Application of high-speed counter-current chromatography coupled with high performance liquid chromatography for the separation and purification of Quercetin-3- O -sambubioside from the leaves of Nelumbo nucifera

Quercetin-3-O-sambubioside [Quercetin-3-O-β-D-xylopyranosyl (1→2)-β-D-glucopyranoside] was separated and purified by semi-preparative high-speed counter-current chromatography (HSCCC) with a two-phase-solvent system composed of ethyl acetate-n-butanol–water (4∶1∶5, v/v) from the leaves of Nelumbo nucifera (Lotus). A total of 5.0mg of the targeted compound with a purity of 98.6% as determined by high performance liquid chromatography (HPLC) was obtained from 100m g of the crude extract cleaned up by AB-8 macroporous resin in a one-step separation. Quercetin-3-O-sambubioside was a novel flavonoid glycoside from the leaves of Nelumbo nucifera, and its chemical structure was identified by means of ESI-MS, 1D NMR and 2D NMR.     

Scale-up purification for rutin hyrdrolysates by high-performance counter-current chromatography coupled with semi-preparative high-performance liquid chromatography

The study made the isolation of three constituents from rutin and exemplified how to achieve quercetin separation and quantification in rapid and scale-up processes by high-performance counter-current chromatography (HPCCC). Meanwhile, we also isolated the other two constituents, kaempferol and isorhamnetin, by semi-preparative high-performance liquid chromatography. After systematic optimization of solvent systems, sample concentration and flow rate on analytical Mini-DE centrifuge, the same conditions were scale-up by 50-fold in preparative Midi-DE centrifuge to purify quercetin, and the maximum sample loading achieved 1.65 g. Eventually, we successfully isolated quercetin, kaempferol and isorhamnetin with the purity of 99.80%, 99.84% and 99.95%, respectively. The process, therefore, provided an efficient method of obtaining sufficient quantities of highly purified quercetin, as well as kaempferol and isorhamnetin.     

Separation and purification of four tannins from Potentilla parvifolia Fisch. (Rosaceae) using high-speed counter-current chromatography

Four tannins were isolated and identified from Potentilla parvifolia using high-speed counter-current chromatography (HSCCC) in this study. A two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (1:11:1.2:11, v/v/v/v) was chosen and yielded 18.8 mg of procyanidin B3, 9.8 mg of procyanidin B6, 9.1 mg of procyanidin B7, and 6.9 mg of 2,3,4,6-tetra-O-galloyl-D-glucose, all at over 95% purity, from 120 mg of the crude sample powder. The identifications were performed with ¹H NMR and ¹³C NMR. This is the first report showing the presence of procyanidin B7 and 2,3,4,6-tetra-O-galloyl-D-glucose in the genus Potentilla, and procyanidin B3 and procyanidin B6 were isolated from P. parvifolia for the first time.     

高效液相色谱法分离2-羟基-3-甲氧基-3,3-二苯基丙酸对映体

建立了重要医药中间体2-羟基-3-甲氧基-3,3-二苯基丙酸对映体的高效液相色谱分离方法。采用手性色谱柱在正相条件下直接拆分2-羟基-3-甲氧基-3,3-二苯基丙酸对映体,考察了固定相种类、流动相中异丙醇比例、柱温、流速等对2-羟基-3-甲氧基-3,3-二苯基丙酸对映体分离的影响。最终确定其拆分条件为:色谱柱为Chiralpak ID(250 mm×4.6 mm,5.0μm),流动相为正己烷-异丙醇-三氟乙酸(97∶3∶0.1,v/v/v),流速为1.0 mL/min,检测波长为220 nm,柱温为25℃;在此条件下2-羟基-3-甲氧基-3,3-二苯基丙酸对映体的分离度为2.3。R构型的检测限、定量限分别为0.5μg/mL,1.0μg/mL,其在1～100μg/mL内线性关系良好(r=0.999 6)。平均回收率为98.4%～104.5%,峰面积的相对标准偏差为1.5%。3批中间体样品中R构型杂质含量均在合格范围内。所建立的分析方法简单快速,重现性好,可用于(S)-2-羟基-3-甲氧基-3,3-二苯基丙酸的异构体检测。     

