药用卤化丁基橡胶塞中2-巯基苯并噻唑的检测

目的 建立高效液相色谱法测定药用卤化丁基橡胶塞中2-巯基苯并噻唑含量的方法。方法 使用丙酮作为萃取剂,对药用卤化丁基橡胶塞中的2-巯基苯并噻唑进行微波萃取。采用ZORBAX SB-C₈（4.6mm×250mm,5 μm）色谱柱,流动相为1%乙腈水溶液和乙腈,梯度洗脱,流速1.0 mL·min^-1,检测波长320 nm,柱温为25 ℃,进样量10 μL。结果 2-巯基苯并噻唑在1.02～102.26 μg·mL^-1的浓度范围内线性良好（r=0.999 9）,检出限达到0.03 µg·mL^-1,精密度、稳定性RSD均小于3%,样品加标回收率为97.8%~108.9%。2种药用卤化丁基橡胶塞中均未检测到2-巯基苯并噻唑。结论 经方法学验证,本法可用于药用卤化丁基橡胶塞中2-巯基苯并噻唑含量的测定。     

GC-MS测定中药提取物中34种抑菌剂与抗氧剂

目的 建立同时测定中药提取物中34种抑菌剂与抗氧剂的GC-MS分析方法。方法 采用Agilent DB-5MS毛细管色谱柱（60 m×320 μm，0.5 μm）；进样口温度：250℃；载气为高纯氦气；流速：2 mL·min^-1；进样量：1 μL；柱温40℃，保持1 min，以5℃·min^-1的速率升至300℃，保持10 min；电离源为电子轰击源（EI）；采用多反应监测模式进行定性及定量分析。结果 34种抑菌剂与抗氧剂在一定浓度范围内线性关系良好，相关系数r均>0.99，各成分在浸膏中回收率为83.7%~124.8%，RSD （n=6）为0.5%~5.2%；在粉末中的回收率为71.3%~119.6%，RSD （n=6）为0.7%~5.6%。结论 该方法的灵敏度、仪器精密度、回收率均能满足中药提取物中抑菌剂与抗氧剂检测的要求。     

GC-MS测定药用LDPE袋中2,2,4,6,6-五甲基庚烷含量及其迁移量

目的 建立药用LDPE袋中2,2,4,6,6-五甲基庚烷含量及其向包装的原料药中迁移量的气相色谱-质谱测定法。方法 采用Stabilwax毛细管色谱柱（30 m×0.25 mm,0.25 μm）;柱温：60℃,离子源温度：240℃,接口温度260℃,流速1.0 mL·min^-1。结果 2,2,4,6,6-五甲基庚烷在0.060 00~120.0 ng内呈良好的线性关系（r=0.999 6）;药用LDPE袋中2,2,4,6,6-五甲基庚烷含量测定的平均回收率为94.3%（RSD=3.8%）。结论 该方法准确、灵敏、简便,适用于对药用LDPE袋中2,2,4,6,6-五甲基庚烷含量及其向包装的原料药中迁移量的测定。     

川菊止痛胶囊的质量标准研究

目的 建立川菊止痛胶囊质量标准。方法 采用薄层色谱法对复方中柴胡、菊花进行定性鉴别。HPLC同时测定制剂中黄芩苷、黄芩素、汉黄芩苷、汉黄芩素及甘草苷含量,色谱柱：Kromasil 100-5C₁₈（250 mm×4.6 mm,5 μm）;流动相：乙腈-0.05%磷酸溶液,梯度洗脱;检测波长为280 nm;流速：1.0 mL·min^-1;进样量：10 μL。结果 薄层色谱斑点清晰且阴性样品无干扰。甘草苷、黄芩苷、黄芩素、汉黄芩苷及汉黄芩素分别在10.7~107 μg·mL^-1、12.12~121.2 μg·mL^-1、11.2~112 μg·mL^-1、10.02~100.2 μg·mL^-1、10.32~103.2 μg·mL^-1内线性关系良好,精密度、重复性、稳定性试验RSD均<1.8%,加样回收率分别为98.34%~101.77%（RSD=1.28%,n=6）、98.69%~101.36%（RSD=1.01%,n=6）、98.46%~101.06%（RSD=0.92%,n=6）、98.67%~101.33%（RSD=1.17%,n=6）、98.43%~100.79%（RSD=0.92%,n=6）。结论 本研究建立的质量标准方法准确、简便,可用于川菊止痛胶囊的质量控制,且所建立的菊花、柴胡薄层鉴别方法可供其他含有二药的复方标准建立作参考。     

