Splitless injection of up to hundreds of microliters of liquid samples in capillary GC: Part 1, concept

If a sample evaporates by flash vaporization in an empty injector insert, the solute material is well mixed with the expanding solvent vapors and the maximum injection volume is determined by the requirement that no vapors must leave the vaporizing chamber. If evaporation occurs from a surface (e.g., of Tenax packing), however, the solvent evaporates first. The site of evaporation is cooled to the solvent's boiling point, and the cool island formed in the hot injector retains solutes of at least intermediate boiling point (visually observed for perylene). Solvent vapors, free from such solutes, may now expand backwards from the injector insert and leave through the septum purge exit. When solvent evaporation is complete, the site of evaporation warms up, causing the high boiling solutes to evaporate and to be carried into the column by the carrier gas. The technique somewhat resembles PTV injection, but is performed using a classical vaporizing injector.     

Splitless injection of large volumes: Improved carrier gas regulation system

The classical vaporizing injector has been modified for splitless injection of large volumes: during solvent evaporation in the packed vaporizing chamber, the carrier gas supply is interrupted and the septum purge outlet fully opened. This prevents vapors penetrating the gas regulation system and keeps the pressure increase in the injector to a minimum.     

Concurrent solvent recondensation large sample volume splitless injection

Concurrent Solvent Recondensation Large Sample Volume (CRS‐LV) splitless injection overcomes the limitation of the maximum sample volume to 1–2 μL valid for classical splitless injection. It is based on control of the evaporation rate in the vaporizing chamber, utilization of a strong pressure increase in the injector resulting from solvent evaporation, and greatly accelerated transfer of the sample vapors from the injector into the inlet of an uncoated precolumn by recondensation of the solvent. The sample vapors are transferred into the column as rapidly as they are formed in the injector (concurrent transfer). 20–50 μL of liquid sample is injected with liquid band formation. The sample liquid is received by a small packing of deactivated glass wool positioned slightly above the column entrance at the bottom of the vaporizing chamber. Solvent evaporation strongly increases the pressure in the injector (auto pressure surge), provided the septum purge outlet is closed and the accessible volumes around the vaporizing chamber are small, driving the first vapors into the precolumn. Transfer continues to be fast because of recondensation of the solvent, obtained by keeping the oven temperature below the pressure‐corrected solvent boiling point. The uncoated precolumn must have sufficient capacity to retain most of the sample as a liquid. The experimental data show virtually complete absence of discrimination of volatile or high boiling components as well as high reproducibility.     

Loop-type interface for concurrent solvent evaporation in coupled HPLC-GC. Analysis of raspberry ketone in a raspberry sauce as an example

Presently, two coupling techniques are used for directly introducing HPLC fractions into capillary GC: The retention gap technique (involving negligible or partially concurrent solvent evaporation) and fully concurrent solvent evaporation. While the former involves use of a conventional on-column injector, it is now proposed that concurrent solvent evaporation technique be carried out using a switching valve with a built-in sample loop. The technique is based on the concept that the carrier gas pushes the HPLC eluent into the GC capillary against its own vapor pressure, generated by a column temperature slightly exceeding the solvent boiling point at the carrier gas inlet pressure. Further improvement of the technique is achieved by flow regulation of the carrier gas (accelerated solvent evaporation) and backflushing of the sample valve (improved solvent peak shape). Concurrent solvent evaporation using the loop-type interface is easy to handle, allows transfer of very large volumes of HPLC eluent (exceeding 1 ml), and renders solvent evaporation very efficient, allowing discharge of the vapors of 1 ml of solvent through the column within 5–10 min.     

