HPLC法同时测定人血浆中帕罗西汀和喹硫平浓度

目的 建立同时测定人血浆中帕罗西汀和喹硫平浓度的高效液相色谱( HPLC)法.方法 采用HPLC法进行测定:Inertsil ODS-C18柱(4.6×150mm,5μm)为色谱柱,流动相为乙腈-磷酸二氢钠(0.05 mol/L磷酸二氢钠)(39∶61),流速为1.0mL/m in,检测波长为210nm,柱温为40℃,以乙酸乙酯-二氯甲烷(体积比75∶25)为萃取剂.结果 帕罗西汀在10～640μ g/L、喹硫平在20 ～ 1200μg/L范围内峰面积与浓度呈良好线性关系,检测限分别为5μg/L,8μg/L.结论 该方法简单、快速、灵敏、准确,可用于临床帕罗西汀与喹硫平的血药浓度监测和药动学研究.     

HPLC测定百炎净合剂中磺胺甲噁唑和甲氧苄啶含量

目的建立用高效液相色谱法(HPLC)对医院制剂百炎净合剂中磺胺甲恶唑(SMZ)和甲氧苄啶(TMP)进行含量测定。方法用色谱柱Waters Spherisorb ODSI 5μm(250*4.6mm)No.2104390911D,以水-三乙胺-乙腈(800:1:200)(以冰醋酸调节pH5.3)为流动相,检测波长为230nm,流速1.0ml/min。结果两种成分能很好分离;SMZ和TMP的线性范围分别为8～64μg/mL(r=0.9992,n=5),1.6～13ug.mL(r=0.9999,n=5);平均回收率分别为99.7%(RSD=1.6%,n=5)和99.6%(RSD=1.8%,n=5)。结论方法简便,快速准确,重现性好,适用于该复方制剂中两种有效成分含量的同时测定。     

左旋加替沙星的高效液相色谱测定方法的建立

目的建立左旋加替沙星的HPLC测定方法。方法色谱柱:UhimateXB-C18柱(5μm,4.6 mm×250 mm);流速:1.0 mL/min;柱温:29℃;流动相:甲醇:0.08 mol/L磷酸二氢钾溶液为40:60,磷酸调节pH为3.87);检测波长:285nm;进样量:20μL。结果左旋加替沙星在浓度为10.3～51.5 μg/mL时线性关系良好,r=0.9995,平均回收率为98.0%;RSD为1.2%(n=6)。保留时间为9.887。结论该方法简单、准确,可作为开发左旋加替沙星制剂的质量控制。     

反相高效液相色谱法测定兔血清中5-氟尿嘧啶的浓度

目的应用反相高效液相色谱法测定兔血清中5-氟尿嘧啶的浓度。方法兔血清样品经乙酸乙酯/异丙醇(84/16)萃取后,水浴氮气吹干,流动相定容后进样。色谱柱为kromasil-C18柱(250mm×4.6mm,5μm),流动相为20mM醋酸铵/乙腈(体积比98.2∶1.8,pH8.05),流速0.8mL/min,紫外检测波长为270nm。结果5-氟尿嘧啶在5～80μg/L浓度范围内呈良好的线性关系。10、20、40μg/L三个浓度的批内RSD分别为11.6%、5.6%、5.4%;批间RSD分别为12.8%、2.7%、6.2%;平均回收率分别为106.7±11.7%、98.0±5.3%、99.3±5.3%。结论本方法可靠、重复性好,具有特异性强、灵敏、精密度与准确度高等特点,满足5-氟尿嘧啶药代动力学研究的要求。     

HPLC法测定感丹中三组分的含量及马来酸氯苯那敏含量均匀度

目的建立高效液相色谱法测定感丹(氨酚伪麻那敏泡腾颗粒)中马来酸氯苯那敏、对乙酰氨基酚、盐酸伪麻黄碱的含量及马来酸氯苯那敏含量均匀度。方法采用Agela Venusil XBP-C_(18)柱(4.6mm×250mm,5μm),流动相为甲醇-0.05mol.L~(-1)磷酸二氢钾溶液(10%磷酸调pH至3.0)(25:75),流速为1.0mL.min~(-1),检测波长为210nm,进样量10μL,柱温为35℃。结果马来酸氯苯那敏浓度在0.16~0.56μg.mL~(-1)之间、对乙酰氨基酚浓度在2.0~7.0μg.mL~(-1)之间、盐酸伪麻黄碱浓度在0.24~0.84μg.mL~(-1)之间均与峰面积呈良好线性关系,r=0.999 0~0.999 8;平均加样回收率分别为101.62%(RSD=1.26%)、97.04%(RSD=0.98%)、95.02%(RSD=2.18%)。结论本法操作简便,结果准确,可作为该制剂的质量控制方法。     

