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Carlhant, D.; Le Bot, M. A.; Guedes, Y.; Riche, C.; Mimouri, F.; Colin, J.; Berthou, F.
doi: 10.1002/bmc.1130030102pmid: 2706358
A method for extracting spiramycin by an octadecylsilica cartridge is described for plasma or vitreous samples. The macrolide antibiotic is then measured by reversed‐phase HPLC with UV detection. The limit of detection is estimated to be 50 ng/mL. The coefficient of variation for the procedure is 6.1% and 5.2% for the range of concentrations 0.2 μg/mL and 10 μg/mL respectively. By this method, pharmokinetic profiles were performed for five adult patients. Spiramycin could be accurately measured in the vitreous humour, allowing the determination of antibiotic at its site of action.
Steffenrud, Steffen; Salari, Hassan
doi: 10.1002/bmc.1130030103pmid: 2706367
A sensitive liquid chromatography method has been developed using electrochemistry for the determination of leukotrienes in biological fluids. Biological specimens are treated with 3,5‐dinitrobenzoyl chloride in acetonitrile which undergoes rapid reaction with hydroxyl groups of non‐peptidic leukotrienes in the presence of pyridine and with amino groups of peptidic leukotrienes in the presence of potassium tetraborate buffer. The resulting dinitroben‐zoate derivatives of leukotrienes are highly electroactive, suitable for reduction or oxidation at moderate potentials by an electrochemical detector. In reductive mode at −0.7 V or oxidative mode at + 1.15 V potentials, the lower limits of detection for leukotriene derivatives were approximately 8±3 pg and 70 ± 16 pg respectively, with a signal‐to‐noise ratio of 5 to 1. This method was applied to the detection of leukotrienes in plasma, nasal and bronchial fluids of patients with asthma.
Shimin, Zhang; Jianxin, Li; Yaomei, Wei; Zhiming, Yang
doi: 10.1002/bmc.1130030104pmid: 2706359
A simplified method for the preparation of wheat germ agglutinin(WGA)‐Sepharose 4B by coupling highly purified WGA, prepared by improved affinity chromatography, with BrCN activated Sepharose 4B in a solution of high carbonate buffer is described. The amount of WGA linked to Sepharose 4B was 82.40% (3.07 mg WGA per ml Sepharose 4B). MN blood group antigens of human erythrocyte membranes purified with WGA‐Sepharose 4B affinity chromatography showed a single band on sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE). The yield of the antigens from 400 mL fresh blood was 32–40 mg. The WGA‐Sepharose 4B column could be used several times without loss of activity.
Kizu, Ryoichi; Hayakawa, Kazuichi; Miyazaki, Motoichi
doi: 10.1002/bmc.1130030105pmid: 2706360
A method for determining cis‐diamminedichloroplatinum(II) (CDDP), an anticancer drug, in plasma and urine by HPLC with UV detection and column‐switching has been developed. Typical conditions were as follows. An apparatus was composed of two columns, two pumps, a UV detector, a sample injector with a 100 μL loop, a switching valve, a column oven and a recorder. A Rheodyne model 7125 sample injector was used as the switching valve. A precolumn (4.6 mm ID × 25 cm) was packed with MCI GEL CK10S (a strong cation exchanger), and an analytical column (4.6 mm ID × 5 cm) was packed with MCI GEL CDR10 (a strong anion exchanger). Both columns were connected in series via the switching valve. The CDDP‐containing fraction of the effluent from the precolumn was loaded to the analytical column by column‐switching and the effluent from the analytical column was monitored at 210 nm. An eluent of 0.3 M sodium dihydrogen phosphate was pumped at a flow rate of 1 mL/min and the columns were maintained at 40°C. CDDP was eluted at about 11 min and the identity of the peak of CDDP on the chromatogram was confirmed by its 3‐dimensional chromatogram and analysis of platinum in the column effluent. Under the conditions described above, a linear relationship was obtained between peak height and concentration of CDDP up to 100 μM. Correlation efficients were 0.998 for plasma and 0.999 for urine. The detection limit was 0.1 μM for CDDP in both plasma and urine (S/N = 3, 0.005 AUFS). The reproducibility was within 3% for 10 determinations. The CDDP concentrations in plasma and urine of a patient with ovarian carcinoma receiving CDDP treatment could be determined.
Elmberger, P. Göran; Eggens, Ivan; Dallner, Gustav
doi: 10.1002/bmc.1130030106pmid: 2706361
Conditions for the isolation and quantitation of dolichyl phosphate, dolichol, cholesterol, and ubiquinone by reversed phase high performance liquid chromatography were investigated. A simple and fast sample preparation procedure using prepacked mini columns was employed. The UV spectra of the fractions obtained were examined and, in the case of dolichol compounds, the maximum absorbance around 205 nm was shown to be linearly dependent on the number of double bonds present in the isoprenolog. The analytical procedure described shows a very broad range of linearity (five orders of magnitude) and detects single dolichyl phosphate isoprenologs in amounts as small as 0.1 ng. The lowest overall recovery, that for dolichyl phosphate, is 77%. Use of isoprenolog 23 and ergosterol as internal standards reduced the variation in the method to 2.5, 4.0 and 5.5% for cholesterol, dolichyl phosphate and dolichol, respectively. The method described was employed to study the lipid composition of rat organs and biological variations in these compositions.
Zhu, Zhengmei; Li, Ruixiang; Cui, Zhaochun; Zhou, Liangmo; Huang, Mingxian
doi: 10.1002/bmc.1130030107pmid: 2706362
A new capillary GC method is described for the compositional analysis of the three main gangliosides isolated from adult human myometrium. The sample was subjected to methanolysis, acetylation and trimethylsilylation which allows all the constituents to be analyzed simultaneously. The predominant ganglioside was found to be GD3, with GM3 and GT1b the next most abundant.
doi: 10.1002/bmc.1130030108pmid: 2706363
A method for the unambiguous characterization of DNA photoproducts has been developed. It does not require radio‐labelled DNA, or specialized techniques. In the preliminary step, UV‐irradiated DNA is hydrolysed to its constituent bases and photoproducts. The photoproducts are then separated from nucleic acid bases using low pressure ion‐exchange chromatography on a Dowex 1 × 8 200–400, and an Amberlite CG–50–H+ column. Further separation and purification of photoproducts is carried out on the HPLC Whatman Partisil ODS2 column, using 15% MeOH. The procedure is simple, reproducible and versatile.
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