LC-MS/MS法测定厄贝沙坦中基因毒性杂质N-亚硝基二甲胺和N-亚硝基二乙胺

建立同时检测厄贝沙坦中N-亚硝基二甲胺(NDMA)和N-亚硝基二乙胺(NDEA)的液质联用(liquid chromatography-tandem mass spectrometry,LC-MS/MS)方法。选择大气压电离串联四极杆质谱(APCI+MRM)模式进行检测,流动相为0.1%甲酸水溶液-0.1%甲酸甲醇溶液梯度洗脱。结果表明该方法专属性良好,标准曲线线性范围分别为1~120 ng·mL^-1(NDMA)和0.4~48 ng·mL^-1(NDEA);检测限浓度分别为0.5 ng·mL^-1(NDMA)和0.2 ng·mL^-1(NDEA);精密度RSD均<3.0%;准确度为96.1%~106.2%,重复性良好;样品的提取回收率分别为102.5%(NDMA)和99.9%(NDEA);对照溶液在进样器和0℃下放置24 h后稳定。该方法简单灵敏准确,可以较好地满足厄贝沙坦原料药生产中基因毒性杂质NDMA和NDEA的检验。     

达卡巴嗪原料药中N-亚硝基二甲胺和N-亚硝基二乙胺的UHPLC-APCI-MS/MS方法的建立

目的:采用UHPLC-APCI-MS/MS法测定达卡巴嗪原料药中的N-亚硝基二甲胺(NDMA)和N-亚硝基二乙胺(NDEA),建立控制该药品遗传毒性杂质的方法。方法:采用Kinetex F5色谱柱(3 mm×100 mm,2.6μm),流动相为0.1%甲酸溶液-甲醇,梯度洗脱,流速0.4 mL·min^-1,柱温40℃,质谱离子化方式为APCI,正离子模式,多重反应监测,检测离子对为m/z 75.1/58.0(NDMA),103.0/47.0(NDEA)。结果:NDMA和NDEA在一定范围内线性关系良好(r>0.990),检出限溶液浓度分别为0.03030、0.003787 ng·mL(-1),精密度、稳定性试验满足检测要求,回收率为97~115%,RSD均小于5%。结论:本文建立的方法准确、快速灵敏、专属性强,可用于控制达卡巴嗪原料药中遗传毒性杂质NDMA和NDEA的含量。     

GC-MS/MS法测定缬沙坦氢氯噻嗪制剂中N-亚硝基二甲胺与N-亚硝基二乙胺

摘要：目的:建立GC-MS/MS法同时测定缬沙坦氢氯噻嗪制剂中基因毒性杂质N-亚硝基二甲胺（NDMA）与N-亚硝基二乙胺（NDEA）含量的方法。方法:使用Thermo TG-WAMS毛细管柱（30 m×0.25 mm, 0.25μm）,采用脉冲不分流进样模式,利用程序升温进行分离,通过EI离子源和SRM模式测定NDMA和NDEA。结果:NDMA和NDEA分别在0.54～108.99 ng·ml-1（r=0.999 9）与0.20～100.00 ng·ml-1（r=0.999 8）浓度范围内线性关系良好,NDMA检出限为0.22 ng·ml-1;NDMA和NDEA加样回收率分别为101.6%,102.0%,RSD分别为2.2%,2.7%（n=9）。结论:该方法专属性好、灵敏度高、准确度高和样品制备简单,可用于缬沙坦氢氯噻嗪制剂中NDMA和NDEA检测。     

