HPLC-MS/MS同时测定尿酸调节类中药及含中草药食品中22种非法添加药物

目的 采用高效液相色谱-串联质谱法同时测定尿酸调节类中药及含中草药食品中22种非法添加药物。方法 样品经含0.1%氨水的甲醇超声提取后,采用Agilent Poroshell 120 Bonus-RP(2.1 mm×100 mm,2.7μm)色谱柱进行分离,以甲醇-乙腈(1∶1)和0.005 mol·L^-1甲酸铵溶液为流动相梯度洗脱。采用电喷雾离子源正、负离子模式,以多反应监测扫描方式检测。结果 22种药物在10.0～200μg·L^-1内线性关系良好,相关系数r²均>0.99,检测限和定量限分别为0.125～0.750 mg·kg^-1和0.416～2.81 mg·kg^-1,平均回收率为80.2%～120.1%,RSD为1.1%～5.9%。收集的37批样品中均未检出非法添加的药物。结论 该方法操作简单、快速、灵敏度高,可实现同时检测尿酸调节类中药及含中草药食品中22种非法添加药物。     

不同成熟度川佛手化学成分及含量比较分析

目的：采用超高液相色谱-四级杆-静电场轨道阱高分辨质谱(UPLC-Q-Orbitrap HRMS)技术及高效液相色谱(HPLC)法,对不同成熟度的川佛手化学成分进行比较分析。方法：UPLC-Q-Orbitrap HRMS法采用Thermo Scientific AccucoreTM C18柱(100 mm×3 mm,2.6μm),以0.1%甲酸水和0.1%甲酸乙腈为流动相进行梯度洗脱,流速0.3m L·min^-1,柱温30℃,进样量3μL,质谱采用电喷雾ESI电离源,正离子模式下扫描采集数据;HPLC采用Thermo Hypersil GOLD C18色谱柱(250 mm×4.6 mm,5μm),流动相为0.05%磷酸水和乙腈进行梯度洗脱,检测波长254 nm。结果：不同成熟度川佛手液质联用测定结果经PCA分析较为分散,无聚集趋势,显示各组之间化学成分总体无明显差异;香叶木苷、橙皮苷、5,7-二甲氧基香豆素、氧化前胡素4种化学成分总含量在青果、青黄果及黄果中范围分别为1.9813～4.8521 mg·g^-1、1.9106～3.8384 mg·...     

庞氏安胎止血标准汤剂HPLC指纹图谱建立及7种成分含量测定

目的：建立庞氏安胎止血标准汤剂HPLC指纹图谱,并对7种成分进行含量测定,为庞氏安胎止血标准汤剂的质量控制和后续研究提供依据。方法：采用YMC-Triart C₁₈(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.1%磷酸水为流动相,检测波长为230 nm,梯度洗脱,流速1.0 mL·min^–1,柱温30℃。采用《中药色谱指纹图谱相似度评价软件》(2012年版)对10批标准汤剂进行相似度评价,并对其进行含量测定。结果：建立了庞氏安胎止血标准汤剂HPLC指纹图谱,相似度均大于0.97,共标定23个共有峰,指认13个共有峰,10批样品中绿原酸、异阿魏酸、甘草苷、菊苣酸、迷迭香酸、黄芩苷、木犀草素的质量分数分别为1.56～3.51 mg·g^–1,3.43～5.28 mg·g^–1,3.42～5.08 mg·g^–1,0.53～0.62 mg·g^–1,0.80～1.34 mg·g^–1,3.51～6.24 mg·g^–1...     

高效液相色谱-三重四级杆质谱快速筛查及定量检测改善睡眠类保健品和中成药中的22个精神类化合物

目的:建立高效液相色谱-三重四级杆质谱(HPLC-MS/MS)法快速筛查及定量分析改善睡眠类保健品及中成药中22个精神类化合物。方法:采用ZORBAX Eclipse Plus-C₁₈ (2.1 mm×50 mm, 1.8μm)色谱柱进行分离,以乙腈-1 mmol·L^-1乙酸铵(含0.1%甲酸)为流动相,梯度洗脱,流速为0.3 mL·min^-1,电喷雾离子源,用多反应监测结合正离子扫描分析。通过比较样品与对照品的色谱保留时间、相对离子丰度比,进行定性分析,并对阳性样品进行定量分析。结果:22个精神类化合物(阿普唑仑、地西泮、米安色林、帕罗西汀、氯丙嗪等)在0.1～400 ng·mL^-1浓度范围呈良好的线性关系,线性相关系数大于0.99,样品回收率在79.1%～109.2%,RSD在1.2%～9.6%,检测下限范围在0.05～0.5 ng·mL^-1。本方法可在13 min内同时完成对保健品和中成药中的22个精神类化合物的分离和筛查。检测12批次样品,结果均未检出22个精神类化合物...     

