Continuum Source Atomic Absorption Research Articles

Development and validation of a LC method for the separation and determination of the anticancer-active FeIII(4-methoxy-salophene) using the new second-generation monolith

LC method with the newly introduced second-generation monolithic silica RP-18e column has been developed for the separation of Fe(III) (salophene) and four methoxy-substituted Fe(III) (salophene) complexes. The method has been validated for the quantitation of Fe(III) (4-OMe-salophene), a highly active anticancer substance in vitro, bound to serum albumin. Our routinely used high-resolution continuum-source atomic absorption spectroscopy method based on the determination of the central iron atom was unsuitable in this case because serum originally contains significant amounts of iron as revealed by a blank sample of serum albumin. The developed LC method depends on detecting the whole complex rather than the bound iron. Two morphologically different first- and second-generation HPLC monolithic columns have been compared for this purpose. The newly introduced second-generation monolithic silica column Chromolith(®) HighResolution RP-18e column (100 × 4.6 mm, Merck) separated the mixture successful within 13 min. A mobile phase consisting of 25 mM phosphate buffer pH 3/methanol (60:40, v/v) was used at a flow rate of 1 mL/min. The dynamic linear working range of the calibration curve for Fe(III) (4-OMe-salophene) was found to be between 1 and 200 μg/mL. Detection and quantitation limits were 0.3 and 1 μg/mL, respectively.

Evaluation of solid sampling high-resolution continuum source graphite furnace atomic absorption spectrometry for direct determination of chromium in medicinal plants

A method for Cr determination in medicinal plants using direct solid sampling graphite furnace high-resolution continuum source atomic absorption spectrometry was developed. Modifiers were dispensable. Pyrolysis and atomization temperatures were 1500°C and 2400°C, respectively. Slopes of calibration curves (50–750pg Cr, R2>0.999) using aqueous and solid standards coincides in 96%, indicated feasibility of aqueous calibration for solid sampling of medicinal plants. Accuracy was checked by analysis of four plant certified reference materials. Results were in agreement at 95% confidence level with certified and non-certified values. Ten samples of medicinal plants were analyzed and Cr contents were in the 1.3–17.7μgg−1 Cr range. The highest RSD (n=5) was 15.4% for the sample Melissa officinalis containing 13.9±2.1μgg−1 Cr. The limit of detection was 3.3ngg−1 Cr.

A Ruthenocene–PNA Bioconjugate — Synthesis, Characterization, Cytotoxicity, and AAS-Detected Cellular Uptake

Labeling of peptide nucleic acids (PNA) with metallocene complexes is explored herein for the modulation of the analytical characteristics, as well as biological properties of PNA. The synthesis of the first ruthenocene-PNA conjugate with a dodecamer, mixed-sequence PNA is described, and its properties are compared to a ferrocene-labeled analogue as well as an acetylated, metal-free derivative. The synthetic characteristics, chemical stability, analytical and thermodynamic properties, and the interaction with cDNA were investigated. Furthermore, the cytotoxicity of the PNA conjugates is determined on HeLa, HepG2, and PT45 cell lines. Finally, the cellular uptake of the metal-containing PNAs was quantified by high-resolution continuum source atomic absorption spectrometry (HR-CS AAS). An unexpectedly high cellular uptake to final concentrations of 4.2 mM was observed upon incubation with 50 μM solutions of the ruthenocene-PNA conjugate. The ruthenocene label was shown to be an excellent label in all respects, which is also more stable than its ferrocene analogue. Because of its high stability, low toxicity, and the lack of a natural background of ruthenium, it is an ideal choice for bioanalytical purposes and possible medicinal and biological applications like, e.g., the development of gene-targeted drugs.

Quantitative determination of sulfur by high-resolution graphite furnace molecular absorption spectrometry

In this paper a new procedure for quantitative determination of sulfur, which is based on the molecular absorption of tin sulfide at a wavelength of 271.578nm, is described. For that purpose a commercially available high-resolution continuum source atomic absorption spectrometer with a transversely heated graphite tube furnace is used. The temperature program is optimized and the measurements are accomplished with a Zr-coated graphite tube without platform.A limit of detection of 0.16ngS absolute is achieved. Furthermore, the accuracy of the procedure is verified by the recovery rate of both an inorganic and an organic compound. Several water samples from rivers and ponds, three mineral waters with a concentration range from 3 to 370mg/LS, and one certified reference material are analyzed, too. The results are consistent with the measured values of ion chromatography and gravimetric analysis. The non-spectral interelement effect of Ni has displayed a signal loss. With a 1:1 relation of metal to analyte a loss of 11% of the absorption signal has appeared. This effect is very seldom in natural environment, so the procedure is robust and there are no spectral interelement effects. In addition, this procedure is faster and more easy-to-handle than other methods at this point of time, and resulting in the improved limit of detection.

