HPLC 法测定人肝微粒体中藏花酸的浓度

目的：建立一种测定人肝微粒体孵育体系中藏花酸浓度的 HPLC 内标法。方法在人肝微粒体温孵体系中加入藏花酸样品进行孵育反应,反应结束后用冰乙腈终止反应。经沉淀法处理样品后,采用 Diamonsil C18色谱柱（4．6 mm×150 mm,5μm）进行分离。流动相：甲醇-2％冰乙酸水溶液（7525,v／v）;流速：1．0 mL·min－1;柱温：30℃;紫外检测波长：440 nm;内标：吲哚美辛。结果藏花酸经孵育反应后质量浓度测定方法的线性范围为0．125～8μg·mL－1,线性关系良好（R 2＝0．9999）。藏花酸低、中、高质量浓度的提取回收率分别为：97．24％、96．58％、98．12％,其 RSD 均小于15％,批内、批间差异＜15％,准确度为85％～115％。结论该 HPLC 法简便、准确,符合生物样品测定的要求,能快速、可靠地检测人肝微粒体中藏花酸的浓度。     

西红花酸对阿霉素所致大鼠心肌线粒体损伤的影响

目的: 研究西红花酸对阿霉素所致大鼠心肌线粒体损伤的保护作用。方法: 建立阿霉素致大鼠心脏毒性模型, 观察西红花酸对心肌线粒体膜电位、线粒体DNA 断裂程度、细胞色素C 氧化酶活性及其亚基IImRNA 表达的影响;测定心肌线粒体超氧阴离子含量及谷胱甘肽过氧化物酶(GSH-PX)活性。结果: 与模型组相比, 西红花酸可明显升高线粒体膜电位, 降低线粒体DNA 断裂程度, 提高细胞色素C氧化酶活性及其亚基IImRNA 表达水平, 显著降低心肌线粒体超氧阴离子含量, 提高GSH-PX 活性。结论: 西红花酸能明显减轻阿霉素所致大鼠心肌线粒体损伤。     

Separation and Purification of Gardecin from Gardenia jasminoides Ellis by High-Speed Countercurrent Chromatography

In the present study, a high-speed counter-current chromatography (HSCCC) was investigated for separation and purification of gardecin on a preparative scale. Hexane–ethyl acetate–methanol–water (0.3:1:0.3:1, v/v) was selected as the optimum solvent system to purify gardecin from a fraction obtained from HPD100 column chromatography fractionation of gardenia. After HSCCC isolation, 20.1 mg of gardecin with purity of 97.4% was obtained from 322 g of dry gardenia fruits. Chemical structure identification of this pigment was carried out by MS, ¹H NMR, and ¹³C NMR. Additionally, antioxidant activity of gardecin in comparison with crocin-1 was investigated. The present results demonstrated that gardecin could be efficiently obtained using HSCCC from this herb and this compound features strong antioxidant activity.     

Screening method for the detection of artificial colours in saffron using derivative UV-Vis spectrometry after precipitation of crocetin

A screening method for the detection of artificial colours (naphthol yellow, tartrazine, quinoline yellow, Sunset yellow, Allura red, amaranth, azorubine, Ponceau 4R and Red 2G) in saffron is described. The method involves removal of crocins by precipitation of crocetin (pH 0.1, 90°C) before adsorption of the artificial colours on polyamide SPE cartridges (pH 2). After washing with methanol, acetone and methanol, elution was done with a methanol:ammonia solution (95:5 v/v), and detection was performed by derivative spectrometry. Sample pretreatment changes the UV-Vis saffron extract profile in such a way that second derivative spectra can be used to identify the presence of added colours. Erythrosine, which was found to be pH dependent, could not be detected under the above conditions. The lowest detectable amount for each colour was strongly dependent on chemical structure. The recovery of carminic acid was very low possibly due to irreversible retention on the polyamide. This procedure can replace the current ISO TLC method (2003) and be used alternatively or in combination with HPLC procedures adopted in the same standard.     

Crocetin from saffron: an active component of an ancient spice

The known properties of saffron (Crocus sativus, L.) and its components have been examined. Recently, hormone like effects in green algae and the anti-cancerogenic and anti-toxic effects, have been observed. In particular, the effects of crocetin, a carotenoids (8,8'-diapo-8,8'-carotenoic acid) present in saffron and characterized by a diterpenic and symmetrical structure with seven double bonds and four methyl groups, have been taken into consideration. It has been found that this compound enhances the oxygen diffusivity through liquids, such as plasma. As a consequence of this property, it has been observed that crocetin increases alveolar oxygen transport and enhances pulmonary oxygenation. It improves cerebral oxygenation in hemorrhaged rats and positively acts in the atherosclerosis and arthritis treatment. It inhibits skin tumor promotion in mice (i.e., with benzo(a)pyrene); it has an inhibitory effect on intracellular nucleic acid and protein synthesis in malignant cells, as well as on protein-kinase-C and prorooncogene in INNIH/3T3 cells. This is most likely due to its anti-oxidant activity. Furthermore, crocetin protects against oxidative damage in rat primary hepatocytes. It also suppresses aflatoxin B1-induced hepatotoxic lesions and has a modulatory effect on aflatoxin, B1 cytotoxicity, and DNA adduct formation on C3H10/T1/2 fibroblast cells. It also has a protective effect on the bladder toxicity, induced by cyclophosphamide. The experiments reported in the scientific literature and the interesting results obtained have been carried out in vitro or on laboratory animals, but not yet on man.     

Effects of mild temperature conditions during dehydration procedures on saffron quality parameters

BACKGROUND: The dehydration procedure is responsible for saffron sensorial properties: colour, taste and aroma. Changes in the compounds responsible for these characteristics have been studied when dehydration processes at high and low temperature are employed. However, the evolution of these changes at mild temperatures is not available in the current bibliography. In this paper the effect of different mild conditions (18–20 °C for 24 h, 40–50 °C for 75 min and 55 °C for 75 min) applied to 45 saffron samples with the same origin was investigated. RESULTS: Crocetin esters, the compounds responsible for saffron colour, increased their content with no significant differences from other processes when high temperatures (55 °C) were used, thus producing a noticeable increment in saffron colouring capability. Similar behaviour was obtained for picrocrocin, the compound responsible for saffron taste, with higher average content at the highest temperature (55 °C) but without significant differences with the inferior conditions (40–50 °C). However, more volatile compounds were generated, especially safranal,at higher temperatures, e.g. 55 °C, during the dehydration procedure. CONCLUSIONS: The results found support the idea for employing mild to high temperatures during the dehydration process of saffron. Copyright © 2010 Society of Chemical Industry     

高效液相色谱法测定糕点中的栀子黄

目的改进GB/T5009.149-2003《食品中栀子黄的测定第一法高效液相色谱法》的方法。方法样品经甲醇:甲酸(6:4,V:V)洗脱,采用Angilent Extend C_(18)反相色谱柱(4.6 mm×250 mm,5μm),以甲醇:0.2%氨水(50:50,V:V)为流动相(等度洗脱),流速为1.0 mL/min,进样量20μL,经配有二极管阵列检测器(DAD)的高效液相色谱仪(high performance liquid chromatograph,HPLC)进行测定。结果本方法检出限为3.2 mg/kg,栀子黄中藏花酸和藏花素峰面积的相对标准偏差分别为1.6%、2.5%,在5~50μg/mL范围内有较好的线性,Y=1.34X-0.54,r~2=0.9998,不同浓度的回收率在91.2%~98.4%之间。结论本方法简单、高效、准确度高,回收率和重现性良好,适用检测的食品类别范围较宽,具有推广应用价值。