Simultaneous esterification of carboxyl and hydroxyl groups with alcohols and heptafluorobutyric anhydride for analysis by gas chromatography

Conditions are described for the detection of tuberculostearic acid (10-methyloctadecanoate; C18 * CH3) in cerebrospinal fluid and serum of patients with tuberculous meningitis. C18. CH3 was found in both the cerebrospinal fluid and serum of patients with tuberculous meningitis at concentrations of 25 to 50 fmol (10-15 mol). The necessary specificity and sensitivity for detection of C18 * CH3 were obtained by extraction under acid conditions with organic solvent, specific functional group esterification with trichloroethanol, cleanup with disposable reverse-phase sorption chromatography columns, analysis on high-resolution polar and nonpolar capillary columns, and detection by a frequency-pulsed electron capture detector. Use of an IBM 9000 computer equipped with CAP software significantly aided comparison between known C18. CH3 standards and C18. CH3 in clinical specimens. Scale expansion and attenuation changes were the major contributions obtained by use of the computer. The data indicate that detection of C18. CH3 by frequency-pulsed electron capture gas-liquid chromatography may be a valuable aid for early detection of tuberculous meningitis.

Section: Methodsmentioning

confidence: 99%

Selective procedures for detecting femtomole quantities of tuberculostearic acid in serum and cerebrospinal fluid by frequency-pulsed electron capture gas-liquid chromatography

Brooks¹,

Daneshvar²,

Fast³

et al. 1987

“…In addition, the HFBA reagent was used to detect alcohols and chloroform-soluble hydroxy acids. The basic chloroform extracts containing amines and the acidic ethyl ether extracts containing hydroxy acids were derivatized with HFBA-pyridine-ethanol to form amides and esters, respectively, as described hefore (4,6). After the TCE and HFBA derivatives of carboxylic acids, alcohols, and amines were prepared, the TCE derivatives were dissolved in 0.3 ml of xylene-ethanol (1:1), and the alcohol and amine HFBA derivatives were dissolved in 0.1 ml of xyleneethanol (1:1).…”

Section: Methodsmentioning

confidence: 99%

Frequency-pulsed electron capture gas-liquid chromatographic analysis of metabolites produced by Clostridium difficile in broth enriched with amino acids

Brooks¹,

Nunez-Montiel²,

Wycoff³

et al. 1984

Four extractions were done on the spent medium, three at pH 2 and one at pH 10, using CHCL3 or ether. Derivatizations were done with trichloroethanol, heptafluorobutyric anhydride, and heptafluorobutyric anhydrideethanol. All samples were analyzed with frequency-pulsed electron capture gas-liquid chromatography. A dedicated computer was used to assist in data analysis. C. difficile produced both short-chain and aromatic acids in Trypticase-yeast-salt broth; hydroxy acids were also detected. p-Cresol, indoleacetic acid, 4methylthio-2-hydroxybutyric acid, and some unidentified alcohols were observed. The basic chloroform extraction contained cadaverine and putrescine. Leucine, norleucine, and isoleucine influenced the production of C5 and C6 acids and alcohols. L-Tyrosine underwent successive degradation to produce p-cresol and arolpatic

“…Carboxylic acids were esterified with trichloroethanol (TCE), amines were derivatized with heptafluorobutyic anhydride, and hydroxy acids were diesterified with heptafluorobutyric anhydride-ethanol as described previously (1,3). Derivatized amines and hydroxy acids were analyzed with a Perkin-Elmer 3920 FPEC GLC on a glass column (2 mm [inside diameter] by 7.6 m) that had been packed with OV-101 on 80/100 mesh Chromosorb W (5,6).…”

mentioning

confidence: 99%

Detection of metabolites by frequency-pulsed electron capture gas-liquid chromatography in serum and cerebrospinal fluid of a patient with Nocardia infection

Brooks¹,

Kasin²,

Fast³

et al. 1987