Determination of NiII(3-OMe-salophene) in MCF7 and HT29 cancer cell lines using HR-CS-AAS and in serum albumin using LC with monolithic silica

A high-resolution continuum source atomic absorption spectrometric method was developed and validated for the determination of NiII(3-OMe-salophene) (a complex with anticancer activity in vitro) in MCF7 and HT29 cancer cell lines. The primarily most sensitive line 232.003nm was selected for analysis. Compared to the standard nickel, the absorbance values obtained for NiII(3-OMe-salophene) complex was at least 93% at the upper end of linear range of the calibration curve. The use of common matrix modifiers including magnesium nitrate, palladium nitrate, ammonium hydrogen phosphate, lanthanum chloride and calcium nitrate brought no significant improvement in the GF AAS measurement. The dynamic linear working range of the calibration curve was found to be between 2.16 and 12.0μgL−1 (ppb). This covers a concentration range of the complex from 0.036μM to 0.204μM. Typical coefficients of variation (n=6) ranged from 0.2% to 6.7% for Ni in NiII(3-OMe-salophene). Detection and quantitation limits were 0.65 and 2.16μgL−1 (ppb), respectively. The proposed method has been successfully applied to the analysis of NiII(3-OMe-salophene) complex in cell lines of breast cancer (MCF7) and colon cancer (HT29). However, being based on the determination of nickel in the salophene complex, the method was unsuitable for the quantitation of NiII(3-OMe-salophene) in serum albumin, which originally contains significant amount of nickel. For this purpose, a high performance liquid chromatographic method with a monolithic silica RP-18e column has been developed to quantitate the complex in serum albumin. The developed chromatographic method depends on detecting the whole complex in serum rather than the bounded nickel. A mobile phase consisting of 25mM phosphate buffer pH 3/methanol (30:70, v/v) was pumped at a flow rate of 1mLmin−1. The eluted complex was monitored at a wavelength of 250nm. The dynamic linear working range of the calibration curve for the developed LC method was found to be between 100 and 20,000μgL−1 (0.23–46.18μM). Detection and quantitation limits were 30 and 100μgL−1 (ppb), respectively.

Multi-element determination of Cu, Fe, Ni and Zn content in vegetable oils samples by high-resolution continuum source atomic absorption spectrometry and microemulsion sample preparation

The aim of this work was to evaluate the microemulsification as sample preparation procedure for determination of Cu, Fe, Ni and Zn in vegetable oils samples by High-Resolution Continuum Source Flame Atomic Absorption Spectrometry (HR-CS FAAS). Microemulsions were prepared by mixing samples with propan-1-ol and aqueous acid solution, which allowed the use of inorganic aqueous standards for the calibration. To a sample mass of 0.5g, 100μL of hydrochloric acid and propan-1-ol were added and the resulting mixture diluted to a final volume of 10mL. The sample was manually shaken resulting in a visually homogeneous system. The main lines were selected for all studied metals and the detection limits (3σ, n=10) were 0.12, 0.62, 0.58 and 0.12mgkg−1 for Cu, Fe, Ni and Zn, respectively. The relative standard deviation (RSD) ranged from 5% to 11 % in samples spiked with 0.25 and 1.5μgmL−1 of each metal, respectively. Recoveries varied from 89% to 102%. The proposed method was applied to the determination of Cu, Fe, Ni and Zn in soybean, olive and sunflower oils.