UHPLC-MS/MS法测定盐酸二甲双胍制剂中2种亚硝胺类基因毒性杂质

目的 建立超高效液相色谱串联质谱(UHPLC-MS/MS)法同时测定盐酸二甲双胍制剂中2种基因毒性杂质N-亚硝基二甲胺(N-nitrosodimethylaminen, NDMA)和N-亚硝基二乙胺(N-nitrosodiethylamine, NDEA)的含量的方法。方法 采用多反应监测(MRM)模式检测，离子源为APCI(+),色谱柱为ACE Excel 3 C₁₈-AR柱(100 mm×4.6 mm, 3μm),以体积分数0.1%甲酸水溶液(A)-质量分数0.1%甲酸甲醇溶液(B)为流动相梯度洗脱，流速0.5 mL·min^-1,进样体积5μL。结果 NDMA和NDEA的线性范围分别为0.99～99μg·L^-1(r>0.999 9)和0.53～53μg·L^-1(r>0.999 9),平均回收率为90.9%～98.9%,定量限分别为0.99和0.53μg·L^-1,检测限分别为0.3和0.2μg·L^-1。用建立的方法测定不同剂型的6批样品，2批...     

GC-MS/MS法测定甲磺酸卡莫司他原料药中N-亚硝基二甲胺

目的建立GC-MS/MS法测定甲磺酸卡莫司他原料药中的亚硝胺类基因毒性杂质N-亚硝基二甲胺(NDMA)。方法采用Agilent DB-SELECT 624UI色谱柱(30 m×0.32 mm×1.80μm)色谱柱,离子源为EI源,采用多反应监测模式,以m/z 74→44.2为定量离子,m/z 74→42.2为定性离子,进行数据采集。结果 NDMA在12.571 2～100.569 6 ng/mL线性良好(r=0.997 6),检测限质量浓度为5.028 5 ng/mL,平均回收率为107.4%,RSD值为2.5%。结论该方法操作简单,灵敏度高,专属性和重复性好,可用于甲磺酸卡莫司他原料药中NDMA的检测。     

GC-MS/MS法同时测定氯沙坦钾原料药及其制剂中6种N-亚硝胺类基因毒性杂质

目的 建立同时测定氯沙坦钾原料药及其制剂中6种N-亚硝胺类基因毒性杂质含量的方法。方法 采用气相色谱-串联质谱（GC-MS/MS）法测定氯沙坦钾原料药、氯沙坦钾片、氯沙坦钾胶囊、氯沙坦钾氢氯噻嗪片中N-亚硝基二甲胺（NDMA）、N-亚硝基二乙胺（NDEA）、N-亚硝基-N-乙基异丙胺（NEiPA）、N-亚硝基二异丙胺（NDiPA）、N-亚硝基二苯胺（NDPA）、N-亚硝基二丁胺（NDBA）6种N-亚硝胺类基因毒性杂质含量。色谱柱为SHIMADZU SH-L-17Sil MS毛细管柱；采用程序升温；进样口温度为250℃；进样量为1μL；载气为氦气，流速为1 mL/min。离子源为电子轰击源，离子源温度为250℃；溶剂延迟时间为3.1 min；采集模式为多反应监测模式。结果 NDMA、NDEA、NEiPA、NDiPA、NDPA、NDBA与其相邻色谱峰之间的分离效果均良好，分离度均大于3.8；其线性范围分别为4.9～486.0、4.9～488.5、4.5～451.5、6.8～683.5、5.2～525.0、5.2～520.0 ng/mL(r≥0.999 8)，定量限分别为4.86、4.88、...     

GC-MS/MS测定替米沙坦片中10种亚硝胺类基因毒性杂质

目的 建立GC-MS/MS同时测定替米沙坦片中N-亚硝基二甲胺(NDMA)、N-亚硝基甲乙胺(NMEA)、N-亚硝基二乙胺(NDEA)、N-亚硝基-N-乙基异丙胺(NEIPA)、N-亚硝基二异丙胺(NDIPA)、N-亚硝基二正丙胺(NDPA)、N-亚硝基二正丁胺(NDBA)、N-亚硝基哌啶(NPIP)、N-亚硝基吡咯烷(NPYR)和N-亚硝基吗啉(NMOR)10种亚硝胺类基因毒性杂质的含量。方法样品经甲醇提取,经Agilent VF-WAXms(30 m×0.25 mm,1μm)毛细管气相色谱柱分离,采用多反应离子监测(MRM)模式进行定量分析。结果 NDMA、NMEA、NDEA、NEIPA、NDIPA、NDPA、NDBA和NPIP在0.2～50 ng·mL^–1线性关系良好,相关系数均为1.000 0;检测限均为0.05 ng·mL^–1,定量限均为0.2 ng·mL^–1,平均回收率分别为103%(RSD=9.2%,n=9),108%(RSD=6.1%,n=9),107%(RSD=5.3%,n=9),106%(RSD=3....     