Screening of four beta-blockers and codeine in urine by on-line coupled RPLC-GC with on-line derivatization

On-line coupled reversed phase liquid chromatography-capillary gas chromatography (RPLC-GC) was used in the separation of four derivatized beta-blockers and codeine in urine. Sample clean-up was accomplished in the LC part and compounds were separated in the GC part. After the LC column the aqueous phase was switched to organic solvent by on-line liquid-liquid extraction and the two phases were separated in a sandwich-type phase separator. The organic extract was then transferred to a loop-type LC-GC interface. Beta-blockers were derivatized on-line in the interface before GC analysis. Concurrent eluent evaporation was used during the introduction of the sample fraction, and excess of solvent vapors was removed via an early vapor exit. The sample pretreatment was minimal; the only manual pretreatment step was the filtration of the urine sample.     

Direct injection of large sample volumes into capillary columns with packed column injector

A direct injection method for large volume samples which avoids severe tailing of the solvent peak has been developed using a packed column injector (up to 100 μl) leading into an ordinary capillary column (0.3 mm i.d.). Modifications are made to the cooler zones of the inlet port and on the carrier gas flow control system. This injection technique is based on the effective use of phase soaking and cold trapping using a retention gap. The large volume of solvent vapor is rapidly purged out of the injector with a higher flow of carrier gas while the solutes trapped at the head of the column are subsequently analyzed with another optimum flow rate. The proposed carrier gas flow regulation system is also compared with conventional split/splitless injection methods.     

PTV splitless injection of sample volumes up to 20 μl

PTV splitless injection cannot compete with on-column injection as far as simplicity, reliability, and accuracy of quantitative analysis is concerned. However, PTV splitless injection is attractive for trace analysis of samples containing high concentrations of involatile sample by-products. Maximum injection volumes are limited by the amount of liquid that can be retained within the PTV injector chamber and are around 20–30 μl injected at once. Solvent evaporation must be carried out in such a way that injector overflow is avoided.     

Splitless injection–development and state of the art. Including a comparison of matrix (“dirt”) effects in conventional and PTV splitless injection

Kurt Grob introduced splitless injection in 1969. He elaborated most of the working guidelines including the techniques required for reconcentrating the broad intial bands, i.e. the solvent effects and cold trapping. He also designed a vaporizing injector suited for splitless injection. Nevertheless, splitless injection is still often carried out using inappropriate conditions, and many of today's vaporizing injectors are not suited for splitless injection. No autosampler is available that introduces the sample at the appropriate position. Conventional splitless injection is compared to PTV splitless injection for the range of samples that cannot be handled by the anyway superior oncolumn injection, i.e. sample with high loads of involatile byproducts. There is a clear preference for PTV splitless injection as matrix effects observed in conventional splitless injection were found to be substantially reduced or even eliminated.     

Development of septum-free injector for gas chromatography and its application to the samples with a high boiling point

A novel apparatus with a simple structure has been developed for introducing samples into the vaporizing chamber of a gas chromatograph. It requires no septum due to the gas sealing structure over the carrier gas supply line. The septum-free injector made it possible to use injection port temperatures as high as 450 degrees C. Repetitive injection of samples with boiling points below 300 degrees C resulted in peak areas with relative standard deviations between 1.25 and 3.28% (n=5) and good linearity (r(2)>0.9942) for the calibration curve. In the analysis of polycyclic aromatic hydrocarbons and a base oil, the peak areas of components with high boiling points increased as the injection port temperature was increased to 450 degrees C.     

AT-column, a novel concentrating technique for large-volume injections in gas chromatography

Nowadays, large-volume injection is widely used for the GC determination of trace analytes, specifically to improve detectability. The most popular injectors for large-volume injections are the programmable temperature vaporisation (PTV) injector and the cold on-column (COC) injector, where each device has its own advantages and limitations. The novel AT-column concentrating technique combines features of two other injection techniques, loop-type large-volume and vapour overflow. AT-column injection is based on solvent evaporation in an empty liner with solvent vapour discharge via the split line. Little or no optimisation is required. The only relevant parameter is the injection temperature which can easily be calculated using the equation of Antoine. As an application, AT-column injection is combined with GC-MS for the trace-level determination of labile analytes and with GC-flame ionisation detection for the analysis of high molecular weight polymer additives. In summary, AT-column is an injection technique that combines the inertness of the COC, and the flexibility and robustness of the PTV large-volume technique.     