A fast and accurate method for the determination of total and soluble fluorine in toothpaste using high-resolution graphite furnace molecular absorption spectrometry and its comparison with established techniques

A fast and reliable method has been developed for the determination of total and soluble fluorine in toothpaste, important quality control parameters in dentifrices. The method is based on the molecular absorption of gallium mono-fluoride, GaF, using a commercially available high-resolution continuum source atomic absorption spectrometer. Transversely heated platform tubes with zirconium as permanent chemical modifier were used throughout. Before each sample injection, a palladium and zirconium modifier solution and a gallium reagent were deposited onto the graphite platform and thermally pretreated to transform them into their active forms. The samples were only diluted and introduced directly into the graphite tube together with additional gallium reagent. Under these conditions the fluoride was stable up to a pyrolysis temperature of 550 °C, and the optimum vaporization (molecule formation) temperature was 1550 °C. The GaF molecular absorption was measured at 211.248 nm, and the limits of detection and quantification were 5.2 pg and 17 pg, respectively, corresponding to a limit of quantification of about 30 μg g −1 (ppm) F in the original toothpaste. The proposed method was used for the determination of total and soluble fluorine content in toothpaste samples from different manufactures. The samples contained different ionic fluoride species and sodium monofluorophosphate (MFP) with covalently bonded fluorine. The results for total fluorine were compared with those obtained with a modified conventional headspace gas chromatographic procedure. Accuracy and precision of the two procedures were comparable, but the proposed procedure was much less labor-intensive, and about five times faster than the latter one.

High-Resolution Continuum Source Atomic and Molecular Absorption Spectrometry—A Review

The present article gives an overview over the historic development, the instrumentation, the special features, and the applications of high-resolution continuum source atomic and molecular absorption spectrometry. The most obvious advantage is the need for only one single lamp for all elements and wavelengths, compared to one lamp for each analyte, as is necessary in line source atomic absorption spectrometry. The visibility of the spectral environment of the analytical line at high resolution greatly helps in method development and to avoid spectral interferences. The advanced simultaneous background correction automatically eliminates lamp flicker noise and continuous background absorption. Fine-structured background absorption may be removed using a reference spectrum and a least-squares algorithm. Because any wavelength between 190 nm and 900 nm can be accessed, the same equipment can be used for molecular absorption spectrometry, making possible the determination of non-metals, such as phosphorus, sulfur, and the halogens. Although commercial equipment for this technique has only been available for a few years, the technique has been accepted extremely well, and the number of applications that have been published using this technique has already reached an impressive number.

High-resolution continuum source graphite furnace atomic absorption spectrometry: Is it as good as it sounds? A critical review

The recent arrival of high-resolution continuum source atomic absorption spectrometry represents a potential revolution in this field, in particular for direct analysis of complex samples. This review tries to illustrate the main advantages of this technology, paying particular attention to the development of direct solid sampling methods. Three solid sampling applications will be discussed, each one of them highlighting one of the main advantages of this technique. The review also intends to clarify some misconceptions on the true potential of the instrumentation that is currently commercially available, such as its performance for multielemental analysis.

Optimization of fluorine determination via the molecular absorption of gallium mono-fluoride in a graphite furnace using a high-resolution continuum source spectrometer

The determination of fluorine using the molecular absorption of gallium mono-fluoride (GaF) at the 211.248 nm rotational line has been optimized using a commercially available high-resolution continuum source atomic absorption spectrometer with a transversely heated graphite tube furnace. The electron excitation spectrum of GaF was generated by adding 500 μg Ga per injection into the graphite tube as molecule forming reagent. Best results were obtained by applying Zr as a permanent modifier and a mixed Pd/Zr modifier, thermally pretreated before each sample injection together with the Ga reagent at 1100 °C. The use of sodium acetate and Ru(III) nitrosyl nitrate as additional modifiers injected together with the sample further improved the performance. This way a maximum pyrolysis temperature of 550 °C could be used, and the optimum molecule forming temperature was 1550 °C. Several drinking water samples, a mineral water sample, and two certified reference materials were analyzed using the standard calibration technique; the absence of potential matrix effects was verified by measuring different dilutions and spiking with known fluorine mass. The results were in good agreement with the certified values and those measured by ion selective electrode; the recovery rate for the spiking experiments was between 97% and 106%. The results show that there was no matrix influence for that kind of samples containing relatively high concentrations of Ca, Mg and chloride, which are known to cause interference in GaF molecule absorption. The limit of detection and the characteristic mass of the method were 5.2 and 7.4 pg F, respectively, and were both about a factor of two better than recently published values.

