中药及保健食品中违禁添加7种降压类化学药物的HPLC-DAD法测定

建立了HPLC法同时测定中药及保健食品中可能违禁添加的7种降压类化学药物.采用C18色谱柱,以0.2%磷酸溶液-乙腈为流动相,梯度洗脱,检测波长220 nm.阿替洛尔、盐酸可乐定、氢氯噻嗪、卡托普利、盐酸哌唑嗪、利血平和硝苯地平的线性范围分别为0.028 4～2.84、0.029～2.9、0.053～5.3、0.054～5.45、0.054～5.4、0.034～3.38和0.031～3.12Bg,回收率分别为99.7%、91.2%、99.5%、107.6%、108.7%、97.8%和106.1%,RSD分别为2.1%、3.5%、2.5%、3.8%、1.0%、4.2%和5.0%.     

气相色谱串联质谱法同时测定黄芪中238种农药残留

 目的 采用气相色谱-串联质谱法建立黄芪中农药多残留的测定方法。方法 样品经改良后QuEChERS方法提取,采用氨丙基固相萃取小柱净化,并采用气相色谱-串联质谱同时定性、定量测定。结果 对黄芪进行了3个添加水平的回收实验,结果表明238种农药的回收率大多在70.0％～120.0％,相对标准偏差小于15％,检测限大多低于0.01 mg·kg^-1。结论 本法操作简单,净化效果好,灵敏度高,重现性好,可用于黄芪样品中农药多残留的日常检测。
     

干眼患者外周血单核细胞培养上清液和泪液中白细胞介素-17和白细胞介素-22的变化

背景 辅助T细胞17(Th17)是一种新发现的CD4+T细胞亚群,动物研究显示,Th17细胞及其效应因子白细胞介素-17(IL-17)在干眼的发病和发展过程中发挥重要作用,但其发病机制还需进一步证实.目的 检测干眼患者泪液及外周血单核细胞(PBMCs)培养上清液中IL-17和IL-22的表达水平,探讨其与干眼临床症状及体征的相关关系. 方法 收集在无锡第二医院眼科确诊的Sj(o)gren综合征(SS)、非Sj(o)gren综合征(NSS)患者各20例,并纳入同期医院内职工和学生健康志愿者20人为正常对照组,3个组间患者年龄、性别比均匹配.所有患者行干眼症状调查量表评分、泪膜破裂时间(BUT)、泪液分泌试验Ⅰ(SⅠt)、角膜荧光素染色检查.收集所有受检者外周血3 ml,采用酶联免疫吸附(ELISA)法检测各组受检者PBMC培养上清液及泪液中IL-17、IL-22的质量浓度并进行比较,并与干眼临床症状和体征进行相关分析. 结果 NSS组、SS组及正常对照组干眼症状调查量表评分依次降低,SS组、NSS组及正常对照组BUT值依次延长,SⅠt值依次增大,角膜荧光素染色评分明显降低,3个组间上述指标的总体比较差异均有统计学意义(症状评分:H=40.81,P＜0.01;BUT:H=40.15,P＜0.01;S Ⅰ t:H=50.07,P＜0.01;角膜荧光素染色评分:H=40.52,P＜0.01).SS组、NSS组及正常对照组PBMCs培养上清液中IL-17质量浓度分别为(964.92±124.83)、(718.85±115.89)、(341.95±85.08) ng/L;泪液中IL-17质量浓度分别为(440.69±126.09)、(364.33±126.85)、(61.16±11.60) ng/L,SS组、NSS组患者PBMCs中和泪液中IL-17质量浓度明显高于正常对照组,3个组间总体比较差异均有统计学意义(F=162.95,P＜0.01;F=75.27,P＜0.01).SS组、NSS组及正常对照组PBMCs培养上清液中IL-22质量浓度分别为(98.77±11.27)、(79.65±11.01)、(32.78±9.34) ng/L,泪液中分别为(22.22±8.96)、(14.92±4.35)、(10.47±2.67) ng/L,表明SS组、NSS组患者PBMCs中和泪液中IL-22质量浓度明显高于正常对照组,3个组间总体比较差异均有统计学意义(F=206.27,P＜0.01;F=19.87,P＜0.01).干眼患者PBMCs培养上清液及泪液中IL-17、IL-22水平与角膜荧光素染色评分、SⅠt有明显关联,但与干眼症状量表评分、BUT均无相关性.结论 干眼患者PBMCs及泪液中IL-17、IL-22的质量浓度明显升高,进一步提示Th17在干眼免疫机制中发挥重要作用.     

