颠茄流浸膏真伪鉴别及含量测定方法研究

目的:对颠茄流浸膏进行定性定量研究,鉴别真伪,提高颠茄流浸膏的质量标准。方法:运用薄层色谱法对颠茄流浸膏进行定性分析;采用高效液相色谱法测定硫酸天仙子胺的含量,色谱柱为Shim-pack VP-ODS(4.6 mm×250 mm,5μm)柱,流动相为0.004%磷酸溶液(含0.25%十二烷基硫酸钠)-乙腈(60∶40),流速1.0 mL.min-1,检测波长210 nm,柱温30℃。结果:薄层色谱方法能有效识别非法添加硫酸阿托品,鉴别特征明显,专属性强;硫酸天仙子胺进样量在0.2888～16.50μg范围内线性良好(r=1.000),平均回收率为103.8%(RSD=1.6%,n=6)。结论:本方法专属性强、灵敏度高、操作简便,对30余批不同生产企业颠茄流浸膏进行鉴别和含量测定,结果有6批次样品检出非法添加硫酸阿托品,该方法可为颠茄流浸膏的质量控制提供依据。     

美国药典（24版）：颠茄提取物片（Belladonna Extract Tablets）

颠茄提取物片中所含颠茄叶生物碱应为标示量的90.0％～110.0％。包装和贮存贮存于遮光、密闭的容器中。美国药典参比标准〈11〉USP 硫酸阿托品标准品,USP 氢溴酸后马托品标准品,USP 氢溴酸东莨菪碱标准品。鉴别取相当于5 mg 颠茄生物碱的片粉用20 mL     

美国药典（24版）：颠茄叶（Belladonna Leaf）

颠茄叶包括茄科植物颠茄 Atropa belladonnaLinné及其变种 A.acumtnata Royle ex Lindley 的干燥叶、开花或结果的枝梢。颠茄叶中生物碱含量不应低于0.35％。包装与贮存贮存于密封容器中,避免目光长时间直射。颠茄叶粉应贮存于遮光容器中。美国药典参比标准〈11〉USP 硫酸阿托品标准品,     

HPLC法测定维U颠茄铝镁胶囊中硫酸阿托品含量

目的建立维u颠茄铝镁胶囊中硫酸阿托品的含量测定方法。方法采用HPLC法,用十八烷基硅胶键合色谱柱,流动相：乙腈-0．01mol／L庚烷磺酸钠溶液（22：78）,检测波长：208nm;流速1．0mL／min,进样量10曲。结果硫酸阿托品存5．0125～100．25μg／mL范围内线性关系良好（r=0．9996）,平均加样回收率为97．4％,RSD为1．1％（n=6）。结论该方法简便快捷,准确、重现性好,灵敏度高,可用于建立维U颠茄铝镁胶囊的定量质量标准。     

RP-HPLC法测定颠茄片中硫酸阿托品的含量

目的建立颠茄片中硫酸阿托品的含量测定方法。方法采用RP-HPLC法,AgilentZORBAXSB-C18(250mm×4.6mm,5μm);流动相:1%三乙胺水溶液(用磷酸调pH至2.8)-乙腈(90∶10);检测波长:218nm;流速:1.0mL·min-1。结果硫酸阿托品进样量在0.5955～25.53μg范围内呈良好的线性关系,平均回收率为98.89%,RSD为1.3%(n=6)。结论该方法快速,准确,专属性强。     

维U颠茄铝胶囊中硫酸阿托品含量测定方法改进

目的改进维U颠茄铝胶囊中硫酸阿托品的含量测定方法。方法采用高效液相色谱（HPLC）法,色谱柱为Thermo ODS C18柱（250mm×4．6mm,5μm）,以三乙胺磷酸溶液（取三乙胺4mL,加水500mL,加磷酸1．8mL,加水至1000mL）-乙腈（85：15）为流动相,检测波长为206nm。结果硫酸阿托品质量浓度线性范围为2．25～13．56μg／mL（r=0．9996）,平均回收率为99．2％,RSD=0．9％（n=6）。结论所建立的方法可准确测定样品中硫酸阿托品含量。     

高效液相法测定颠茄磺苄啶片中硫酸阿托品含量

目的：建立测定颠茄磺苄啶片中硫酸阿托品含量的高效液相色谱（HPLC）法。方法：以十八烷基硅烷键合硅胶为填充剂,0.8%三乙胺溶液（用磷酸调节pH值至2.9）-乙腈（85∶15）为流动相,检测波长为210nm,柱温为室温。结果：硫酸阿托品在12.56～200.96μg/mL范围内线性关系良好;r=0.9998;平均回收率为96.79%（n=6）,RSD为1.4%。结论：该方法简便、快速、准确并且专属性强。     

