A simplified method for detecting isoniazid compliance in patients receiving antituberculosis chemotherapy.
The objective of this study was to develop a new simplified method using thin-layer chromatography (TLC) for determining isoniazid (INH) compliance in patients receiving antituberculosis chemotherapy. TLC was performed on silica gel plates using a standard solution of INH and acetylisoniazid (AcINH) and ethyl acetate-methanol (70:30 v/v) as the developing solvent. The spots of compound were detected by iodine. In the human study, fractional urine samples were collected over 24 hours from 4 healthy human subjects genotyped for NAT2* and to whom 400 mg of INH were administered orally. These samples were used for TLC analysis. The results of TLC were compared with those of high-performance liquid chromatography (HPLC). This method indicated good separation between INH and AcINH in standard solutions. The detection limits for INH and AcINH (applied volume; 20 microl of standard solution) were 2.2 nmole and 5 nmole, respectively, as detected by iodine. In the human study, the INH spot in urine was not detected on the TLC plate, except in one sample over the 0- to 4-hour period from 1 volunteer. However, the AcINH spot was detected in all urine samples from all volunteers. The total experimental time from application of the urine sample to analysis on TLC was 30 minutes. The results suggest that this method for detecting AcINH on TLC is an excellent, convenient, and simple method for determining INH compliance in patients receiving standard antituberculosis chemotherapy regimen or INH preventative therapy, regardless of the patient's NAT2* genotype.
Hashiguchi
Masayuki
M
Department of Medication Use Analysis and Clinical Research, Meiji Pharmaceutical University, Kiyose, Tokyo, Japan.
Ohno
Keiko
K
Sakuma
Atsuko
A
Hino
Fumio
F
Tanaka
Takanori
T
Ohtsuji
Masahiko
M
Matsumoto
Naoto
N
Yanase
Kumiko
K
Urae
Akinori
A
Hosogai
Yuki
Y
Sato
Naomi
N
Yazaki
Asami
A
Matsuda
Kaoru
K
Yamazaki
Katsuji
K
Rikihisa
Tadaaki
T
D000107:Acetylation; D000328:Adult; D000995:Antitubercular Agents / Q000008:administration & dosage / Q000627:therapeutic use* / Q000652:urine; D001191:Arylamine N-Acetyltransferase / Q000235:genetics; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004247:DNA / Q000235:genetics; D005260:Female; D005838:Genotype; D006801:Humans; D007538:Isoniazid / Q000008:administration & dosage / Q000627:therapeutic use* / Q000652:urine; D007564:Japan; D008297:Male; D010349:Patient Compliance / Q000706:statistics & numerical data*; D015203:Reproducibility of Results; D014397:Tuberculosis, Pulmonary / Q000188:drug therapy*
D000995:Antitubercular Agents; D004247:DNA; D001191:Arylamine N-Acetyltransferase; C478900:NAT2 protein, human; D007538:Isoniazid