高效液相色谱法测定赤芍中芍药苷的含量

目的 建立赤芍中芍药苷的含量测定方法,探讨不同溶液提取对芍药苷含量的影响.方法 采用甲醇和70%乙醇超声提取样品,用Agileni1100型高效液相色谱仪,Agilent EeliPse XDB C18(4.6mm×250mm,5靘)色谱柱,柱温:25℃,流动相:V(乙腈):V(0.1%磷酸溶液)=14:86,流速:1.0ml/min,检测波长:230nm,进样体积10μL.结果 芍药苷进样量在0.27μg～1.60μg范围内与峰面积呈良好线性关系,R=0.9998;平均回收率为100.34%,RSD=0.46%.乙醇提取液中芍药苷的平均含量为0.0700μg/ml(n=3),甲醇提取液中芍药苷的平均含量为0.0462μg/ml(n=3).结论 采用70%乙醇作为溶液提取赤芍芍药苷得率高;本实验所建立的方法重复性良好,准确度较高,可用于赤芍中芍药苷的质量控制.     

HPLC法同时测定益心舒片中丹参酮ⅡA的含量

目的：建立采用 HPLC法测定益心舒片中丹参酮ⅡA含量的方法。方法采用 KromasilC18色谱柱(4.6 mm×250 mm,5 um),以甲醇原水(82：18)水梯度洗脱,流速为1 mL/min,柱温30℃；紫外检测器波长为270 nm。结果丹参酮ⅡA在2.556~51.12μg/ml范围内呈良好的线性关系(r=0.9998)。其平均回收率为97.58%,RSD为0.6%(n=9)。结论本方法灵敏高、简便、重复性好,可用于益心舒片中丹参酮ⅡA的含量测定。     

HPLC法测定利培酮片有关物质的优化

目的:建立HPLC梯度洗脱法测定利培酮片的有关物质。方法:采用Sepax GP-C18(4.6×100mm,3μm)为色谱柱;以流动相A:水-乙腈-三氟乙酸(80:19.5:0.1)(用氨水调节pH值至3.0)与流动相B:水-甲醇-三氟乙酸(61:39:0.1)(用氨水调节pH值至3.0),进行梯度洗脱;柱温35℃,流速1.0 mL·min~(-1),检测波长275nm。结果:利培酮在0.5088μg·ml~(-1)~1017.6μg·ml~(-1)浓度范围内与相应峰面积线性关系良好;检测限(S/N=3)为0.5088ng·mL~(-1)。结论:优化后的方法更有利于利培酮片杂质的检出。     

HPLC法测定六味地黄胶囊中马钱苷含量

目的：建立六味地黄胶囊中马钱苷含量测定方法。方法采用Phenomenex色谱柱(250mm×4.6mm,5μm),流动相为乙腈-0.1%磷酸(8：92);流速1.0ml·min-1,柱温30℃,检测波长为236nm。结果马钱苷在11.25μg/ml~180.0μg/ml范围内,呈良好的线性关系(r=0.9993),定量限为7.88ng·ml-1,回收率为97.1%,RSD为1.2%(n=9)。结论本方法简便、准确。方法的稳定性、重复性良好,可作为六味地黄胶囊质量分析控制方法。     

