Gas chromatographic determination of midazolam in low-volume plasma samples.
A gas chromatographic method for the sensitive determination of midazolam in plasma volumes as low as 40 microliters was developed, utilizing clinazolam as the internal standard. After liquid-liquid extraction at basic pH into 1-chlorobutane-dichloromethane (96:4) a 2- to 4-microliters portion of the reconstituted extract was injected under electronic pressure control onto a 12 m x 0.2 mm I.D. methyl silicone capillary column, and was exposed to a three-step temperature program from 120 to 310 degrees C, to separate the analytes from the plasma constituents. The compound of interest was identified and quantified by means of a mass-selective detector. The assay was linear from 10 to 500 ng/ml using 40 microliters of plasma (limit of quantification: 10 ng/ml) and was linear from 0.25 to 100 ng/ml using 500 microliters of plasma (limit of quantification: 0.25 ng/ml). The intra-day precision for the 40-microliters aliquots varied from 2.2 to 6.6%, the corresponding accuracy from -7.4 to -4.4%; the inter-day precision ranged from 5 to 7.2% and the corresponding accuracy from -7.2 to -5.1%.
Methods for the measurement of benzodiazepines in biological samples.
A review of methods for the measurement of benzodiazepines in biological specimens published over the last five years is presented. A range of immunoassay procedures using EIA, ELISA, FPIA, agglutination or kinetic interaction of microparticles, or RIA methods are now available. Cross reactivities to benzodiazepines are variable such that no one kit will recognise all benzodiazepines and their relevant metabolites at concentrations likely to be encountered during therapeutic use. Prior hydrolysis of urine to convert glucuronide metabolites to immunoreactive substances improves detection limits for many benzodiazepines. Several radioreceptor assays have now been published and show good sensitivity and specificity to benzodiazepines and offer the advantage (over immunoassay) of being able to detect these drugs with equal sensitivity. Solvent extraction techniques using a variety of solvents were still popular and offer acceptable recoveries and lack of significant interference from other substances. A number of papers describing solid phase extraction procedures were also published. Direct injection of specimens into a HPLC column with back flushing were also successfully described. Seventy two chromatographic methods using HPLC, LC-MS, GC and GC-MS methods were reviewed. HPLC was able to achieve detection limits for many benzodiazepines using UV or DAD detection down to 1-2 ng/ml using 1-2 ml of urine or serum (blood). ECD detectors gave detection limits better than 1 ng/ml from 1 ml of specimen, which was an order of magnitude lower than for NPD. EI-MS offered similar sensitivity, whilst NCI-MS was capable of detection down to 0.1 ng/ml. Methods suitable for the separation of enantiomers of benzodiazepines have been described using HPLC. Electrokinetic micellar chromatography has also been shown to be capable of the analysis of benzodiazepines in urine.
The crystal structure of stilpnomelane. Part II. The full cell
Summary The structural formula of stilpnomelane averaged from 37 representative literature analyses and based on the determined structure is (Ca,Na,K)4(Ti0·1Al2·3Fe35·5Mn0·8Mg9·3)[Si63Al9](O,OH)216·nH2O. Ferrostilpnomelanes have up to 12 R 3+ cations in the octahedral sheet, ferristilpnomelanes commonly have between 24 and 30 R 3+ cations, rarely up to 37. The two groups can be distinguished by accurate X-ray powder diffraction, ferrostilpnomelanes having d 001 less and a greater than have ferristilpnomelanes. The unit cell is triclinic, a = b∼21·8 Å, γ = 120°, d 001∼12·2 Å, P . Hexagonal groups or ‘islands’ of 24 silicon-oxygen tetrahedra co-ordinate to the octahedral sheet, the centres of the ‘islands’ are displaced by 5a/12, 8b/12 on either side of the octahedral sheet as a result of warping of this sheet to permit articulation with the smaller tetrahedral sheet. The 24-tetrahedra ‘islands’ are linked laterally by 6-member rings of inverted tetrahedra, which in turn link at their apices to like rings connecting the next sheet of ‘islands’. Thus a single tetrahedral layer is 4 tetrahedra thick and has symmetry 6/m. The 6-member rings of tetrahedra act as hinges permitting up to 0·5 Å extension or contraction of the tetrahedral sheet to accommodate variations in the octahedral sheet dimensions with varying R 2+/R 3+ ratio.