鸡蛋参的总生物碱含量测定

鸡蛋参是桔梗科(Codcnopsis Conno-lrulacca Karz)植物的干燥块根,别名牛尾参、补血草。主产于四川甘孜州道孚、甘孜,西藏南部和云南等地。据记载,该药可冶疗感冒咳嗽等。是民问常用的藏药之一。经化学成分预试,主要含生物碱,植物甾醇和醣类。用重量法测得总生物碱的含量约为0.2%。总生物碱含量测定精密称取鸡蛋参干燥粉末10g,置250ml园底烧瓶中,加石油醚(30°～60℃)100ml,在60℃水浴上加热回流1小时,过滤,弃去滤液,残渣挥干石油醚后,加95%乙醇100ml,在60℃水浴上加热回流2小时,过滤,残渣反复用95%乙醇适当提取至无生物碱反应为止。合并滤液,减压浓缩至稠羔状。用2%盐酸液20、15、10ml分次提取,合并盐酸液,置分液漏斗中,加氯仿20ml充分振摇,静置后,弃去氯仿液,盐酸液用氢氧化铵硷化,调pH到8-9。用氯仿20、15、15ml分次提取,合并氯仿液。置60℃干燥至恒重的蒸发皿中。在水浴上蒸去氯仿,残渣在60℃干燥至恒重。按下式计算总生物碱的含量: 总生物硷的含量%=(提取残渣重/取样量结果见表。     

PEG6000为载体的熊果酸固体分散体水分测定的研究

目的采用干燥失重法建立以PEG6000为载体辅料制备的熊果酸固体分散体的水分测定方法。方法先将样品在较低的温度（52℃）下干燥至大部分水分除去后,然后逐渐升温至105℃干燥至恒重。另应用甲苯法测定样品的水分。结果在低温（52℃）下可以将70%以上的大部分水分除去,在105℃极易恒重。测定结果同甲苯法无显著性差异。结论干燥失重法不需要特殊设备,简单易行、重复性好,可用于该制剂水分的测定。     

制备条件对甲酸棉酚失酸的影响

本文通过测定贮存不同时间的甲酸棉酚的酸含量、熔点和 IR 光谱,研究了制备条件对甲酸棉酚失酸的影响。实验结果表明,在本实验条件下制备的六个甲酸棉酚样品均有不同程度的失酸,最小失酸速度的重结晶条件是:重结晶温度为0℃,加入的甲酸量与甲酸棉酚饱和乙醚溶液量之比为3:10(V:V)。     

硫酸庆大霉素水分测定方法的改进

硫酸庆大霉素具引湿性,其纯度、稳定性、理化性质等均与含水量的多少密切相关.为了控制水分含量,中国药典的含水量测定方法为干燥失重.在105℃的条件下干燥至恒重.在干烤过程中,温度、恒重时间、放置时间对供试品有影响,从而不能充分地反映出该品种的含水量.因此,特进行了干燥失重和水分测定法的比较.     

一苯胺棉酚—棉酚中的一种杂质

本文介绍了棉酚中的一种杂质(苯胺法精制)一苯胺棉酚的研究。该化合物文献中未见报道。实验数据符合组成式C₃₈H₃₅O₇N;质谱分子离子峰593,紫外光谱于439～440 nm有最大吸收(E_1com^1% 329±6 CHCl₃);红外光谱在1610 cm^-1有醛基和1590 cm^-1碳氮吸收峰;核磁共振谱表明有—CHO和—HC=N质子峰,其峰面积比为1:1;熔点272～275℃(分解);硅胶薄层上Rf值界于二苯胺棉酚和棉酚之间。一苯胺棉酚的存在,干扰分光光度法测定棉酚含量的准确性。利用薄层层析法可检查棉酚中一苯胺棉酚的存在与否。本文还介绍了一苯胺棉酚的合成方法。     

