Recent advances of online coupling of sample preparation techniques with ultra high performance liquid chromatography and supercritical fluid chromatography

Ultra high performance liquid chromatography and supercritical fluid chromatography techniques are favored because of their high efficiency and fast analysis speed. Although many sample preparation techniques have been coupled with common liquid chromatography online, the online coupling of sample preparation with the two popular chromatography techniques have gained increasing attention owing to the increasing requirements of efficiency and sensitivity. In this review, we have discussed and summarized the recent advances of the online coupling of sample preparation with ultra high performance liquid chromatography and supercritical fluid chromatography techniques. The main sample preparation techniques that have been coupled with ultra high performance liquid chromatography online are solid‐phase extraction and in‐tube solid‐phase microextraction, while solid‐phase extraction and supercritical fluid extraction are the main techniques that have been coupled with supercritical fluid chromatography online. Especially, the strategies for online coupling of sample preparation with chromatography techniques were summarized. Typical applications and growing trends of the online coupling techniques were also discussed in detail. With the increasing demands of improving the efficiency, throughput, and analytical capability toward complex samples of the analysis methods, online coupling of sample preparation with chromatography techniques will acquire further development.     

Simultaneous sample preparation for high-performance liquid chromatographic determination of Vitamin A and β-carotene in emulsified nutritional supplements after solid-phase extraction

Determination of small amounts of the fat-soluble species Vitamin A (VA) (2.5 μg/g) and β-carotene (9 μg/g) from emulsified nutritional supplements containing 50 kinds of co-existing compounds and a fat content between 2000 and 8000 times higher was performed by solid-phase extraction (SPE) and high-performance liquid chromatography (HPLC) with fluorescence detection set at ex. 350 nm and em. 480 nm, and visible detection at 450 nm using an Inertsil ODS 80A (5 μm) analytical column. Mobile phases of methanol-ethanol (50:50) and acetonitrile-ethanol (70:30) were used for the both vitamins. A Bond Elut C₁₈ cartridge was chosen for SPE after comparison with eight other types of SPE cartridge. Retention time of VA and β-carotene was 7 and 8 min, respectively, giving a limit of detection of ca. 0.1 ng per injection at a signal-to-noise ratio 3:1. Recoveries of VA and β-carotene were over 90% by the standard addition method. Relative standard deviation of VA and β-carotene were ca. 2.9 (n=5) and 2.3% (n=5), respectively.     

Generation and characterization of polyclonal antibodies against microcystins-Application to immunoassays and immunoaffinity sample preparation prior to analysis by liquid chromatography and UV detection

Polyclonal antibodies against microcystin-LR (MC-LR), a cyclic heptapeptide toxin, were generated in rabbits using MC-LR-BSA. An enzyme-linked immunosorbent assay (ELISA) was developed for the characterization of the antibodies and their potential use for analytical purposes. The concentration of MC-LR that inhibits 50% of antibody-antigen binding (IC₅₀) was 0.5 μg L⁻¹ for the indirect ELISA format and 0.9 μg L⁻¹ for the direct ELISA, using MC-LR-horseradish peroxidase conjugate. The limit of detection corresponding to IC₈₀ was found to be 0.06 μg L⁻¹, well below the Word Health Organization level for drinking water of 1 μg L⁻¹. The direct competitive ELISA was applied to water samples and was shown useful for screening purposes. The developed anti-microcystin antibodies were immobilized on solid supports for use in selective solid phase extraction (SPE) systems, prior to liquid chromatography (LC) quantification. An immunoaffinity cartridge (IAC), a Sepharose^®-based cartridge incorporating 2 mg of antibodies allowed the selective and quantitative recovery of a mixture of 0.2 μg of MCs showing potential use in sample preparation of real matrices. When applied to water and green algae samples, average recoveries from Sepharose^®-based cartridges were in the range of 86-113% for water samples and 85-92% for blue-green algae samples. Selectivity of the IAC clean-up was proven by comparison with non-specific solid phase extraction using octadecylsilica (ODS) sorbent. Results obtained using LC/UV after IAC clean-up agreed well with results obtained using liquid chromatography and mass spectrometry detection (LC/MS and LC/MS/MS) after SPE-C18 clean-up, allowing therefore to validate the resulting technique.     