酸性染料比色法测定颠溴合剂中颠茄酊的含量

颠溴合剂为我院协定处方的自制制剂，临床用于解除平滑肌痉挛，减少胃液分泌。而现行药品三级标准中含颠茄酊的制剂，多未规定其含量测定。本文采用酸性染料比色法[1]，以硫酸阿托品为工作标准品，测定颠澳合剂中颠茄酊的含量，方法简单易行，结果较为满意。1．仪器与试药：751－GW分光光度计（上海分析仪器厂）；硫酸阿托品对照品（中国药品生物制品检定所）；颠茄配（广州星群药业股份有限公司）；颠澳合剂（本院制剂室）；氯仿（AR）。2．方法与结果：2．1处方颠沛配12ml，澳化钾10g，枸橼酸钠0．1g，10％尼泊金乙酯0．26ml，水加至100…     

高效液相色谱法测定维U颠茄铝镁片中硫酸阿托品含量

目的用高效液相色谱法测定维U颠茄铝镁片中硫酸阿托品的含量。方法采用Phenomenex C18色谱柱(250 mm×4.6mm,5μm),以0.5%三乙胺溶液(0.2%四氢呋喃,用冰醋酸调pH至6.10)-乙腈(80∶20)为流动相,检测波长为210 nm,柱温为室温。结果硫酸阿托品质量浓度在24.18～290.16μg/mL范围内与峰面积线性关系良好(r=0.9998),平均回收率为97.21%,RSD=1.1%(n=5)。结论该法简便、快速、准确且专属性强,可用于产品的质量控制。     

颠茄流浸膏和颠茄浸膏非法勾兑快速筛查

目的建立专属的颠茄流浸膏和颠茄浸膏特征图谱检测方法,筛查不同企业生产的颠茄流浸膏和颠茄浸膏非法勾兑情况。方法采用Welch XB C18（4.6 mm×250 mm,5 μm）色谱柱;流动相为甲醇 0.05%磷酸梯度洗脱;柱温：30 ℃;流速：1.0 mL•min－1;检测波长344 nm。结果检测不同企业生产的颠茄流浸膏和颠茄浸膏,发现约84%的产品使用了颠茄草投料,部分产品除检出硫酸阿托品峰外未检出任何其他色谱峰,部分产品检出色谱峰但与颠茄草特征成分不符。结论所建立的颠茄草特征成分高效液相色谱检测方法能有效识别产品是否为颠茄草投料,专属性强,可快速有效的筛查颠茄流浸膏和颠茄浸膏非法勾兑品。     

Influence of atropine on the acute toxicity of isometamidium.

The effectiveness of atropine in blocking the acute toxic effects of the antitrypanosomal drug isometamidium (ISMM) was evaluated in mice and goats using lethality as the primary index. The median lethal dose (LD50) of ISMM in nonatropinized mice was 45.3 mg/kg bodyweight (SE +/- 5.3 mg/kg bodyweight), whereas the LD50 of ISMM in mice pre-treated with atropine was 71.7 mg/kg bodyweight (SE +/- 13.2 mg/kg bodyweight). The increase in LD50 was about 60%. The ratio of the slope of the dose-response curve for ISMM in non-atropinized mice to that in atropinized mice was about 4:1. The results showed that atropine was effective in considerably reducing the lethal effect of ISMM in mice. In contrast, experiments in goats showed markedly different results; atropine was not effective in reducing the lethality of ISMM. It was concluded that in mice atropine has some beneficial action in cases of overdosage or poisoning by ISMM. Treatment with atropine should be attempted as an antidotal measure in humans accidentally poisoned with ISMM.     