Quantification of Cell Labeling with Micron-Sized Iron Oxide Particles Using Continuum Source Atomic Absorption Spectrometry

Detection of cells after transplantation is necessary for quality control in regenerative medicine. Labeling with micron-sized iron oxide particles enables noninvasive detection of single cells by magnetic resonance imaging. However, techniques for evaluation of the particle uptake are challenging. The aim of this study was to investigate continuum source atomic absorption spectrometry (CSAAS) for this purpose. Porcine liver cells were labeled with micron-sized iron oxide particles, and the iron concentration of the cell samples was investigated by a CSAAS spectrometer equipped with a Perkin-Elmer THGA graphite furnace. The weak iron line at 305.754 nm provides only about 1/600 sensitivity of the iron resonance line at 248.327 nm and was used for CSAAS measurements. Iron concentrations measured from labeled cells ranged from 5.8 +/- 0.3 to 25.8 +/- 0.9 pg Fe/cell, correlating to an uptake of 8.2 +/- 0.5 to 25.7 +/- 0.8 particles/cell. The results were verified by standardized morphometric evaluation. CSAAS enabled rapid quantification of particle load from small quantities of cells without extensive preparation steps. Thereby, CSAAS could be used for quality control in a clinical setting of cell transplantation.

Investigation of the feasibility to use Zeeman-effect background correction for the graphite furnace determination of phosphorus using high-resolution continuum source atomic absorption spectrometry as a diagnostic tool

The determination of phosphorus by graphite furnace atomic absorption spectrometry at the non-resonance line at 213.6 nm, and the capability of Zeeman-effect background correction (Z-BC) to deal with the fine-structured background absorption due to the PO molecule have been investigated in the presence of selected chemical modifiers. Two line source atomic absorption spectrometers, one with a longitudinally heated and the other with a transversely heated graphite tube atomizer have been used in this study, as well as two prototype high-resolution continuum source atomic absorption spectrometers, one of which had a longitudinally arranged magnet at the furnace. It has been found that Z-BC is capable correcting very well the background caused by the PO molecule, and also that of the NO molecule, which has been encountered when the Pd + Ca mixed modifier was used. Both spectra exhibited some Zeeman splitting, which, however, did not cause any artifacts or correction errors. The practical significance of this study is to confirm that accurate results can be obtained for the determination of phosphorus using Z-BC. The best sensitivity with a characteristic mass of m 0 = 11 ng P has been obtained with the pure Pd modifier, which also caused the lowest background level. The characteristic mass obtained with the mixed Pd+Ca modifier depended on the equipment used and was between m 0 = 9 ng P and m 0 = 15 ng P, and the background signal was higher. The major problem of Z-BC remains the relatively restricted linear working range.

Use of high resolution continuum source atomic absorption spectrometry as a detector for chemically generated noble and transition metal vapors

Vapor generation and atomization conditions in a heated quartz tube to detect Ag, Cd, Co, Cu, Ni and Zn using High Resolution Continuum Source AAS (HRCSAAS), were optimized. Vapors were generated after mixing acidified solutions containing 8-hydroxiquinoline (oxine) with sodium tetrahydroborate. Afterwards, they were swept to the heated quartz cell by an argon flow. Reaction loop size and temperature of the quartz cell were optimized for each element. A temperature of 960 °C was selected as a compromise value to detect most of the metals. Afterwards, a Plackett–Burmann design was proposed to select which parameters were most important. Type of acid and its concentration were the most statistical significant variables. Optimum conditions for sequential detection of Cd, Cu, Ni and Zn were: 1 mg L − 1 Co as catalyst, 250 mg L − 1 oxine, 0.6 M nitric acid, 1.75% (w/w) sodium tetrahydroborate (prepared in 0.4 (w/v)% NaOH), a reaction loop of 250 µL, and a 25 L h − 1 carrier Ar flow. Ag and Co were each detected in their own optimized conditions. Analytical performance of the system was evaluated in connection with a selected pixel number, and spectral correction was used to eliminate NO absorption bands interference in Zn detection. Detection limits were in the range of 1.5–18 μg L − 1 for Ag, Cu, Cd and Zn, whereas sensitivity was worst for Co (169 μg L − 1) and Ni (586 μg L − 1). Atomization in a quartz cell of Co and Ni volatile species, generated by an addition of sodium tetrahydroborate to an acidified solution of the analytes, was reported for the first time in this paper. Precision expressed as RSD(%) had values lower than 10% except for Ni.