离子色谱法测定氯化琥珀胆碱原料及注射液中的氯化胆碱和氯化琥珀酰单胆碱

目的 采用离子色谱法测定氯化琥珀胆碱原料及注射液中的氯化胆碱和氯化琥珀酰单胆碱。方法 采用Dionex RFICTM Ionpac CS17色谱柱（4 mm×250 mm）,以甲烷磺酸溶液为流动相进行梯度洗脱,流速为1.0 mL·min^-1,采用抑制电导检测器进行测定。结果 氯化胆碱、氯化琥珀酰单胆碱浓度分别在18.0~562.8 μg·mL^-1（r=0.999 9）和14.4~449.8 μg·mL^-1（r=0.999 9）内线性关系良好,定量限均为0.01 μg·mL^-1。原料中氯化胆碱、氯化琥珀酰单胆碱回收率分别为103.4%~105.7%和102.4%~107.1%;注射液中氯化胆碱、氯化琥珀酰单胆碱回收率分别为95.6%~100.2%和94.4%~103.5%。结论 该方法专属性强、准确、简便、快速,适用于氯化琥珀胆碱原料及制剂中的有关物质测定。     

HPLC测定早产儿咖啡因血药浓度

目的 建立HPLC测定早产儿咖啡因血药浓度的方法。方法 色谱柱为Insertil ODS-3（250 mm×4.6 mm,5 μm）,甲醇-水（28：72）为流动相,流速1 mL·min^-1,检测波长274 nm,柱温30℃,进样量20 μL。结果 咖啡因血药浓度在2.5~60 mg·L^-1内呈良好的线性关系（r²=0.999 9,n=9）,定量限为0.125 mg·L^-1。提取回收率为82.5%~89.7%,方法回收率为97.2%~103.0%;日内与日间精密度RSD均<10%。随机检测5名负荷剂量给药的患儿次日咖啡因谷浓度为（15.8±2.1） mg·L^-1,另5例患儿给药7 d后检测咖啡因谷浓度为（24.3±4.6） mg·L^-1,均在有效线性范围内。结论 本方法灵敏、可靠,可作为咖啡因血药浓度的常规监测方法。     

UPLC-MS/MS测定减肥食品中8种非法添加化学成分

目的 建立一种简便有效检测减肥食品中非法添加化学成分的UPLC-MS/MS检测方法。方法 采用UPLC-MS/MS法,以Eclipse Plus C₁₈（2.1 mm×50 mm,1.8 μm）色谱柱分离,串联四级杆质谱仪检测,多反应监测模式进行定性定量分析,以流动相A（0.02 mol·L^-1乙酸铵水溶液）-B（甲醇）进行梯度洗脱;柱温：40℃;流速：0.3 mL·min^-1,进样量2 μL。样品以甲醇为溶剂超声提取,快速定性定量检测非法添加在减肥食品中的麻黄碱、咖啡因、呋塞米、芬氟拉明、酚酞、N-单去甲基西布曲明、N,N-双去甲基西布曲明、西布曲明8种化学成分。结果 8种减肥类化学成分质谱检测的线性范围宽,相关性R² ≥ 0.995;方法检测限为0.042~0.175 μg·mL^-1,定量限为0.126~0.525 μg·mL^-1,方法精密度RSD（n=6）为1.7%~4.7%;3个浓度水平的平均回收率为90.3%~105.1%。结论 本方法专属性强、操作简单、方便快捷,可作为食品中非法添加减肥类化学成分的有效检测方法。     