HPLC-MS/MS测定绿豆中黄曲霉毒素G_2、G_1、B_2、B_1

目的建立绿豆中黄曲霉毒素G2、G1、B2、B1的测定方法。方法样品经70%甲醇提取、HLB柱净化后,用高效液相色谱串联三重四极杆质谱进行分析测定。结果黄曲霉毒素G2、B2在0.75~30 pg范围内与峰面积呈良好的线性关系,黄曲霉毒素G1、B1在2.5~100 pg范围内与峰面积呈良好的线性关系,r>0.999。回收率在61.5%~107.0%之间。结论本法简便快速、结果准确、重现性好,可用于绿豆中黄曲霉毒素的测定。     

GC测定灵芝中24种有机磷农药残留量

 目的研究灵芝中24种有机磷农药残留量的测定方法。方法样品经冰浴超声提取,凝胶渗透色谱(GPC)净化后,采用不分流进样,用双柱双火焰光度检测器(FPD)气相色谱同时定性、定量测定。结果样品3水平添加时的24种有机磷农药回收率均在75.28%～117.18%,相对标准偏差在3.19%～15.57%,符合农药残留分析的要求。结论本法净化效果好,重现性好,可用于灵芝中24种有机磷农药残留量的检测。     

脑卒中患者偏瘫的早期康复护理

目的:探讨脑卒中偏瘫患者应用早期综合性康复效果训练后肢体运动功能的恢复。方法:将2003—06~2004—10在我院神经科96例脑卒中偏瘫患者随机分为观察组和对照组各48例,两组患者在年龄、性别、病变部位、病情程度、药物治疗方法及统计学处理方法均无明显差异,具有可比性,对照组沿袭常规护理,观察组采用早期康复、心理康复、中医针灸、中药药熨穴位法、残肢功能锻炼及家庭追踪指导康复服务,观察脑卒中患者偏瘫早期康复护理的疗效及变化。结果:早期康复护理训练后,两组肢体运动功能训练及日常生活能力的差别具有显著性意义(P<0.05)。结论:早期康复护理对脑卒中偏瘫患者的康复疗效极佳,有利于提高运动功能和生活能力。     

HPLC-MS/MS法测定中药材枳壳中展青霉素

目的:建立中药材枳壳中展青霉素的HPLC-MS/MS测定方法。方法:样品经果胶酶酶解,用水提取、经过展青霉素免疫亲和柱净化后,用高效液相色谱-三重四级杆串联质谱进行分析测定。结果:展青霉素在5.08 ng/ml～254.0 ng/ml范围内与峰面积呈良好的线性关系,线形方程为Y=2.17×103X-222,r=0.9977,回收率在80.0%～101.1%之间。在枳壳中最低检测限为0.02μg/kg。结论:本法灵敏、快速、准确、专属性强,可用于中药枳壳中展青霉素的测定。     

熊胆粉提取物中熊去氧胆酸和鹅去氧胆酸的HPLC-ELSD测定

建立THPLC-ELSD法测定熊胆粉提取物中的熊去氧胆酸和鹅去氧胆酸.采用C18色谱柱,以0.05%三氟乙酸-乙腈梯度洗脱,蒸发光散射检测器,漂移管温度95℃.熊去氧胆酸和鹅去氧胆酸分别在2.51～12.54μg和0.40～2.02μg范围内线性关系良好,回收率为98.8%和99.8%,RSD为1.4%和2.1%.     