Uniformly sized molecularly imprinted polymer for atropine and its application to the determination of atropine and scopolamine in pharmaceutical preparations containing Scopolia extract

A uniformly sized molecularly imprinted polymer (MIP) for atropine has been prepared. The MIP was prepared using 2-(trifluoromethyl) acrylic acid and ethylene glycol dimethacrylate as a functional monomer and cross-linker, respectively, by a multi-step swelling and thermal polymerization method. The selectivity factor, which is defined as the ratio of the retention factors (k) on the molecularly imprinted and non-imprinted polymers, k(imprinted)/k(non-imprinted), was 2.2 for atropine on the MIP. The obtained MIP was applied for the determination of tropane alkaloids (atropine and scopolamine) in a commercial gastrointestinal drug by a column-switching HPLC system, consisting of an MIP material as a pre-column, and a conventional cation-exchange analytical column. An interference peak was observed at the retention time of atropine derived from pre-column. However, since the peak area was less than 0.5% the peak area of atropine of a standard solution under the analytical conditions of this study (0.2 microg of atropine was loaded), this interference was negligible in the determination of atropine. On the other hand, no interference peak was observed at the retention time of scopolamine. Calibration curves of atropine and scopolamine showed good linearity in the range of 0.02-0.9 microg/ml (r=0.9999) and 0.003-0.09 microg/ml (r=0.9998), respectively. The mean recoveries of atropine and scopolamine from a placebo pharmaceutical preparation sample were 98.9 and 99.9%, respectively. The intra-day precision (measured by relative standard deviation, R.S.D. (%)) of both ingredients was less than 2.0%. The optimized column-switching system was applied successfully to the determination of atropine and scopolamine in a commercial gastrointestinal drug.     

Stability of atropine sulfate prepared for mass chemical terrorism

BACKGROUND: Preparedness for chemical terrorism includes the procurement of the appropriate pharmacological antagonists. A large emphasis has been placed on having a sufficient quantity of atropine available to treat patients exposed to acetylcholinesterase inhibitors such as sarin. Severe exposures may necessitate the administration of large amounts of atropine and dictate the need to prepare significant quantities of extemporaneously compounded atropine solution to respond to mass numbers of casualties over the first 24-48 hours postexposure. OBJECTIVE: The objective of this project was to determine the stability of a 1 mg/mL atropine solution prepared in multidose IV solutions of 0.9% sodium chloride over a 72-hr period stored at varying temperatures. METHODS: Atropine sulfate solution 1 mg/mL in 0.9% sodium chloride was prepared from sterile pharmaceutical-grade atropine sulfate powder. Multidose bags of atropine sulfate (100 mL) were stored at controlled temperatures of 4 degrees C to 8 degrees C, 20 degrees C to 25 degrees C, and 32 degrees C to 36 degrees C for 3 days and covered with an amber occlusive cover to minimize exposure to light. Six samples from each bag were drawn at 6, 12, 24, 48, and 72 h after preparation and compared with a time zero control sample. The samples were assayed using United States Pharmacopeia/National Formulary (USP/NF) high-performance liquid chromatography (HPLC) methods for atropine sulfate injection. The USP standard of 95% for atropine sulfate stability was used as the primary endpoint. RESULTS: Atropine sulfate 1 mg/mL in 0.9% sodium chloride was stable for at least 72hr at 4 degrees C to 8 degrees C (percent initial concentration ranging from 96.5% to 103.4%), 20 degrees C to 25 degrees C (percent initial concentration ranging from 98.7% to 100.2%), and 32 degrees C to 36 degrees C (percent initial concentration ranging from 98.3% to 102.8%). Because the IV bags were protected from light during this study, we recommend this practice after preparing the atropine solution. CONCLUSIONS: The amount of atropine necessary to treat hundreds to thousands of victims of a chemical attack is immense. The extemporaneous preparation of atropine solution from pharmaceutical-grade powder eliminates concerns about the storage of excessive quantities of atropine. A 1 mg/mL solution is stable for at least 3 days, allowing for use during the most critical treatment periods after exposure.     

Interaction of atropine, toxogonin and fosfakol

Protective action of atropine and toxogonin against phosphacol poisoning has been studied in experiments on white mice. The protective effect has been shown to depend on the magnitude of the doses combined. The most powerful is the combination that provides for the effect potentiation (30-200 mg/kg atropine, 5-70 mg/kg toxogonin). When toxogonin was applied in a dose of 100 mg/kg and higher, atropine addition lowered the ability to prevent phosphacol poisoning, with this effect being more powerful the higher the dose. Between the zones of potentiation and antagonism there is a zone within which the prophylactic actions of both drug doses are summed up.     

