Comparison of the response of four aerosol detectors used with ultra high pressure liquid chromatography

The responses of four different types of aerosol detectors have been evaluated and compared to establish their potential use as a universal detector in conjunction with ultra high pressure liquid chromatography (UHPLC). Two charged-aerosol detectors, namely Corona CAD and Corona Ultra, and also two different types of light-scattering detectors (an evaporative light scattering detector, and a nano-quantity analyte detector [NQAD]) were evaluated. The responses of these detectors were systematically investigated under changing experimental and instrumental parameters, such as the mobile phase flow-rate, analyte concentration, mobile phase composition, nebulizer temperature, evaporator temperature, evaporator gas flow-rate and instrumental signal filtering after detection. It was found that these parameters exerted non-linear effects on the responses of the aerosol detectors and must therefore be considered when designing analytical separation conditions, particularly when gradient elution is performed. Identical reversed-phase gradient separations were compared on all four aerosol detectors and further compared with UV detection at 200 nm. The aerosol detectors were able to detect all 11 analytes in a test set comprising species having a variety of physicochemical properties, whilst UV detection was applicable only to those analytes containing chromophores. The reproducibility of the detector response for 11 analytes over 10 consecutive separations was found to be approximately 5% for the charged-aerosol detectors and approximately 11% for the light-scattering detectors. The tested analytes included semi-volatile species which exhibited a more variable response on the aerosol detectors. Peak efficiencies were generally better on the aerosol detectors in comparison to UV detection and particularly so for the light-scattering detectors which exhibited efficiencies of around 110,000 plates per metre. Limits of detection were calculated using different mobile phase compositions and the NQAD detector was found to be the most sensitive (LOD of 10 ng/mL), followed by the Corona CAD (76 ng/mL), then UV detection at 200 nm (178 ng/mL) using an injection volume of 25 μL.     

Supercritical fluid chromatography and two-dimensional supercritical fluid chromatography of polar car lubricant additives with neat CO(2) as mobile phase

Car lubricant additives are added to mineral or synthetic base stocks to improve viscosity and resistance to oxidation of the lubricant and to limit wear of engines. Their total amount in the commercial lubricant varies from a few percents to 20-25%. As they belong to various chemical classes and are added to a very complex medium, the base stock, their detailed chromatographic analysis is very difficult and time consuming as it should involve sample treatment and preparative scale separations in order to simplify the sample. The aim of this work is to determine the feasibility of the separation of low molecular weight lubricant additives using various packed columns with pure CO(2) as a mobile phase to enable implementation of flame ionisation detection as universal detector. This is part of a hypernated system including more sophisticated specific detectors, such as AED, FTIR or MS to obtain detailed structural information of compounds. This paper is devoted to the comparison of some stationary phases supposed to provide hydrocarbon group type separation (silica and normal phase) or separations on alkyl-bonded silica in non-aqueous mode of some selected classes of additives in test mixtures or in base stocks. Adsorption chromatography allows partial separation of additives from the base stocks while the direct elution of test additives can only be obtained on reversed phase supports having a very efficient silanol group protection so the interaction of the more polar compounds is much reduced. A two-dimensional scheme of analysis is also described. It combines adsorption chromatography to separate most of the polar additives from the base stock and alkyl-bonded silica for more detailed separation of the additives. However, overlapping between groups of compounds and the lack of resolution between some additives and the base stock should be addressed by the implementing of selective detectors.     

Isocratic Separation of Some Food Additives by Reversed Phase Liquid Chromatography

In recent years, concern about food safety has been growing. The use of food additives in different countries is limited by specific regulations. Therefore, analytical methods that simultaneously determine artificial sweeteners and preservatives are advantageous. High performance liquid chromatography has been the most popular choice for the determination of food additives. In this study, reversed-phase liquid chromatography was developed for the separation of α-aspartame, sodium saccharin, acesulfame-K, vanillin, sorbic acid and benzoic acid. The effects of the proportion of the organic modifier on the chromatographic separation were investigated in order to separate these additives. The results showed that optimum chromatographic separation for these compounds takes place when the acetonitrile content in the mobile phase was 15% containing ammonium acetate buffer (0.005 M). It was concluded that the best separation was obtained with YMC-ODS pack column by using this mobile phase at pH 4.0. All additives were separated within 40 min. The RP-HPLC-UV method was validated in terms of LOD/LOQ, linearity, recovery and repeatability. This method was used for the determination of α-aspartame, acesulfame-K and benzoic acid in cola and instant powder drinks.     

Standardization of a multi-wavelength UV detector for liquid chromatography-based toxicological analysis.

