液相色譜—串聯(lián)質(zhì)譜法在體內(nèi)有毒動植物成分檢測中的應用
發(fā)布時間：2018-08-14 08:41
【摘要】： 在我國民間,許多動植物作為藥用和食用,有的甚至作為“滋補品”食用,這些動植物往往含有毒性成分,甚至是劇毒成分,其治療量與中毒量接近,因用藥不當、盲目和超量服用可造成中毒和死亡。在司法鑒定實踐中,由此導致的各類誤服、自殺、投毒、醫(yī)療糾紛等的案件時有發(fā)生。由于有毒動植物所含成分復雜多樣、毒性大、含量低以及在人體內(nèi)代謝迅速、且中毒無特異性臨床表現(xiàn)等問題,使復雜生物體系中有毒動植物成分的鑒定成為長期以來未能有效解決的技術難題,對有毒動植物中毒的法醫(yī)學鑒定、臨床診治以及食品安全監(jiān)測缺乏有效的檢測手段和科學的評判依據(jù)。 現(xiàn)代分析技術的發(fā)展使得復雜生物體系中有毒動植物成分的檢測和中毒鑒定成為可能。近年來,液相色譜—質(zhì)譜聯(lián)用法在法醫(yī)和臨床毒物分析領域的應用得到迅速發(fā)展。具有液相色譜高效的在線分離能力和質(zhì)譜的高選擇性、高靈敏度的檢測能力的液相色譜-質(zhì)譜聯(lián)用法,可以同時得到化合物的保留時間、分子量以及特征結構碎片等豐富的信息,尤其是串聯(lián)質(zhì)譜可從復雜基質(zhì)中確認目標物,采用選擇離子監(jiān)測或多反應監(jiān)測模式,更可大大提高分析的專一性和靈敏度,完全有望解決復雜生物體系中痕量有毒動植物成分的檢測,并在分析目標物不明確或系統(tǒng)中可能存在數(shù)種不同性質(zhì)的目標物時,用一個分析系統(tǒng)同時檢測多種不同性質(zhì)的目標物。同時液相色譜-質(zhì)譜聯(lián)用法分析目標物范圍廣,前處理簡單,不需衍生化,體現(xiàn)出比氣相色譜-質(zhì)譜聯(lián)用法更靈敏、有效的檢測手段,從而對復雜生物體系中的有毒動植物成分的鑒定和確認提供科學依據(jù)。 本課題以血液、尿液和組織為主要生物檢材,研究、探索、建立了22種常見有毒生物堿成分系統(tǒng)篩選方法,烏頭堿、新烏頭堿、次烏頭堿、秋水仙堿和河豚毒素的定性定量方法,并對烏頭堿、新烏頭堿、次烏頭堿和秋水仙堿的定量分析方法進行了全面、系統(tǒng)的方法有效性驗證,解決了LC-MS/MS分析中的樣品處理、色譜分離及優(yōu)化、質(zhì)譜條件的選擇與優(yōu)化等關鍵技術問題,同時對它們在急性中毒豚鼠的體內(nèi)分布進行探討,并成功地將所建方法應用于司法鑒定實踐,為涉毒案件的審理提供了科學證據(jù)。 具體研究內(nèi)容如下: 一、建立了基于LC-MS/MS-MRM技術的血液中22種有毒生物堿成分的篩選體系和確證方法。目標物涵蓋了在實際檢案工作中常見的有毒生物堿。所建方法簡便、快速、靈敏度高、專一性強,色譜分離采用兩根不同性能的色譜柱,使目標物的分離、檢測靈敏度、檢測所需時間等方面得到互補,整個過程可在16min內(nèi)完成。最低檢測限除羥基喜樹堿稍高(20ng/mL)外,絕大多數(shù)的LOD足以滿足法醫(yī)毒物學和臨床藥物學中中毒或治療濃度目標物的檢測要求。 二、烏頭生物堿的中毒、檢測及應用 1.建立了測定血液、尿液和組織中烏頭堿、新烏頭堿和次烏頭堿的LC-MS/MS-MRM分析方法,并對其定量方法進行了全面系統(tǒng)的方法有效性驗證,內(nèi)容包括考察方法的線性范圍、最低檢出限、精密度、回收率、準確度、基質(zhì)效應、穩(wěn)定性等,取得了滿意的結果。所建方法簡便、快速、靈敏,適用于體內(nèi)痕量烏頭堿、新烏頭堿和次烏頭堿的檢測。 2.研究了烏頭堿在急性中毒豚鼠的體液和組織中的分布。取豚鼠8只,以1.0-4.0mg/kg劑量灌胃給藥致豚鼠烏頭堿急性中毒,解剖后取體液和組織進行烏頭堿測定。烏頭堿在體液和組織中的含量從大到小依次為:膽汁＞血液＞尿液＞胃＞肺＞心＞脾＞腎＞胰＞肝。血液可作為烏頭堿中毒、死亡分析的首選生物檢材。 3.將所建方法成功地應用于鑒定實踐,對生物檢材中痕量烏頭生物堿的成分進行定性定量分析,并對多起烏頭植物中毒尸體中體液和組織中的烏頭堿、新烏頭堿和次烏頭堿的含量進行測定,首次報道了新烏頭堿和次烏頭堿在尸體組織中分布,為涉及烏頭中毒案件的死因判定和案件的審理提供了明確的方向和科學的證據(jù)。 三、秋水仙堿的中毒、檢測及應用 1.建立了測定血液、尿液和組織中秋水仙堿的LC-MS/MS-MRM分析方法,并對其定量方法進行了全面系統(tǒng)的方法學驗證,內(nèi)容包括方法的線性范圍、最低檢出限、精密度、回收率、準確度、基質(zhì)效應、穩(wěn)定性等,所得結果符合法醫(yī)毒物分析的要求。所建方法簡便、快速、靈敏,最低檢出限可低至0.05ng/mL,能夠滿足法醫(yī)毒物分析和臨床毒物分析的要求。 2.研究了秋水仙堿在急性中毒豚鼠體內(nèi)的分布。取豚鼠5只,以4.0mg/kg劑量灌胃給藥致豚鼠秋水仙堿急性中毒,解剖后取體液和組織進行秋水仙堿含量測定。秋水仙堿在體液和組織中的含量從大到小依次為:膽汁＞尿液＞脾＞胃＞肺＞腎＞心＞胰＞肝＞腎上腺＞睪丸,心血中秋水仙堿的含量最低。尿液可作為秋水仙堿分析的首選生物檢材。 3.將所建方法成功地應用于鑒定實踐,對中毒尸體的生物檢材及野生植物中秋水仙堿的成分進行定性定量分析,為涉及有毒植物中毒案件的死因判定提供了明確的方向和證據(jù)。 