online in August 2006 with corrections to the Cover Date.     

Determination of fluoride in sea-water by molecular absorption spectrometry of aluminium monofluoride after removal cation and anion interferences

Three procedures are proposed for the determination of trace levels of fluoride in sea-water, based on the formation of aluminium monofluoride in an electrothermal graphite furnace, followed by measurement of its molecular absorption at 227.45 nm. They involve the use of dilution, a matrix modifier, or a matrix modifier and an ion-exchange resin, and are all acceptably sensitive and specific for fluoride. Interferences from cations and anions are removed by a simple 20-fold dilution of the sample. At 10-fold sample dilution, chloride interference can be removed by adding 0.3M ammonium nitrate together with 0.01M aluminium + 0.01M strontium as a matrix modifier. The same matrix modifier is valid for use with 5-fold sample dilution and a cation-exchange step to avoid matrix affects from cations and chloride. The detection limit is about 8-10 ng/ml fluoride and the determination limit is 20 ng/ml. The precision of peak-height measurement at 0.2 mug/ml is 5-7%.     

恒温式滴定量热计连续测定溶液稀释热的研究

本文提出, 以不同的电热功率, 对稀释后的系统分别进行升温的方法, 求出K值, 进而计算出热容C.     

生化需氧量(BOD_5)测定中稀释倍数的简捷确定法

在生化需氧量 (BOD5)的测定中 ,稀释倍数的估计是否适当至关重要。该文根据废水生物处理可行性的判断标准———BOD5/CODcr比值的波动范围 ,将其分为三个区间 ,经过数学推导 ,得出各区间的稀释倍数 ,即n1=0 .57COD/d ,n2 =1.3COD/d ,n3=1.2 3COD/d ,其中d为当天稀释水的溶解氧。这三个稀释倍数可覆盖BOD5整个波动范围。由此 ,根据化学需氧量 (CODcr)可以简捷地确定出稀释倍数 ,从而提高了BOD5测定的有效性和准确性     

On-line dilution with sequential injection analysis: a system for monitoring sulphate in industrial effluents

 On-line dilution with sequential injection analysis has been evaluated for sulphate monitoring in industrial effluents using a dilution coil in the conduits of the manifold system and a dilution step as part of the timing sequence. The manifold of the SIA system with the dilution coil is more complex than the system including the dilution step. The former method needs more complicated programming as well. Shorter analysis time favoured the dilution with the dilution step, but the limited linear range of this method is a large drawback. The large linear range of the SIA system with dilution coil makes it very suitable to be used as a process analyser. The proposed SIA system is suitable to monitor sulphate in industrial effluents in the range 50–5000 mg/L with a dilution coil, in the range 50–1000 mg/L with a timing sequence of 6.5 s and in the range 750–5000 mg/L with a timing sequence of 15 s using a dilution step; all three ranges with a RSD <4.5%. Received: 11 March 1996/Revised: 1 July 1996/Accepted: 4 July 1996     

Thermische und spektroskopische Untersuchungen zur Wassersorption an Alkalihalogeniden

Investigations on the extension of the diffuse reflectance spectroscopy by means of the dilution method of KORTÜM show the decisive influence of water sorption on the dilution substances, particularly on their absolute reflectance. Water sorption is investigated on alkali halides (NaCl and LiF) as dilution substances by gravimetric, thermal desorption and reflectance methods. Sorption gravimetry gives data on the quantities of adsorbed substances and the time function of adsorption processes, thermal desorption on chemical binding of adsorbed molecules and reflectance spektroscopy on the molar absorptivity of adsorbed water molecules.     

Henry’s constants and infinite dilution activity coefficients of propane,propene, butane,isobutene, 1-butene,isobutane, trans-2-butene and 1,3-butadiene in 1-propanol at T=(260 to 340) K

The Henry’s constants and the infinite dilution activity coefficients of propane, propene, butane, isobutane, 1-butene, isobutene, trans-2-butene and 1,3-butadiene in 1-propanol at T=(260 to 340) K are measured by a gas stripping method. The rigorous formula for evaluating the Henry’s constants from the gas stripping measurements is used for these highly volatile mixtures. The accuracy of the measurements is about 2% for Henry’s constants and 3% for the estimated infinite dilution activity coefficients. In the evaluations for the infinite dilution activity coefficients, the nonideality of solute is not negligible especially at higher temperatures and the estimated uncertainty in the infinite dilution activity coefficients include 1% for nonideality.     

Combating matrix effects in LC/ESI/MS: The extrapolative dilution approach

Liquid chromatography electrospray mass spectrometry - LC/ESI/MS—a primary tool for analysis of low volatility compounds in difficult matrices - suffers from the matrix effects in the ESI ionization. It is well known that matrix effects can be reduced by sample dilution. However, the efficiency of simple sample dilution is often limited, in particular by the limit of detection of the method, and can strongly vary from sample to sample.In this study matrix effect is investigated as the function of dilution. It is demonstrated that in some cases dilution can eliminate matrix effect, but often it is just reduced. Based on these findings we propose a new quantitation method based on consecutive dilutions of the sample and extrapolation of the analyte content to the infinite dilution, i.e. to matrix-free solution.The method was validated for LC/ESI/MS analysis of five pesticides (methomyl, thiabendazole, aldicarb, imazalil, methiocarb) in five matrices (tomato, cucumber, apple, rye and garlic) at two concentration levels (0.5 and 5.0 mg kg⁻¹). Agreement between the analyzed and spiked concentrations was found for all samples. It was demonstrated that in terms of accuracy of the obtained results the proposed extrapolative dilution approach works distinctly better than simple sample dilution.The main use of this approach is envisaged for (a) method development/validation to determine the extent of matrix effects and the ways of overcoming them and (b) as a second step of analysis in the case of samples having analyte contents near the maximum residue limits (MRL).     