Recent advances in application and progress of advanced materials as adsorbents in sample preparation for plant growth regulators

Plant growth regulators are a class of physiologically active substances that could modify or regulate basic physiological processes in the plant and defense against abiotic and biotic stresses, including natural plant growth regulators and synthetic ones. Different from natural plant growth regulators with low content and high cost of extraction in plants, synthetic ones can be produced in large-scale production and widely used in agriculture for increasing and securing yield and quality of the harvested produce. However, like pesticides, the abuse of plant growth regulators will have negative impacts on human beings. Therefore, it is important to monitor plant growth regulators residues. Due to the low concentration of plant growth regulators and complex matrices of food, it is necessary to isolate and extract plant growth regulators by appropriate adsorbents in sample preparation for obtaining satisfactory results. In the last decade, several advanced materials as adsorbents have shown superiority in sample preparation. This review briefly introduces the recent application and progress of advanced materials as adsorbents in sample preparation for extraction of plant growth regulators from the complex matrix. In the end, the challenge and outlook about the extraction of plant growth regulators of these advanced adsorbents in sample preparation are presented.     

废弃线路板化学分析采样、制样及检测方法

介绍废弃线路板中有回收价值元素和有害物质分析的采样、制样及检测技术。样品经过分类采样、剪切破碎和高温灰化制样,采用样品全分析或副批混合样分析。通过提高称样量、多次测定求平均值的办法,火试金富集-重量法测定贵金属金、银。湿法王水溶解样品,碘量法测定主体元素铜。电感耦合等离子体发射光谱(ICP-AES)法测定其它杂质元素,被测元素质量浓度在0~10μg/mL范围内与光谱强度呈良好线性关系,相关系数均大于0.9998。测定结果的相对标准偏差小于10%(n=5),加标回收率为97.0%~102.5%。该方法简单、快速,有效地解决了线路板样品不均匀而难采样,以及硬度、韧性强制样难,金属易包裹难分解的技术瓶颈,测定结果准确,具有代表性。该方法适用于废弃线路板化学成分分析,其它废弃电子产品检测可参考此方法。     

Pressurized liquid extraction and GC‐MS analysis for simultaneous determination of seven components in Cinnamomum cassia and the effect of sample preparation

A pressurized liquid extraction and GC‐MS method was developed for simultaneous quantitative determination of the seven components, including cinnamaldehyde, copaene, cinnamic acid, coumarin, 2‐methoxycinnamaldehyde, 2‐methoxycinnamic acid and safrole in Cinnamomum cassia. The results showed that methanol and ethanol was not available for extraction of cinnamaldehyde and 2‐methoxycinnamaldehyde due to aldol reaction. The developed method was validated to be sensitive, accurate and simple, and was successfully employed for the analysis of 15 samples of C. cassia. The contents of the investigated components were significantly variant and cinnamaldehyde is the most abundant compound, but safrole was not detected in all samples.     

Sieve-based device for MALDI sample preparation. I. Influence of sample deposition conditions in oligonucleotide analysis to achieve significant increases in both sensitivity and resolution

Spraying of oligonucleotide-matrix solutions through a stainless steel (ss) sieve (38 microm, 450 mesh) leads to the formation, on the matrix-assisted laser desorption/ionization (MALDI) sample holder, of uniformly distributed microcrystals, well separated from each other. When the resulting sample holder surface is irradiated by laser, abundant molecular species form, with a clear increase in both intensity and resolution with respect to values obtained by 'Dried Droplet', 'Double Layer', and 'Sandwich' deposition methods. In addition, unlike the usual situation, the sample is perfectly homogeneous, and identical spectra are obtained by irradiating different areas. On one hand, the data indicate that this method is highly effective for oligonucleotide MALDI analysis, and on the other, that it can be validly employed for fully automated MALDI procedures.     

