高效液相法同时测定大鼠脑组织中积雪草苷和羟基积雪草苷浓度

目的:建立高效液相色谱(HPLC)同时测定脑组织中积雪草苷和羟基积雪草苷浓度的方法。方法:大鼠尾静脉注射积雪草苷、羟基积雪草苷和积雪草总苷,30 min后用生理盐水进行心脏灌注,去除脑组织中残留血液,取脑组织,经甲醇沉淀蛋白后,利用HPLC测定脑组织中积雪草苷和羟基积雪草苷浓度。结果:积雪草苷和羟基积雪草苷在0.5-12μg/mL线性关系良好,定量下限为0.5μg/mL,日内和日间精密度(RSD)均小于15%,准确度(RE)在-5.25%-9.16%,提取回收率在70.54%-102.61%。结论:该方法符合生物样品测定要求,可以同时测定大鼠脑组织中积雪草苷和羟基积雪草苷的浓度。     

草苁蓉中氨基酸和营养元素含量的测定

采用日立L-8800型氨基酸自动分析仪和岛津AA-6300原子吸收分光光度仪,测定了草苁蓉中的氨基酸与营养元素的质量分数。结果表明,草苁蓉氨基酸总量为6.55%,必需氨基酸含量占氨基酸总量的36.18%。营养元素钾、钠、钙、镁、铁、锌、锰、铜质量分数分别为20.436 mg.g-1、2.275 mg.g-1、0.820 mg.g-1、0.994 mg.g-1、0.236 mg.g-1、0.036 mg.g-1、0.034 mg.g-1、0.016 mg.g-1。     

HPLC法测定杨梅叶中杨梅苷的含量

目的:建立HPLC法测定杨梅叶中杨梅苷含量的方法。方法:采用Diamonsil C18(150 mm×4.6 mm,5μm)柱,流动相乙腈-0.1%磷酸水溶液梯度洗脱,流速1.0 mL/min,检测波长358 nm,柱温30℃。结果:杨梅苷在0.088-0.880 mg/mL内与峰面积呈良好线性关系,A=36113C-401.56,r=0.9994;平均回收率为102.4%,RSD=1.9%(n=9)。测得杨梅叶中杨梅苷含量为1.43%。结论:该测定方法准确,可靠,对于杨梅的进一步开发利用具有重要的参考意义。     

HPLC法同时测定鸡矢藤中熊果酸和齐墩果酸的含量

目的：建立高效液相色谱法同时测定鸡矢藤中熊果酸和齐墩果酸的含量。方法：色谱柱为DiamonsilTM C18（250×4.6 mm,5滋m）,流动相为甲醇-0.05mol.L-1磷酸溶液（93∶7）为流动相,检测波长210 nm,进样量10滋L。结果：熊果酸在21.28滋g.mL-1～212.8滋g.mL-1范围内线性关系良好（r=0.9997）,平均回收率为97.79%（RSD=1.30%）;齐墩果酸在24.02滋g.mL-1～240.2滋g.mL-1范围内线性关系良好（r=0.9996）,平均回收率为98.36%（RSD=1.11%）。结论：该方法简便、灵敏、准确,可用于鸡矢藤药材的质量评价。     

HPLC法同时测定酱腌菜中常用防腐剂和甜味剂的研究

目的：建立一种高效液相色谱法同时测定酱腌菜中苯甲酸、山梨酸、脱氢乙酸、糖精钠和安赛蜜的分析方法。方法：样品经超声提取,沉淀蛋白,高速离心后,过0.22 μm微孔滤膜,上机测定。流动相为甲醇和20 mmol/L乙酸铵溶液,等度洗脱,经ZORBAX SB-C18色谱柱分离,于230 nm波长下紫外检测器检测。结果：5种添加剂在15 min内能很好的分离,浓度在1.00~150 mg/L范围内线性很好（R2>99.99%）;平均加标回收率为83.0%~99.9%,精密度为0.06%~5.47%;方法的检出限（S/N=3）和定量限（S/N=10）分别为0.221~1.15 mg/kg、0.736~3.82 mg/kg;方法间比对数据结果呈现高度一致性。结论：该方法简单易操作,准确可靠,能够满足酱腌菜中常用防腐剂和甜味剂的检验需要。     