棉酚的极谱研究

本文介绍了棉酚的极谱测定方法。棉酚在柠檬酸—磷酸氢二钠缓冲液中可产生一还原波,在pH值为2.60时,其E_(1/2)为-1.14 V(SCE)。实验证明该波为受扩散控制的单电子可逆波,推测为棉酚芳醛基之一的单电子还原。所建立的方法较简便、快速、灵敏。     

关于《中国药典》2005年版二部“干燥失重测定法”中对减压干燥新增温度规定的若干思考

在《中国药典》2000年版中，当测定颗粒剂的【干燥失重】时规定：“除另有规定外，照干燥失重测定法测定，含糖颗粒剂宜在80℃真空干燥，减失重量不得超过2．0％”。但附录“干燥失重测定法”中并未规定干燥温度。为消除此缺陷，2005年版药典在颗粒剂【干燥失重】测定中增加了温度的规定：“…于105℃干燥至恒重”。[第一段]     

基层医院临床生化检验技术 （续20）

第三节 血氨测定1 原理 血液采集后,立即加入钨酸蛋白沉淀剂中,使蛋白质沉淀,氨与硫酸则生成硫酸铵而游离于血滤液中,再以酚-次氯酸钠显色反应测定含量。2 试剂2.1 0.3mol/L钨酸钠溶液 称取钨酸钠(Na_2WO_4·2H_2O,AR,M=329.87)20g,加无氨蒸馏水溶解并加至200ml。2.2 0.5mol/L 硫酸溶液2.3 显色剂Ⅰ 精确称取酚(C_6H_5OH,AR,M:94.11)10g,及亚硝基铁氰化钠50mg,加无氨蒸馏水溶解并加至200ml。置4℃冰箱保存,可长年稳定。2.4 显色剂Ⅱ 称取氢氧化钠5g,次氯酸钠0.5g(或安替福民溶液8ml),溶于无氨蒸馏水并加至200ml。置4℃冰箱保存。2.5 硫酸铵标准贮存液(10mmol/L)精确称取干燥至恒重的硫酸铵(AR,M:132.14)132.14mg,溶于无氨蒸馏水并加至200ml。加氯仿数毫升,置4℃冰箱保存,可长年稳定。     

氚标记( )与(-)棉酚在大鼠主要脏器的亚细胞分布及共价结合

给大鼠腹腔及睾内注射氚标记的( )与(-)棉酚后第7、18天,对主要脏器中各亚细胞组分的总放射活性及共价结合的放射活性进行了动态观察。结果表明,(-)棉酚在心肌线粒体共价结合放射活性较明显高于( )棉酚;( )及(-)棉酚在睾丸细胞膜、微粒体共价结合的放射活性随时间增高,且(-)棉酚较为明显。     

改良酚/氯仿法提取人全血DNA的研究

目的对传统酚/氯仿法进行改良,以期提取到高纯度的DNA,提高回收率。方法采用改良酚/氯仿法提取人全血基因组DNA,与传统酚/氯仿法在DNA纯度、质量浓度和聚合酶链反应(PCR)扩增短串联重复序列(STR)位点等方面进行比较。结果改良前后酚/氯仿法提取到的DNA纯度比较,差异无统计学意义(P>0.05);改良酚/氯仿法提取DNA的量明显高于酚/氯仿法,二者比较,差异有统计学意义(P<0.05)。改良后酚/氯仿法提取2μL全血DNA仍可准确进行CFS1PO分型。结论改良酚/氯仿法提取DNA具有较高纯度和较高回收率的特点,可能在微量检材DNA提取中有一定应用价值。     

红外光谱法和X-射线粉末衍射法研究棉酚的多晶型

棉酚是棉子中的一种色素,具有广泛生理活性。棉酚、醋酸棉酚、甲酸棉酚曾初步试用于男性节育,但有一定毒副作用,影响了临床进一步试用。目前国内外学者仍在进一步研究。棉酚的结构式如下:     