二维柱切换高效液相色谱法测定美白类化妆品中氢醌和苯酚

目的:建立二维柱切换高效液相色谱法测定化妆品中氢醌、苯酚的分析方法。方法:二维色谱柱均为C₁₈色谱柱(250 mm×4.6 mm, 5μm),捕获柱为2条串联的C₁₆色谱柱(20 mm×4.0 mm, 5μm),10 mmol·L^-1磷酸二氢钾溶液(用磷酸调节pH3.5)-甲醇为流动相,梯度洗脱,流速1.0 mL·min^-1,柱温35℃,检测波长280 nm,进样量2μL。结果:氢醌和苯酚线性范围分别为5.570～111.5μg·mL^-1(r=0.999 7)和5.410～108.1μg·mL^-1(r=0.999 6),方法提取回收率分别为91.9%～92.6%和94.4%～94.7%,精密度试验的RSD分别为3.9%～4.1%和0.9%～2.1%,检测下限分别为0.70μg·g^-1和0.69μg·g^-1。市场随机抽取40批美白类化妆品样品(包含乳剂、霜剂、水剂等)均单次进样即可快速完成样品分析,无基质干扰。结论...     

HPLC-MS/MS测定化妆品中14种荧光增白剂

目的建立HPLC-MS/MS定量测定化妆品中14种荧光增白剂的分析方法。方法样品经N,N-二甲基甲酰胺超声提取,选用Waters ACQUITY UPLC?BEH C₁₈(2.1 mm×50 mm,1.7μm)色谱柱,以甲醇-0.1%氨水水溶液梯度洗脱,流速0.3 mL·min^–1,柱温40℃;质谱法采用三重四级杆质谱,电喷雾离子源,在正/负离子、多反应监测模式下进行扫描。结果14种荧光增白剂在定量范围内线性关系良好,相关系数均>0.99,定量限为0.01～20μg·g^–1,检出限为0.004～8μg·g^–1;在3个添加水平下的平均回收率为85.4%～108.9%,相对标准偏差为0.3%～7.2%。结论该方法灵敏度高,专属性强,操作简单方便,适用于化妆品中14种荧光增白剂的含量测定。     

HPLC测定重组Exendin-4-FC融合蛋白注射液中抗氧剂及其降解物

目的 建立HPLC对重组Exendin-4-FC融合蛋白注射液中11种抗氧剂及其降解物含量测定方法。方法 采用饱和硫酸铵沉淀蛋白,离心后上清液转移至经甲醇活化后的C₁₈固相萃取小柱中,分别用4 mL甲醇、5 mL乙酸乙酯依次洗脱小柱,洗脱液用甲醇-乙酸乙酯(2∶3)混合溶剂定容后过0.22μm PTFE疏水滤膜,经HPLC分析,外标法定量。色谱条件：Kinetex^?XB-C₁₈ 100?(100 mm×4.6 mm,2.6μm)色谱柱,检测波长230 nm,柱温箱30℃,进样量5μL,流速0.4 mL·min^–1,流动相0.1%甲酸-甲醇(A)-0.1%甲酸水溶液(B),运行时间45 min。结果 11种目标物在2.5～35μg·mL^–1内线性关系良好(R²≥0.99)。在25,10,5μg·mL^–1 3个浓度水平下,平均加样回收率为88.1%～106.5%,RSD为0.10%～9.05%。6份样品重复性RSD为2.01%～4.77%...     

液质联用技术用于醋酸阿托西班注射液中杂质结构鉴定

目的:采用高效液相色谱(HPLC)、超高效液相色谱-四极杆串联静电场轨道阱高分辨质谱(UPLC-Q Orbitrap MS)技术对醋酸阿托西班注射液中杂质结构进行鉴定。方法:HPLC法采用Inertsil ODS-2 C₁₈色谱柱(250 mm×4.6 mm, 5μm),流动相A为乙腈-甲醇-三氟乙酸溶液(pH 3.2)(15∶10∶75),流动相B为乙腈-甲醇(60∶40),梯度洗脱，流速1.2 mL·min^-1,检测波长220 nm。UPLC-Q Orbitrap MS法采用BEH300 C₁₈色谱柱(150 mm×2.1 mm, 1.7μm),流动相为0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B),梯度洗脱，流速0.2 mL·min^-1;采用电喷雾离子源，选择正离子模式进行Full MS/dd-MS²扫描。结果:对5家企业各1批醋酸阿托西班注射液进行了有关物质测定，以面积归一化计算，含量>0.1%的杂质峰个数分别为9、10、13、9和6,总杂含量分别为0.35...     