The performance of a multi-wavelength UV detector for automated drug identification following liquid chromatographic separation was evaluated. The ability of selected wavelength ratios to distinguish two closely related drugs was considered at different concentrations. Calibration of the detector based on wavelength ratios was then utilized to standardize two different detectors and to evaluate instrument-to-instrument variation of a series of detectors. Reproducibility of the second-derivative zero intercept for these drug spectra was also evaluated. Standardization of detector performance by reference to these two parameters permitted the transfer of UV spectral libraries stored on one instrument to another without compromising the reliability of qualitative data.     

Simultaneous use of fluorescence,ultraviolet, and electrochemical detectors in high performance liquid chromatography-separation and identification of phenolic antioxidants and related compounds

Isocratic conditions are described for the separation, identification and quantitation of selected phenolic antioxidants and related compounds by high performance liquid chromatography on a reverse phase microparticulate ODS column. Mobile phases containing 0.05 M lithium perchlorate in aqueous methanol are used. Three detection systems: ultraviolet; fluorescence; and electrochemical detectors are connected in tandem. The use of these three detection systems in sequence as additional means for separation, identification, and quantitation is demonstrated. Other advantages of the system include speed of analyses and improved detection. The ? values and detector responses at 2 settings per detector for 12 compounds are reported.     

黄芪药材的指纹图谱研究方法的建立

用反相高效液相色谱－紫外－质谱联用技术对黄芪药材进行指纹图谱研究，为阐明不同产地药材的异同性，建立黄芪质量的国际统一标准奠定基础，黄芪的总提取物各类成分得到很好分离，紫外和质谱两种检测器可对不同特性化合物的检测进行互补，获得相对充分的指纹图谱信息。     

Experimental implementation of identification-based optimizing control of a simulated moving bed process

Experimental implementation of an optimizing controller based on identified model for the separation of nucleosides in a laboratory scale simulated moving bed (SMB) unit is reported in this study. The manipulative variables are the three external and one internal flow rates while the outputs are productivity, solvent consumption, and purities of extract and raffinate streams averaged over a switching period. The feedback information is the concentration profile of extract and raffinate measured online using two ultraviolet (UV) detectors. Experimental results show that the designed controller is able to operate the SMB units under optimal condition fulfilling the purity requirements. Besides, the controller demonstrated excellent performance in terms of rejecting disturbances that may occur during SMB operations.     

Electrochemical Detection of Retinoids Using Normal Phase HPLC

Abstract

Non-aqueous electrochemical (EC) detection of 13-cis-retinoic acid, all-trans retinoic, acitretin and vitamin A palmitate in non-aqueous solvents are reported.

Non-aqueous (EC) detection allows for normal-phase chromato-graphy of these compounds prior to detection. The normal phase system used a mobile phase of HEX/THF/AcOH for the separation of all four compounds. The stationary phase was either silica or PVA-sil. The lipophilic salts, t-butylammoniumtetrafluoroborate or t-butyl-ammoniumhexafluorophosphate necessary for EC detection were added post-column.

The limit of detection (LOD) for EC detection of these compounds is approximately 1 ng on column compared with an LOD by UV absorption of 2 ng on column.

The linear detection for these compounds with the EC detector was about two orders of magnitude.     

True and false reversal of the elution order of barbiturates on a bonded cellulose-based chiral stationary phase

A set of racemic N-alkylated barbiturates were investigated for their direct enantioselective HPLC separation on a new chiral stationary phase (CSP) containing cellulose tris(3,5-dimethylphenylcarabamate) immobilized onto silica gel (Chiralpak IB) and its coated version (Chiralcel OD). They were online detected by UV and optical rotation detectors to trace the elution order of their enantiomers. Surprisingly, examples of false and true reversal of the elution order of enantiomers of barbiturates were observed and reported.     

High performance liquid chromatographic strategy for the analysis of exopolysaccharides extracted from pathogenic bacteria.

A high performance liquid chromatographic (HPLC) method has been developed to permit the rapid comparison of acidic polysaccharides of diverse compositions and the sensitive determination of their constituents. It is based on two combined analyses of the polysaccharide hydrolysates--a separation of the released compounds by ion-moderated partition chromatography with UV detection at two wavelengths and a separation of the sugar dansylhydrazine derivatives by reversed phase chromatography. The former permits identification and quantitation of uronic and carboxylic acids, the latter permits more sensitive and specific determination of the neutral aldoses. Some bacterial exopolysaccharides have been used to demonstrate the validity of this HPLC procedure for the chemical characterization of uronic acid-containing polysaccharides. This method appears to be useful for studying capsular polysaccharides, which are involved in the evasion of phagocytosis by pathogenic bacteria.     

