目的建立高效液相色谱-二极管阵列检测器（HPLC-DAD）法同时测定右归丸（熟地黄、附子、肉桂等）中6种成分的含有量。方法分析采用Waters X Bridge-C18色谱柱（4.6 mm×250 mm,5μm）;流动相甲醇-1%冰醋酸,梯度洗脱;柱温30℃;体积流量1.0 m L/min。结果绿原酸、莫诺苷、松脂醇二葡萄糖苷、马钱苷、阿魏酸和肉桂酸均呈良好的线性关系（r≥0.999）,平均加样回收率（n=6）为99.7%~102.2%（RSD〈3%）。结论该方法简便快速,准确可靠,重复性好,可用于右归丸的质量控制。
Objective Osteoarthritis（OA） is a kind of degenerative arthritis, involving degradation of joints, including articular cartilage and subchondral bone. Bushen-Huoxue Herb Couple（BHHC） is a Chinese herb pair with invigorating the kidney and promoting blood circulation. In our study, a Gas Chromatography-Mass Spectrometer（GC-MS） based metabolomics was developed to achieve a systematic observation of the progression of OA and evaluate the efficacy of BHHC in OA rats. Methods Forty Sprague-Dawley rats（half males and half females） were randomly divided into five groups, including normal group, OA model group, sham-operated group, BHHC group, positive medicine（PM） group after anterior cruciate ligament reconstruction and meniscectomy on rats. Urine samples for analysis were collected on week 0, 2, 4, 6, 8, 10, 12, and 14 during the experiment. Results The histological results showed that BHHC had a strong action of repairment and reconstruction of joint damage. By combining with partial least squares discriminant analysis（PLS-DA）, 14 biomarkers were identified in urine samples. The biomarkers were mainly involved 10 metabolism pathways, such as tricarboxylic acid cycle metabolism, acid amino metabolism, tryptophan metabolism, lipid metabolism and phenylalanine metabolism. Using these potential biomarkers as a screening index, it indicated that BHHC could reverse the pathological process of OA through regulating amino acid metabolism, lipid metabolism, nucleotide metabolism and energy metabolism. Conclusions This study demonstrates that a metabolomics is a useful tool for identifying potential OA biomarkers and investigating the underlying mechanisms of Traditional Chinese Medicine in OA treatment.
Hua YinLing WangQing-lin LiJian-hua ZhangLin ZhangXuan Wang