GC-MS/MS测定头孢菌素类药物中8种N-亚硝胺类基因毒性杂质

摘要：目的:建立GC-MS/MS法测定头孢孟多酯钠、头孢地嗪钠、盐酸头孢替安中8种N-亚硝胺类杂质（N-亚硝基二甲胺、N-亚硝基二乙胺、N-亚硝基二丙胺、N-亚硝基二丁胺、N-亚硝基甲基乙基胺、N-亚硝基吡咯烷、N-亚硝基哌啶、N-亚硝基吗啉）。方法:采用VF-WAX ms色谱柱（30 m×0.25 mm,膜厚0.25μm）,程序升温,进样口温度250℃,流速:1.0 ml·min-1;质谱采用EI源,电压为70 eV,离子源温度250℃、四极杆温度150℃,采用MRM模式对8种杂质进行定量测定;以0.1 mol·L-1氢氧化钠溶液溶解样品,以二氯甲烷进行萃取。结果:在上述条件下,8个N-亚硝胺杂质及相邻色谱峰之间分离效果良好,在所测浓度范围内线性关系良好,定量限为0.002 1～0.003 3μg·g-1,检出限为0.000 6～0.001 1μg·g-1;8种N-亚硝胺杂质的低、中、高浓度的平均回收率为85.5%～110.0%,RSD≤4.8%（n=3）。结论:建立的GC-MS/MS测定3种头孢菌素类药物中8种N-亚硝胺杂质的方法,准确度好,灵敏度高,简便可靠,对仪器污染小,可用于头孢孟多酯钠、头孢地嗪钠、盐酸头孢替安中8个N-亚硝胺基因毒性杂质的质量控制。     

HS-GC-MS/MS测定富马酸丙酚替诺福韦中微量的基因毒性杂质N-亚硝基二甲胺

目的 采用顶空进样气相色谱三重四级杆质谱联用（HS-GC-MS/MS）法测定富马酸丙酚替诺福韦中微量的基因毒性杂质N-亚硝基二甲胺（NDMA）。方法 采用三重四极杆GC-MS/MS，Agilent VF-WAX ms（30 m×0.25 mm，1 μm）色谱柱，载气：氦气；恒流模式1.0 mL·min^-1；程序升温，进样口温度230℃，顶空温度130℃；质谱采用电子轰击电离源（EI），电离能量为70 eV，离子源温度230℃，多反应监测（MRM）模式进行检测，溶剂为N-甲基吡咯烷酮（NMP）。进行专属性、系统适用性、检测限与定量限、线性与范围、准确度、精密度、溶液稳定性、耐用性考察。结果 NDMA与相邻色谱峰之间分离效果良好；NDMA在7.0～105.0 ng·mL^-1线性关系良好，检测限为3.5 ng·mL^-1，定量限为7.0 ng·mL^-1；NDMA低、中、高质量浓度（56、70、84 ng·mL^-1）回收率为95.6%～109.3%，RSD为4.0%（n=9）；重复性试验NDMA质量浓度RSD为6.5%，中间精密度RSD为6.1%；对照品溶液室温放置24 h稳定，供试品溶液室温放置90 h内溶液稳定；保持其他条件不变，分别改变进样口温度（230、225、235℃）、离子源温度（230、225、235℃）、载气体积流量（1.0、0.9、1.1 mL·min^-1）、顶空温度（130、128、132℃），方法耐用性良好。结论 所建立的方法准确度好、灵敏度高、简便可靠，对仪器污染小，可用于富马酸丙酚替诺福韦中NDMA的质量控制。     

