Identification of 18 mycobacterial species was performed by analysis of profiles obtained by using gas-liquid chromatography. Organisms were saponified in methanolic NaOH, and the reaction mixture was treated with BF3 in methanol and extracted with a hexane-chloroform mixture. An identification scheme was developed from 128 stock strains and tested against a collection of 79 clinical isolates. By using gas-liquid chromatographic profiles alone, 58% of specimens were correctly identified to species level, and an additional 41% were correctly identified to a group of two or three organisms. Use in a clinical laboratory over a 2-month period proved chromatography to be as accurate as and more rapid than concurrent biochemical testing. Of 81 isolates tested, 64% were identified to species level by chromatography alone. An additional 35% were differentiated to the same groups of two or three organisms as found in our analysis of stock strains. These groups consisted of: Mycobacterium tuberculosis, M. bovis, and M. xenopi; M. avium complex, M. gastri, and M. scrofulaceum; or M. fortuitum and M. chelonei. Identification to species level from these groups could usually be done by colonial morphology alone and could always be done by the addition of one selected biochemical test. This study demonstrated the practical application of gas-liquid chromatography in the identification of mycobacteria in a clinical laboratory. In particular, all strains of M. gordonae and M. kansasii were identified to species level. M. tuberculosis was definitively identified in 85% of cases. When it could not be definitely identified, the only alternatives were M. bovis and M. xenopi, both of which are rare causes of infection.

Identification of routine mycobacterial isolates by gas-liquid chromatography profile analysis was performed on 335 strains received at the Mayo Clinic over a 10-month period. Comparison of identification by gas-liquid chromatography versus conventional biochemical profiles was made. The two methods agreed on the identification of 320 isolates, with gas-liquid chromatography profiling making eight errors and biochemical profiling making four errors. In three cases, discrepancies could not be resolved.

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A liquid chromatographic method is described for analysis of aerobic actinomycetes for isomers of diaminopimelic acid. One or two colonies of organism were hydrolyzed with 6.0 mol of HCI per liter at 121°C for 15 min. The hydrolysate was neutralized and buffered with an NaOH solution (3 mol/liter) containing 0.15 mol of sodium borate per liter. Precolumn derivatization with dansyl chloride was used to forn a fluorescent product for detection. Analysis was performed by reversed-phase, ion-pair chromatography. The L-diaminopimelic acid isomer was detected in all 10 strains of Streptomyces tested, and the meso-diaminopimelic acid isomer was detected in all 10 strains of Nocardia tested. Liquid chromatography was compared simultaneously with thin-layer chromatography in the analysis of three strains of aerobic actinomycetes. Liquid chromatography required less growth of the organisms, and analysis was completed within 1 h, compared with the 3 to 5 days required by thin-layer chromatography.

We describe a liquid-chromatographic procedure for detection in urine of all thiazide drugs currently used clinically. Urine is treated initially with NaBH4 to convert any chlorothiazide present to hydrochlorothiazide. The urine is acidified with NaH2PO4 (1.0 mol/L, pH 5), and thiazides are extracted with ethyl acetate. Interfering substances are then removed in two washes with 0.1 mol/L Na2HPO4 at pH 8. The ethyl acetate is evaporated and the residue redissolved in mobile phase. Thiazides are assayed by reversed-phase chromatography, with detection by ultraviolet absorption. Analytical recovery of thiazides ranged from 53 to 93%. Urines from 48 patients were so studied, and the results were compared with results by the currently used spectrophotometric method. The two methods agreed for 56% of samples. Evaluation of the discrepancies by review of patients’ histories clearly showed liquid chromatography to have correctly identified seven of eight positive urines that the spectrophotometric method failed to detect. Ultraviolet scanning incorrectly identified as positive two samples, whereas liquid chromatography did not falsely identify any urines as positive. Our method was more sensitive and more specific than the spectrophotometric method.