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Coccolithophores are an important group of calcifying marine phytoplankton. Although coccolithophores are not silicified, some species exhibit a requirement for Si in the calcification process. These species also possess a novel protein (SITL) that resembles the SIT family of Si transporters found in diatoms. However, the nature of Si transport in coccolithophores is not yet known, making it difficult to determine the wider role of Si in coccolithophore biology. Here, we show that coccolithophore SITLs act as Na + -coupled Si transporters when expressed in heterologous systems and exhibit similar characteristics to diatom SITs. We find that CbSITL from Coccolithus braarudii is transcriptionally regulated by Si availability and is expressed in environmental coccolithophore populations. However, the Si requirement of C. braarudii and other coccolithophores is very low, with transport rates of exogenous Si below the level of detection in sensitive assays of Si transport. As coccoliths contain only low levels of Si, we propose that Si acts to support the calcification process, rather than forming a structural component of the coccolith itself. Si is therefore acting as a micronutrient in coccolithophores and natural populations are only likely to experience Si limitation in circumstances where dissolved silicon (DSi) is depleted to extreme levels., (© 2022 The Authors. Environmental Microbiology published by Applied Microbiology International and John Wiley & Sons Ltd.)

In the California Current Ecosystem, upwelled water low in dissolved iron (Fe) can limit phytoplankton growth, altering the elemental stoichiometry of the particulate matter and dissolved macronutrients. Iron-limited diatoms can increase biogenic silica (bSi) content >2-fold relative to that of particulate organic carbon (C) and nitrogen (N), which has implications for carbon export efficiency given the ballasted nature of the silica-based diatom cell wall. Understanding the molecular and physiological drivers of this altered cellular stoichiometry would foster a predictive understanding of how low Fe affects diatom carbon export. In an artificial upwelling experiment, water from 96 m depth was incubated shipboard and left untreated or amended with dissolved Fe or the Fe-binding siderophore desferrioxamine-B (+DFB) to induce Fe-limitation. After 120 h, diatoms dominated the communities in all treatments and displayed hallmark signatures of Fe-limitation in the +DFB treatment, including elevated particulate Si:C and Si:N ratios. Single-cell, taxon-resolved measurements revealed no increase in bSi content during Fe-limitation despite higher transcript abundance of silicon transporters and silicanin-1. Based on these findings we posit that the observed increase in bSi relative to C and N was primarily due to reductions in C fixation and N assimilation, driven by lower transcript expression of key Fe-dependent genes., (© 2022. The Author(s).)

Measuring diet choice in grazing animals is challenging, complicating the assessment of feed efficiency in pasture-based systems. Furthermore, animals may modify their intake of a forage species depending on its nutritive value and on their own physiological status. Various fecal markers have been used to estimate feed intake in grazing animals. However, plant-wax markers such as n-alkanes (ALK) and long-chain alcohols may provide reliable estimates of both dietary choices and intakes. Still, their use in beef cattle has been relatively limited. The present study was designed to test the reliability of the ALK technique to estimate diet choices in beef heifers. Twenty-two Angus-cross heifers were evaluated at both post-weaning and yearling age. At each age, they were offered both red clover and fescue hay as cubes. Following 3-week acclimation periods, daily intake of each forage species was assessed daily for 10 days. During the final 5 days, fecal grab samples were collected twice daily. The ALK fecal concentrations were adjusted using recovery fractions compiled from literature. Diet composition was estimated using two statistical methods. Post-weaning, dietary choices were reliably estimated, with low residual error, regardless of the statistical approach adopted. The regression of observed on estimated red clover proportion ranged from 0.85±0.08 to 1.01±0.09 for fecal samples collected in the p.m. and for daily proportions once averaged, respectively. However, at yearling age, the estimates were less reliable. There was a tendency to overestimate the red clover proportion in diets of heifers preferring fescue, and vice versa. This was due to greater variability in ALK fecal concentrations in the yearling heifers. Overall, the ALK technique provided a reliable tool for estimating diet choice in animals fed a simple forage diet. Although further refinements in the application of this methodology are needed, plant-wax markers provide opportunities for evaluating diet composition in grazing systems in cattle.