UHPLC-MS/MS法测定盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔

目的 建立盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔的UHPLC-MS/MS检测方法。方法 Waters ACQUITY UPLC® CSHTM C₁₈色谱柱(150 mm×3.0 mm, 1.7 μm),10 mmol•L^-1甲酸铵的水溶液（含0.1%甲酸）作为流动相A,乙腈溶液（含0.1%甲酸）作为流动相B,梯度洗脱,流速为0.5 mL•min^-1,柱温为50 ℃,进样器温度为5 ℃,进样体积为10 μL,采用多反应监测（MRM)模式,对盐酸普萘洛尔缓释片中的N-亚硝基普萘洛尔进行定量检测。结果 N-亚硝基普萘洛尔在1～20 ng•mL^-1范围内具有良好的线性关系。低、中、高3个浓度的加样回收率（n=3）在98.4%～103.2%之间,RSD≤2.7%。检测限和定量限分别为0.09 ng•mL^-1和0.3 ng·mL^-1。检出盐酸普萘洛尔缓释片中基因毒性杂质N-亚硝基普萘洛尔含量为1.8 μg•g^-1。结论 该方法灵敏度高、专属性强,可用于测定盐酸普萘洛尔缓释片中的N-亚硝基普萘洛尔,为盐酸普萘洛尔缓释片的质量控制提供参考。     

Effects of N-cyclopentyladenosine and caffeine on sleep regulation in the rat

To study the role of adenosine in sleep regulation, the adenosine A₁ receptor agonist N⁶-cyclopentyladenosine (CPA) and the antagonist caffeine were administered to rats. Intraperitoneal (i.p.) CPA 1 mg/kg but not 0.1 mg/kg, suppressed rapid-eye-movement (REM) sleep and enhanced electroencephalographic (EEG) slow-wave activity (power density 0.75–4.0 Hz) in non-REM sleep. The latter effect was remarkably similar to the response to 6-h sleep deprivation. The effects persisted when CPA-induced hypothermia was prevented. Caffeine (10 and 15 mg/kg i.p.) elicited a dose-dependent increase in waking followed by a prolonged increase of slow-wave activity in non-REM sleep. The combination of caffeine (15 mg/kg) and sleep deprivation caused less increase in slow-wave activity than sleep deprivation alone, indicating that caffeine may reduce the buildup of sleep pressure during waking. The results are consistent with the involvement of adenosine in the regulation of non-REM sleep.     

褪黑激素(N-乙酰基-5-甲氧基色胺)的合成

This paper reports the synthesis of melatonin(N-acetyl-5-methoxytryptamine).The structure of all the compounds synthesized were characterized by elemental analysis,IR, MS and ¹HNMR spectra.     

N-(4-取代氨基-4-氧代丁酰基)-N-取代甘氨酸的合成

血管紧张素转化酶抑制剂(ACEI)是近年发展的一类新型抗高血压和治疗心力衰竭的有效药物。有关研究工作还在世界范围内广泛进行。 本研究组前曾报道一类具有降压活性的N-取代甘氨酸的合成。作者等根据Cushman等的血管紧张素转化酶(ACE)与底物的作用模型和构效关系,在保持3个主要作用部分(即ACE     

N-甲基-8,9-次甲二氧基-2-羧基氯化菲啶季铵盐的合成

Lycobetaine (AT-1840, I) showed antitumor activity both in vitro and in vivo . Structure activity studies had concluded that in order to keep the activity of I, the intramolecular betaine structure must be kept while the fission of the five membered N-containing ring would not significantly decrease its activity. But one of the analogues of compound I, N-methyl-2-methyl-8,9-methylenedioxy phenanthridinium chloried (II) also showed activity without a betaine structure. It was assumed that N-methyl-2-carboxyl-8,9-methylenedioxy-phenanthridinium chloride (III) was the active metabolite of compound II in vivo . In the present paper, the synthesis of compound III is reported. The procedure of synthesis is shown in the scheme. Preliminary pharmacological tests showed that compound III was inactive both in vitro (P388) and in vivo (Ehrlich ascites carcinoma).     