Quality control in transplantation of micron-sized iron oxide particle labelled liver cells by using continuum source atomic absorption spectrometry

Quality control in transplantation of micron-sized iron oxide particle labelled liver cells by using continuum source atomic absorption spectrometry

Internal standardization and least-squares background correction in high-resolution continuum source flame atomic absorption spectrometry to eliminate interferences on determination of Pb in phosphoric acid

In this work the least-squares background correction (LSBC) and internal standardization (IS) techniques were combined to eliminate spectral and transport interferences in the determination of Pb in phosphoric acid by high-resolution continuum source atomic absorption spectrometry (HR-CS AAS). Blanks, samples and reference solutions [0.10–1.00 mg L − 1 Pb in 1% (v/v) HNO 3] were spiked with 4.00 mg L − 1 Co used as internal standard. For absorbance measurements at the wavelength integrated absorbance equivalent to 9 pixels, correlations between the ratio of absorbance of Pb to absorbance of Co and the analyte concentration were close to 0.9992. Relative standard deviations of measurements varied from 0.6 to 4% and 1 to 7% ( n = 12) without and with IS/LSBC techniques, respectively. Recoveries for Pb spikes were in the 96–104% and 76–180% range with and without IS/LSBC, respectively. The limit of detection improved with IS/LSBC techniques. Accuracy of the proposed method was checked for the determinations of Pb in commercial phosphoric acid samples and results obtained with IS were better than those without IS.

Investigation of artifacts caused by deuterium background correction in the determination of phosphorus by electrothermal atomization using high-resolution continuum source atomic absorption spectrometry

The artifacts created in the measurement of phosphorus at the 213.6-nm non-resonance line by electrothermal atomic absorption spectrometry using line source atomic absorption spectrometry (LS AAS) and deuterium lamp background correction (D 2 BC) have been investigated using high-resolution continuum source atomic absorption spectrometry (HR-CS AAS). The absorbance signals and the analytical curves obtained by LS AAS without and with D 2 BC, and with HR-CS AAS without and with automatic correction for continuous background absorption, and also with least-squares background correction for molecular absorption with rotational fine structure were compared. The molecular absorption due to the suboxide PO that exhibits pronounced fine structure could not be corrected by the D 2 BC system, causing significant overcorrection. Among the investigated chemical modifiers, NaF, La, Pd and Pd + Ca, the Pd modifier resulted in the best agreement of the results obtained with LS AAS and HR-CS AAS. However, a 15% to 100% higher sensitivity, expressed as slope of the analytical curve, was obtained for LS AAS compared to HR-CS AAS, depending on the modifier. Although no final proof could be found, the most likely explanation is that this artifact is caused by a yet unidentified phosphorus species that causes a spectrally continuous absorption, which is corrected without problems by HR-CS AAS, but which is not recognized and corrected by the D 2 BC system of LS AAS.

Feasibility of peak volume, side pixel and multiple peak registration in high-resolution continuum source atomic absorption spectrometry

In high-resolution continuum source atomic absorption spectrometry (HR-CS AAS) with a pixel detector, such as a charge-coupled device array for signal registration, the absorbance A not only depends on the absorption coefficient, the length of the absorbing layer and the number of absorbing atoms therein, but also on the spectral interval over which the signal is recorded, i.e., the spectral bandwidth per pixel and the number of pixels evaluated. Although the problem of different (absorption and emission) line widths is known for several decades already to exist in conventional line source AAS, it is usually disregarded. By choosing a certain number of pixels in HR-CS AAS a defined wavelength interval can be selected over which the absorbance is recorded. As the numerical values obtained this way are not directly comparable with the conventional absorbance, it is necessary to define new terms and symbols for this kind of signal evaluation. With a steady-state signal the individual pixel absorbance values can be added or integrated, resulting in the unit-free wavelength-selected absorbance (WSA, symbol A Σ), or the wavelength-integrated absorbance (WIA, symbol A λ ) having a wavelength unit, such as picometers (pm). Similarly, with transient signals one can add-up or integrate (over wavelength) the individual integrated (in time) absorbance values of the selected pixels to obtain the volume under the absorbance peak. This results in the peak volume selected absorbance (PVSA, symbol A Σ,int), and the peak volume integrated absorbance (PVIA, symbol A λ ,int), with the units second (s) and second times picometer (s pm), respectively. For comparison purposes, however, the integrated absorbance values, i.e., WIA or PVIA, should be used since they are instrument-independent. Several elements with narrow and broad absorption lines and with multiplets, respectively, have been chosen to investigate the feasibility of using different signal registration schemes. Especially the evaluation of WSA and PVSA with respect to sensitivity, linearity of the calibration curves and limits of detection was focused on.