HPLC法检测醋酸奥曲肽注射剂中5种抑菌剂

目的：检测醋酸奥曲肽注射剂中是否添加苯酚、苯甲醇、三氯叔丁醇、对羟基苯甲酸甲酯及对羟基苯甲酸丙酯5种抑菌剂。方法：采用C18（L）（4.6 mm×250 mm,5 μm）色谱柱,流动相A为水,流动相B为甲醇,梯度洗脱,流速为1.0 mL·min^-1,柱温30℃,检测波长为220 nm及256 nm。结果：苯酚、苯甲醇、三氯叔丁醇、对羟基苯甲酸甲酯及对羟基苯甲酸丙酯各峰均分离良好;苯酚在1.29~258.60 μg·mL^-1范围内线性关系良好（r=1.000）,平均回收率为100.0%（n=9）;苯甲醇在2.72~544.20 μg·mL^-1范围内线性关系良好（r=0.9997）,平均回收率为100.3%（n=9）;三氯叔丁醇在9.95~1990.00 μg·mL^-1范围内线性关系良好（r=1.000）,平均回收率为100.9%（n=9）;对羟基苯甲酸甲酯在0.27~53.80 μg·mL^-1范围内线性关系良好（r=1.000）,平均回收率为100.5%（n=9）;对羟基苯甲酸丙酯在0.26~52.00 μg·mL^-1范围内线性关系良好（r=1.000）,平均回收率为99.4%（n=9）;91批醋酸奥曲肽注射剂中均未检出5种抑菌剂。结论：该方法准确、灵敏、简便,可用于醋酸奥曲肽注射剂中苯酚、苯甲醇、三氯叔丁醇、对羟基苯甲酸甲酯及对羟基苯甲酸丙酯5种抑菌剂的检查。     

HPLC测定沅江产芦笋中3种酚酸和6种黄酮类成分含量

目的 建立同时测定沅江产芦笋中原儿茶酸、绿原酸、香草酸、柚皮苷、芦丁、槲皮素、木犀草素、山奈酚、芹菜素含量的HPLC。方法 采用Diamonsil C₁₈色谱柱（250 mm×4.6 mm,5 μm）,以甲醇-0.1%甲酸水溶液为流动相,梯度洗脱,双波长检测（酚酸：283,327 nm;黄酮：283,335 nm）,流速1 mL·min^-1,柱温35℃,进样量为10 μL。结果 3种酚酸及6种黄酮分别在25 min和50 min内分离,线性范围为14.8~500 μg·mL^-1（r=0.999 0~0.999 7）,检测限0.006 3~0.053 9 μg·mL^-1,定量限0.021 1~0.179 7 μg·mL^-1,加样回收率为98.73%~102.44%,RSD为1.27%~2.20%（n=6）。结论 该方法准确灵敏、重复性好,可用于检测芦笋中9种成分含量。     

GC-MS/MS测定中药中18种多氯联苯

目的 建立GC-MS/MS测定中药材中18种多氯联苯（PCBs）的方法。方法 采用超声提取,浓硫酸净化,内标法定量;采用EI源,在多反应监测（MRM）模式下进行检测。结果 18种PCBs在0.5~50 ng·L^-1时线性关系良好,相关系数（r）为0.999 8~1.000 0,方法的检出限为0.013~0.080 μg·kg^-1,定量限为0.054~0.328 μg·kg^-1。在2,5,10 μg·L^-1下平均回收率分别为106.7%~118.7%,93.5%~114.7%和89.9%~115.6%,相对标准偏差分别为1.6%~8.0%,2.7%~4.6%和1.0%~3.1%。结论 本方法样品处理简单、快速,灵敏度高,准确性好,可用于中药材中18种PCBs的测定。     

Interaction of estrone and estradiol with DNA and protein of liver and kidney in rat and hamster in vivo and in vitro