芳香醇类化合物高效液相色谱法的分离研究

采用高效液相色谱法对15种芳香醇及结构相近化合物进行了分离研究。在优化的色谱条件下,8种对映体和4种非手性芳香醇同系物得到了基线分离。与单纯的紫外检测器相比,旋光检测器可方便地得到手性对映体的左旋或右旋特性。有关分离机理得到了合理的解释。     

固相萃取-高效液相色谱法测定化妆品中的丙烯酰胺

建立了保湿防晒系列化妆品中丙烯酰胺含量的高效液相色谱测定方法.采用丙酮超声提取样品中的待测物,提取液经浓缩吹干后用水溶解,经固相萃取小柱净化后用反相高效液相色谱法测定.丙烯酰胺在0.01 mg/L～5.0 mg/L与对应峰面积呈良好线性关系(R=0.999 5).在添加量为0.05 mg/kg,0.50 mg/kg和1.00 mg/kg时,平均回收率在81.3%～83.8%,变异系数在2.3%～3.0%,方法检出限为0.05 mg/kg.结果表明,该法操作简便、灵敏、准确,适用于化妆品中丙烯酰胺含量的测定.     

Separation and purification of six isoflavones from Iris tectorum Maxim by macroporous resin-based column chromatography coupled with preparative high-performance liquid chromatography

ABSTRACT

A rapid and robust preparation method of six isoflavones from ethanol aqueous extract of Iris tectorum Maxim (I. tectorum) was established by using macroporous resin column chromatography and preparative high-performance liquid chromatography (Prep-HPLC). After separation by AB-8 resins, total flavonoids content increased from 10.60% in the crude extract to 54.20% with a recovery yield of 75.12%. Subsequently, the extracts were purified by Prep-HPLC, and the purities were more than 82.2% after assessment by analytical HPLC and characterization by mass spectrometry. The established method is expected to be used for preparing available quantities of pure isoflavones from I. tectorum.     

高效液相色谱-串联质谱测定豆芽中6-苄氨基嘌呤方法的研究

建立了豆芽中的6-苄氨基嘌呤的高效液相色谱-串联质谱法（HPLC-MS/MS）定量定性分析方法.样品经酸化甲醇2次提取后,在高效液相色谱-串联质谱仪（HPLC-MS/MS）选择反应监测（MRM）模式下测定.采用质谱定性,外标法定量.色谱柱为资生堂MGⅡ-C18（5μm2.1 mm ×150mm）色谱柱,以0.1％甲酸水-0.1％甲酸甲醇为流动相进行梯度洗脱,流速0.3 mL/min.在优化的实验条件下,得到了较宽的线性范围和较低的定量检出限：线性范围为0.1～20.0μg/L,线性相关系数在0.999以上,定量限为：7.3 μg/kg.方法的回收率和重现性较好,回收率为98.3％～106.9％之间,相对标准偏差（RSD）在1.16％～3.56％之间.该方法操作简单高效、灵敏度高、结果准确可靠,可应于豆芽中6-苄氨基嘌呤的定量及确证分析.     

高效液相色谱质谱联用鉴别口服疫苗中硫柳汞降解产物

目的采用高效液相色谱质谱联用的方法鉴别口服疫苗中硫柳汞的降解产物。方法采用C18色谱柱,甲醇-醋酸铵缓冲溶液(pH 4.5)梯度洗脱,以紫外和质谱检测器进行检测,对某口服疫苗中的未知色谱峰进行鉴别。结果疫苗样品中两个色谱峰的保留时间及其一级二级质谱图与硫柳汞的降解产物硫代水杨酸以及2,2’-二硫代二苯甲酸基本一致。结论高效液相色谱质谱联用可鉴别口服疫苗中的硫柳汞降解产物。     

高效液相色谱法在评估化妆品气垫海绵中的应用

采用高效液相色谱法测定气垫CC霜中二乙氨羟苯甲酰基苯甲酸己酯、甲氧基肉桂酸乙基己酯和水杨酸乙基己酯3种防晒剂在海绵内和挤出料体中的含量,通过比较海绵吸附性的差异来评估海绵质量。样品经甲醇提取,以Zorbax SB C_(18)色谱柱为固定相,柱温30℃,以乙腈-水(体积比90∶10)为流动相等度洗脱,经二极管阵列检测器测定。二乙氨羟苯甲酰基苯甲酸己酯和甲氧基肉桂酸乙基己酯在1~100 mg·L~(-1)范围内线性良好,水杨酸乙基己酯在5~500 mg·L~(-1)范围内线性关系良好,样品加标回收率为91.8%~95.7%,6组平行样测定得出3种防晒剂峰面积相对标准偏差为1.51%~4.65%。结果表明,丁腈橡胶海绵和聚氨酯海绵对防晒剂的吸附差异较大。丁腈橡胶海绵的吸附率在16.5%~53.4%,而聚氨酯海绵的最大吸附率为7.5%。     

