Ability of anaesthetists to identify a marked lumbar interspace.

Anaesthetists' ability to identify correctly a marked lumbar interspace was assessed in 100 patients undergoing spinal magnetic resonance imaging scans. Using ink, one anaesthetist marked an interspace on the lower spine and attempted to identify its level with the patient in the sitting position. A second anaesthetist attempted to identify the level with the patient in the flexed lateral position. A marker capsule was taped over the ink mark and a routine scan performed. The actual level of markers ranged from one space below to four spaces above the level at which the anaesthetist believed it to be. The marker was one space higher than assumed in 51% of cases and was identified correctly in only 29%. Accuracy was unaffected by patient position (sitting or lateral), although it was impaired by obesity (p = 0.001) and positioning of the markers high on the lower back (p < 0.001). The spinal cord terminated below L(1) in 19% of patients. This, together with the risk of accidentally selecting a higher interspace than intended for intrathecal injection, implies that spinal cord trauma is more likely when higher interspaces are selected.

High performance liquid chromatographic determination of vitamin D in fortified milks, margarine, and infant formulas.

Fortified milks were saponified overnight at room temperature with 1% ethanolic pyrogallol and KOH. The digest was extracted with hexane after adding water and ethanol, and the extract was washed consecutively with 5% KOH, water, and 55% aqueous ethanol to remove polar lipids. After evaporation, the residue was first chromatographed on a column of 5 micrometer silica. A fraction containing vitamin D was collected, evaporated, and rechromatographed on a reverse phase column for the separation and quantitation of vitamins D2 and D3. Recovery was 96-99% and the coefficient of variation was 3% (8 replicates). Infant formula was diluted and then saponified and extracted as in the analysis of milk. Margarine was saponified by shaking overnight with 1% ethanolic pyrogallol and 80% KOH. Water and ethanol were added to the digest before extraction. Extracts from formula and margarine were chromatographed as milk except, before HPLC, the extract was dissolved in isopropanol-hexane (1 + 99) and passed through 5 cm alumina in a Pasteur pipet, and the concentration of isopropanol in the first high performance liquid chromatographic (HPLC) solvent system was halved to improve the separation of vitamin D from other absorbing lipids. Usually several peaks were obtained during the final HPLC analysis, and the identification of vitamins D2 and D3 was less certain than in the analysis of milk. The coefficients of variation for formula and margarine were 6% (5 replicates) and 9% (6 replicates), respectively.

High performance liquid chromatographic determination of vitamin A in margarine, milk, partially skimmed milk, and skimmed milk.

Saponification and subsequent evaporation of extracts can cause losses of vitamin A during analysis and thus cause erratic results. These steps were therefore avoided by measuring retinyl palmitate and beta-carotene directly in hexane extracts of milk and margarine. Extracts were purified before high performance liquid chromatography (HPLC) by washing with aqueous alcohol. Retinyl palmitate was measured at 325 nm after chromatography on LiChrosorb Si60, 5 micrometer, using ethyl ether-hexane (2+98) as the solvent system. Milk (2 mL) was shaken with absolute ethanol (5 mL) and hexane (5 mL). Water (3 mL) was added and, after mixing and centrifugation, 100 microL hexane hexane layer was injected. Margarine (5 g) was dissolved in hexane (100 mL). After 5 mL of the hexane solution was washed with 60% ethanol and centrifuged, a 50 microL aliquot was injected. The results of the retinyl palmitate estimation in milk agreed with those obtained by a fluorometric method; the results in margarine agreed with those obtained by saponification, extraction, and HPLC. The coefficients of variation on analysis of 10 replicate samples of milk and margarine were 3 and 4.5%, respectively. The analysis of margarine for vitamin A was completed by measuring beta-carotene with a detector set at 453 nm; the coefficient of variation of this measurement was 3% (10 replicates).

High pressure liquid chromatographic determination of vitamin D in fortified milk.

Vitamin D was determined in fortified milk by high pressure liquid chromatography (HPLC) after preliminary purification involving saponification at room temperature and low pressure chromatography on hydroxyalkoxyprophyl Sephadex (HAPS). The saponified milk (50 ml) was extracted with ether, the extract was chromatographed in hexane on a 60 X 1 cm column of HAPS, and the absorption of the eluate at 254 nm was recorded. The fraction containing vitamin D was collected by using the position of characteristic peaks in the chromatogram as a guide. An aliquot of this fraction was chromatographed in an HPLC system on a 25 cm X 3.2 mm column of LiChorosorb Si 60 5 micrometer silicic acid, using 0.6% isopropanol in hexane (50% water-saturated) as the mobile phase. The absorption was set at 265 nm. Vitamin D was cleanly separated from other absorbing materials, and the amount could be calculated from the peak area. Recovery of a added vitamin D was greater than 97%.

Reverse phase high pressure liquid chromatography of vitamin A in margarine, infant formula, and fortified milk.

Retinal was determined in margarine (50 mg), infant formula (1 ml), and fortified milk (1 ml) by saponification in centrifuge tubes, extraction of the unsaponifiable lipid with hexane, and high pressure liquid chromatography (HPLC) in 90% methanol on a 25 cm x 3.2 mm column containing 10 micrometer LiChrosorb reverse phase. beta-Carotene was determined using the same column and 99% methanol as eluant. The vitamins in the eluate were identified and measured from their absorption at 325 (retinol) and 453 nm (beta-carotene), using a computing integrator. Retinol and carotene were prominent peaks in the chromatograms from properly fortified samples and were satisfactorily separated from other materials. In a survey of 12 different margarines for retinol, the results from liquid chromatography agreed with those of a variety of spectrophotometric and fluorometric procedures but they were obtained with greater ease and they could be interpreted more confidently. The recovery of retinol from oil was better than 99%. The coefficients of variation was 6.8% for 12 replicate analyses of a margarine for retinol and 5.4% for 10 replicate analyses of a margarine for beta-carotene.