Vapor pressure measurements in carrier gas containing ligand vapor using the transpiration technique

The vapor pressures of 1,1,1-trifluoro-2,4-pentanedionates were measured in 0.1–10 mm Hg using the transpiration technique with helium or helium containing the ligand vapor as carrier gas. The injection chamber of a gas chromatograph equipped with a ligand vapor generator was used. The vapor pressure of bis(1,1,1-trifluoro-2,4-pentanedionato)nickel(II) was measured for the first time, stabilized in ligand vapor atmosphere. The vapor pressures of standard samples (naphthalene and benzoic acid) agreed well with previously reported values.     

Large volume, low discrimination GC injection into a modified splitless injector

Summary A standard GC split/splitless injector was sealed with an airlock. The carrier gas and the sample were introduced through this valve. Such a configuration efficiently prevents an injector overflow. Injections up to 50 μL were made. An almost quantitative analyte and solvent transfer was observed, with only a minimal discrimination, of even volatile analytes. The use of an early vapor exit permitted a high initial liner flow and therefore a fast sample transfer.     

Injection by Programmed Temperature Vaporization Injection (PTV) of Gaseous Samples for Gas Chromatography – Atomic Emission Spectrometry (GC-AED)

Programmed temperature vaporization injection (PTV) coupled to gas chromatography and atomic emission detector (AED) has been studied for large volume injection of gaseous samples. As examples of the effectiveness of the technique, the results of the analysis of a series of headspace samples of foods such as garlic and onion, and of landfill gases are presented. The volumes of gaseous samples reconcentrated varied from a few milliliters up to liters depending on analyte dilution, through focusing onto a sorbent trap, then rapid liberation into the GC-AED system by programmed thermal desorption. Despite the high carrier gas flow rates associated with direct PTV-GC, AED performance and sensitivity were unaffected. The detailed elemental information obtained from the PTV-GC-AED analyses was confirmed using a PTV coupled to a gas chromatograph with ion trap detector mass spectrometer as detector (PTV-GC-ITD/MS).     

Determination of solvent recondensation or evaporation in the GC-precolumn by measurement of temperature changes on the column wall

Temperature measurements on the column outer well were used for detecting recondensation or evaporation of solvent inside the precolumn during injection or on-line transfer of large solvent volumes. This facilitates the choice of the most critical parameter of these techniques, i.e. oven temperature. When using the vaporizer/precolumn solvent split/gas discharge system, the dew point of the solvent is determined, either to just prevent solvent recondensation or to limit it to the capacity of the precolumn to retain liquid. In concurrent eluent evaporation through the loop type LC-GC interface, temperature measurement enables the determination of whether or not the flooded zone exceeds a given limit. Fanally, when solvent trapping is used (on-column injector/partially concurrent solvent evaporation evaporation or vaporizer/partial recondensation), temperature measurement near the front end of the flooded zone is used as a signal for accurate closure of the vapor exit shortly before the end of solvent evaporation.     

Injector-internal thermal desorption from edible oils performed by programmed temperature vaporizing (PTV) injection