[6,7-³H] Estrone (E) and [6,7-³H]estradiol-17 (E₂) have been synthesized by reduction of 6-dehydroestrone and 6-dehydroestradiol with tritium gas. Tritiated E and E₂ were administered by oral gavage to female rats and to male and female hamsters on a dose level of about 300 g/kg (54 mCi/kg). After 8 h, the liver was excised from the rats; liver and kidneys were taken from the hamsters. DNA was purified either directly from an organ homogenate or via chromatin. The radioactivity in the DNA was expressed in the units of the Covalent Binding Index, CBI = (mol chemical bound per mol DNA-P)/(mmol chemical administered per kg b.w.). Rat liver DNA isolated via chromatin exhibited the very low values of 0.08 and 0.09 for E and E₂, respectively. The respective figures in hamster liver were 0.08 and 0.11 in females and 0.21 and 0.18 in the males. DNA isolated from the kidney revealed a detectable radioactivity only in the female, with values of 0.03 and 0.05 for E and E₂, respectively. The values for male hamster kidney were < 0.01 for both hormones. The minute radioactivity detectable in the DNA samples does not represent covalent binding to DNA, however, as indicated by two sets of control experiments. (A) Analysis by HPLC of the nucleosides prepared by enzyme digest of liver DNA isolated directly or via chromatin did not reveal any consistent peak which could have been attributed to a nucleoside-steroid adduct. (B) All DNA radioactivity could be due to protein contaminations, because the specific activity of chromatin protein was determined to be more than 3,000 times higher than of DNA. The high affinity of the hormone to protein was also demonstrated by in vitro incubations, where it could be shown that the specific activity of DNA and protein was essentially proportional to the concentration of radiolabelled hormone in the organ homogenate, regardless of whether the animal was treated or whether the hormone was added in vitro to the homogenate.Carcinogens acting by covalent DNA binding can be classified according to potency on the basis of the Covalent Binding Index. Values of 10³–10⁴ have been found for potent, 10² for moderate, and 1–10 for weak carcinogens. Since estrone is moderately carcinogenic for the kidney of the male hamster, a CBI of about 100 would be expected. The actually measured limit of detection of 0.01 places covalent DNA binding among the highly unlikely mechanisms of action. Similar considerations can be made for the liver where any true covalent DNA binding must be below a level of 0.01. It is concluded that an observable tumor induction by estrone or estradiol is unlikely to be due to DNA binding.Paper presented at the Satellite Symposium of the European Society of Toxicology, Rome, March 29, 1983     

Metabolic interaction between imipramine and carbamazepine in vivo and in vitro in rats

Summary The pharmacokinetic consequences of the combination of carbamazepine with imipramine in male Wistar rats have been investigated. It was found that a 2-week treatment with the combination resulted in the increase of the concentrations of the parent compounds and a simultaneous decrease in their metabolites in blood plasma i.e. carbamazepine inhibited imipramine demethylation in the side chain while imipramine inhibited carbamazepine 10,11-epoxidation. The velocity of imipramine 2-hydroxylation and 10,11-epoxy-carbamazepine hydration did not seem to be changed by the combination. On the basis of studies in vitro it is concluded that the observed metabolic interaction between carbamazepine and imipramine is due to the competition of the drugs for the active centre of cytochrome P 450 and to a certain qualitative alteration of the enzyme by imipramine as can be deducted from the decrease of carbamazepine binding to the cytochrome. Send offprint requests to K. J. Netter     

Buprenorphine and naloxone alone and in combination in opioid-dependent humans

Subjective, physiological and behavioral effects of subcutaneously administered hydromorphone (6 mg), naloxone (0.2 mg), buprenorphine (0.2 and 0.3 mg), and two buprenorphine-naloxone combinations (buprenorphine 0.2 mg plus naloxone 0.2 mg and buprenorphine 0.3 mg plus naloxone 0.2 mg) were assessed under double-blind conditions in six opioid-dependent volunteers. Physiologic measures and subject- and observer-rated behavioral responses were measured before dosing and for 120 min after drug administration. Hydromorphone decreased pupil diameter and respiration, increased blood pressure and increased scores on subjective measures indicating opioid-like effects. Buprenorphine given alone had no significant effect on any variable measured. Naloxone given alone produced opioid abstinence-like effects which were measurable on subject- and observer-rated behavioral measures and physiological measures. Buprenorphine in combination with naloxone somewhat attenuated the naloxone-precipitated withdrawal response. Overall, the naloxone-buprenorphine combinations produced effects which were qualitatively similar to the effects of naloxone alone, suggesting a low potential for abuse of the combination product by opioid-dependent individuals.Supported by a grant from Reckitt and Colman Pharmaceutical Division and USPHS Grants DA-00050 and DA-04089 from the National Institute on Drug Abuse     