A mix of adaptive strategies enable diatoms to sustain rapid growth in dynamic ocean regions, making diatoms one of the most productive primary producers in the world. We illustrate one such strategy off coastal California that facilitates continued, high, cell division rates despite silicic acid stress. Using a fluorescent dye to measure single-cell diatom silica production rates, silicification (silica per unit area) and growth rates we show diatoms decrease silicification and maintain growth rate when silicon concentration limits silica production rates. While this physiological response to silicon stress was similar across taxa, in situ silicic acid concentration limited silica production rates by varying degrees for taxa within the same community. Despite this variability among taxa, silicon stress did not alter the contribution of specific taxa to total community silica production or to community composition. Maintenance of division rate at the expense of frustule thickness decreases cell density which could affect regional biogeochemical cycles. The reduction in frustule silicification also creates an ecological tradeoff: thinner frustules increase susceptibility to predation but reducing Si quotas maximizes cell abundance for a given pulse of silicic acid, thereby favouring a larger eventual population size which facilitates diatom persistence in habitats with pulsed resource supplies., (© 2018 Society for Applied Microbiology and John Wiley & Sons Ltd.)

The metabolic activity and growth of phytoplankton taxa drives their ecological function and contribution to biogeochemical processes. We present the first quantitative, taxon-resolved silica production rates, growth rates, and silica content estimates for co-occurring diatoms along two cross-shelf transects off the California coast using the fluorescent tracer PDMPO (2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)methoxy)phenyl)oxazole), and confocal microscopy. Taxon contribution to total diatom community silica production was predominantly a function of the surface area of new frustule that each taxon created as opposed to cell abundance or frustule thickness. The influential role of surface area made large diatoms disproportionately important to community silica production over short time scales (<1 d). In some cases, large taxa that comprised only ~15% of numerical cell abundance accounted for over 50% of total community silica production. Over longer time scales relevant to bloom dynamics, the importance of surface area declines and growth rate becomes the dominant influence on contribution to production. The relative importance of surface area and growth rate in relation to silica production was modeled as the time needed for a smaller, faster-growing taxon to create more surface area than a larger, slower-growing taxon. Differences in growth rate between the taxa effected the model outcome more than differences in surface area. Shifts in relative silica production among taxa are time restricted by finite resources that limit the duration of a bloom. These patterns offer clues as to how taxa respond to their environment and the consequences for both species succession and the potential diatom contribution to elemental cycling.

The vacuum ultraviolet detector (VUV) is a new non-destructive mass sensitive detector for gas chromatography that continuously and rapidly collects full wavelength range absorption between 120 and 240 nm. In addition to conventional methods of quantification (internal and external standard), gas chromatography - vacuum ultraviolet spectroscopy has the potential for pseudo-absolute quantification of analytes based on pre-recorded cross sections (well-defined absorptivity across the 120-240 nm wavelength range recorded by the detector) without the need for traditional calibration. The pseudo-absolute method was used in this research to experimentally evaluate the sources of sample loss and gain associated with sample introduction into a typical gas chromatograph. Standard samples of benzene and natural gas were used to assess precision and accuracy for the analysis of liquid and gaseous samples, respectively, based on the amount of analyte loaded on-column. Results indicate that injection volume, split ratio, and sampling times for splitless analysis can all contribute to inaccurate, yet precise sample introduction. For instance, an autosampler can very reproducibly inject a designated volume, but there are significant systematic errors (here, a consistently larger volume than that designated) in the actual volume introduced. The pseudo-absolute quantification capability of the vacuum ultraviolet detector provides a new means for carrying out system performance checks and potentially for solving challenging quantitative analytical problems. For practical purposes, an internal standardized approach to normalize systematic errors can be used to perform quantitative analysis with the pseudo-absolute method., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Ocean acidification is a threat to many marine organisms, especially those that use calcium carbonate to form their shells and skeletons. The ability to accurately measure the carbonate system is the first step in characterizing the drivers behind this threat. Due to logistical realities, regular carbonate system sampling is not possible in many nearshore ocean habitats, particularly in remote, difficult-to-access locations. The ability to autonomously measure the carbonate system in situ relieves many of the logistical challenges; however, it is not always possible to measure the two required carbonate parameters autonomously. Observed relationships between sea surface salinity and total alkalinity can frequently provide a second carbonate parameter thus allowing for the calculation of the entire carbonate system. Here, we assessed the rigor of estimating total alkalinity from salinity at a depth <15 m by routinely sampling water from a pier in southern California for several carbonate system parameters. Carbonate system parameters based on measured values were compared with those based on estimated TA values. Total alkalinity was not predictable from salinity or from a combination of salinity and temperature at this site. However, dissolved inorganic carbon and the calcium carbonate saturation state of these nearshore surface waters could both be estimated within on average 5% of measured values using measured pH and salinity-derived or regionally averaged total alkalinity. Thus we find that the autonomous measurement of pH and salinity can be used to monitor trends in coastal changes in DIC and saturation state and be a useful method for high-frequency, long-term monitoring of ocean acidification., Competing Interests: The authors have declared that no competing interests exist.