Deproteinization with ZnSO4–Ba(OH)2 reduces the photodegradation of montelukast during plasma sample preparation for HPLC analysis

Montelukast (MKT), a leukotriene receptor antagonist, degrades when it is exposed to light. The analysis of MKT content in blood plasma by high-pressure liquid chromatography requires several sample preparation steps including deproteinization. This study aimed to evaluate MKT photodegradation in blood plasma samples and optimize a deproteinization method to reduce MKT photodegradation, and thereby improve analytical quality. We evaluated the stability of MKT in water and plasma in real time using high-pressure liquid chromatography and optimized a sample deproteinization procedure by comparing the effectiveness of several deproteinization methods. When exposed to light, MKT photodegraded quickly. Although MKT photodegradation was slightly slower than that in water, a half portion (55%) of the MKT in plasma degraded within 2 h when exposed to light. The rate of MKT photodegradation was dramatically reduced by sample deproteinization using ZnSO₄–Ba(OH)₂, but it was accelerated by deproteinization through precipitation using methanol or acetonitrile. These results suggest that precautions should be taken when preparing plasma samples for the analysis of MKT, and that deproteinization of such samples using ZnSO₄–Ba(OH)₂ can reduce the risk of analytical error arising from MKT photodegradation.     

Microchip based sample treatment device interfaced with ICP-MS for the analysis of transition metals from environmental samples

A downscaled solid phase (SPE) device applicable for sample preparation prior to ICP-MS monitoring, have been constructed making use of the lab on a chip concept. Standard photolithography and wet chemical etching were used to fabricate glass microfluidic devices accommodating three microchannels, each of them incorporating a defined section that could be packed with SPE materials; selective chelating resin. The microfluidic device was interfaced with the ICP-MS instrument throughout a low flow rate concentric nebuliser using a Teflon connector, and coupled with a flow injection manifold delivering samples and reagents via a manually operated splitting valve. The feasibility of the miniaturized prototype to perform SPE of trace metals was proved by analyzing trace metals, Cd, Co, and Ni, in seawater reference materials; CASS-2 and SLEW-1. The obtained result was in good agreement with the certified values. The device could be used as a remote miniaturized sample treatment for field work.     

Recent advances and applications of molecularly imprinted polymers in solid‐phase extraction for real sample analysis

Sample pretreatment is essential for the analysis of complicated real samples due to their complex matrices and low analyte concentrations. Among all sample pretreatment methods, solid‐phase extraction is arguably the most frequently used one. However, the majority of available solid‐phase extraction adsorbents suffer from limited selectivity. Molecularly imprinted polymers are a type of tailor‐made artificial antibodies and receptors with specific recognition sites for target molecules. Using molecularly imprinted polymers instead of conventional adsorbents can greatly improve the selectivity of solid‐phase extraction, and therefore molecularly imprinted polymer‐based solid‐phase extraction has been widely applied to separation, clean up and/or preconcentration of target analytes in various kinds of real samples. In this article, after a brief introduction, the recent developments and applications of molecularly imprinted polymer‐based solid‐phase extraction for determination of different analytes in complicated real samples during the 2015‐2020 are reviewed systematically, including the solid‐phase extraction modes, molecularly imprinted adsorbent types and their preparations, and the practical applications of solid‐phase extraction to various real samples (environmental, food, biological, and pharmaceutical samples). Finally, the challenges and opportunities of using molecularly imprinted polymer‐based solid‐phase extraction for real sample analysis are discussed.     

在线样品前处理大体积进样离子色谱法直接测定海水中亚硝酸盐、硝酸盐和磷酸盐

采用戴安公司谱睿(Pre)在线样品除氯技术,结合OnGuard Ba柱去除硫酸盐,建立了离子色谱直接测定海水中亚硝酸盐、硝酸盐和磷酸盐的方法。该方法以IonPac AG23为富集柱,高容量IonPac AS23为分离柱,淋洗液自动发生装置在线产生KOH溶液进行梯度淋洗,抑制电导检测。实验结果表明: 样品稀释5～10倍时,直接进样不会干扰目标物测定。当流速为1 mL/min、进样量为500 μL时,海水中NO~2-N、NO~3-N、PO3~4-P的方法检出限分别为0.3、0.4、0.2 μg/L,线性范围分别为10～500 μg/L、14～680 μg/L、3.4～170 μg/L,线性相关系数r均大于0.9990。测得人工海水样品中目标物的加标回收率为92%～106%,相对标准偏差(RSD, n=6)为1.2%～7.7%。该方法一次进样可在13 min内完成分析,具有操作简单快捷、无污染等优点,能满足近海海水中NO~2、NO~3、PO3~4的定量分析要求。     