Application of high performance liquid chromatography to the analysis of some non-volatile coffee components

High performance liquid chromatography (HPLC) was applied to the analysis of caffeine, trigonelline, nicotinic acid and sucrose in Arabica and Robusta coffee. Green and roasted coffee samples were used in this study and the degradation of sucrose and trigonelline, with the formation of nicotinic acid, was followed during roasting. Caffeine did not undergo significant degradation with only 5.4% being lost under severe roasting. Sucrose was degraded rapidly during processing with light roasting producing a 97% loss and dark roasting degrading it completely. Loss of trigonelline was strongly dependent on the degree of roasting, being higher in the Robusta coffee. Trigonelline degradation was associated with nicotinic acid formation both in the Arabica and Robusta coffees as a consequence of the roasting process. Trigonelline and sucrose were determined simultaneously by partition chromatography and detection with the mass detector. Determination of caffeine was carried out using reversed phase chromatography and nicotinic acid by ion-pair reversed phase chromatography. Detection in both cases was achieved using an ultraviolet detector at 272 nm or 254 nm, respectively. HPLC showed adequate precision and accuracy for routine analyses. In addition, the methods used were more rapid and simple than traditional procedures. HPLC appears to be a suitable technique for quality control in the coffee industry, and for fundamental investigation on the mechanisms involved in the roasting process.     

高效液相色谱法用于毒死蜱测量不确定度的评定

采用高效液相色谱法分析毒死蜱的含量,在标准曲线的绘制、标准样品的使用、测定结果的输出等方面建立数学模型,监控检测结果的质量,对实验室的检验系统测量不确定度进行评定,从而确认了测量数据的准确性和可靠性。     

Magnetic molecularly imprinted polymer nanoparticles coupled with high performance liquid chromatography for solid‐phase extraction of carvedilol in serum samples

Herein, we report a magnetic molecularly imprinted polymers (m‐MIPs) using Fe₃O₄ as a magnetic component, carvedilol as a template molecule for the solid‐phase extraction (MISPE) as the sample clean‐up technique combined with high‐performance liquid chromatography (HPLC) and for the controlled release of carvedilol at different pH values of 1.0 (simulated gastric fluid), 6.8 (simulated intestinal fluid), and 7.4 (simulated biological fluid). The adsorption kinetics was modeled with the pseudo‐first‐order and pseudo‐second‐order kinetics, and the adsorption isotherms were fitted with Langmuir and Freundlich models. The performance of the m‐MIPs for the controlled release of carvedilol was assessed and results indicated that the magnetic MIPs also have potential applications in controlled drug release. Furthermore, the m‐MIPs were applied to the extraction of carvedilol from human blood plasma samples. Carvedilol can be quantified by this method in the 2–350 μg L^?1 concentration range. The limit of detection and limit of quantification in plasma samples are 0.13 and 0.45 μg L^?1. The results from HPLC showed good precision (3.5% for 50.0 μg L^?1) and recoveries (between 85 and 93) using m‐MIP from human plasma samples. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 41209.     

制备色谱分离纯化桑叶提取物的研究

选取桑叶中高附加值的活性成分异槲皮苷,以异槲皮苷制备率和纯度为评价指标,应用制备色谱技术,考察流动相体系及比例、流速和进样量等影响因素对异槲皮苷分离纯化的影响,通过分析、检测制备样品成分,开展高纯度异槲皮苷制备的效益分析。结果表明,异槲皮苷分离纯化的最优条件为:V_(乙腈)∶V_水=20∶80,进样量10 mL,流速20 mL/min时,异槲皮苷制备率26.36%,纯度为98.78%;同时可得绿原酸制备率30.37%,纯度为99.48%,芦丁制备率19.51%,纯度为98%和紫云英苷制备率11.47%,纯度为98%,为桑叶资源化及高值利用提供实验支撑。     

