从蚕砂中制备脱镁叶绿酸的工艺研究

目的 :优化以蚕砂为原料制备脱镁叶绿酸工艺条件 ,提高产出率。方法 :以叶绿素提取率为指标 ,对脱镁叶绿酸制备的乙醇法和丙酮法工艺进行比较 ,用正交试验法对丙酮法的工艺条件进行优化。结果 :脱镁叶绿酸的最佳制备条件为蚕砂粉碎度60目 ,加1/10倍药材量的水软化5h以上 ,加3倍药材量的溶媒 ,提取温度70℃ ,回流时间60min ,提取次数2次 ,酸化时 pH调至2 5。结论 :软化水用量、丙酮量对叶绿素提取率影响较大 ,经优化后可提高丙酮法制备脱镁叶绿酸的产出率4 6倍     

脱镁叶绿环类衍生物的分离,制备及鉴定

自蚕砂糊状叶绿素分得的叶绿素 a(1),于不同条件下经酸降解、脱羧焦化、甲酯化、氧化,在各步中可得脱镁叶绿素 a(2)、脱镁叶绿酸 a(3),焦脱镁叶绿酸 a(4)、甲基脱镁叶绿酸 a(5)、甲基焦脱镁叶绿酸a(6)、10-羟基甲基脱镁叶绿酸 a(7)和红紫素7-内酯二甲酯(8)。7和8的 ~1HNMR 和 MS 数据为首次报道。     

蚕沙粗品叶绿素酸降解产物的分离与鉴定

从蚕沙粗品叶绿素酸降解产物中分离制得在肿瘤中有选择性潴留作用,光动力学活性强于癌光啉的脱镁叶绿甲酯一酸 a 类组分混合物。经 HPLC 制备分离,~1H-NMR 和 MS 证明,它们为脱镁叶绿甲酯一酸 a(58%),脱镁叶绿甲酯一酸 a′(11%)和焦脱镁叶绿酸 a(31%)三种成分。     

荧光光谱法测定小鼠血浆中异丙酚的浓度

目的应用荧光光谱法测定小鼠血浆中异丙酚的浓度.方法在含不同量异丙酚的小鼠血浆中加入磷酸缓冲液及环己烷/正丁醇,振荡、离心后取上清液测荧光强度(发射波长299 nm,激发波长270 nm).结果异丙酚投入量(0.1～10 mg·L-1)与荧光强度(INT)间有良好的线性关系(r=0.999 6).结论该方法灵敏度高,线性范围大,操作简便,专一性好,可用于临床异丙酚血药浓度的监测.     

蚕沙叶绿素提取及合成脱镁叶绿酸工艺改进研究

目的探讨不同提取工艺对蚕沙中提取叶绿素及制取叶绿素衍生物脱镁叶绿酸的影响,以提高蚕沙叶绿素及叶绿素衍生物的制备量。方法单因素实验设计,以蚕沙软化时间(含水量)、不同提取溶剂为考察条件,优化蚕沙叶绿素提取工艺,并探讨浓盐酸脱镁时间对叶绿素合成脱镁叶绿酸制备率的影响。结果蚕沙软化时间2 h(含水量26%)、丙酮∶乙醇体积比1∶1做溶剂为提取叶绿素的最佳条件,提取率提高到1.43%。以蚕沙叶绿素粗品合成叶绿素衍生物脱镁叶绿酸,浓盐酸脱镁搅拌反应72 h制备率达670 mg.g 1。结论通过改良工艺,提高了蚕沙叶绿素及其衍生物脱镁叶绿酸的提取制备率。     

脱镁叶绿酸的制备及其光敏抗菌活性

以初步纯化的叶绿素为原料,利用叶绿素酶水解、选择性酸降解两种方法制备脱镁叶绿酸。高效液相色谱分析显示两种方法得到的产物图谱相似,酸降解产物中脱镁叶绿酸A的含量较高。抗菌实验表明脱镁叶绿酸对革兰阳性菌具有光敏抑菌效果,对革兰阴性菌和酵母无明显作用。     