Trace determination of parabens in cosmetics and personal care products using fabric‐phase sorptive extraction and high‐performance liquid chromatography with UV detection

A simple, fast, and sensitive analytical protocol using fabric‐phase sorptive extraction followed by high performance liquid chromatography with ultraviolet detection has been developed and validated for the extraction of five parabens including methylparaben, ethylparaben, propylparaben, butylparaben, and benzylparaben. In the present work, sol‐gel polyethylene glycol coated fabric‐phase sorptive extraction membrane is used for the preconcentration of parabens (polar) from complex matrices. The use of fabric‐phase sorptive extraction membrane provides a high surface area which offers high sorbent loading, shortened equilibrium time, and overall decrease in the sample preparation time. Various factors affecting the performance of fabric‐phase sorptive extraction, including extraction time, eluting solvent, elution time, and pH of the sample matrix, were optimized. Separation was performed using a mobile phase consisting of water:acetonitrile (63:37; v/v) at an isocratic elution mode at a flow rate of 0.9 mL/min with wavelength at 254 nm. The calibration curves of the target analytes were prepared with good correlation coefficient values (r² > 0.9955). The limit of detection values range from 0.252 to 0.580 ng/mL. Finally, the method was successfully applied to various cosmetics and personal care product samples such as rose water, deodorant, hair serum, and cream with extraction recoveries ranged between 88 and 122% with relative standard deviation <5%.     

Molecularly imprinted stir bar sorptive extraction coupled with high performance liquid chromatography for trace analysis of sulfa drugs in complex samples

A novel sulfamethazine molecularly imprinted polymer (MIP)-coated stir bar for sorptive extraction of eight sulfa drugs from biological samples was prepared. The MIP-coating was about 20 μm thickness with the relative standard deviation (RSD) of 6.7% (n = 10). It was characterized by scanning electron microscope, infrared spectrum, thermogravimetric analysis, and solvent-resistant investigation, respectively. The non-imprinted polymer (NIP)-coating was used for comparison. The adsorptive capacity and selectivity of MIP-coating were evaluated in detail. The MIP-coating showed higher adsorption capability and selectivity than the NIP-coating. The saturated adsorption amount of the MIP-coating was 4.6 times over that of the NIP-coating in toluene. Sulfamethazine could be detected after the MIP-coated stir bar sorptive extraction even at a low concentration of 0.2 μg/L. The MIP-coating also exhibited selective adsorption ability to analogues of the template. A method for the determination of eight sulfa drugs in biological samples by MIP coated stir bar sorptive extraction coupled with high performance liquid chromatography (HPLC) was developed. The extraction conditions, including extraction solvent, extraction time, desorption solvent, desorption time and stirring speed, were optimized. The linear ranges were 1.0-100 μg/L and 2.0-100 μg/L for eight sulfonamides, respectively. The detection limits were within the range of 0.20-0.72 μg/L. The method was successfully applied to simultaneous multi-residue analysis of eight sulfonamides in spiked pork, liver and chicken samples with the satisfactory recoveries.     