Microcolumn LC enantioseparation of chiral compounds using diol silica gel functionalized with vancomycin crystalline degradation products

Vancomycin crystalline degradation products (CDPs) have been introduced as one of the newest and most interesting derivatives of vancomycin for enantiomer separation of a wide variety of chiral compounds. In this attempt, a chiral stationary phase (CSP) has been prepared using diol silica gel based on vancomycin CDPs which led to a new chiral selector with new functionality of functional groups on a microcolumn LC. Different kinds of mobile phases were examined to realize the behavior of the chiral selector in separation of atropine, fluoxetine, amlodipine, mandelic acid, alanine, and phenylalanine which were separated successfully on this column. Good results were obtained by using a polar mobile phase containing water, methanol, and acid additives for separation of chiral acidic compounds and amino acid samples. Considerable results were obtained for analysis of basic compounds by using polar organic mobile phase (POP) containing methanol, acid and base additives. These results can be associated with the presence of the carboxylic acid groups present in new CSP by using a diol silica gel.     

Comprehensive two-dimensional HPLC to study the interaction of multiple components in Rheum palmatum L. with HSA by coupling a silica-bonded HSA column to a silica monolithic ODS column

A mode of comprehensive 2-D LC was developed by coupling a silica-bonded HSA column to a silica monolithic ODS column. This system combined the affinity property of the HSA column and the high-speed separation ability of the monolithic ODS column. The affinity chromatography with HSA-immobilized stationary phase was applied to study the interaction of multiple components in traditional Chinese medicines (TCMs) with HSA according to their affinity to protein in the first dimension. Then the unresolved components retained on the HSA column were further separated on the silica monolithic ODS column in the second dimension. By hyphenating the 2-D separation system to diode array detector and MS detectors, the UV and molecular weight information of the separated compounds can also be obtained. The developed separation system was applied to analysis of the extract of Rheum palmatum L., a number of low-abundant components can be separated on a single peak from the HSA column after normalization of peak heights. Six compounds were preliminarily identified according to their UV and MS spectra. It showed that this system was very useful for biological fingerprinting analysis of the components in TCMs and natural products.     

Gas chromatographic applications with the dielectric barrier discharge detector

With gas chromatography, there are many more choices for detectors when compared to other separation disciplines in analytical chemistry. The presence of sensitive and selective detectors aids in easing the separation requirements imposed on the capillary column. The current gas phase detectors, however, do not completely fulfill contemporary analytical needs. One example is in the area of ultratrace analysis of permanent gases for semiconductor industry. Another example is in the area of environmental/industrial hygiene monitoring for compounds such as 1,3-butadiene or vinyl chloride. The dielectric barrier discharge detector, a new highly sensitive detector with tuneable selectivity, has recently been innovated and commercialized. In this paper, the principle of operation of the detector, along with critical challenging industrial applications such as the analysis of oxygenated compounds, sulfur-containing compounds, and other compounds of industrial significance is presented.     

Rapid SPE-HPLC determination of the 16 European priority polycyclic aromatic hydrocarbons in olive oils

Polycyclic aromatic hydrocarbons (PAHs) are a large class of organic compounds. It has been established that the main source of exposure to these compounds for human beings is through food, particularly fats and oils, due to the lipophilic nature of these polycyclic compounds. The aim of this work was to optimise and validate a method involving SPE and HPLC for rapid determination of the 16 European Union (EU) priority PAHs (required by the Recommendation 2005/108/EC) in vegetable oils. Two spectrofluorometric detectors and a UV-Visible detector in series were used to identify and quantify the target compounds. Linearity, recoveries, LOD, and LOQ were found to be in agreement with the performance criteria for benzo[a]pyrene (BaP) analysis as required by the Commission Directive 2005/10/EC, and satisfactory for all the compounds of interest, except for cyclopenta[c,d]pyrene, which presented a very low signal in the UV. Optimised chromatographic conditions for the separation of 25 PAHs, comprising both EPA and EU priority PAHs plus benzo[e]pyrene and benzo[b]chrysene, have been also proposed.     

HPLC separation of flavonols, flavones and oxidized flavonols with UV-, DAD-, electrochemical and ESI-ion trap MS detection

The cation-induced or electrochemical oxidation of flavonols has been reported to yield 2-(hydroxybenzoyl)-2-hydroxy-3(2H)-benzofuranone. Two new gradient reversed phase HPLC methods are presented which allow the determination of those oxidized flavonols simultaneously with flavonols and flavones. UV and electrochemical detection are used because of their high sensitivity. Qualitative detection together with quantification of all compounds is achieved with photodiode-array detection. An electrospray ionization ion trap mass spectrometric method is presented for unique identification of the benzofuranones after HPLC separation.     