新型5-HT2A选择性配体N-取代哌啶-4-苯硫醚和砜类衍生物的合成及其生物活性

目的为寻找新型的5-HT_2A受体选择性配体,设计合成了一系列二芳烷基哌啶类化合物的含硫衍生物。方法以2,3-二甲氧基硫酚为原料,经烃化、氧化和水解等反应合成3个N-取代哌啶-4-苯硫醚和砜类化合物,所有目标化合物结构均经元素分析、¹HNMR谱、质谱和红外光谱确证,并测定其对5-HT_2A,5-HT_2C,5-HT₆和5-HT₇受体及其他一些中枢神经递质受体的体外亲和力。结果3个目标化合物(2a-2c)及5个中间体均为新化合物。体外受体竞争结合试验结果表明2a-2c均有较高的5-HT_2A受体选择性。结论此类化合物对5-HT_2A受体的选择性较高,值得进一步研究。     

Opiate physical dependence and N-methyl-d-aspartate receptors

The present review focused the involvement of N-methyl-d-aspartate (NMDA) receptors in morphine physical dependence. The increased levels of extracellular glutamate, NMDA receptor ζ subunit (NR1) mRNA, NMDA receptor 1 subunit (NR2A) protein, phosphorylated Ca²⁺/calmodulin kinase II (p-CaMKII) protein, c-fos mRNA, c-Fos protein, are observed in the specific brain areas of mice and/or rats showing signs of naloxone-precipitated withdrawal. In preclinical and clinical studies, a variety of NMDA receptor antagonists and pretreatment with an antisense oligonucleotide of the NR1 have been reported to inhibit the development, expression and/or maintenance of opiate physical dependence. In contrast to data obtained in adult animals, NMDA receptor antagonists are neither effective in blocking the development of opiate dependence nor the expression of opiate withdrawal in neonatal rats. In the NMDA receptor-deficient mice, the NR2A knockout mice show the marked loss of typical withdrawal abstinence behaviors precipitated by naloxone. The rescue of NR2A protein by electroporation into the nucleus accumbens of NR2A knockout mice reverses the loss of abstinence behaviors. The activation of CaMKII and increased expression of c-Fos protein in the brain of animals with naloxone-precipitated withdrawal syndrome are prevented by NMDA receptor antagonists, whereas the increased levels of extracellular glutamate are not prevented by them. These findings indicate that glutamatergic neurotransmission at the NMDA receptor site contributes to the development, expression and maintenance of opiate dependence, and suggest that NMDA receptor antagonists may be a useful adjunct in the treatment of opiate dependence.     

N-tosyl- -phenylalanyl-chloromethylketone reduces hypoxic–ischemic brain injury in rat pups

N-tosyl- -phenylalanyl-chloromethylketone (TPCK) in vitro blocks apoptotic pathways leading to cell death. We wished to see if TPCK would reduce brain injury in vivo. Seven-day-old rat pups had the right carotid artery ligated and then received either vehicle or TPCK (5 to100 mg/kg i.p.). They were then given 8% oxygen for 2.25 h. Twenty-two days later, the cerebral hemispheres were weighed to determine the reduction in size in the right hemisphere. TPCK decreased the reduction in right hemisphere weight from 15±3% (vehicle, n=20), to 4±2% (10 mg/kg, n=19, P<0.01). TPCK reduced the number of cells staining for DNA breaks 3 days after injury from 1729±275 mm⁻² (vehicle, n=8) to 550±236 mm⁻² (10 mg/kg TPCK, n=9, P<0.01), decreased the amount of DNA fragmentation 3 days after injury by gel electrophoreses (20 mg/kg, n=16, P<0.01) and eliminated the increase in nitric oxide metabolites 6 h after injury (vehicle 1.5±0.4, n=10; and 20 mg/kg TPCK 0.0±0.1 nM/mg protein, n=10, P<0.001). TPCK pretreatment in the newborn rat model of hypoxic–ischemic brain injury reduces DNA fragmentation, nitric oxide production and brain injury.     