Development of a quantitative method for the analysis of cocaine analogue impregnated into textiles by Raman spectroscopy

Cocaine trafficking in the form of textile impregnation is routinely encountered as a concealment method. Raman spectroscopy has been a popular and successful testing method used for in situ screening of cocaine in textiles and other matrices. Quantitative analysis of cocaine in these matrices using Raman spectroscopy has not been reported to date. This study aimed to develop a simple Raman method for quantifying cocaine using atropine as the model analogue in various types of textiles. Textiles were impregnated with solutions of atropine in methanol. The impregnated atropine was extracted using less hazardous acidified water with the addition of potassium thiocyanate (KSCN) as an internal standard for Raman analysis. Despite the presence of background matrix signals arising from the textiles, the cocaine analogue could easily be identified by its characteristic Raman bands. The successful use of KSCN normalised the analyte signal response due to different textile matrix background interferences and thus removed the need for a matrix‐matched calibration. The method was linear over a concentration range of 6.25–37.5 mg/cm² with a coefficient of determination (R²) at 0.975 and acceptable precision and accuracy. A simple and accurate Raman spectroscopy method for the analysis and quantification of a cocaine analogue impregnated in textiles has been developed and validated for the first time. This proof‐of‐concept study has demonstrated that atropine can act as an ideal model compound to study the problem of cocaine impregnation in textile. The method has the potential to be further developed and implemented in real world forensic cases.     

Impurity profiling of atropine sulfate by microemulsion electrokinetic chromatography

An oil-in-water microemulsion electrokinetic chromatography (MEEKC) method has been developed and validated for the determination of atropine, its major degradation products (tropic acid, apoatropine and atropic acid) and related substances from plants material (noratropine, 6-hydroxyhyoscyamine, 7-hydroxyhyoscyamine, hyoscine and littorine). Separation of atropine and all impurities was optimized by varying the voltage, the nature of the oil droplet and the buffer, as well as the organic modifier (methanol, 2-propanol or acetonitrile) and the surfactant type and concentration. The optimum O/W microemulsion background electrolyte (BGE) solution consists of 0.8% (w/w) octane, 6.62% (w/w) 1-butanol, 2.0% (w/w) 2-propanol, 4.44% (w/w) SDS and 86.14% (w/w) 10 mM sodium tetraborate buffer pH 9.2. In order to shorten the analysis time a voltage gradient was applied. The validation was performed with respect to specificity, linearity, range, limit of quantification and detection, precision, accuracy and robustness. The established method allowed the detection and determination of atropine sulfate related substances at impurity levels given in the European Pharmacopoeia. Good agreement was obtained between the established MEEKC method and the traditional RP-HPLC method.     

Stability of milrinone and epinephrine, atropine sulfate, lidocaine hydrochloride, or morphine sulfate injection

The stability of both drug components of admixtures of milrinone and epinephrine, atropine sulfate, lidocaine hydrochloride, morphine sulfate, calcium chloride, or sodium bicarbonate injections was studied. Duplicate solutions of admixtures of milrinone injection 1 mg/mL and epinephrine injection 1:10,000, atropine sulfate injection 1 mg/mL, lidocaine hydrochloride injection 1%, morphine sulfate injection 8 mg/mL, calcium chloride injection 10%, or sodium bicarbonate injection 7.5% were prepared and stored in glass containers at 22-23 degrees C under fluorescent light. Samples were taken immediately and after 20 minutes for assay by high-performance liquid chromatography (HPLC). Milrinone at initial concentrations of 0.10-0.73 mg/mL showed no degradation in any of the solutions during the study period, nor was any degradation observed for lidocaine, morphine, atropine, or epinephrine. Milrinone 0.10-0.73 mg/mL is compatible with atropine sulfate, lidocaine hydrochloride, epinephrine, calcium chloride, or sodium bicarbonate in glass containers stored for 20 minutes at room temperature. These results support the use of milrinone in combination with these agents immediately after the preparation of admixtures.     