维拉帕米片的含量测定及均匀度检查

本文建立了测定维拉帕米片含量及均匀度检查的荧光分光光度法。片剂中维拉帕米用0.1N 盐酸溶媒,作荧光测定,其激发波长为285nm,发射波长为350nm;样品浓度与荧光强度的线性范围为5～25μg/ml。在此条件下其它成份及赋型剂对测定无干扰。平均回收率为99.86±0.99(SD)%,变异系数0.99%。用本法和紫外法测定同一样品,结果基本一致。     

抗凝药物华法林的荧光光谱测定方法研究

目的:基于华法林钠的荧光光谱特性,以水-乙醇混合溶剂为介质,研究建立了一种测定华法林钠的荧光光谱分析方法。方法:水-乙醇的最适宜配比为3:7,该体系的最大激发波长(λ_(ex))和发射波长(λ_(em))分别为308 nm 和386 nm,测定系列标准溶液在测定波长下的荧光强度,并计算样品中华法林钠的含量。结果:在所选的最佳试验条件下,华法林钠浓度在0.3～7.1μg·mL~(-1)范围内与荧光强度具有良好的线性关系,检出限为0.1μg·mL~(-1),回收率为96.7%～104.9%。结论:本方法可应用于药物华法林钠含量的测定。     

响应面分析优化荧光衍生法测定人参中二氧化硫残留量的研究

目的:优化荧光衍生化反应条件,建立荧光分光光度法测定人参中二氧化硫的残留量。方法:采用三维荧光光谱扫描的方法确定试验的最佳激发波长与发射波长;利用响应面分析法(response surface methodology)对影响荧光强度的因素进行优化,在单因素试验的基础上选取实验因素与水平,根据中心组合(Box-Behnken)试验设计原理,采用四因素三水平的响应面分析法确定最佳荧光衍生反应条件。在最佳激发波长与发射波长处测定其荧光强度。结果:确定最佳激发波长为320 nm,发射波长为390 nm。最佳荧光衍生反应条件:缓冲溶液p H为6.7,用量为2.0 m L;1 mmol·L-1的邻苯二甲醛溶液2.1 m L;1.0 m L浓度为11.0 mmol·L-1乙酸铵溶液;静止反应时间为50 min。在该反应条件下,荧光强度与二氧化硫对照品加入量在0.1~0.6μg范围内线性关系良好,r=0.999 2;平均回收率101.7%,RSD为4.1%。测得市售生晒参样品中二氧化硫残留量分别为57.6、39.8、101.8μg·g-1;新采挖鲜人参样品中二氧化硫残留量分别为35.6μg·g-1(集安参地)、33.9 g·g-1(通化参地)、42.6μg·g-1(抚松参地)。结论:该法简便易行,精密度、重复性良好,准确度较高,可为人参及人参相关加工产品中二氧化硫残留量的测定提供参考。     

二氢卟吩e6衍生物的简便制备

蚕沙叶绿素粗提物经酸降解得到脱镁叶绿酸a粗品，不经分离，直接与亲核试剂如甲醇、乙胺发生开环反应，一步得到二氢卟吩e6衍生物。     

A spectrofluorometric method for the determination of ajmaline in plasma.

1 Ajmaline was found to have maximum fluorescence at neutral pH with 300 nm excitation and 365 nm emission wavelengths (corrected). 2 The fluorescence intensity had a linear relationship to concentration up to 50 microgram ml-1 and the recovery of ajmaline after extraction from plasma was 92.5 +/- 3%. 3 Extraction of drug-free plasma and of samples containing known concentrations of ajmaline showed that drug levels in the range found clinically could be measured accurately by fluorimetry. 4 Serial plasma ajmaline concentrations were measured in a subject after intravenous injection of ajmaline (50 mg). The rates of plasma clearance of the drug were found to be similar to those obtained in previous studies.     