4-Methylcyclohexanemethanol (4-MCHM) is a saturated higher alicyclic primary alcohol that is used in the froth flotation process for cleaning coal. In early 2014, a large spill of crude chemical (containing primarily 4-MCHM) to the Elk River near Charleston, WV contaminated the local water supply. Carbon filters at the affected water treatment facility quickly became saturated, and the contaminated water was distributed to nearby homes and businesses. Sorption of 4-MCHM to granular activated carbon (GAC) was studied in the laboratory using head space (HS) analysis via gas chromatography with a flame ionization detector (GC-FID). Sorption to raw coal was also investigated, since this material may be of interest as a sorbent in the case of an on-site spill. As expected, sorption to both materials increased with decreased particle size and with increased exposure time; although exposure time proved to be much more important in the case of GAC than for coal. Under similar conditions, GAC sorbed more 4-MCHM than raw coal (e.g., 84.9 vs. 63.1 mg/g, respectively, for 20 × 30 mesh particles exposed to 860 mg/L 4-MCHM solution for 24 h). Desorption from both materials was additionally evaluated. Interestingly, desorption of 4-MCHM on a mass per mass basis was also higher for GAC than for raw coal. Overall, results indicated that GAC readily sorbs 4-MCHM but can also readily release a portion of the chemical, whereas coal sorbs somewhat less 4-MCHM but holds it tightly., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Diatoms require silicic acid to construct ornately detailed cell walls called frustules. The growth and geographic distribution of diatoms is often controlled by the availability of silicic acid. Analytical methods exist to assess diatom community biogenic silica (bSiO 2 ) production, but partitioning production among taxa has been largely qualitative. We present a method for the quantitative analysis of taxa-specific silica production through labeling diatoms with the fluorescent dye PDMPO [2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)methoxy)phenyl)oxazole]. To make PDMPO a quantitative tool: diatom frustules were solubilized to assess the total diatom community incorporation by quantitation of PDMPO fluorescence using a fluorometer, and laser confocal microscopy was used to quantify the fluorescence of PDMPO in single diatom cells. We created a fluorescence standard to intercalibrate the raw fluorescence signals of the fluorometer and microscope and to determine the fluorescence per mole of PDMPO. PDMPO incorporation was converted to silica production using diatom bSiO 2 :PDMPO incorporation ratios which varied systematically with silicic acid concentration. Above 3 μM Si(OH) 4 , bSiO 2 :PDMPO was constant and PDMPO incorporation was converted to silica production using a mole ratio of 2,916 as determined from cultures. Below 3 μM, the ratio was a linear function of [Si(OH) 4 ] (bSiO 2 :PDMPO = 912.6 × [Si(OH) 4 ]), as determined using data from two oceanographic cruises. Field evaluation of the method showed that total community PDMPO incorporation generally agreed to within 30% of radioisotope-determined silica production. This PDMPO method has the potential to be a powerful tool for understanding physiology, silicification and resource competition among diatom taxa.