四、生物檢材中河豚毒素的檢測及應用 建立了測定血液、尿液和組織中河豚毒素的LC-MS/MS-MRM測定方法,考察了方法的線性范圍及最低檢出限。并將所建方法應用于食用河豚魚中毒的實際案例中,取得了滿意的效果。 本研究的創(chuàng)新點及研究的意義如下: 1.建立的有毒生物堿成分篩選分析方法簡便、靈敏,避免了生物檢材中該類物質(zhì)用傳統(tǒng)方法檢測可能出現(xiàn)的漏檢、錯檢現(xiàn)象,整個分析可在16min內(nèi)完成。篩選目標物涵蓋了司法鑒定中常見的有毒生物堿成分,且LC-MS/MS篩選體系可隨著目標物的增加而不斷擴充,從而實現(xiàn)對更多的目標物進行篩選。該方法為首創(chuàng)方法。 2.全面系統(tǒng)地闡述了生物檢材中定量方法的有效性驗證,規(guī)范了方法的評價指標,通過對生物檢材中烏頭堿、新烏頭堿、次烏頭堿和秋水仙堿的檢測方法的有效性驗證,證明了所建方法準確、靈敏、專屬、重現(xiàn)和可靠。 3.建立了生物檢材中烏頭生物堿LC-MS/MS-MRM分析方法。首次報道了烏頭中毒尸體中與烏頭堿共存的新烏頭堿和次烏頭堿的體內(nèi)分布,為涉及有毒植物中毒案件的死因判定提供了明確的方向和證據(jù)。 4.建立了生物檢材中秋水仙堿LC-MS/MS-MRM分析方法。所建方法足夠靈敏,最低檢出限可低至0.05ng/mL,能夠滿足法醫(yī)毒物分析和臨床毒物分析的要求。 5.建立了生物檢材中河豚毒素的LC-MS/MS-MRM測定方法,并首次報道了河豚魚中毒尸體中河豚毒素在各組織中的分布情況。 研究成果填補了體內(nèi)有毒動植物檢測及評判的空白,使我國的復雜生物體系中常見有毒動植物成分分析技術躍升到國際先進水平,將為有毒動植物的科學研究、中毒鑒定、臨床診斷及救治、食品安全監(jiān)測等提供技術平臺。
[Abstract]:In our country, many animals and plants are used as medicinal and edible, and some even as "tonic" food. These animals and plants often contain toxic ingredients, even highly toxic ingredients. Their treatment dose is close to that of poisoning. Drug, suicide, poisoning, medical disputes and other cases occur from time to time. The identification of toxic animal and plant components in complex biological systems has long been an unsolved technical problem due to the complex and diverse components, high toxicity, low content, rapid metabolism in the human body, and no specific clinical manifestations of poisoning. There is no effective means and scientific basis for forensic identification, clinical diagnosis, treatment and food safety monitoring of toxic animal and plant poisoning.
The development of modern analytical techniques makes it possible to detect and identify toxic plant and animal components in complex biological systems. In recent years, the application of liquid chromatography-mass spectrometry (LC-MS) in forensic and clinical toxicological analysis has been developed rapidly. It has high performance in on-line separation of liquid chromatography and high selectivity and high sensitivity in mass spectrometry. Liquid chromatography-mass spectrometry (LC-MS) can simultaneously obtain abundant information such as retention time, molecular weight and characteristic structure fragments of compounds. Especially, tandem mass spectrometry can identify the target from