HPLC-MS/MS法测定化妆品中非法添加的他克莫司和吡美莫司

目的:采用高效液相色谱串联飞行时间质谱法高通量筛查化妆品中的非法添加成分，并建立高效液相色谱串联三重四极杆质谱法测定化妆品中他克莫司和吡美莫司的方法。方法:通过高效液相色谱串联飞行时间高分辨质谱法对市售儿童化妆品进行风险物质高通量初步筛查，发现1批他克莫司阳性样品，然后运用高效液相色谱串联三重四极杆质谱法进行定性定量测定。经实验条件优化，以含17.5 mmol·L^-1乙酸-5 mmol·L^-1乙酸铵的水溶液(A)和含17.5 mmol·L^-1乙酸-5 mmol·L^-1乙酸铵的甲醇溶液(B)为流动相，采用Waters ACQUITY UPLC BEH C₁₈色谱柱，进行梯度洗脱分离，柱温45℃,流速0.3 mL·min^-1,然后经三重四极杆质谱正离子多反应监测模式扫描。结果:他克莫司和吡美莫司质量浓度在0.5～50 ng·mL^-1范围内线性关系良好，检出限均为10 ng·g^-1,定量限均为25 ng·g^-...     

多波长HPLC-DAD法同时测定美白类化妆品中8种甘草化学成分

目的 建立多波长高效液相色谱-二极管阵列检测器(HPLC-DAD)法同时测定美白类化妆品中8种甘草化学成分的含量。方法 样品经70%甲醇溶液提取、过滤后,经Phenomenex C₁₈色谱柱(4.6 mm×250 mm, 5μm)分离,流动相为乙腈-0.1%磷酸,梯度洗脱,231、249和365 nm波长同时检测,流速1.0 mL·min^-1,柱温30℃。结果 8种成分分离度良好,在各进样浓度范围内线性关系良好,相关系数r均大于0.998;4种不同化妆品基质的平均加样回收率(n=3)为88.3%～96.2%,相对标准偏差(RSD)为0.9%～3.5%。结论 本方法准确可靠、灵敏度高,可用于同时测定化妆品中8种甘草化学成分。     

Analgesia induced by brief footshock is inhibited by 5-hydroxytryptamine but unaffected by antagonists of 5-hydroxytryptamine or by naloxone

Exposure to footshock (1 mA) for 30 sec induced a marked analgesia that was enhanced by pretreatment with the 5HT synthesis inhibitor, p-chlorophenylalanine, and attenuated by the 5HT releasing drugs p-chloroamphetamine and fenfluramine, by the 5HT re-uptake inhibitor, fluoxetine and by the 5HT agonists, 5-methoxy-N,N-dimethyltryptamine and MK212. However, agonists, quipazine and trifluoromethylphenylpiperazine, with greated reported affinities for 5HT binding sites on rat brain membranes than MK212 were without effect as were the antagonists metergoline, methysergide, cyproheptadine, mianserine and methiothepin. The specific opioid antagonist naloxone was also without effect. The results in general indicate that analgesia induced by brief footshock (1 mA, 30 sec) is inversely related to 5HT availability but thereis little evidence of involvement of known 5HT receptors.     

Synergism by caffeine and by cocaine of the motor control deficit produced by midazolam

To evaluate the effects of caffeine and cocaine on the impairment of discriminative motor control produced by midazolam, rats were trained to hold a force transducer operated with a paw so that it remained between upper and lower limits of a force band for a continuous 1.5-s period to deliver each food pellet. Acute doses of 3 mg/kg midazolam SC impaired motor performance. Except for one animal, caffeine (10-40 mg/kg IP) had little or no effect on performance, while cocaine (3.75-22.5 mg/kg IP) produced dose-related impairment. When each dose of caffeine was combined with 3 mg/kg midazolam, a marked synergism in motor performance impairment occurred. Cocaine plus midazolam produced mainly an additive synergism. The conspicuous synergistic action of caffeine on the motor control deficit produced by midazolam contrasts with the typical antagonism found between the benzodiazepines and methylxanthines when performance is evaluated by psychomotor tests not requiring fine motor control.     

Deamination of beta-phenylethylamine by monoamine oxidase--inhibition by imipramine

The oxidative deamination of tyramine (Tyr), 5-hydroxytryptamine (5-HT), and β-phenylethylamine (PEA) by mitochondrial preparations of rabbit lung and brain was inhibited by imipramine. This tricyclic iminodibenzyl antidepressant drug was most effective in decreasing the deamination of PEA: at 1 × 10^?4M imipramine, deamination of PEA, Tyr and 5-HT was inhibited by approximately 70, 45 and 45 per cent, respectively, when either lung or brain mitochondrial monoamine oxidase (MAO) preparations were used. Imipramine-induced inhibition of MAO was shown to be of a mixed type based on Lineweaver-Burk plots, but was found to be completely reversible. The desmcthyl and didesmethyl derivatives of imipramine were equally as effective as the parent drug in inhibiting the deamination of PEA, whereas the N-oxide analog of imipramine was less effective as an inhibitor of this reaction. These results support the premise that the action of imipramine as a clinically effective antidepressive agent may be related to its inhibitory effect on the specific form of MAO which deaminates PEA.     

