【摘要】：目的：探討鷓鴣茶多酚的提取純化工藝并對其進(jìn)行質(zhì)量控制研究；研究鷓鴣茶多酚防治去勢大鼠骨質(zhì)疏松的作用，并初步探討其作用機(jī)制；建立鷓鴣茶藥材的指紋圖譜，并進(jìn)行模式識別研究。 方法：以鷓鴣茶多酚提取率為指標(biāo)考察提取工藝，采用單因素和正交實(shí)驗(yàn)確定鷓鴣茶多酚的最佳提取工藝；通過AB-8、NKA-9、DM-130、HPD-600、HPD-826五種樹脂的靜態(tài)實(shí)驗(yàn)優(yōu)選最佳純化樹脂，通過動態(tài)實(shí)驗(yàn)確定最佳的吸附、解吸條件。 對鷓鴣茶多酚進(jìn)行質(zhì)量控制。采用薄層色譜法對鷓鴣茶多酚進(jìn)行定性鑒別，并采用一測多評方法進(jìn)行沒食子酸與表兒茶素的含量測定。 探討鷓鴣茶多酚防治去勢大鼠骨質(zhì)疏松的作用。將大鼠隨機(jī)分為6組，即假手術(shù)組、模型組、鷓鴣茶多酚低、中、高劑量組和戊酸雌二醇陽性對照組。除假手術(shù)組大鼠外，其他組大鼠均行卵巢切除術(shù)，手術(shù)4w后開始給藥。給藥8w后，10%水合氯醛麻醉，，腹主動脈取血，按試劑盒方法測定E2、SOD、MDA、ALP（堿性磷酸酶）；并計(jì)算子宮指數(shù)、胸腺指數(shù)、脾臟指數(shù)、股骨骨密度等指標(biāo)。 建立鷓鴣茶藥材的指紋譜，并進(jìn)行模式識別研究。采用戴安Acclaim120C18色譜柱（250mm×4.6mm，5μm）；以乙腈-0.03%三氟乙酸梯度洗脫；檢測波長：280nm；流速1.0ml/min；柱溫30℃；信號采集時間70分鐘。結(jié)果：提取工藝確定為：取鷓鴣茶藥材適量，粉碎，加15倍量40%乙醇回流提取3次，每次45min，合并濾液，過濾，減壓回收乙醇并濃縮至相對密度約1.1（60℃），噴霧干燥（進(jìn)口溫度105℃，出口溫度95℃，蠕泵轉(zhuǎn)速7r/min，風(fēng)機(jī)轉(zhuǎn)速1900r/min），即得鷓鴣茶提取物。 通過靜態(tài)試驗(yàn)選出AB-8大孔吸附樹脂為最佳樹脂，純化工藝確定為：取鷓鴣茶提取物過AB-8大孔吸附樹脂，上樣量為12mg/ml樹脂，上樣流速為1BV/h，上樣藥液pH值為3。先用水洗至洗脫液無色，再用4BV80%乙醇洗脫，洗脫流速為2BV/h，收集醇洗脫液，減壓回收乙醇并濃縮至相對密度約1.0（60℃），低溫冷凍干燥(-50℃，400Pa)，得到鷓鴣茶多酚。 建立了鷓鴣茶多酚的質(zhì)量標(biāo)準(zhǔn)草案：采用薄層色譜法鑒別出兒茶素、EGCG、ECG、EGC、沒食子兒茶素、沒食子酸等6種成分，斑點(diǎn)清晰，重現(xiàn)性好，可作為鑒別方法。利用HPLC采用一測多評法測定沒食子酸與表兒茶素含量，該方法準(zhǔn)確可靠，重現(xiàn)性好，可作為鷓鴣茶多酚的質(zhì)控方法。 鷓鴣茶多酚可以用于防治去勢大鼠骨質(zhì)疏松�？稍黾庸敲芏取⑸哐錝OD、E2含量，降低MDA、ALP含量，升高子宮指數(shù)等指標(biāo)。 在建立的鷓鴣茶藥材HPLC指紋圖譜中，共標(biāo)記15個共有峰，指認(rèn)出沒食子酸、EGC、兒茶素、EC、EGCG、ECG等6種成分；并進(jìn)行聚類分析與主成分分析，最終將鷓鴣茶藥材分為3類。 結(jié)論：鷓鴣茶多酚提取純化工藝簡便可行，其質(zhì)量穩(wěn)定可控，可以用于防治去勢大鼠骨質(zhì)疏松癥；建立鷓鴣茶藥材的指紋圖譜，并進(jìn)行了模式識別研究，為開發(fā)這一民間藥用資源提供了科學(xué)依據(jù)。
[Abstract]:Objective: to study the extraction and purification of polyphenols from Partridge tea and its quality control, to study the effect of tea polyphenols on preventing and treating osteoporosis in ovariectomized rats, and to explore its mechanism. To establish the fingerprint of Partridge tea and to study the pattern recognition. Methods: the extraction process of polyphenols from Partridge tea was investigated by single factor and orthogonal experiment. The best purified resin was selected by static experiment of five kinds of AB-8,NKA-9,DM-130,HPD-600,HPD-826 resins, and the optimum conditions of adsorption and desorption were determined by dynamic experiment. The quality control of tea polyphenols from Partridge was carried out. The content of Gallic acid and epigallocatechin was determined by TLC. To investigate the effect of tea polyphenols on preventing and treating osteoporosis in ovariectomized rats. Rats were randomly divided into six groups: sham-operated group, model group, partridge tea polyphenol group, middle and high dose group and estradiol valerate positive control group. Ovariectomies were performed in all rats except sham-operated rats. After 8 weeks of administration, 10% chloral hydrate was used to anesthetize abdominal aorta, blood was collected from abdominal aorta, ALP (alkaline phosphatase) was determined by kit method, and the indexes of uterus index, thymus index, spleen index and bone mineral density of femur were calculated. The fingerprint spectrum of Partridge tea was established and the pattern recognition was studied. A Acclaim120C18 column (250mm 脳 4.6 mm ~ 5 渭 m);) with acetonitrile-0.03% trifluoroacetic acid gradient elution was used. The detection wavelength was 280 nm, the flow rate was 1.0 ml / min, the column temperature was 30 鈩

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