complex matrices, and selective ion monitoring or multi-reaction monitoring mode can greatly improve the specificity and sensitivity of the analysis. It is hopeful to solve the problem of detecting trace toxic animal and plant components in complex biological systems, and to use an analytical system for simultaneous detection of a variety of different kinds of targets when the target is not clear or there may be several different kinds of targets in the system. Simple and non-derivatization methods are more sensitive and effective than gas chromatography-mass spectrometry, thus providing a scientific basis for the identification and identification of toxic animal and plant components in complex biological systems.
A systematic screening method for 22 common toxic alkaloids, including aconitine, neoaconitine, hypoaconitine, colchicine and tetrodotoxin, was established. The quantitative analysis methods of aconitine, neoaconitine, hypoaconitine and colchicine were also studied. All-round and systematic validation of the method has solved the key technical problems in LC-MS/MS analysis, such as sample processing, chromatographic separation and optimization, and the selection and optimization of mass spectrometry conditions. At the same time, the distribution of the methods in acute poisoning guinea pigs has been discussed, and the established methods have been successfully applied in forensic practice for the trial of drug-related cases. Li provides scientific evidence.
The specific research contents are as follows:
Firstly, a screening system and confirmation method of 22 toxic alkaloids in blood based on LC-MS/MS-MRM technology were established. The target substances covered the common toxic alkaloids in the actual detection work. The method was simple, rapid, sensitive and specific. Two chromatographic columns with different performance were used for chromatographic separation and detection of the target substances. The detection sensitivity and time required are complementary, and the whole process can be completed within 16 minutes. Except for the slightly higher hydroxycamptothecin (20ng/mL), the vast majority of LOD is sufficient to meet the requirements of forensic toxicology and clinical pharmacology for the detection of poisoning or therapeutic concentration targets.