醋酸棉酚对子宫和卵巢的影响

Gossypol has been employed in gynecologic clinics for the treatment of endometriosis and functional uterine bleedings with encouraging results. In the present communication the effects of gossypol on the uterus and ovary were studied both experimentally and clinically by histologic and histochemical techniques. It was found that treatment of female rats with gossypol acetic acid at 30 mg/kg/d, six days a week, for 8 weeks caused marked atrophy of the endometrium and a reduction of the glycogen content of the myometrium. Vaginal smears showed a picture of atypical estrous cycle. Simultaneous administration of 1 mg/kg/d of estriol reduced the effect of gossypol acetic acid though the influence of gossypol treatment was still discernible. Treatment of 16 patients suffering from endometriosis or functional uterine bleeding with gossypol acetic acid for 2-3 months induced changes similar to those observed in animal experiments, namely, atrophy of the endometrium and decrease in endometrium glycogen. In 3 other patients treated with gossypol acetic acid for 4～5 months there was a slight increase in Sudan Black B staining and a decrease in alkaline phosphatase, acid phosphatase, PAS staining and RNA of the endometrium. There was also a change of biphasic basal body temperature into monophasic and a lowered estrogen level as revealed by vaginal cytology.One may speculate from the above results that gossypol may act on the uterus directly as well as indirectly through inhibition of the secretory function of the ovary and/or through antagonism against the effect of estrogens in the female. Hence, it may be desirable to conduct further studies on the effect of gossypol on the pituitary-gonadal axis.     

(-)和(+)棉酚对雄大鼠生育力的影响

Racemic, (-) and (+) gossypol, provided by the Department of Organic Chemistry of our institute, was suspended in 2.5% tween 80 solution. Adult male Wistar rats 190～220 g in weight were allotted to 5 groups. Animals in group 1 received 2.5% tween 80 solution as control. Rats in group 2 were treated with racemic gossypol at the dosage of 30 mg/kg for 2 weeks. Animals in group 3 and 4 were given 15 mg/kg of (-) gossypol for 2 weeks and 30 mg/kg of (-) gossypol for 1 week respectively. Rats in group 5 were treated with (+) gossypol at the dosage of 30 mg/kg for 2 weeks.Four weeks from the beginning of gossypol treatment the rats were cohabited with adult females for 7 days. Then the motility of the sperms in the cauda epididymides was estimated The female rats were examined for pregnancy 7 days later.Treatment with (-) gossypol at 30 mg/kg caused significant decreases in body weight of the rats (P<0.05). One of the five rats died 7 days after the last administration, while (+) gossypol and racemic gossypol at the dosage employed had no effect on the body weight. (+) Gossypol at 30 mg/kg for 2 weeks had no effect on the motility of the sperms in the cauda epididymides and no effect on the fertility of the animals: nor was there any effect on the weights of the testis, epididymis, prostate and seminal vesicle. The sperms of the cauda epididymides were found to be dead in the groups treated with 15 and 30 mg/kg of (-)gossypol. Raccmic gossypol given for 2 weeks at 30 mg/kg caused loss of fertility of the male rats which confirmed our previous findings.(?)t may be postulated that (+) gossypol has no antifertility effect nor toxicity at the dosage employed. (-) Gossypol is the active stereoisomer of racemic gossypol.     

棉酚的极谱研究

本文介绍了棉酚的极谱测定方法。棉酚在柠檬酸—磷酸氢二钠缓冲液中可产生一还原波,在pH值为2.60时,其E_1/2为-1.14 V(SCE)。实验证明该波为受扩散控制的单电子可逆波,推测为棉酚芳醛基之一的单电子还原。所建立的方法较简便、快速、灵敏。     

高效液相色谱法分析棉酚

本文制定了棉酚的高效液相色谱分析法,选用ODS柱及流动相THF—H₂O—H₃PO₄(60:40:0.08),紫外检测λ_254nmo此法分离能力强且结果稳定。与目前常用于棉酚分析的方法作了比较。本方法适用于棉酚(包括甲酸棉酚和乙酸棉酚)的质量控制,并有可能用于棉酚的药代动力学研究。     