HPLC法同时测定鸡矢藤中熊果酸和齐墩果酸的含量

目的:建立高效液相色谱法同时测定鸡矢藤中熊果酸和齐墩果酸的含量。方法:色谱柱为DiamonsilTM C18(250×4.6 mm,5滋m),流动相为甲醇-0.05mol.L-1磷酸溶液(93∶7)为流动相,检测波长210 nm,进样量10滋L。结果:熊果酸在21.28滋g.mL-1～212.8滋g.mL-1范围内线性关系良好(r=0.9997),平均回收率为97.79%(RSD=1.30%);齐墩果酸在24.02滋g.mL-1～240.2滋g.mL-1范围内线性关系良好(r=0.9996),平均回收率为98.36%(RSD=1.11%)。结论:该方法简便、灵敏、准确,可用于鸡矢藤药材的质量评价。     

HPLC法测定感冒退热颗粒中连翘苷的含量

采用DiamonsiltmC18(4 6mm× 2 5 0mm ,5 μm)色谱柱 ;以乙腈 水 (2 0∶80 )为流动相 ,流速为 1mL/min ,柱温为 30℃ ,检测波长为 2 77nm ,以外标法用HPLC测定了感冒退热颗粒中连翘苷的含量。连翘苷在0 10 2 9～ 1 6 4 6 4 μg范围内呈线性关系 ,其在制剂中的平均回收率 (n =6 )为 99 5 5 %。     

RP-HPLC法同时测定颐脑解郁颗粒中栀子苷和五味子醇甲北大核心CSCD

本实验建立了颐脑解郁颗粒中栀子苷和五味子醇甲的高效液相色谱测定方法。实验中采用Eclipse XDB-C18色谱柱（250 mm×4.6 mm,5 μm）,流动相甲醇-水,梯度洗脱,体积流量1.0 mL/min,柱温25 ℃,检测波长0-20 min为238 nm,20-40 min为250 nm。结果表明栀子苷在0.2864-2.8640 μg范围内线性关系良好,r=0.9997,平均回收率为98.82%,RSD为1.53%;五味子醇甲在0.1062-1.0620 μg范围内线性关系良好,r = 0.9999,平均回收率为 99.23%,RSD为2.29%;3批样品中栀子苷的平均含量为8.53 mg/g,五味子醇甲的平均含量为2.82 mg/g。该方法简便准确,专属性强,重复性好,可有效控制颐脑解郁颗粒的质量。     

苦玄参药材中苦玄参苷ⅠA和ⅠB的含量分析

采用HPLC法对不同产地的苦玄参药材主成分苦玄参苷ⅠA和ⅠB的含量进行分析,发现苦玄参苷ⅠB含量高于苦玄参苷ⅠA;色谱条件:Waters C18(4.6mmi.d.×250mm,5μm)色谱柱,流动相∶乙睛∶水=36?64(体积比),流速为0.8mL/min,检测波长为264nm。苦玄参苷ⅠA、ⅠB分别在0.05～0.50mg/mL和0.072～0.720mg/mL与峰面积有良好的线性关系。苦玄参苷ⅠA的线性回归方程Y=13658X-92.72(r=0.9993),平均回收率为100.58%(n=5),相对标准偏差(RSD)为1.86%;苦玄参苷ⅠB的线性回归方程Y=5258.9X-41.01(r=0.9994),平均回收率为101.15%(n=5),相对标准偏差(RSD)为1.31%。该研究为制定苦玄参药材质量检测标准提供了简便、快速、准确的方法。     

草苁蓉根、茎水溶性多糖BRT的结构特征

本文以长白山区珍贵野生药用植物草苁蓉为研究对象 .草苁蓉又名“不老草” ,具有滋补强壮、益寿延年之功及补肾壮阳、润肠止血之效 ,为国家三级重点保护植物[1] .近年的研究发现 ,草苁蓉醇提物不仅可以清除体内的游离基 ,而且还可以显著增强机体的免疫能力 ,同时对草苁蓉化学成分的研究也在逐步深入[2 ] ,但对于草苁蓉多糖的系统研究尚未见报道 .为了更全面地认识和利用草苁蓉这一珍贵的植物资源 ,同时也为探讨多糖的结构与功能的关系 ,本文对草苁蓉根、茎的水溶性多糖ＢＲＴ组分进行了结构测定方面的研究 .1　材料和方法1 1　材料为本研究…     