HPLC荧光检测法测定人血浆中的特拉唑嗪

目的采用HPLC荧光检测法测定人血浆中的特拉唑嗪。方法采用Apollo C18色谱柱(150 mm×4.6 mm,5μm),流动相为乙腈-50 mmol·L-1磷酸二氢钾缓冲液(22∶78),流速1.1 mL·min-1,柱温30℃,进样量10μL,荧光检测器激发波长为238 nm、发射波长为370 nm。以哌唑嗪为内标,血浆样本碱化后经乙醚-二氯甲烷(3∶2)提取后进样。结果特拉唑嗪的线性范围为0.5～125.0μg·L-1(r≥0.9983);日内、日间RSD均小于10%;平均提取回收率为88.0%±4.4%。结论所用方法简便、准确、灵敏,无杂质干扰,适用于临床上盐酸特拉唑嗪口服制剂的血药浓度监测及药动学研究。     

Fluorimetric assay of tetracycline mixtures.

A simple and precise fluorimetric method is described for the simultaneous assay in plasma of a mixture containing chlortetracycline, demethylchlortetracycline and tetracycline. Assay within the therapeutic ranges of 0-5 mg litre(-1) is achieved by formation of strongly fluorescent aluminum/tetracycline complexes, without prior extraction or separation of the individual antibiotics. This is performed by determinations at the peak excitation and fluorescence emission wavelengths of each tetracycline chelate. The method can be similarly applied to the assay of oxytetracycline, rolitetracycline and minocycline.     

Fluorescence studies of beta-adrenergic ligand binding to alpha 1-acid glycoprotein with 1-anilino-8-naphthalene sulfonate, isoprenaline, adrenaline and propranolol.

The present study shows that ANS (1-anilino-8-naphthalene sulfonate), propranolol, isoprenaline, adrenaline and dopamine have common binding sites on AAG (alpha 1-acid glycoprotein). A fluorescence technique was employed to characterize the interaction between the ligands and AAG at 20-22 degrees. The binding of ANS to AAG caused increased fluorescence intensity at emission and excitation wavelengths of 400 and 470 nm. In this situation, propranolol displaced ANS in a concentration-dependent mode with an apparent dissociation constant of 6.2 +/- 0.01 microM, whereas isoprenaline did not reduce the ANS-AAG fluorescence. However, in the presence of AAG, catecholamines caused a marked increase of fluorescence at excitation and emission wavelengths of 250 and 325 nm, respectively. These wavelengths were employed to characterize the binding of isoprenaline, adrenaline and propranolol to AAG. Two subsets of binding sites were demonstrated. The Kd values were 0.87 +/- 0.03 and 25.1 +/- 10.7 microM for ANS, 0.76 +/- 0.09 and 133 +/- 30.4 microM for propranolol, 140 +/- 14 and 2.18 +/- 0.58 mM for isoprenaline, 137 +/- 24 and 14.8 +/- 0.1 mM for adrenaline, respectively. AAG had identical high affinity binding capacity for these ligands (n approximately 1). However, the second class of binding sites showed ligand-dependent binding capacity: n = 1 for ANS, n approximately 10 for propranolol, n approximately 15 for adrenaline, n approximately 20 for isoprenaline, respectively. ANS, propranolol, dopamine and adrenaline caused concentration-dependent inhibition of isoprenaline binding to AAG with apparent dissociation constants of 5.1 +/- 1.8 microM, 6.4 +/- 1.1 microM, 0.57 +/- 0.13 mM and 1.5 +/- 0.46 mM, respectively.     

蛹油α-亚麻酸乙酯胶丸治疗脂肪肝的临床疗效观察

目的：探讨蛹油α-亚麻酸乙酯胶丸治疗脂肪肝的疗效。方法:将120例脂肪肝患者随机分为两组,一组60例给予基础治疗(对照组),另一组60例在基础治疗的基础上加用蛹油α-亚麻酸乙酯胶丸治疗(治疗组)。结果:对照组有效率68.3%,治疗组有效率95.0%,两组有效率比较有显著性统计学差异(P<0.01),未见不良反应。结论:在基础治疗基础上加用蛹油α-亚麻酸乙酯胶丸治疗脂肪肝疗效显著。     