Augmentation by serrapeptase of tissue permeation by cefotiam

Cefotiam (CTM) is a new cephalosporin with a broad spectrum of activity against both Gram-positive and Gram-negative microorganisms. Cephalosporins are widely used for prophylaxis of infections in patients undergoing thoracotomy. Augmentation by serrapeptase on tissue permeation of CTM was examined in 35 thoracotomy patients with lung cancer. The subjects were divided into two groups according to the method of the administration of CTM. Group I consisted of 17 subjects, each of whom received a single dose of 2 g of CTM alone by an instillation for 30 minutes. Group II consisted of 18 subjects, each of whom received a combination of CTM and serrapeptase; serrapeptase was given 2 tablets (10 mg) each time for three times/day until the day before surgery, and then CTM was administered by the same procedure. The following results were obtained: Individual difference was observed for the permeation of CTM into tissues. Pathologic differences also affected the permeation. Nevertheless, the CTM levels in pulmonary tissues reached about a half of those in the blood in both the single dose group and the combination group, hence sufficient concentrations exceeding MIC80 for main microorganisms that caused infections in the lung were obtained. The concentrations of CTM in inflammatory tissues have showed lower levels than those of normal tissues in both CTM single dose and the combination groups. Decrease of blood flow volume may have contributed to the reduction in levels of CTM in the inflammatory tissues. The ratio of the concentration of the drug in pulmonary tissues to that in the blood was 29.1 +/- 2.5% in the single dose group, and 44.2 +/- 6.0% in the combination group, the latter showing quite a significant increase (P less than 0.05). Combined administrations of CTM and serrapeptase deserves more trials in the case when surgical treatments of the lung are performed. An antiinflammatory effect of serrapeptase in the respiratory system is expected, and in addition, the combined use of CTM and serrapeptase should stimulate permeation of the antibiotic into tissues.     

Interference by cephalosporins with creatinine measurement by desk-top analyzers

Summary We have carried out a study to evaluate the interference by cephalosporins with the measurement of creatinine by desk-top analyzers. The cephalosporins evaluated at concentrations of 0–250 mg/l were cefazolin sodium, cefoxitin sodium, cefotaxime sodium, and ceftazidime pentahydrate. The instruments evaluated were DT60 (Kodak, Rochester, USA), Seralyzer (Ames Division, Miles Laboratories, IN, USA), and Vision (Abbott Labs, Chicago, USA). All studies were done in plasma.None of the cephalosporins showed any interference with the DT60 analyzer. With the Vision and Seralyzer no interference was seen with cefotaxime or cefazolin. With cefazolin an increase of 10–20 µmol/l creatinine was seen for every 20 mg/l of drug; with cefoxitin there was an increase of 50–80 µmol/l of creatinine for every 100 mg/l of drug.Erroneous creatinine values may be found in patients taking cefazolin and cefoxitin and may lead to inappropriate clinical management.     

Antagonism of morphine by beta-melanocyte-stimulating hormone and by tetracosactin

Using the decerebrate—spinal Lloyd preparation morphine depressed evoked mono- and polysynaptic reflex activity, β-melanocyte-stimulating hormone enhanced monosynaptic reflex activity, and tetracosactin had no effect. When morphine injection was preceded either by β-melanocyte-stimulating hormone or by tetracosactin a statistically significant depression was not observed. The stimulant actions of β-melanocyte-stimulating hormone did not appear to account for its capacity to antagonize morphine. The fall of blood pressure which follows the administration of morphine in this preparation was not antagonized by the prior administration of either polypeptide.     

Immunomodulation by non-absorbable antibiotics given by the intragastric route

To test the role of bacterial fractions released from intestinal flora during immunomodulation by antimicrobial agents, BALB/c mice were treated with the non-absorbable antibiotics polymyxin B or teicoplanin by the intragastric route. The composition of faecal microbiota and the capacity of spleen cells to proliferate in response to B-cell and T-cell mitogens were assessed at several times during the treatment. Both antibiotics lowered the count of some bacteria of the intestinal flora and induced significant modifications in spleen cell ability to proliferate in response to mitogens. Thus, the active fractions released from intestinal bacteria during antibiotic treatments may be able to induce immunomodulating effects.