制备色谱分离纯化桑叶提取物的研究

选取桑叶中高附加值的活性成分异槲皮苷,以异槲皮苷制备率和纯度为评价指标,应用制备色谱技术,考察流动相体系及比例、流速和进样量等影响因素对异槲皮苷分离纯化的影响,通过分析、检测制备样品成分,开展高纯度异槲皮苷制备的效益分析。结果表明,异槲皮苷分离纯化的最优条件为:V_(乙腈)∶V_水=20∶80,进样量10 mL,流速20 mL/min时,异槲皮苷制备率26.36%,纯度为98.78%;同时可得绿原酸制备率30.37%,纯度为99.48%,芦丁制备率19.51%,纯度为98%和紫云英苷制备率11.47%,纯度为98%,为桑叶资源化及高值利用提供实验支撑。     

A mathematical model for the prediction of triglyceride molecular species by high performance liquid chromatography

The physicochemical and theoretical relationships between the traditional equivalent carbon number (ECN) and a proposed elution theory for triglyceride molecular species that has been expressed as a matrix model were demonstrated by multiple regression analysis. It was concluded that the ECN expression has two independent variables, that is, the total acyl carbon number (CN) and the total double bonds (DB), and one dependent variable, the relative retention time (RRT). In the proposed elution equation, there are six independent variables, including the carbon numbers and number of double bonds in each acyl group to be considered.     

高效液相色谱-二极管阵列检测法测定马来酸和乙醛酸

建立了高效液相色谱-二极管阵列法同时测定乙醛酸和马来酸的高效液相色谱方法。色谱条件为:Shim-pack VP.ODS柱(250 mm×4.6 mm I.D.,5μm);二极管阵列检测器;流动相为甲醇/磷酸盐缓冲溶液(NaH2PO4浓度为0.1 mol/L,H3PO4调节至pH=2.98),体积比为5∶95,流速为1.0 mL/min,检测波长为195 nm,柱温为40℃。线性范围分别为79.6～26 660 mg/L和9.3～7 864 mg/L,检出限分别为39.4 mg/L和4.7 mg/L;日内、日间测定的相对标准偏差(RSD):乙醛酸为0.22%和0.46%,马来酸为0.02%和0.03%。     

高效液相色谱法测定微量季铵盐的改进方法

利用季铵盐与间硝基酚反应生成溶于有机溶剂的络合物,该络合物可以用紫外检测器进行定量分析,建立了一种高效液相色谱测定微量季铵盐的改进方法.该方法的线性范围为0.60～99.40 mg/L,最低检出限可达0.4mg/L,平均回收率为94.97％～104.02％,相对标准偏差为3.41％.该方法稳定、简便且灵敏度高,适于测定...     

反相高效液相色谱DAD法测定尿囊素及其相关物的含量

为了确定高效液相色谱法测定尿囊素及其相关物含量的方法,采用流动相超声溶解样品,用ZORBAX SB-Aq分离,流动相为A(甲醇与0.2mol/L磷酸水溶液体积比1:9)-B(超纯水),梯度洗脱,在190nm波长下检测,外标法定量,测定尿囊素及其相关物的含量。结果表明:尿素质量浓度在0.2～5mg/mL内呈良好的线性关系,检测限为20μg/mL;甘脲在0.1～2.5mg/mL内呈良好的线性关系,检测限为2μg/mL;缩二脲在0.2～5mg/mL内呈良好的线性关系,检测限为4μg/mL。该方法快速、灵敏、准确,可同时测定尿囊素、尿素、甘脲及缩二脲的含量。     

3D printing of NiZn ferrite architectures with high magnetic performance for efficient magnetic separation

Magnetic materials with desired architectures and high performance have become increasingly important for applications. In this work, NiZn ferrites with mesh structures have been successfully fabricated from preliminary precursors by three-dimensional (3D) printing accompanied by the solid-state reaction. The influence of compositions on structural and magnetic properties of NiZn ferrites were systematically investigated. It was obvious that the saturation magnetization was significantly enhanced to 77 emu/g for NiZn ferrites with a zinc concentration of 0.4, which was a much higher value compared to Ni ferrites. This is attributed not only to the well-known cationic distribution, but also to the grain growth with extraordinarily high crystallinity. Consequently, NiZn ferrites with intricate mesh structures were utilized for magnetic separation, and the distributions of magnetic flux density were simulated. Overall, the fabrication of NiZn ferrites by 3D printing is attractive for scaled-up applications, and also paves the promising avenue for magnetic separation.