Two, poisoning and detection and application of Aconitum alkaloids
1. A LC-MS/MS-MRM method for the determination of aconitine, neoaconitine and hypaconitine in blood, urine and tissues was established, and its quantitative method was validated by a comprehensive and systematic method, including the linear range, minimum detection limit, precision, recovery rate, accuracy, matrix effect, stability and so on. The method is simple, rapid and sensitive for the determination of trace aconitine, neoaconitine and hypaconitine in vivo.
2. The distribution of aconitine in body fluid and tissues of acute poisoning guinea pigs was studied. Eight guinea pigs were given aconitine by intragastric administration at a dose of 1.0-4.0 mg/kg. After dissection, the contents of aconitine in body fluid and tissues were determined. The order of aconitine content in body fluid and tissues was: bile > blood > urine > stomach > lung > Blood can be used as the first choice of biological analysis for death analysis.
3. The established method was successfully applied to the identification practice. The contents of trace aconitine in biological samples were qualitatively and quantitatively analyzed. The contents of aconitine, neoaconitine and hypaconitine in body fluids and tissues of several aconitine poisoned corpses were determined. The contents of neoaconitine and hypaconitine in corpse tissues were reported for the first time. The distribution provides clear direction and scientific evidence for the determination of the cause of death and the trial of cases involving Aconitum poisoning.
Three, colchicine poisoning, detection and Application
1. A LC-MS/MS-MRM method for the determination of colchicine in blood, urine and tissues was established, and its quantitative method was systematically and systematically validated, including linear range, minimum detection limit, precision, recovery rate, accuracy, matrix effect, stability and so on. The method is simple, rapid, sensitive and the minimum detection limit can be as low as 0.05ng/mL. It can meet the requirements of forensic toxicology analysis and clinical toxicology analysis.
2. The distribution of colchicine in acute poisoned guinea pigs was studied. Five guinea pigs were given colchicine by intragastric administration at a dose of 4.0 mg/kg. The contents of colchicine in body fluid and tissues were measured after dissection. The order of colchicine content in body fluid and tissues was bile > urine > spleen > stomach > lung > kidney. The content of colchicine in heart blood is the lowest. Urine can be used as the first choice for colchicine analysis.
3. The method has been successfully applied to the identification of poisoned plants. The qualitative and quantitative analysis of colchicine in the biological samples of poisoned corpses and wild plants has provided clear direction and evidence for the determination of the cause of death in poisoned plants.
Four, the detection and application of tetrodotoxin in biological samples.
A method for the determination of tetrodotoxin in blood, urine and tissues by LC-MS/MS-MRM was established. The linear range and the minimum detection limit of the method were investigated. The method was applied to the practical cases of tetrodotoxin poisoning in edible puffer fish and satisfactory results were obtained.
The innovation and significance of this study are as follows:
1. The established screening and analysis method for toxic alkaloids is simple and sensitive, avoiding the missed and mistaken detection phenomena that may occur in biological samples by traditional methods. The whole analysis can be completed within 16 minutes. The screening target covers the common toxic alkaloids in forensic identification, and the LC-MS/MS screening system can follow the target. With the increase and expansion of objects, more targets can be screened. This method is a pioneering method.
2. The validity of the quantitative method in biological samples was expounded systematically and comprehensively, and the evaluation index of the method was standardized. The validity of the method for the determination of aconitine, neoaconitine, hypoaconitine and colchicine in biological samples was proved to be accurate, sensitive, exclusive, reproducible and reliable.
3. A LC-MS/MS-MRM method for the determination of aconitine in biological samples was established. The distribution of neoaconitine and hypaconitine in aconitine-poisoned corpses was reported for the first time, which provided clear direction and evidence for the determination of the cause of death in poisoned plants.