HPLC separation of flavonols, flavones and oxidized flavonols with UV-, DAD-, electrochemical and ESI-ion trap MS detection

The cation-induced or electrochemical oxidation of flavonols has been reported to yield 2-(hydroxybenzoyl)-2-hydroxy-3(2H)-benzofuranones. Two new gradient reversed phase HPLC methods are presented which allow the determination of those oxidized flavonols simultaneously with flavonols and flavones. UV and electrochemical detection are used because of their high sensitivity. Qualitative detection together with quantification of all compounds is achieved with photodiode-array detection. An electrospray ionization ion trap mass spectrometric method is presented for unique identification of the benzofuranones after HPLC separation.     

离子液体作高效液相色谱流动相添加剂分离测定芳香胺

建立了以离子液体作反相高效液相色谱流动相添加剂分离测定邻苯二胺、苯胺和对甲苯胺3种芳香胺的方法。实验以C18反相色谱柱为分离柱,采用紫外检测方法,考察了检测波长、甲醇含量、咪唑离子液体烷基链长度、离子液体溶液浓度等条件对分离和测定的影响,并与其它分离测定芳香胺的方法进行了比较。优化的色谱条件为:以甲醇/1-丁基-3-甲基咪唑四氟硼酸盐水溶液(3.0mmol/L,乙酸调节pH 3.5)=30/70(V/V)为流动相;检测波长254 nm;流速1.0mL/min;柱温30℃。在此条件下,3种芳香胺达到基线分离,在6.5 min之内分离完全;在1～40 mg/L范围内,线性回归方程的相关系数达到0.99以上;检出限为0.07～0.41 mg/L。将本方法应用于废水的测定,加标回收率在92.3%～96.7%之间,相对标准偏差小于3.5%。     

Size-exclusion chromatography with on-line ultraviolet, proton nuclear magnetic resonance and mass spectrometric detection and on-line collection for off-line Fourier transform infrared spectroscopy.

The coupling of HPLC with UV detection and on-line NMR spectroscopy and mass spectrometry combined with a dedicated interface for the collection of the chromatographic eluent for subsequent Fourier transform (FT) IR has been investigated using a number of polymer additives as model compounds. Size-exclusion chromatography was performed using deuterated chloroform as eluent with the separation monitored on-line by UV detection at 254 nm and on-flow 1H-NMR and MS. The effluent from the NMR probe was directed to a dedicated HPLC interface where it was deposited on a germanium plate for subsequent FT-IR. NMR and MS spectra were successfully obtained for 2,6-di-tert.-butyl-4-methylphenol, octadecyl-3-(3,5-di-tert.-butyl-4-hydroxyphenyl) propionate (Irganox 1076) and diisooctyl phthalate on-line and FT-IR spectra for all three compounds were obtained off-line. Practical problems encountered with this multiple hyphenation are described.     

Novel imidazolium stationary phase for high-performance liquid chromatography

A new imidazolium anion-exchange phase immobilized on silica is synthesized. HPLC separations of common inorganic anions (IO3-, Cl-, NO2-, Br-, NO3-, I-, SCN-) have been performed using a HPLC column (200 mm x 4.6 mm I.D.) packed with this stationary phase, with a phosphate buffer solution as the mobile phase and UV detection at 200 nm. The effects of pH and concentration of eluent on the separation of anions have been studied. Chromatographic parameters are calculated and the results show that the new stationary phase is of significant potential for the analysis of these anions. Successful separations of some ordinary organic anions have also been achieved with the said stationary phase. Meaningfully, organic and inorganic anions can be determined simultaneously and satisfactorily with several neutral compounds using the column. The separation of some organic compounds including hydroxybenzenes, bases and amines by this stationary phase with only water as the eluent has been investigated.     

手性固定相AD、AS和OD的拆分性能

手性固定相-高效液相色谱法在手性药物、手性农药等的分离分析中应用广泛。本文采用3种多糖衍生物的手性固定相(即EnantioPak AD、AS和OD)对20种手性化合物开展手性分离研究,进而探讨样品分子结构、多糖骨架和衍生基团对手性分离的影响。结果表明,除化合物13外,其余化合物在EnantioPak AD上均实现基线分离,分离度多在2.0以上,在正己烷-醇流动相中加入酸碱添加剂可改善和优化酸性或碱性化合物的分离效果;芳香醇(化合物13~16)随着侧链碳数增加在色谱柱上的保留减弱,其分离度呈现增加的趋势;对比8种化合物在3种手性固定相上的分离结果可知,EnantioPak AD表现出更优的分离性能。这为深入研究和了解多糖手性固定相、拓展其手性分离应用提供了参考。