Consistency in catecholamine and cortisol excretion in males and females

Data from a series of experiments performed on 24 female and 24 male subjects were used to evaluate the consistency in urinary catecholamine and cortisol excretion. Data were available from 8 laboratory situations of varying activity level and content, spaced at intervals of maximum 3 months. Correlational analyses showed that for cortisol, interindividual consistency was higher for measures obtained on the same day than for measures obtained on different days. Interindividual consistency was generally high in catecholamine and cortisol excretion during non-stressful situations in both sexes. During experimental stress, however, consistency was as high as during nonstress for males, while it was lower for females. Analysis of variance components confirmed these results and showed that in males variation due to interindividual differences was high during both baseline and experimental-stress situations, while in females it was high during baseline situations only. During experimental stress, variation for females was due primarily to interaction. It is suggested that the males showed a more generalized stress response over situations than the females.     

Nicotine metabolism and urinary elimination in mouse: in vitro and in vivo

This study aimed at elucidating the in vivo metabolism of nicotine both with and without inhibitors of nicotine metabolism. Second, the role of mouse CYP2A5 in nicotine oxidation in vitro was studied as such information is needed to assess whether the mouse is a suitable model for studying chemical inhibitors of the human CYP2A6. The oxidation of nicotine to cotinine was measured and the ability of various inhibitors to modify this reaction was determined. Nicotine and various inhibitors were co-administered to CD2F1 mice, and nicotine and urinary levels of nicotine and four metabolites were determined. In mouse liver microsomes anti-CYP2A5 antibody and known chemical inhibitors of the CYP2A5 enzyme blocked cotinine formation by 85–100%, depending on the pre-treatment of the mice. The amount of trans-3-hydroxycotine was five times higher than cotinine N-oxide, and ten times higher than nicotine N-1-oxide and cotinine. Methoxsalen, an irreversible inhibitor of CYP2A5, significantly reduced the metabolic elimination of nicotine in vivo, but the reversible inhibitors had no effect. It is concluded that the metabolism of nicotine in mouse is very similar to that in man and, therefore, that the mouse is a suitable model for testing novel chemical inhibitors of human CYP2A6.     

Strain-dependency in motor activity and in concentration and turnover of catecholamines in synchronized rats

Circadian rhythm in motor activity was studied with an Animex motimeter in six strains of rats (ACI, BH, BS, DA, LEW, TNO) synchronized by a 12 hr light: 12 hr dark cycle. ANOVA revealed significant interstrain differences in motor activity as well as in the concentration and turnover of central noradrenaline and dopamine. Strain-dependent differences were also found with regard to tyrosine hydroxylase inhibition on motor activity. However, no significant interstrain correlations were found between endogenous concentration and/or turnover rates of the catecholamines and motor activity in normal and drug-treated rats.     

Pharmacokinetics and tolerability of artesunate and amodiaquine alone and in combination in healthy volunteers

Objectives  The WHO recommends artemisinin-based combination therapies for treatment of uncomplicated falciparum malaria. At least 15 African countries have adopted artesunate plus amodiaquine as treatment policy. As no pharmacokinetic data on this combination have been published to date, we investigated its pharmacokinetic interactions and tolerability in healthy volunteers in Africa. Methods  In a randomized, three-phase, cross-over study, amodiaquine (10 mg/kg) and artesunate (4 mg/kg) were given as single oral doses to 15 healthy volunteers. Artesunate was given to all volunteers on day 0. On day 7 they received either amodiaquine or amodiaquine plus artesunate and the alternative regimen on day 28. The pharmacokinetics of artesunate and amodiaquine and their main active metabolites dihydroartemisinin and desethylamodiaquine were compared following monotherapy and combination therapy using analysis of variance. Results  Thirteen volunteers completed the study, and pharmacokinetic parameters could be determined for twelve volunteers. When given in combination, the mean AUC was lower for dihydroartemisinin [ratio 67% (95% CI 51–88%); P = 0.008] and desethylamodiaquine [ratio 65% (95% CI 46–90%); P = 0.015] when compared with monotherapy. Adverse events of concern occurred in four volunteers (27%): grade 3 transaminitis (n = 1), neutropaenia (n = 2), and hypersensitivity (n = 1). Conclusion  The total drug exposure to both drugs was reduced significantly when they were given in combination. The clinical significance of these interactions is unclear and must be studied in malaria patients. The frequency and nature of adverse events among the healthy volunteers were of concern, and suggest laboratory monitoring would be needed in malaria patients treated with artesunate plus amodiaquine.     