4. A LC-MS/MS-MRM method for the determination of colchicine in biological samples was established. The method is sensitive enough and the detection limit can be as low as 0.05 ng/mL. It can meet the requirements of forensic toxicology analysis and clinical toxicology analysis.
5. A method for the determination of tetrodotoxin in biological samples by LC-MS/MS-MRM was established. The distribution of tetrodotoxin in various tissues of tetrodotoxin poisoned corpses was reported for the first time.
The research results fill the blank of toxic animal and plant detection and evaluation in vivo, and make the component analysis technology of common toxic animal and plant in complex biological system of our country leap to the international advanced level. It will provide a technical platform for the scientific research of toxic animal and plant, poisoning identification, clinical diagnosis and treatment, food safety monitoring and so on.
【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：D919.1
本文編號：2182278
[Abstract]:In our country, many animals and plants are used as medicinal and edible, and some even as "tonic" food. These animals and plants often contain toxic ingredients, even highly toxic ingredients. Their treatment dose is close to that of poisoning. Drug, suicide, poisoning, medical disputes and other cases occur from time to time. The identification of toxic animal and plant components in complex biological systems has long been an unsolved technical problem due to the complex and diverse components, high toxicity, low content, rapid metabolism in the human body, and no specific clinical manifestations of poisoning. There is no effective means and scientific basis for forensic identification, clinical diagnosis, treatment and food safety monitoring of toxic animal and plant poisoning.
The development of modern analytical techniques makes it possible to detect and identify toxic plant and animal components in complex biological systems. In recent years, the application of liquid chromatography-mass spectrometry (LC-MS) in forensic and clinical toxicological analysis has been developed rapidly. It has high performance in on-line separation of liquid chromatography and high selectivity and high sensitivity in mass spectrometry. Liquid chromatography-mass spectrometry (LC-MS) can simultaneously obtain abundant information such as retention time, molecular weight and characteristic structure fragments of compounds. Especially, tandem mass spectrometry can identify the target from complex matrices, and selective ion monitoring or multi-reaction monitoring mode can greatly improve the specificity and sensitivity of the analysis. It is hopeful to solve the problem of detecting trace toxic animal and plant components in complex biological systems, and to use an analytical system for simultaneous detection of a variety of different kinds of targets when the target is not clear or there may be several different kinds of targets in the system. Simple and non-derivatization methods are more sensitive and effective than gas chromatography-mass spectrometry, thus providing a scientific basis for the identification and identification of toxic animal and plant components in complex biological systems.
A systematic screening method for 22 common toxic alkaloids, including aconitine, neoaconitine, hypoaconitine, colchicine and tetrodotoxin, was established. The quantitative analysis methods of aconitine, neoaconitine, hypoaconitine and colchicine were also studied. All-round and systematic validation of the method has solved the key technical problems in LC-MS/MS analysis, such as sample processing, chromatographic separation and optimization, and the selection and optimization of mass spectrometry conditions. At the same time, the distribution of the methods in acute poisoning guinea pigs has been discussed, and the established methods have been successfully applied in forensic practice for the trial of drug-related cases. Li provides scientific evidence.
The specific research contents are as follows:
Firstly, a screening system and confirmation method of 22 toxic alkaloids in blood based on LC-MS/MS-MRM technology were established. The target substances covered the common toxic alkaloids in the actual detection work. The method was simple, rapid, sensitive and specific. Two chromatographic columns with different performance were used for chromatographic separation and detection of the target substances. The detection sensitivity and time required are complementary, and the whole process can be completed within 16 minutes. Except for the slightly higher hydroxycamptothecin (20ng/mL), the vast majority of LOD is sufficient to meet the requirements of forensic toxicology and clinical pharmacology for the detection of poisoning or therapeutic concentration targets.
Two, poisoning and detection and application of Aconitum alkaloids
1. A LC-MS/MS-MRM method for the determination of aconitine, neoaconitine and hypaconitine in blood, urine and tissues was established, and its quantitative method was validated by a comprehensive and systematic method, including the linear range, minimum detection limit, precision, recovery rate, accuracy, matrix effect, stability and so on. The method is simple, rapid and sensitive for the determination of trace aconitine, neoaconitine and hypaconitine in vivo.