A pre-column derivatization high-performance liquid chromatography method for simultaneous determination of short-chain and medium-chain fatty acids in a fecal sample

Short-chain and medium-chain fatty acids have plentiful biological functions, which play a crucial role in the diagnosis and therapy of many diseases. Herein, a new method for simultaneous quantifying 17 short-chain and medium-chain fatty acids with high-performance liquid chromatography coupled with an ultraviolet detector was developed and the pre-column derivatization by indole-3-acetic acid hydrazide was performed to improve the separation and detection. The conditions of the derivatization reaction were systematically investigated. Subsequently, the method was validated and the results showed a satisfactory linearity (linear regression coefficients > 0.9969), the limit of detection (4.0×10⁻³–1.9×10⁻² μmol/L), precision (0.9%–7.3% for intra-day and 2.0%–9.8% for inter-day), recovery (90.0%–109.1% with relative standard deviation <7.7%) and stability (0.1%–3.3% for standard solution and 0.2%–3.9% for fecal sample). Finally, the established method was successfully applied to quantify short-chain and medium-chain fatty acids in the feces of healthy control and diabetic rats. Eleven kinds of short-chain and medium-chain fatty acids were detected and six of them showed a significant difference between the control group and the model group.     

Construction of a portable sample preparation device with a magnetic poly(methacrylic acid‐co‐ethylene dimethacrylate) monolith as the extraction medium and its application in the enrichment of UV filters in water samples

A portable sample preparation device with a magnetic polymer monolith as the extraction medium was constructed. The monolith was synthesized by polymerizing methacrylic acid and ethylene dimethacrylate around a cylindrical magnet. In this way, the monolith with a magnetic core could be readily attached to the extraction device by magnetism. The constructed device was evaluated for the enrichment of UV filters in water samples, followed by high‐performance liquid chromatographic analysis. The extraction efficiency for the targets was satisfactory with no matrix interference. Good linearities were obtained for the UV filters with the correlation coefficients >0.9986. The limits of detection and quantification for the UV filters were 0.3–0.8 and 1.0–2.4 ng/mL, respectively. The recoveries of the UV filters from the spiked water samples at the concentration of 100 ng/mL were 95.3–101.7%, with relative standard deviations <10%. Accordingly, the proposed portable device was demonstrated to be suitable for on‐site simultaneous sampling, purification, and preconcentration within a single step.     

Comparison of sample preparation methods combined with gas chromatography with electron‐capture detection for the analysis of multipesticide residues in lotus seeds

Sample preparation is always the major bottleneck in analytical chemistry for the determination of pesticide residues. Different sample preparation methods have been proposed due to the wide variety of pesticides used and the inherent complexity of the matrices. In this study, different sample preparation methods including SPE, matrix solid‐phase dispersion, the quick, easy, cheap, efficient, rugged, and safe method, and a one‐step completion method were compared and evaluated for extracting pesticides from lotus seeds. Analysis was carried out using GC with electron‐capture detection. The results showed that good recoveries for tested pesticides were obtained by using Florisil in the four methods, and the extraction efficiency of the one‐step completion method was superior to the other three methods. The one‐step completion method was confirmed to have good linearity, reproducibility, stability, and recovery for the detection of 36 pesticides in lotus seed samples. The data collected from this study are expected to prove useful in regulating the concentration of the residues in lotus seeds, as well as in protecting human health from the hazards posed by these residues.     

The role of sample preparation in multidimensional gas chromatographic separations for non‐targeted analysis with the focus on recent biomedical,food, and plant applications

In this review, we consider and discuss the affinity and complementarity between a generic sample preparation technique and the comprehensive two‐dimensional gas chromatography process. From the initial technical development focus (e.g., on the GC×GC and solid‐phase microextraction techniques), the trend is inevitably shifting toward more applied challenges, and therefore, the preparation of the sample should be carefully considered in any GC×GC separation for an overreaching research. We highlight recent biomedical, food, and plant applications (2016–July 2020), and specifically those in which the combination of tailored sample preparation methods and GC×GC–MS has proven to be beneficial in the challenging aspects of non‐targeted analysis. Specifically on the sample preparation, we report on gas‐phase, solid‐phase, and liquid‐phase extractions, and derivatization procedures that have been used to extract and prepare volatile and semi‐volatile metabolites for the successive GC×GC analysis. Moreover, we also present a milestone section reporting the early works that pioneered the combination of sample preparation techniques with GC×GC for non‐targeted analysis.     