Evaluation of pharmacokinetic interactions between bicyclol and co-administered drugs in rat and human liver microsomes in vitro and in rats in vivo

1. Bicyclol is a new synthetic anti-hepatitic drug and primarily metabolized by CYP3A. The aim of this study was to evaluate the pharmacokinetic interactions between bicyclol and co-administered drugs including metformin, pioglitazone, atorvastatin, fenofibrate, Cyclosporin A (CsA), and tacrolimus in rat and human liver microsomes (RLMs/HLMs) in vitro and in rats in vivo.

2. The depletion rate of bicyclol in RLMs was significantly inhibited by 44.8% and 35.5% after preincubation with pioglitazone and fenofibrate while the metabolite formation rate of bicyclol in HLMs was inhibited by 26.1% and 23.9% after preincubation and coincubation with tacrolimus, and by 20.2% after preincubation with CsA. Conversely, preincubation and coincubation with bicyclol significantly inhibited the depletion rate of pioglitazone in RLMs by 34.1% and 27.1%, respectively, and the formation rate of para- and ortho-hydroxy atorvastatin in RLMs and HLMs by 20.6–36.2%. There were no significant pharmacokinetic interactions between bicyclol and pioglitazone in rats after a single or multiple oral treatment.

3. As the selected inhibitory drug concentrations in vitro were significantly higher than those in clinical settings and the maximum inhibition rate did not exceed 50%, the clinically significant interaction between bicyclol and these co-administered drugs in humans is predicted less likely to happen.

     

Fluvastatin efficacy and tolerability in comparison and in combination with cholestyramine

The aim of this study was to investigate the new synthetic HMG-CoA reductase inhibitor, fluvastatin, for efficacy, safety and tolerability in comparison to cholestyramine. One hundred fifty one primary hypercholesterolaemic patients participated in this double-blind, parallel-group, randomized study. During the first 12 weeks of the study, fluvastatin (20 mg and 40 mg daily) was compared with cholestyramine (16 g per day). In the subsequent, 6-week part of the study, the comparative efficacy, safety and tolerability of 20 mg fluvastatin, combined with cholestyramine (4 g, 8 g, or 16 g) were assessed.Fluvastatin (40 mg) reduced LDL cholesterol by 28.0%, triglycerides by 10.5% and increased HDL cholesterol by 3.7%. Cholestyramine (16 g) reduced LDL cholesterol by 35.0%, but raised triglycerides and HDL cholesterol by 12.3% (p<0.01) and 3.7% respectively.The combination of fluvastatin 20 mg and cholesty-ramine (4 g, 8 g and 16 g) induced the following reductions in LDL cholesterol: 30.4%, 35.6% and 46.6% respectively. There was no significant change in triglycerides in either group although HDL cholesterol was raised by 4.9%, 8.3% and 7.2% respectively. One patient treated with fluvastatin and two treated with cholesty-ramine were withdrawn from the study due to elevation of liver transaminases. The most frequent subjective adverse effects in both treatment groups were mild, transient gastrointestinal complaints.Thus, fluvastatin was effective as a lipid-lowering agent; the effect was further enhanced when fluvastatin was combined with cholestyramine.     

Nicotine-induced hypophagia and hypodipsia in deprived and in hypothalamically stimulated rats

Nicotine, in doses of 0.15 and 0.45 mg/kg, induced hypophagia and hypodipsia in 20 hrs-deprived rats and elevated the threshold currents for hypothalamically induced feeding and drinking in satiated rats. These effects were blocked by pretreatment with 0.5 mg/kg mecamylamine, but not by pretreatment with 3.0 or 9.0 mg/kg hexamethonium. These results indicate a centrally-located mechanism for the hypophagic and hypodipsic effect of single injections of nicotine.