Sulfur hexafluoride (SF6) is widely used as a tracer gas because of its detectability at low concentrations. This attribute of SF6 allows the quantification of both small-scale flows, such as leakage, and large-scale flows, such as atmospheric currents. SF6's high detection sensitivity also facilitates greater usage efficiency and lower operating cost for tracer deployments by reducing quantity requirements. The detectability of SF6 is produced by its high molecular electronegativity. This property provides a high potential for negative ion formation through electron capture thus naturally translating to selective detection using negative ion chemical ionization mass spectrometry (NCI-MS). This paper investigates the potential of using gas chromatography (GC) with NCI-MS for the detection of SF6. The experimental parameters for an ultra-trace SF6 detection method utilizing minimal customizations of the analytical instrument are detailed. A method for the detection of parts per trillion (ppt) level concentrations of SF6 for the purpose of underground ventilation tracer gas analysis was successfully developed in this study. The method utilized a Shimadzu gas chromatography with negative ion chemical ionization mass spectrometry system equipped with an Agilent J&W HP-porous layer open tubular column coated with an alumina oxide (Al2O3) S column. The method detection limit (MDL) analysis as defined by the Environmental Protection Agency of the tracer data showed the method MDL to be 5.2 ppt., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Analytical performance characteristics of a new vacuum ultraviolet (VUV) detector for gas chromatography (GC) are reported. GC-VUV was applied to hydrocarbons, fixed gases, polyaromatic hydrocarbons, fatty acids, pesticides, drugs, and estrogens. Applications were chosen to feature the sensitivity and universal detection capabilities of the VUV detector, especially for cases where mass spectrometry performance has been limited. Virtually all chemical species absorb and have unique gas phase absorption cross sections in the approximately 120-240 nm wavelength range monitored. Spectra are presented, along with the ability to use software for deconvolution of overlapping signals. Some comparisons with experimental synchrotron data and computed theoretical spectra show good agreement, although more work is needed on appropriate computational methods to match the simultaneous broadband electronic and vibronic excitation initiated by the deuterium lamp. Quantitative analysis is governed by Beer-Lambert Law relationships. Mass on-column detection limits reported for representatives of different classes of analytes ranged from 15 (benzene) to 246 pg (water). Linear range measured at peak absorption for benzene was 3-4 orders of magnitude. Importantly, where absorption cross sections are known for analytes, the VUV detector is capable of absolute determination (without calibration) of the number of molecules present in the flow cell in the absence of chemical interferences. This study sets the stage for application of GC-VUV technology across a wide breadth of research areas.

Fast high-performance liquid chromatography is becoming routine in laboratories that require high throughput or for combinatorial libraries. Reduced analysis time is commonly achieved by using shorter columns and higher flow rates. Shorter columns require smaller particles in order to maintain efficiency. However, smaller particles increase backpressure, which limits both column length and higher flow rates for typical LC pumps. This disadvantage has been addressed by the emergence of monolithic liquid chromatographic columns (1). Unlike particle-base columns, monolithic columns consist of a continuous rod-shaped porous network with a bimodal pore distribution. In this study, a commercially available 50- x 4.6-mm silica-based octadecyl silane monolithic column (Chromolith SpeedROD RP18e, EM Science, Gibbstown, NJ) was used to separate a seven-component test mixture with a wide range of polarity. The primary goals of this investigation were: (a) to study the chemistry (selectivity) of the new silica-based monolithic columns and (b) to study their run-to-run and column-to-column performance (retention times and peak areas). The selectivity (alpha factor) is a ratio of partition coefficients and, if comparable for a variety of solutes, would mean that methods could be readily transferred between particulate and monolithic columns.

This work serves as a follow-up to Part I of experiments designed to determine the underlying principles in the formation of pseudomolecular, or adduct, ions during electrospray ionization. Aromatic acids were studied by flow injection analysis in the negative ionization mode of electrospray ionization mass spectrometry. Part I dealt with common acidic anti-inflammatory pharmaceuticals. such as ibuprofen and related analogues. Part II deals with functionally less complex molecules, namely benzoic acid (BA) and substituted benzoic acids. Halide-substituted molecules are investigated to deduce the effect of electron-withdrawing substituents (bromo-, chloro-, and fluoro-) and ring position (ortho-, meta- and para-) on the response of a traditional deprotonated molecular ion ([M-H]-) and a sodium-bridged dimer ion ([2M-2 H+Na]-). Amino-substituted benzoic acids are also analyzed in order to study the effect of an additional ionizable group on the molecule, and para-tert.-butyl-BA is analyzed to study the effect of increased hydrophobicity, as they relate to the formation of pseudomolecular ions. This study shows that solution character [octanol-water partition coefficient (or log P) and pKa] of the model compounds controls the relative efficiency of formation of [M-H]- and [2M-2H+Na]- ions. However the relative gas phase character (gas phase basicity and proton affinity) also has a significant effect on the formation of the sodium-bridged dimer ion. For the halide-substituted species, placement of the electron-withdrawing atom at the meta-position gives the greatest enhancement in sensitivity. Observations also show that as the structural complexity of the model compound increases, predictions relating analyte acidity to sodium-bridged dimer ion formation give way to a stronger dependence between log P values and ionization efficiency. Supporting this hypothesis is the nearly ten-fold enhancement in signal for tert.-butyl BA relative to BA. due to the greater hydrophobicity, and consequently, increased surface activity in an electrosprayed droplet of the analyte molecule.