黄芪注射液中药用辅料聚山梨酯80两种含量测定方法的比较研究

目的比较采用两种方法测定黄芪注射液中聚山梨酯80含量的优缺点。方法分别采用高效液相色谱-蒸发光散射检测法(HPLC-ELSD)和紫外-可见分光光度法(UV)进行检测;采用分子凝胶色谱柱;以乙腈-水(90:10)为流动相;流速为0.6 mL.min-1;漂移管温度110℃,载气流速为2.3 L.min-1。结果两种方法测得的结果之间存在系数关系,HPLC-ELSD法测得结果约为UV法的0.65;与黄芪注射液中聚山梨酯80理论投料量进行比较,HPLC-ELSD法测定结果比UV法更接近理论值。结论 HPLC-ELSD法简便、准确,更有利于黄芪注射液中聚山梨酯80的含量控制。     

全氟辛烷磺酸对大鼠仔鼠学习记忆能力的影响

Objective To study the effects of prenatal and postnatal perfluorooetane sulfonate (PFOS) exposure on spatial learning and memory, N-methyl-D-aspartate receptor 2B (NR2B) mRNA and protein level in frontal cortex and hippoeampus of rat pups and to explore the mechanism of developmental neurotoxicity induced by PFOS. Methods Twenty-eight pregnant mrs were randomly divided into three groups in proportion of 3: 2: 2,including control group (C), low dose group (L) and high dose group (H) by means of randomized number table, which respectively received 0,7.2, 14.4 mg/kg PFOS feed from pregnancy day 0 to postnatal day(PND)30 by free feedings. The animal models of prenatal and postnatal non-exposure (CC), prenatal exposure (LC and HC), postnatal exposure (CL and CH), and prenatal and postnatal exposure (LL and HH) to PFOS were established by cross-fostering method. The spatial learning and memory were measured by water maze experiment,the NR2B mRNA levels in frontal cortex of rat pups was determined with semi-quantitative RT-PCR, NR2B protein express in cerebral cortex (frontal and tempera]cortex) and hippocampus (CAI, CA3, CA4 and DG regions) of rat pups was detected by immunohistochemistry. Results The escape latency of CL, CH, LL and HH groups pups in water maze experiment were (99. 83±25.77) s, (111.30±17. 82) s, (106. 40±18. 71) s, (107.70±16. 85) s, and longer as compared with CC group[(54.90±26.69)s](q value were 4.349,4.773,6.026 and 5.641, respectively,P<0. 01). The number of errors of HH group rat pups entering dead end was (22. 30±7.56) at the training day 4,and it was significantly higher than that of CC group (9. 80±4. 64) (q=5. 173,P<0. 01). The NR2B mRNA levels of frontal cortex of pups in HC group at PND1 ,and LC group, HC group and HH group at PND14 were (0. 167±0. 008), (0. 364±0. 035), (0. 341±0. 030) and (0. 328±0. 045) respectively,which were significantly lower than CC group (0.271±0.060) and (0.465±0.067) (q values were 3. 547, 3. 739, 4. 597 and 5. 006, respectively, P<0.05) . The results of immunohistochemistry indicated that NR2B protein express of the hippocampus CA1 region of pups in LC group was (0.091±0.005),and showed significant lower than CC group which was (0. 123±0.009) at PND1 (q=5. 209 ,P<0. 05). At PND14, the effect of PFOS extended to cerebral cortex and hippocampus regions. At PND28, the effects of PFOS were showed in hippocampus CA1, CA3 and temporal cortex regions. Conclusion Prenatal and postnatal exposure to PFOS should result in the spatial learning and memory damage,and the mechanism might be possibly involved in the decrease of NR2B level in cerebra lcortex and hippocampal formation regions.