Progress in direct solid sampling analysis using line source and high-resolution continuum source electrothermal atomic absorption spectrometry

The literature about direct solid sample analysis of the past 10-15 years using electrothermal atomic absorption spectrometry has been reviewed. It was found that in the vast majority of publications aqueous standards were reported as having been used for calibration after careful program optimization. This means the frequently expressed claim that certified reference materials with a matrix composition and analyte content close to that of the sample have to be used for calibration in solid sample analysis is not confirmed in the more recent literature. There are obviously limitations, and there are examples in the literature where even calibration with certified reference materials did not lead to accurate results. In these cases the problem is typically associated with spectral interferences that cannot be corrected properly by the systems available for conventional line source atomic absorption spectrometry, including Zeeman-effect background correction. Using high-resolution continuum source atomic absorption spectrometry, spectral interferences become visible owing to the display of the spectral environment at both sides of the analytical line at high resolution, which makes program optimization straightforward. Any spectrally continuous background absorption is eliminated automatically, and even rapidly changing background absorption does not cause any artifacts, as measurement and correction of background absorption are truly simultaneous. Any kind of fine-structured background can be eliminated by "subtracting" reference spectra using a least-squares algorithm. Aqueous standards are used for calibration in all published applications of high-resolution continuum source atomic absorption spectrometry to direct solid sample analysis.

Determination of aluminum in highly concentrated iron samples: Study of interferences using high-resolution continuum source atomic absorption spectrometry

High-resolution continuum source atomic absorption spectrometry (HR-CS AAS) has been used to investigate spectral and non-spectral interferences found with a conventional line source atomic absorption spectrometer in the determination of aluminum in pharmaceutical products containing elevated iron and sugar concentrations. A transversely heated graphite furnace was used as the atomizer in both spectrometers. The two most sensitive aluminum lines at 309.3 nm and 396.2 nm were investigated and it was found that an iron absorption line at 309.278 nm, in the vicinity of the aluminum line at 309.271 nm, could be responsible for some spectral interference. The simultaneous presence of iron and the organic components of the matrix were responsible for radiation scattering, causing high continuous and also structured background absorption at both wavelengths. The aluminum and iron absorption could not be separated in time, i.e., the iron interference could not be eliminated by optimizing the graphite furnace temperature program. However, an interference-free determination of aluminum was possible carrying out the measurements with HR-CS AAS at 396.152 nm after applying least squares background correction for the elimination of the structured background. Analytical working range and other figures of merit were determined and are presented for both wavelengths using peak volume registration (center pixel ± 1) and the center pixel only. Limits of detection and characteristic masses ranged from 1.1 to 2.5 pg and 13 to 43 pg, respectively. The method was used for the determination of the aluminum contamination in pharmaceutical formulations for iron deficiency treatment, which present iron concentrations from 10 to 50 g l − 1 . Spike recoveries from 89% to 105% show that the proposed method can be satisfactorily used for the quality control of these formulations.

Investigation of phosphorus atomization using high-resolution continuum source electrothermal atomic absorption spectrometry

The atomization of phosphorus in electrothermal atomic absorption spectrometry has been investigated using a high-resolution continuum source atomic absorption spectrometer and atomization from a graphite platform as well as from a tantalum boat inserted in a graphite tube. A two-step atomization mechanism is proposed for phosphorus, where the first step is a thermal dissociation, resulting in a fast atomization signal early in the atomization stage, and the second step is a slow release of phosphorus atoms from the graphite tube surface following the adsorption of molecular phosphorus at active sites of the graphite surface. Depending on experimental conditions only one of the mechanisms or both might be active. In the absence of a modifier and with atomization from a graphite or tantalum platform the second mechanism appears to be dominant, whereas in the presence of sodium fluoride as a modifier both mechanisms are observed. Intercalation of phosphorus into the graphite platform in the condensed phase has also been observed; this phosphorus, however, appears to be permanently trapped in the structure of the graphite and does not contribute to the absorption signal.