中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (28): 5177-5180.doi: 10.3969/j.issn.1673-8225.2011.28.012

• 骨组织构建 bone tissue construction • 上一篇    下一篇

骨组织中吡嗪酰胺浓度的高效液相色谱测定

石  华，戈朝晖，刘  斌，王自立   

  1. 宁夏医科大学附属医院骨科，宁夏回族自治区银川市  750004
  • 收稿日期:2011-01-19 修回日期:2011-06-08 出版日期:2011-07-09 发布日期:2011-07-09
  • 通讯作者: 王自立，教授，主任医师，宁夏医科大学附属医院骨科，宁夏回族自治区银川市 750004 WangZlnx@126.com
  • 作者简介:石华★，男，1980年生，宁夏回族自治区银川市人，汉族，宁夏医科大学在读硕士，主要从事脊柱外科工作。 shihuayisheng@126.com 并列第一作者：戈朝晖，硕士，副教授，副主任医师。 myovid@126. com
  • 基金资助:

    宁夏回族自治区科技攻关项目(K200512)，课题名称：超短程化疗下的脊柱结核病椎部分切除术的临床研究。

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*

PDF

270

被引次数

2

摘要：

背景：测定结核病化疗药物在病变椎体组织中的生物分布对脊柱结核的治疗具有重要价值。
目的：建立吡嗪酰胺在骨组织中药物浓度的高效液相色谱分析法并测定其在脊柱结核病椎组织中的浓度。
方法：取10例脊柱骨折患者前路椎体次全切除减压的椎骨制备成空白骨匀浆，高效液相色谱法测定吡嗪酰胺骨组织药物浓度。取18例脊柱结核患者(2HRZ/H2R2Z2化疗方案)的病椎硬化骨、亚正常骨、坏死组织以及髂骨为骨组织标本，采用高氯酸蛋白沉淀法获得骨匀浆吡嗪酰胺上清，对方法的专属性、回收率与精密度以及线性范围进行评价。
结果与结论：吡嗪酰胺在265 nm波长下有较大吸收，保留时间7.48 min，无干扰峰出现。在0.048~3.120 μg/g骨匀浆浓度范围内吡嗪酰胺峰面积与骨样本浓度线性关系良好(r=0.999 91)，绝对回收率在89.18%~93.75%，方法回收率为96.30%~100.45%，日内、日间精密度分别为4.26%~8.78%和5.12%~9.01%。应用此方法测得吡嗪酰胺在病椎硬化壁为最低抑菌浓度，壁外亚正常骨及对照髂骨达到有效治疗浓度，壁内核心病灶未检测到分布。提示吡嗪酰胺在脊柱结核病椎中存在明显的组织分布差异，硬化壁是其经正常椎体向核心病灶穿透的组织屏障。　

关键词: 高效液相色谱, 脊柱, 结核, 吡嗪酰胺, 分析

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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