Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (28): 5177-5180.doi: 10.3969/j.issn.1673-8225.2011.28.012

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Determination of pyrazinamide content in bone tissue by high performance liquid chromatogray

Shi Hua, Ge Zhao-hui, Liu Bin, Wang Zi-li   

  1. Department of Orthopedics, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Received:2011-01-19 Revised:2011-06-08 Online:2011-07-09 Published:2011-07-09
  • Contact: Wang Zi-li, Professor, Chief physician, Department of Orthopedics, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China WangZlnx@126.com
  • About author:Shi Hua★, Studying for master’s degree, Department of Orthopedics, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China shihuayisheng@126.com Ge Zhao-hui, Master, Associate professor, Associate chief physician, Department of Orthopedics, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China myovid@126.com Shi Hua and Ge Zhao-hui contributed equally to this paper and were considered as co-first authors.
  • Supported by:

    Key Technologies Research & Development Program of Ningxia Hui Autonomous Region, No. K200512*

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Abstract:

BACKGROUND: Pyrazinamide is a primary drug for intensive chemotherapy and its tissue distribution is greatly significant for therapy of spinal tuberculosis.
OBJECTIVE: To establish a high performance liquid chromatogray method to determine pyrazinamide content in bone tissue of patients with spinal tuberculosis.
METHODS: The vertebrae obtained from 10 patients with spinal fracture who underwent anterior cervical corpectomy were prepared into blank bone homogenate and pyrazinamide content in bone tissue was determined by high performance liquid chromatogray. Bone tissue samples including sclerotic bone, subnormal bone, necrotic tissue and ilium were harvested from 18 patients with spinal tuberculosis who received 2HRZ/H2R2Z2 chemotherapy. Supernatant of pyrazinamide bone homogenate was prepared by perchloric acid precipitation. The specificity, recovery rate, precision, and linear range of the high performance liquid chromatogray method were evaluated.
RESULTS AND CONCLUSION: Pyrazinamide was greatly absorbed at a wavelength of 265 nm and retained for 7.48 minutes, without presence of interference peak. Within the range of 0.048-3.120 μg/g bone homogenate, there was a good linear relationship between pyrazinamide peak area and bone homogenate concentration (r = 0.999 91), absolute recovery rate was 89.18%-93.75%, method recovery rate was 96.30%-100.45%, inter-day and intra-day relative standard deviation was 4.26%-8.78% and 5.12%-9.01%, respectively. The pyrazinamide content in the sclerotic wall of the vertebrae was approximate to the minimal inhibitory content and its content may reach the effective therapeutic content in the subnormal bone outside the sclerotic bone and self-contrast ilium. But pyrazinamide was hardly detectable in the sclerotic foci in the compromised vertebrae. The established method is accurate, precise, and reproducible, and it is effective for detection of pyrazinamide content in bone tissue. The pyrazinamide content varys greatly in different tissues of spinal tuberculosis, and the sclerotic wall is a tissue barrier to prevent pyrazinamide penetration into tuberculosis focus from normal vertebrae.

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