Determination of serine/threonine protein phosphatase type 2B PP2B in lymphocytes by HPLC

Current cyclosporin (CsA) and tacrolimus (FK506) monitoring methods are based on blood concentrations determination, but the optimal sampling times are not clearly defined. An alternative pharmacodynamic approach has recently been introduced, based on assaying lymphocytes activity of calcineurin (PP2B), a phosphatase that is partially inhibited by CsA and FK506. However, the princeps method uses large amounts of radioactive [32P]substrate, raising a number of safety issues. Here we describe an alternative method for PP2B activity determination, based on HPLC coupled with spectrophotometric detection. A 19-amino-acid peptide is phosphorylated by a protein kinase, and further dephosphorylated by lymphocyte PP2B in the presence of okadaic acid. The two peptides are separated by using reverse-phase chromatography with a detection wavelength of 205 nm. After lymphocyte isolation by density-gradient centrifugation, PP2B activity is derived from the dephosphorylated peptide concentration measured during the first 6 min of the enzymatic reaction. This technique gives reproducible results and good analytical sensitivity with 10(6) lymphocytes. With an average isolation rate of 59.6%, only 7 ml of blood is required, making the method suitable for lymphopenic patients. Moreover, PP2B activity is stable in blood samples kept for 24h at room temperature and in isolated lymphocytes stored for 48 h at -20 degrees C. We validated the method by comparing median PP2B activity in 10 healthy volunteers (285.0+/-46.5 pmol/min/10(6)lymphocytes) and in 10 liver transplant patients (147.8+/-71.0 pmol/min/10(6)lymphocytes) (p<0.001). The relation between calcineurin activity and tacrolimus blood concentrations was also studied.     

Purification and physicochemical properties of different polysaccharide fractions from the water extract of Boschniakia rossica and their effect on macrophages activation

Today more and more attentions had been attracted by many nutritionists and pharmacologists on polysaccharides from natural plants or animals due to their significant biological activities. In this research three polysaccharides (BRR-W1, BRR-WA1 and BRR-WA2) were isolated and purified from the water extract of Boschniakia rossica by DEAE Sepharose Fast Flow and Sepharose 6 Fast Flow column chromatography. Chemical and physical characteristics of three polysaccharides were investigated by a combination of chemical and instrumental analysis methods. The assays of their effect on macrophages activation were also investigated in vitro, including phagocytosis of macrophages, detections for NO production and TNF-α secretion. The results indicated that the effect of polysaccharides on macrophages activation was influenced by their respective physicochemical properties.     

HPLC同时测定雪胆属植物中3种活性成分的含量

建立了高效液相色谱法同时测定雪胆属植物中雪胆乙素、竹节参皂苷Ⅳa和雪胆甲素含量的方法,可用于11种雪胆属植物中3种活性成分的同时测定。采用InertSustain C18(250 mm×4.6 mm,5μm)色谱柱;流动相为0.1%磷酸水溶液(A)-乙腈(B),梯度洗脱(0~20 min,5%~20%B;20~60 min,20%~30%B;60~80 min,30%~64%B);流速1.0 mL/min;柱温35℃;检测波长203 nm。经方法学考察,本方法具有准确快速、精密度高、重复性好的特点,雪胆乙素、竹节参皂苷Ⅳa和雪胆甲素分别在0.075~3.000μg、0.810~32.400μg和0.620~24.800μg范围内与峰面积呈现良好的线性关系(r≥0.999 5)。定量分析测得11种雪胆属植物中雪胆乙素含量在0.479~1.904 mg/g范围内,彭县雪胆中含量最高;竹节参皂苷Ⅳa含量在1.098~18.58 mg/g范围内,峨眉雪胆中含量最高;雪胆甲素含量在3.079~17.56 mg/g范围内,十一叶雪胆中含量最高。本方法可为雪胆属优质药用资源开发及综合利用提供参考。     