Direct gas chromatographic (GC) analysis of thermally labile N-carbamates is studied by using fast GC with cold on-column injection. With the greatly reduced injection temperature, short column lengths, high flow rate, and fast temperature-programming rate, the exposure of carbamates to high temperature is reduced and degradation can be avoided. Nine N-carbamates in EPA Method 531 are eluted without thermal decomposition. The relative standard deviation percentage for peak areas average 1.9% for all of the carbamates analyzed. A conventional GC instrument is employed to simplify the experiments. GC-mass spectrometry is used to monitor the decomposition peaks.

The retention of ionogenic bases in liquid chromatography is strongly dependent upon the pH of the mobile phase. Chromatographic behavior of a series of substituted aniline and pyridine basic compounds has been studied on C18 bonded silica using acetonitrile-water (10:90) as the eluent with different pHs and at various concentrations of the acidic modifier counter anions. The effect of different acidic modifiers on solute retention over a pH range from 1.3 to 8.6 was studied. Ionized basic compounds showed increased retention with a decrease of the mobile phase pH. This increase in retention was attributed to the interaction with counter anions of the acidic modifiers. The increase in retention is dependent on the nature of the counter anion and its concentration in the mobile phase. It was shown that altering the concentration of counter anion of the acidic modifier allows the optimization of the selectivity between basic compounds as well as for neutral and acidic compounds.

The effect of alkyl chain length on adsorbent pore volume and void volume of HPLC columns is described. The results provide evidence that alkyl chains attached on silica surface are densely packed. A correlation of a decrease of pore volume with an increase of the alkyl modifier chain length was found. Effective molecular volume of bonded chains was found to be similar to the molecular volume of corresponding liquid alkanes. An absence of noticeable penetration of acetonitrile, methanol, or tetrahydrofuran molecules between bonded chains at any water-organic eluent composition was found.

Liquid chromatographic systems with very weak excessive analyte-adsorbent interactions have been studied. These systems consisted of a homologous series of n-alkanes as both analytes and mobile phases with a C18 reversed-phase adsorbent. A linear decrease of the analyte retention volume with an increase of the number of analyte carbon atoms was found. Corresponding increases of analyte retention with an increase in the number of eluent carbon atoms was also discovered. An explanation of these two effects on the basis of adsorption theory is proposed. A good correlation of column hold-up volume calculated by interpolation of the retention dependencies for above mentioned systems with that measured by the minor disturbance method has been shown. A study of the temperature dependencies of these alkane systems has shown entropy-governed retention dependencies.

A fast HPLC method for the determination of the oxygen heterocyclic compounds of citrus essential oils was developed. Five different oils were analyzed under identical conditions, by reversed-phase HPLC with photodiode array detector, for a direct comparison of the composition of their oxygen heterocyclic fraction. Analysis time was 7 min. The oils analyzed were lemon, bergamot, mandarin, sweet orange, and bitter orange. The method developed is good for rapid screening or fingerprinting of these essential oils; a slightly slower method is recommended for higher resolution and better quantitative results.

Changes in histamine, putrescine, and cadaverine concentrations in bluefish filets (Pomatomus saltatrix) stored at 5, 10, and 15 degrees C were determined using high-performance liquid chromatography. An organoleptic assessment was conducted simultaneously with the biogenic amine analyses. The histamine levels found in fresh bluefish obtained from wholesale seafood distributors ranged between <1 ppm and 99 with an average of 39 ppm. Putrescine and cadaverine were not found in fresh bluefish. Fish fillets stored at each of the three temperatures developed histamine. The greatest accumulation of histamine was observed in fish stored at 15 degrees C, which developed histamine levels as high as 2,200 ppm. Putrescine levels increased at each temperature during storage. Cadaverine was present only in uninoculated bluefish stored at 15 degrees C. Histamine achieved higher levels in bluefish pieces inoculated with Morganella morganii, which demonstrates that bluefish support bacterial histamine formation. Histamine levels at each temperature exceeded the 50-ppm advisory level established by the Food and Drug Administration before 100% sensory rejection. Standard plate counts increased during storage of fish at all temperatures, but the correlation between histamine levels and standard plate count was not significant.