体积排阻高效液相色谱法测定酸溶性Ⅰ型胶原蛋白含量的研究

目的 建立酸溶性I型胶原蛋白含量测定的体积排阻高效液相色谱法（size-exclusion high-performance liquid chromatography,SE-HPLC）并进行方法确认，为快速、准确测定酸溶性I型胶原蛋白含量提供技术参考。方法 通过优化流动相组成、流速及柱温等因素来改善峰形，提高柱效，确定最佳检测条件，采用含DAD检测器的Agilent 1260高效液相色谱仪进行检测，检测波长为210 nm，外标法定量并进行方法学研究。结果 选择300 mmol/L磷酸钠-150 mmol/L氯化钠（pH=6.0）为流动相，设定流速为0.4 mL/min，柱温为20℃时，酸溶性I型胶原蛋白在体积排阻色谱柱Agilent Bio SEC-5(300?，4.6 mm×300 mm,5μm)(1?=0.1 nm）上的分离能获得最佳柱效。方法学研究结果表明，酸溶性I型胶原蛋白在1.5～120.0μg/mL范围内，与峰面积呈良好的线性关系（r=0.999 9），定量限为0.6μg/mL。用建立的方法连续进样6次，检测供试品溶液的I型胶原蛋白含量，相对标准偏差（relative standard deviation,RSD）为1.1%；加标回收率为92.8%～101.3%。结论 该方法简便、快速，准确度、精密度和稳定性良好，可用于制备医疗器械产品的酸溶性I型胶原蛋白含量的测定。     

Localization of 5-HT2A, 5-HT3, 5-HT5A and 5-HT7 receptor-like immunoreactivity in the rat cerebellum

Although serotonin (5-hydroxytryptamine, 5-HT) is known to exert a modulatory action on cerebellar function, our current knowledge of the nature of receptor subtypes mediating serotonergic activity in this part of the brain remains fragmentary. In this study, we report the presence and distribution of 5-HT3, 5-HT5A and 5-HT7 receptor-like immunoreactivity in the rat cerebellum using immunofluorescence histochemistry. 5-HT3 immunoreactivity was found in fibers sparsely distributed throughout the cerebellum. Most of them were seen in the cerebellar cortex as fine varicose 5-HT3-positive axonal processes. 5-HT5A immunoreactivity, on the other hand, was observed in neuronal somata of the cerebellar cortex and deep cerebellar nuclei. Based upon cell morphology and the use of cell-specific markers, Purkinje cells, molecular layer interneurons and Golgi cells were found to be 5-HT5A immunopositive. In addition, the use of cell-specific markers allowed us to identify previously reported large 5-HT2A-positive cells in the granular layer as being Golgi cells. Finally, 5-HT7 immunoreactivity was observed only in Purkinje cells. Corroborating previous radioligand-binding, in situ hybridization and immunohistochemical studies, our data relate serotonin receptor subtypes to specific cerebellar cell types and may consequently contribute to the elucidation of serotonergic actions in the cerebellum.     

Case 5

A 53-year-old man with a heavy exposure to asbestos 25 years earlier was admitted to the East Orange Veterans Administration Medical Center with a rapidly enlarging chest wall mass of 4-5 weeks duration.

The mass, overlying the ninth and tenth ribs in the infraaxillary area, was firm. On X-ray the ribs appeared eroded and the mass was seen to extend into the pleural space. A chest wall resection was performed, and the tumor was sharply dissected from the diaphragm to which it was firmly adherent. Grossly, it measured 7.5 X 5.0 X 3.0 cm and was yellow-tan with homogeneous and nodular areas The inferior aspect of the ninth rib was obviously involved and the tumor extended between the pleura and intercostal muscle.

The patient did not receive further therapy and was asymptomatic 7 months after surgery.     

Case 5

A 53-year-old man with a heavy exposure to asbestos 25 years earlier was admitted to the East Orange Veterans Administration Medical Center with a rapidly enlarging chest wall mass of 4-5 weeks duration.

The mass, overlying the ninth and tenth ribs in the infraaxillary area, was firm. On X-ray the ribs appeared eroded and the mass was seen to extend into the pleural space. A chest wall resection was performed, and the tumor was sharply dissected from the diaphragm to which it was firmly adherent. Grossly, it measured 7.5 X 5.0 X 3.0 cm and was yellow-tan with homogeneous and nodular areas The inferior aspect of the ninth rib was obviously involved and the tumor extended between the pleura and intercostal muscle.

The patient did not receive further therapy and was asymptomatic 7 months after surgery.     