HPLC法同时测定千斤拔属植物7种黄酮的含量

目的:测定千斤拔属7种植物中的7种黄酮类的含量,为千斤拔类药材质量控制提供依据。方法:色谱条件为Hedera C18色谱柱(250 mm×4.6 mm,5.0μm):流速1.2 mL/min,流动相为0.5%HAc水和乙腈梯度洗脱,柱温35℃,检测波长263 nm。结果:七种黄酮类成分在90 min内达到现行分离,线性关系良好(,R2>0.9995),日内、日间精密度、稳定性和重复性的RSD≤3.0%。该法成功的应用于千斤拔药材7种黄酮的评均回收率分别为96.94%～105.29%(RSD≤2.98%)结论:本方法准确、快速、重现性好,不同品种的千斤拔药材质量存在差异,可以为研究千斤拔药材的质量标准提供依据。     

青海地区不同品种青稞中B族维生素含量分布

利用HPLC法对青海不同产地的15个不同品种青稞中6种B族维生素质量分数进行了测定.结果表明:15个不同品种的青稞,除叶酸(未检出)、VB12(含量较少)外,其余4种所测B族维生素含量均较丰富.其中称多产的二道眉黑青稞VB总量最高,可达0.10842 mg·g-1,民和产的长芒黄脉青稞VB总量最低,为0.03892 m...     

高效液相色谱法测定五味子乙素含量

采用高效液相色谱法,测定了五味子果实中五味子乙素的含量,检测波长为254 nm,标准曲线线性关系R²为0.9907,相对标准偏差为0.48%,平均加样回收率为99.14%。结果表明,此方法准确度较高,为五味子生药及成品药中五味子乙素含量测定提供了一种切实可行的方法。     

湘西蛇足石杉中石杉碱甲、乙和丙含量的测定

采用超声萃取法提取湘西蛇足石杉总生物碱,用高效液相色谱法同时测定其石杉碱甲、乙和丙含量,并分析其在植株不同部位的分布。结果表明:湘西4个样地蛇足石杉中石杉碱甲、乙和丙含量基本一致,分别达到0.5‰、0.3‰和0.04‰;但植株不同部位三种石杉碱含量差异显著,其中石杉碱甲和乙的分布均为叶>茎>根,石杉碱丙则是根大于叶和茎。由此可知,湘西蛇足石杉具有资源优势,石杉碱在植株的分布具有明显的部位选择性;采用HPLC可同时检测石杉碱甲、乙和丙,方法简单快速、准确可靠。     

Determination of acteoside in Cistanche deserticola and Boschniakia rossica and its pharmacokinetics in freely-moving rats using LC-MS/MS

A sensitive LC-MS/MS method with a simple solid-phase extraction for the determination of acteoside in rat plasma and tissue homogenates was established for the investigation of bioavailability and brain distribution in freely-moving rats. Acteoside in Cistanche deserticola and Boschniakia rossica was also determined. Acteoside and internal standard were separated on a RP-select B column (125mmx4.6mm i.d., particle size 5microm). The mobile phase consisted of 35% methanol and 65% acetic acid-water (1:100, v/v) at a flow-rate of 1mL/min. Acteoside and the internal standard were monitored using the multiple-reaction monitoring (MRM) mode at m/z transitions of 623-->161 and 609-->301, respectively. The acteoside content was 38.4+/-2.4mg/kg (n=3) for B. rossica, which is obviously lower than 21134.2+/-805.5mg/kg (n=3) of C. deserticola. The protein binding in rat plasma was 75.5+/-1.8%. The brain distribution result indicated that acteoside was evenly distributed in brain tissues (brain stem, cerebellum, the rest of the brain, cortex, hippocampus and striatum) which was about 0.45-0.68% of that in plasma (4.5+/-0.5microg/mL) after 15min of acteoside administration (10mg/kg, i.v.). After acteoside was given (3mg/kg, i.v.; 100mg/kg, p.o.), the oral bioavailability (AUC(p.o.)/dose(p.o.))/(AUC(i.v.)/dose(i.v.)) was only 0.12%.