A new and simple method of analysis for polyamide composition is described. Polymer samples were decomposed to their corresponding monomers by hydrobromic acid (HBr) and the decomposed products were analyzed by capillary zone electrophoresis (CZE). Amines from polyamides were derivatized with fluorescamine and detected by UV at 280 nm. Aliphatic acids were detected by direct UV at 205 nm. Sample treatment is much simpler than published methods, and electropherograms show a good separation of decomposed products under the proper conditions.

The vitamin B6 status of seemingly healthy adolescent girls was determined using several accepted and proposed parameters in an effort to establish guidelines for status evaluation. High-performance liquid chromatography-derived plasma B6 vitamers (pyridoxal phosphate, PLP; pyridoxine phosphate. PNP; pyridoxamine phosphate, PMP; pyridoxal, PL; pyridoxine, PN; and pyridoxamine, PM) and 4-pyridoxic acid (4-PA) concentrations and urinary 4-PA levels of 28 white adolescent females, 12-15 years, having radiomonitored plasma PLP concentrations and coenzyme stimulation of erythrocyte alanine aminotransferase activities indicative of adequate status were determined. Mean vitamin B6 and protein intakes were 1.48 mg and 78.3 g. Ranges for plasma B6 vitamer and 4-PA concentrations (nmol/l) were: PLP, 40.9-122.2; PNP, non-detectable (ND)-16.1; PMP, ND-8.1; PL, ND-15; PN, ND-21.9; PM, ND-17.8; and 4-PA, ND-55.7. PLP was the only vitamer found in plasma of all subjects. Urinary 4-PA concentrations ranged from 0.11 to 2.50 mumol/mmol of creatinine. B6 vitamer values of these girls should be of use in the establishment of normal ranges for vitamin B6 status parameters.

High-resolution liquid-chromatographic methods developed for analyzing nucleotide pools at the nanogram level in four representative species of ascomycetes (Penicillium citrinum, Aspergillus niger, Fusarium moniliforme, and Cladosporium herbarum) were used to study polysaccharide biosynthesis. Nucleotides extracted from the mycelial mat were preseparated from interfering polysaccharides, glycoproteins, and nucleic acids on a column of Biogel P-2. Resolution of 18 nucleotides from each fungal species was accomplished on AS-Pellionex-SAX, pellicular anion-exchanger by using a high-pressure liquid chromatograph. Nucleotides were identified by comparing peak retention-times, by differential u.v. absorption with two detectors in series at selected wavelengths, and by acid or enzymic hydrolysis with product identification by liquid chromatography. Pyrimidine bases exceeded purines by at least three fold, and uridine nucleotides often constituted 60-80 mole percent of the total nucleotides; extractable cytidine nucleotides were negligible. Uridine 5'-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl disphosphate) is the preponderant nucleotide throughout the growth cycles of all four species, amounting to 30-60% of all nucleotides present. For all four fungal species, a burst of nucleotide formation was observed after the first 48h (15-30 mumol/g tissue), with fluctuations that eventually fell to 0.1 mumol/g on the tenth day.

A rapid and sensitive procedure is described for the analysis of all the B6 vitamers and 4-pyridoxic acid in human plasma utilizing reversed-phase high-performance liquid chromatography with ultraviolet and fluorometric detection. Pyridoxal phosphate values obtained by radiometric and chromatographic, ultraviolet and fluorometric, assays were highly correlated as were pyridoxine phosphate values determined using both detectors. Plasma B6 vitamer and 4-pyridoxic acid concentrations of 22 men fed diets containing 0.75-0.98 mg of vitamin B6 daily for eight weeks were in the range of reported values; pyridoxal phosphate was their predominant plasma B6 vitamer. This methodology should be useful in the assessment of vitamin B6 requirements.