N-Nitroso-N-Methyldodecylamine and N-Nitroso-N-Methyltetradecylamine in household dishwashing liquids

Eleven household dishwashing liquids and four household surface cleaners were analysed for N-nitroso-N-methyldodecylamine and N-nitroso-N-methyltetradecylamine by gas chromatography with detection using a Thermal Energy Analyzer. Both nitrosamines were found in three of the dishwashing detergents and one of the surface cleaners. [1-¹⁴C]-N-Nitroso-N-methyldodecylamine was used to determine recoveries, which were between 65 and 88%. Levels of N-nitroso-N-methyldodecylamine ranged from 112 to 661 ppb and those of N-nitroso-N-methyltetradecylamine from 46 to 151 ppb. A simple method was developed to screen the products for N,N-dimethyldodecylamine-N-oxide, a surfactant ingredient suspected of being the source of these nitrosamines. By application of this method it was established that all of the products formulated with this amine oxide contained these two nitrosamines, whereas in products that did not contain this ingredient, these nitrosamines were not detected.     

W3D固体分散体的制备及其抗对乙酰氨基酚诱导的急性肝损伤的保护作用

目的 将4-(5’-二甲氨基)-萘磺酰氧基苯并噁唑酮(W3D)制成固体分散体，并研究其对对乙酰氨基酚(N-acetyl-para-aminophenol，APAP)诱导的急性肝损伤(acute liver injury, ALI)的保护作用。方法 采用溶剂法将原料药W3D及辅料PVPk30按1:7反应得W3D固体分散体。采用差示扫描量热(DSC)和X-粉末射线衍射(XRD)手段表征固体分散体的物相变化，同时对其饱和溶解度、溶出重现性进行考察。建立对乙酰氨基酚诱导的ALI模型，分别灌胃给予CMC-Na、阳性药N-乙酰半胱氨酸(NAC 12.5 mg·kg-1)、W3D原料药(12.5 mg·kg-1)、W3D固体分散体(12.5、6.25和3.125 mg·kg-1)，24 h后处死小鼠，取出肝脏，计算肝脏指数；HE染色观察肝组织病理变化，微孔板法检测血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)的含量及肝组织中超氧化物歧化酶(SOD)活力、微量还原型谷胱甘肽(GSH)含量，TBA法检测丙二醛(MDA)的含量，ELISA法检测血清中肿瘤坏死因子(TNF-α)、白介素6(IL-6)的含量，免疫组化检测肝组织MD2蛋白的表达。结果 W3D固体分散体为无定形形态，溶出度10 min即可达70 %；W3D可降低APAP诱导的ALI小鼠肝脏指数，减少肝组织血管周围细胞胞核固缩、碎裂或溶解等现象，剂量依赖性地减少血清中ALT、AST、TNF-α、IL-6的含量，升高肝组织中GSH含量及SOD活力，降低MDA含量而呈现出优于临床用药NAC的肝保护活性；W3D亦可降低ALI小鼠肝组织中MD2蛋白的含量。结论 W3D固体分散体可通过抑制氧化应激和炎症而发挥抗APAP诱导的ALI作用，其作用机制可能与降低MD2蛋白表达有关。     

N-硬脂酰酪氨酸的理化性质研究

目的 表征N-硬脂酰酪氨酸及其盐的理化常数和血浆蛋白结合率。方法 采用摇瓶法及HPLC-UV法测定N-硬脂酰酪氨酸及其盐在不同介质和pH值下的溶解度、表观脂水分配系数,采用平衡透析法测定血浆蛋白结合率。结果 N-硬脂酰酪氨酸盐的溶解度和血浆蛋白结合率明显优于其原型药物;N-硬脂酰酪氨酸的溶解度为（27.62±0.43）mg/L,其二钾盐的溶解度为（51.70±0.29）mg/L;N-硬脂酰酪氨酸及其二钾盐的血浆蛋白结合率分别为20.96%～23.11%、35.41%～40.38%。结论 N-硬脂酰酪氨酸成盐后其理化性质得到明显改善。