The pharmacological properties of anisodamine

Anisodamine is a naturally occurring atropine derivative that has been isolated, synthesized and characterized by scientists in the People's Republic of China. Like atropine and scopolamine, anisodamine is a non-specific cholinergic antagonist exhibiting the usual spectrum of pharmacological effects of this drug class. It appears to be less potent and less toxic than atropine and displays less CNS toxicity than scopolamine. Anisodamine has been shown to interact with and disrupt liposome structure which may reflect its effects on cellular membranes. Experimental evidence implicates anisodamine as an anti-oxidant that may protect against free radical-induced cellular damage. Its cardiovascular properties include depression of cardiac conduction and the ability to protect against arrhythmia induced by various agents. Anisodamine is a relatively weak alpha(1) adrenergic antagonist which may explain its vasodilating activity. Its anti-thrombotic activity may be a result of inhibition of thromboxane synthesis. The T(1/2) of anisodamine in humans is about 2-3 h. Numerous therapeutic uses of anisodamine have been proposed including treatment of septic shock, various circulatory disorders, organophosphorus (OP) poisoning, migraine, gastric ulcers, gastrointestinal colic, acute glomerular nephritis, eclampsia, respiratory diseases, rheumatoid arthritis, obstructive jaundice, opiate addiction, snake bite and radiation damage protection. The primary therapeutic use of anisodamine has been for the treatment of septic shock. Several mechanisms have been proposed to explain its beneficial effect though most mechanisms are based upon the assumption that anisodamine ultimately acts by an improvement of blood flow in the microcirculation. Preliminary studies suggest another important therapeutic use of anisodamine is for the treatment of OP poisoning. Additional research is needed to delineate further the clinical usefulness of anisodamine relative to other anti-muscarinic drugs such as atropine and scopolamine.     

Pharmacological antagonism of lethal effects induced by O-isobutyl S-[2-(diethylamino)ethyl]methylphosphonothioate

O-Isobutyl S-[2-(diethylamino)ethyl]methylphosphonothioate (VR) is a structural isomer of a more widely known chemical warfare agent O-ethyl S-[2(diisopropylamino)ethyl]methylphosphonothioate (VX). VR has the potential of being used as military threat/sabotage/terrorist agent. The development of a sound medical countermeasure will undoubtedly enhance not only our medical readiness and ability in VR casualty management, but also our defense posture against the deployment of VR in both combat and politically volatile environments. Acute exposure to a lethal dose of VR has been shown to cause cholinergic hyperfunction, incapacitation, seizures, convulsions, cardiorespiratory depression and death. In this study, pharmacological antagonism of VR-induced cardiorespiratory failure and lethality was investigated in guinea pigs chronically instrumented for concurrent recordings of electrocorticogram, diaphragmatic EMG, Lead II ECG, heart rate and neck skeletal muscle EMG. Thirty (30) min prior to intoxication with a 2 x LD50 dose of VR (22.6 micrograms/kg, s.c.), animals were pretreated with pyridostigmine (0.026 mg/kg, i.m.). Immediately after VR intoxication, animals were given pralidoxime chloride (2-PAM; 25 mg/kg, i.m.) and atropine sulfate (2, 8 or 16 mg/kg, i.m.). In animals that displayed seizures and convulsions, diazepam (5 mg/kg, i.m.) was administered 10 min following the onset of epileptiform activities. Responses to pretreatment/therapy modality were evaluated at 24 h post-VR. All animals survived the 2 x LD50 VR challenge. With the exception of an increased heart rate in response to atropine, the myocardial and diaphragmatic (respiratory) activity profiles appeared normal throughout the course of intoxication and recovery. Animals receiving 2 mg/kg atropine all developed fasciculations, seizures, signs of excessive mucoid/salivary secretion, and needed diazepam adjunct therapy. One-half (50%) of the animals receiving 8 mg/kg atropine developed seizure activities and were given diazepam, whereas the other half only showed a brief period of increase in CNS excitability. No fasciculations, seizures or convulsions were noted in animals receiving 16 mg/kg atropine. In summary, although lethality can be prevented with the pretreatment/therapy modality containing 2 mg/kg atropine and diazepam adjunct, a complete CNS and cardiorespiratory recovery from 2 x LD50 of VR requires a minimum of 8 mg/kg atropine.     

A severe organophosphate poisoning requiring the use of an atropine drip

A 68-year-old male attempted suicide by drinking three ounces of concentrated Cygon 2-E (23.4% dimethoate). He was immediately brought to the hospital, responded to standard treatment (ipecac, activated charcoal, 2-PAM, atropine), and was transferred from the ICU to general care 24 hours after the exposure. Within eight hours of the transfer, he relapsed and was moved to the CCU, where he required five milligrams of atropine every ten minutes for 24 hours, before being started on an atropine drip. The patient was maintained on the atropine drip (0.5-2.4 mg/kg/hr) for five weeks. He required a total atropine dose of 30 grams, the largest amount ever reported to have been administered to a human. Although S-ChE activities gradually increased they were not found to be helpful in determining when the drip could be safely stopped. Control of hypersecretions served as the best monitoring parameter for titration of the drip rate. The patient recovered completely with the exception of a detectable sensorineural hearing deficit, a slight, nonspecific personality change, and minimal spastic rigidity thought to be secondary to several anoxic episodes.