Development of a solid-phase extraction with capillary liquid chromatography tandem mass spectrometry for analysis of estrogens in environmental water samples

Capillary liquid chromatography (cLC) hyphenated with tandem mass spectrometry (MS-MS) was used to separate and quantitate trace concentrations of five estrogens in aqueous samples. New C(18)-based sorption materials bound to the silica support by monomeric and polymeric mechanisms were compared and tested for solid-phase extraction (SPE) of selected analytes with respect to optimization of their preconcentration yield. Application of an endcapped, monomer-bound preconcentration Discovery DSC-18Lt column under the optimized conditions provides yields in the range from 95 to 100% with a high repeatability (n=3, RSD≤7.2%). Using the electrospray ionization in the positive mode (ESI+), the cLC-MS-MS system (the Zorbax SB C18 capillary column and a binary mobile phase of acetonitrile and water containing 0.1% formic acid in both the components) was optimized to attain a sufficient retention of the early eluting estriol, a satisfactory resolution of the analytes and the maximum sensitivity of the determination. Both the isocratic and gradient elution were used and the optimized gradient method permitted analyses of aqueous environmental samples in 14 min within a linearity range from 6.1 to 25.0 (LOQ of analytes) to 500 ng/L and with a very good linearity (r>0.9981) for all the estrogens studied. The detection limits are in the range from 3.0 to 6.8 ng/L (1 μL injection volume). Six environmental water samples were analyzed and the studied estrogens were found in the Vltava river sample collected in Prague (13.2 ng/L for 17β-estradiol) and in the inlet to the wastewater treatment plant in Prague, at an overall concentration of 371.4 ng/L.     

Study of blood collection and sample preparation for analysis of vitamin D and its metabolites by liquid chromatography–tandem mass spectrometry

The analysis of vitamin D status, with special emphasis on 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D, is gaining interest in clinical studies due to the classical and non-classical effects attributed to this prohormone. In this research, the influence of the two steps preceding determination (viz. sample collection and preparation) on the quantitative analysis of vitamin D and its more important metabolites has been studied. Two preparation approaches, deproteination and solid-phase extraction (SPE), have been evaluated in terms of sensitivity to delimit their application, thus establishing that detection of 1,25-dihydroxyvitamin D cannot be addressed by protein precipitation. Concerning sample collection, serum and plasma reported high accuracy (above 83.3%) for vitamin D and metabolites, while precision, expressed as relative standard deviation, was below 12.9% for all analytes in both samples. Statistical analysis revealed that serum and plasma provided similar physiological levels for vitamin D₃, 24,25-dihydroxyvitamin D₃ and 25-hydroxyvitamin D₃, while significantly different levels were obtained for 1,25-dihydroxyvitamin D₃, always higher in plasma than in serum. Sample collection and treatment have proved to be significant in the analysis of vitamin D and its relevant metabolites.     

Sample preparation and current applications of liquid chromatography for the determination of non-structural carbohydrates in plants

This article summarizes the current methods of determination of non-structural carbohydrates (NSCs) in plant samples based on liquid chromatography (LC). NSCs comprise several types of carbohydrates: sugar alcohols (e.g., sorbitol), monosaccharides (e.g., glucose and fructose), disaccharides (e.g., sucrose), oligosaccharides (e.g., raffinose) and polysaccharides [e.g., starch and polyfructans (e.g., inulin)]. NSCs are important in plant metabolism and have to be strictly distinguished from all sorts of structural carbohydrates (e.g., polysaccharide cellulose) that make up the backbone of the plants. Consequently, preservation of structural carbohydrates is a crucial step during sample preparation for NSC determination and is therefore addressed.Sugar alcohols, monosaccharides, disaccharides and those oligosaccharides that are easily soluble in polar solvents can be analyzed directly by high-performance LC. They are also referred to as free carbohydrates (FCs).However, polysaccharides are generally submitted to hydrolyzation into monomers prior to their quantitative analysis. This can be done either chemically, using acids, or enzymatically - both methods are discussed. For identification and quantification of the NSCs after LC separation, the following detectors are used: pulsed amperometry, refractive index, evaporate light scattering and finally, mass spectrometry.