Injector-internal thermal desorption is a promising technique for the analysis of a wide range of food components (e.g., flavors) or food contaminants (e.g., solvent residues, pesticides, or migrants from packaging materials) in edible oils and fats or fatty food extracts. Separation from the fatty matrix occurs during injection. Using programmed temperature vaporizing (PTV) injection, the oily sample or sample extract was deposited on a small pack of glass wool from which the components of interest were evaporated and transferred into the column in splitless mode, leaving behind the bulk of the matrix. Towards the end of the analysis, the oil was removed by heating out the injector and backflushing the precolumn. The optimization dealt with the gas supply configuration enabling backflush, the injector temperature program (sample deposition, desorption, and heating out), separation of the sample liquid from the syringe needle and positioning it on a support, deactivation of the support surface, holding the plug of fused silica wool by a steel wire, and the analytical sequence maintaining adsorptivity at the desorption site low. It was performed for a mixture of poly(vinyl chloride) (PVC) plasticizers in oil or fatty food. Using MS in SIM, the detection limit was below 0.1 mg/kg for plasticizers forming single peaks and 1 mg/kg for mixtures like diisodecyl phthalate. For plasticizers, RSDs of the concentrations were below 10%; for the slip agents, oleamide and erucamide, it was 12%. The method of incorporating PTV injection was used for about one year for determining the migration from the gaskets of lids for glass jars into oily foods.     

Automated sample cleanup using on-line coupling of size exclusion chromatography to high resolution gas chromatography

A fully automated on-line sample cleanup system based on the coupling of size exclusion chromatography to high resolution gas chromatography is described. The transfer technique employed is based on fully concurrent solvent evaporation using a loop-type interface, early vapor exit and co-solvent trapping. Optimization of the LC-GC transfer was done visually via an all-glass oven door. To circumvent the problem of mixing within the injection loop, an adaptation was made to the standard loop-type interface. The determination of a series of additives in a polymer matrix is presented as one example of the vast range of applications opened up by this technique.     

The influence of the syringe needle on the precision and accuracy of vaporizing GC injections

The discrimination of the high boiling components of a C9 to C44 alkane test mixture in a vaporizing injector (analysed with stream splitting on a capillary column) is mainly due to a selective elution of the sample out of the syringe needle. Different methods of handling the syringe needle are tested. The discrimination is quantitated and correlated with the material left in the needle. The poorest method of injection was found to retain the sample in the needle when the latter is introduced into the vaporizer. The sample should be pulled back into the barrel of the syringe and the needle allowed to warm up in the injector before the sample is transferred into it. The solvent flush technique may be preferred for components sensitive to the hot metal surface of the needle.     

Programmable Temperature Vaporizer (PTV) Applied to the Triglyceride Analysis of Milk Fat

The PTV (Programmable Temperature Vaporizer) is a split/splitless injector which allows the sample to be introduced at a relatively low temperature, thus affording accurate and reproducible sampling. After injection the PTV is rapidly heated to transfer the vaporized components into the capillary column. This type of injector has proved to be an efficient tool for the evaluation of fatty acids, essential oils, and pesticides in food analysis. In this work the suitability of PTV for the analysis of milk fat purity by the Official EU method was evaluated. This method is based on the gas chromatographic determination of triglycerides only according to their total number of carbon atoms followed by the application of formulae deriving from multiple linear regressions. The analysis is currently carried out with a packed column or a short capillary column and an on-column injection system. Several samples of pure milk fat and mixtures of milk fat with foreign fat were analyzed with the same capillary column and by using both PTV and on-column injection systems. The results show that the gas chromatographic profile obtained by PTV is comparable with that obtained by the on-column injector, while repeatability and reproducibility data meet the requirements indicated in the Official Method. Therefore, this study demonstrates that it is possible to use the PTV instead of the on-column injector to determine the purity of milk fat with this method.     

Multipurpose cold injector for high resolution gas chromatography

The multipurpose cold injector described in this paper represents a solution for an universal sampling system for high resolution gas chromatography. The system is modular and is built around the Carlo Erba cold on-column injector provided with secondary cooling. An auxiliary module, easily fixable on the bottom of the on-column injector, creates a temperature programmable vaporizing chamber making the system also suitable for cold split-splitless injections or solvent venting prior to the sample transfer into the capillary. The system can be operated manually or in a fully automatic mode using the auto-sampler for cold on-column injections. The experimental data illustrate its benefits and limitations.