On-line dilution with sequential injection analysis: a system for monitoring sulphate in industrial effluents

 On-line dilution with sequential injection analysis has been evaluated for sulphate monitoring in industrial effluents using a dilution coil in the conduits of the manifold system and a dilution step as part of the timing sequence. The manifold of the SIA system with the dilution coil is more complex than the system including the dilution step. The former method needs more complicated programming as well. Shorter analysis time favoured the dilution with the dilution step, but the limited linear range of this method is a large drawback. The large linear range of the SIA system with dilution coil makes it very suitable to be used as a process analyser. The proposed SIA system is suitable to monitor sulphate in industrial effluents in the range 50–5000 mg/L with a dilution coil, in the range 50–1000 mg/L with a timing sequence of 6.5 s and in the range 750–5000 mg/L with a timing sequence of 15 s using a dilution step; all three ranges with a RSD <4.5%. Received: 11 March 1996/Revised: 1 July 1996/Accepted: 4 July 1996     

Henry’s law constant for environmentally significant compounds

Many nonvolatile organic compounds, e.g., polyaromatic hydrocarbons (PAHs), are readily stripped during aerobic biodegradation. This is because of the high infinite dilution activity coefficient resulting from forces generated by the water-organic interactions at the molecular level. Several models have been proposed for air-stripping based on the Henry’s law constant. By definition, the Henry’s law constant is the infinite dilution activity coefficient multiplied by the pure component vapor pressure. In this article, a gas saturation technique was used to measure the very low vapor pressures exhibited by these nonvolatile compounds. Literature values of other PAHs have been tabulated and are presented. For determining infinite dilution activity coefficients, a differential ebulliometery apparatus has been constructed. In this technique, the boiling point difference between pure water and a water-organic solution is measured very precisely. Thermodynamics is then used to calculate the infinite dilution activity coefficient. The method’s accuracy has been tested using the phenol-water system.     

Quantitative detection of adenosine in urine using silver enhancement of aptamer-gold nanoparticle aggregation and progressive dilution

We implement the progressive dilution strategy to bring assays based on gold nanoparticles to a quantitative level. This is demonstrated by the detection of adenosine in urine by combining progressive dilution with the silver enhancement of aptamer-gold nanoparticle aggregation, giving good accuracy, high selectivity, and an unlimited dynamic range above LOD.     

Comparison of the processes induced by nitrogen dilution on the photodissociation of silane and disilane at 193 nm

We compare the influence of the dilution of silane and disilane in nitrogen during laser photodissociation to produce silicon at 193 nm, at room temperature in a static reaction chamber. The experimental results show that the conversion of the reactant gas and its deposition yield can be controlled by varying adequately the extent of dilution. So, two total pressure regions have been observed, independent of the dilution: below 40–50 Torr, the variations of stable species concentration are very important but above these values the variation in the dilution rate has practically no effects on their concentrations. In the first region, during the silane photodissociation at the initial reactant pressure below 5 Torr, the conversion of silane increases with increasing dilution, and at higher initial reactant pressure the conversion of silane tends to rise only a little. In contrast, at any initial reactant pressure, the conversion of disilane during its photodecomposition decreases with increasing dilution. In the second region, the concentration of each stable gaseous species tends to reach a pressure stationary-state. For both the silane and disilane photodissociation, the deposition yield of silicon increases with decreasing the initial reactant gas pressure and it reaches a pressure stationary-state above 50% dilution; but in all the cases, it is greater in disilane photolysis than that of silane. A simple kinetic model is proposed for which the computed results predict the time-evolution of gas composition and amount of silicon deposited.     

气体扩散法测氨中几种流动注射扩展线性范围方式的比较及发酵液中氨态氮的测定

本文以气体扩散法测氨为例，比较了三种流动注射扩展线性范围方法的性能；讨论了影响分析精度的因素。把微升进样在线稀释技术同气体扩散光度法结合，测定了农药发酵液中氨态氮的含量。方法的回收率为９３—１０６％，每小时可分析６０—８０个样。     

Enthalpies of Dilution of D-p-Hydroxyphenylglycine in Buffer Solutions at Different pH

The effect of pH on the dilution enthalpies of D-p-hydroxyphenylglycine in phosphate buffer solutions has been investigated by isothermal titration microcalorimetry at 298.15 K. The corresponding homogeneous enthalpic interaction coefficients have been calculated according to the excess enthalpy concept. The results show that the enthalpies of dilution of D-p-hydroxyphenylglycine in phosphate buffer solutions at different pH are all positive. The overall trend is that enthalpies of dilution become more positive with an increase of pH, but there is a minimum of the enthalpy of dilution at pH = 7.0. The enthalpic pair interaction coefficients, h ₂, all have negative values. The results are interpreted from the point of view of solute-solute and solute-solvent interactions involved in the solvent effects.