Inhibiting the C5-C5a receptor axis

Activation of the complement system is a major pathogenic event that drives various inflammatory responses in numerous diseases. All pathways of complement activation lead to cleavage of the C5 molecule generating the anaphylatoxin C5a and, C5b that subsequently forms the terminal complement complex (C5b-9). C5a exerts a predominant pro-inflammatory activity through interactions with the classical G-protein coupled receptor C5aR (CD88) as well as with the non-G protein coupled receptor C5L2 (GPR77), expressed on various immune and non-immune cells. C5b-9 causes cytolysis through the formation of the membrane attack complex (MAC), and sub-lytic MAC and soluble C5b-9 also possess a multitude of non-cytolytic immune functions. These two complement effectors, C5a and C5b-9, generated from C5 cleavage, are key components of the complement system responsible for propagating and/or initiating pathology in different diseases, including paroxysmal nocturnal hemoglobinuria, rheumatoid arthritis, ischemia-reperfusion injuries and neurodegenerative diseases. Thus, the C5-C5a receptor axis represents an attractive target for drug development. This review provides a comprehensive analysis of different methods of inhibiting the generation of C5a and C5b-9 as well as the signalling cascade of C5a via its receptors. These include the inhibition of C5 cleavage through targeting of C5 convertases or via the C5 molecule itself, as well as blocking the activity of C5a by neutralizing antibodies and pharmacological inhibitors, or by targeting C5a receptors per se. Examples of drugs and naturally occurring compounds used are discussed in relation to disease models and clinical trials. To date, only one such compound has thus far made it to clinical medicine: the anti-C5 antibody eculizumab, for treating paroxysmal nocturnal hemoglobinuria. However, a number of drug candidates are rapidly emerging that are currently in early-phase clinical trials. The C5-C5a axis as a target for drug development is highly promising for the treatment of currently intractable major human diseases.     

Mast Cells and 5-HT: Uptake of labelled 5-hydroxytryptamine (5-HT) and 5-hydroxytryptophan in relation to storage of 5-HT in individual rat mast cells

The capacity of individual rat peritoneal mast cells to take up and concentrate radioactively labelled 5-hydroxytryptamine (5-HT) and 5-hydroxytryptophan (5-HTP) was studied by quantitative cyto-chemical methods. By means of consecutive application of microfluorometric, autoradiographic and microinterferometric methods, the following parameters could be determined for each cell: 1. Relative amount of endogenous 5-HT. 2. Concentration of accumulated isotopes. 3. Dry mass. 4. Concentration of endogenous 5-HT. 5. Total amount of accumulated isotopes. The calculated correlations of these variables to each other showed general heterogeneity of the population, variations between the different cells being very large. However, on the whole, increasing dry mass of the mast cells was accompanied by an increase in 5-HT content, but with a decrease in concentrations of endogenous 5-HT and accumulated radioactive 5-HTP. Similarly, cells having large stores of endogenous 5-HT showed a higher total uptake, but a lower cellular concentration of exogenous 5-HT and 5-HTP, than cells that were poor in endogenous 5-HT. The results indicated that the uptake of ³H-5-HTP was proportional to the total area of the cell membrane, while the degree of 3-HT uptake was determined by other factors. When the concentrations of radioactivity in the incubation medium was compared to that of the living mast cell, it was found that the averagr cell was able to concentrate exogenous 5-HTP and 5-HT around 4 and 30 times, respectively.     

Partial monosomy 5p and partial trisomy 5q due to paternal pericentric inversion of chromosome 5

Summary A male infant with partial monosomy 5p and partial trisomy 5q due to paternal pericentric inversion of chromosome 5 (46,XY,rec(5), dup q,inv(5)(p15.1q35.1)pat) is reported together with the oral findings. The phenotype was chiefly the cri-du-chat syndrome. Severe retardation of mental and motor development, microencephaly, cardiac malformation, crying and facial appearance unique to the cri-du-chat syndrome were observed. Perioral and intraoral findings included thin upper lip, downturning corners of mouth, micrognathia, shallow palate, and cleft of soft palate. Anterior deciduous teeth were small and canine deciduous teeth were conic. The row of deciduous teeth showed a flat arch-like shape that was very wide but short in length. No abnormality was noted in the number of deciduous teeth or the timing of eruption.