醋酸棉酚长期给药的进一步观察

给成年雄性大鼠每日服醋酸棉酚10 mg/kg,每周服6天。服6个月后进行病理和组织化学观察。结果说明给醋酸棉酚大鼠各主要脏器(心、肝、脾、肺、肾、肾上腺)的形态都没有明显变化。组织化学观察说明,给棉酚大鼠的肝脏油红“O”染色、G-6-P酶、ATP酶、ALP酶、ACP酶、糖原、RNA和DNA肾脏G-6-PDH、油红“O”染色、ATP酶、ACP酶、ALP酶;肾上腺3β-甾体脱氢酶、油红“O”染色、苏丹黑染色等与对照组比较都没有明显差别。     

醋酸棉酚——PVP固体分散体研究及体外杀精子活性的观察

本文对溶剂法制备醋酸棉酚—PVP固体分散体作了改进。以二氯甲烷为溶剂在45℃常压下完成,结果与Waller方法一致,并具有更大的安全性。经溶出实验筛选确定以1∶7醋酸棉酚—PVP配比可比Waller等人的1∶4配比提高药物溶出速率8倍,不同比例的固体分散体释药曲线均服从Q—t^1/2规律,且速度常数随固体分散体中PVP比例增加而线性递增。利用X射线衍射和¹HNMR研究了固体分散机制,认为是形成了间充型固体溶液,且PVP与醋酸棉酚之间有氢键结合。1∶7棉酚—PVP抑动精子实验结果表明10^-3M(0.052%)时20秒钟可抑制全部精子的向前运动。棉酚和醋酸棉酚结果一致,无统计学差异。PVP与生理盐水结果一致,表明PVP本身无抑制精子运动作用。     

雷公藤多甙与棉酚合用对雄大鼠生育力的影响

本研究探讨雷公藤多甙(GTW)与棉酚合用对雄大鼠生育力的影响。SD雄大鼠用棉酚及GTW各6 mg/kg/d,灌胃给药,每周6日,共11周。10只用药动物全部不育,附睾精子密度和活率也明显下降,而体重增长如常,性行为存在。睾丸光镜下结构绝大多数无明显异常,血清睾酮水平正常,副性腺重量无明显变化。停药6周后生育力恢复。在相同剂量下,单用GTW或棉酚均无抗生育效果。表明两药合用有相加作用,为减少棉酚和GTW副作用提供一条可能途径。     

Regulation of antiapoptotic MCL-1 function by gossypol: mechanistic insights from in vitro reconstituted systems

Small-molecule drugs that induce apoptosis in tumor cells by activation of the BCL-2-regulated mitochondrial outer membrane permeabilization (MOMP) pathway hold promise for rational anticancer therapies. Accumulating evidence indicates that the natural product gossypol and its derivatives can kill tumor cells by targeting antiapoptotic BCL-2 family members in such a manner as to trigger MOMP. However, due to the inherent complexity of the cellular apoptotic network, the precise mechanisms by which interactions between gossypol and individual BCL-2 family members lead to MOMP remain poorly understood. Here, we used simplified systems bearing physiological relevance to examine the impact of gossypol on the function of MCL-1, a key determinant for survival of various human malignancies that has become a highly attractive target for anticancer drug design. First, using a reconstituted liposomal system that recapitulates basic aspects of the BCL-2-regulated MOMP pathway, we demonstrate that MCL-1 inhibits BAX permeabilizing function via a “dual-interaction” mechanism, while submicromolar concentrations of gossypol reverse MCL-1-mediated inhibition of functional BAX activation. Solution-based studies showed that gossypol competes with BAX/BID BH3 ligands for binding to MCL-1 hydrophobic groove, thereby providing with a mechanistic explanation for how gossypol restores BAX permeabilizing function in the presence of MCL-1. By contrast, no evidence was found indicating that gossypol transforms MCL-1 into a BAX-like pore-forming molecule. Altogether, our findings validate MCL-1 as a direct target of gossypol, and highlight that making this antiapoptotic protein unable to inhibit BAX-driven MOMP may represent one important mechanism by which gossypol exerts its cytotoxic effect in selected cancer cells.