高效液相荧光法测定人尿液中儿茶酚胺含量

目的：建立高效液相色谱－荧光检测（ HPLC－FD）法同时测定人尿液中儿茶酚胺（去甲肾上腺素、肾上腺素、多巴胺）含量,用于嗜铬细胞瘤的诊断。方法自制活性氧化铝,在弱碱性Tris缓冲液条件下吸附尿液样本中儿茶酚胺,使用 ZORB-AX－SB （4．6 mm ×250 mm,5μm）色谱柱,以甲醇－20 mmol· L－1磷酸二氢钾溶液为流动相;流速：0．8 mL· min－1,柱温25℃;激发波长（EX）为286 nm,检测波长（EM）为318 nm。结果重酒石酸去甲肾上腺素、肾上腺素、盐酸多巴胺在1．56～1000μg· L－1浓度范围内线性关系良好（r＝0．9995、0．9997、0．9998）;日内及日间精密度RSD在0．31％～8．02％之间;高、中、低三个浓度的平均方法回收率在94．22％～108．69％之间。结论该方法准确,稳定性较好,线性范围广,为嗜铬细胞瘤的诊断及围手术期治疗提供依据。     

气相色谱法测定蛹油中α-亚麻酸乙酯的含量

目的：用气相色谱内标法测定蛹油α－亚麻酸乙酯制剂中α－亚麻酸乙酯的含量。方法：采用气相色谱法，DEGS为固定相，涂布浓度为10％，载体为Chromosorb W AW DMCS80-100目的气相柱。结果：平均回收率为97.87%，RSD为1.52%。结论：该法操作简便、准确、重现性、稳定性好，可作为蛹油α－亚麻酸乙酯的含量测定方法。     

Fluorescence determination of sulphobutylether-beta-cyclodextrin in human plasma by size exclusion chromatography with inclusion complex formation

A selective method for the determination of sulphobutylether-beta-cyclodextrin (SBECD) in human plasma has been developed and validated over the range 4-200 microg ml(-1). SBECD is extracted from plasma using end-capped cyclohexyl solid phase extraction cartridges. This is followed by high performance size exclusion chromatography with a mobile phase consisting of 1-naphthol (0.1 mM) in methanol-potassium nitrate (0.2 M) (1:9 v/v), 1 ml min(-1). The high aqueous content of the mobile phase quenches the fluorescence of 1-naphthol. However, the naphthol forms an inclusion complex with SBECD. The non-polar 'bucket' environment of the inclusion region restores the fluorescence, which is measured at excitation and emission wavelengths of 290 and 360 nm, respectively, when SBECD elutes from the column.     

Digital techniques for luminescence detection in liquid chromatography with an intensified linear photodiode array

The use of a rapid-scanning fluorescence detector in liquid chromatography is examined in the context of its potential contribution in pharmaceutical and biomedical analysis. The data generated by an intensified linear photodiode array detector is presented as an isometric plot of (I_em,λ_em,t) at a defined excitation wavelength. Techniques examined for peak homogeneity assessment include: emission spectral normalisation at points through the chromatographic peak profile, second order differentiation (d²I/dt²) and the fluorescence emission ratio chromatogram, generated by calculating the ratio of emission intensifies at two defined emission wavelengths, at all points in the time domain elution profile. These techniques are illustrated with reference to some polynuclear aromatic hydrocarbons and to the β-blocker drug atenolol and its related impurities. Future developments of this new detector technology in LC are also considered.     

HPLC-FLU法测定人血清中硫酸吲哚酚浓度及其在血液透析病人中的应用

目的：建立测定人血清中硫酸吲哚酚浓度的高效液相色谱-荧光检测法,了解血液透析和血液透析滤过对硫酸吲哚酚的清除效果。方法：血清样品经乙腈沉淀蛋白;采用Hypersil BDS C18色谱柱（250mm×4.6mm,5μm）;流动相为乙腈-水-三氟乙酸（15∶85∶0.2,V/V/V）,流速为1mL/min;入射波长为280nm,发射波长为390nm。分别测定尿毒症患者透析前后血清中硫酸吲哚酚浓度。结果：线性范围为0.5~80.0μg/mL,线性方程为Y=2.438×104X＋6.964×103,r=0.9986;日内、日间精密度均小于15%,方法回收率为92.8%~111.4%,提取回收率为90.9%。血液透析和血液透析滤过后患者血清中硫酸吲哚酚的下降率分别为（19.6±7.3）%和（25.3±7.0）%。结论：本法简便、快速、灵敏、准确,可用于测定人血清中硫酸吲哚酚浓度。