2. The distribution of aconitine in body fluid and tissues of acute poisoning guinea pigs was studied. Eight guinea pigs were given aconitine by intragastric administration at a dose of 1.0-4.0 mg/kg. After dissection, the contents of aconitine in body fluid and tissues were determined. The order of aconitine content in body fluid and tissues was: bile > blood > urine > stomach > lung > Blood can be used as the first choice of biological analysis for death analysis.
3. The established method was successfully applied to the identification practice. The contents of trace aconitine in biological samples were qualitatively and quantitatively analyzed. The contents of aconitine, neoaconitine and hypaconitine in body fluids and tissues of several aconitine poisoned corpses were determined. The contents of neoaconitine and hypaconitine in corpse tissues were reported for the first time. The distribution provides clear direction and scientific evidence for the determination of the cause of death and the trial of cases involving Aconitum poisoning.
Three, colchicine poisoning, detection and Application
1. A LC-MS/MS-MRM method for the determination of colchicine in blood, urine and tissues was established, and its quantitative method was systematically and systematically validated, including linear range, minimum detection limit, precision, recovery rate, accuracy, matrix effect, stability and so on. The method is simple, rapid, sensitive and the minimum detection limit can be as low as 0.05ng/mL. It can meet the requirements of forensic toxicology analysis and clinical toxicology analysis.
2. The distribution of colchicine in acute poisoned guinea pigs was studied. Five guinea pigs were given colchicine by intragastric administration at a dose of 4.0 mg/kg. The contents of colchicine in body fluid and tissues were measured after dissection. The order of colchicine content in body fluid and tissues was bile > urine > spleen > stomach > lung > kidney. The content of colchicine in heart blood is the lowest. Urine can be used as the first choice for colchicine analysis.
3. The method has been successfully applied to the identification of poisoned plants. The qualitative and quantitative analysis of colchicine in the biological samples of poisoned corpses and wild plants has provided clear direction and evidence for the determination of the cause of death in poisoned plants.
Four, the detection and application of tetrodotoxin in biological samples.
A method for the determination of tetrodotoxin in blood, urine and tissues by LC-MS/MS-MRM was established. The linear range and the minimum detection limit of the method were investigated. The method was applied to the practical cases of tetrodotoxin poisoning in edible puffer fish and satisfactory results were obtained.
The innovation and significance of this study are as follows:
1. The established screening and analysis method for toxic alkaloids is simple and sensitive, avoiding the missed and mistaken detection phenomena that may occur in biological samples by traditional methods. The whole analysis can be completed within 16 minutes. The screening target covers the common toxic alkaloids in forensic identification, and the LC-MS/MS screening system can follow the target. With the increase and expansion of objects, more targets can be screened. This method is a pioneering method.
2. The validity of the quantitative method in biological samples was expounded systematically and comprehensively, and the evaluation index of the method was standardized. The validity of the method for the determination of aconitine, neoaconitine, hypoaconitine and colchicine in biological samples was proved to be accurate, sensitive, exclusive, reproducible and reliable.
3. A LC-MS/MS-MRM method for the determination of aconitine in biological samples was established. The distribution of neoaconitine and hypaconitine in aconitine-poisoned corpses was reported for the first time, which provided clear direction and evidence for the determination of the cause of death in poisoned plants.
4. A LC-MS/MS-MRM method for the determination of colchicine in biological samples was established. The method is sensitive enough and the detection limit can be as low as 0.05 ng/mL. It can meet the requirements of forensic toxicology analysis and clinical toxicology analysis.
5. A method for the determination of tetrodotoxin in biological samples by LC-MS/MS-MRM was established. The distribution of tetrodotoxin in various tissues of tetrodotoxin poisoned corpses was reported for the first time.
The research results fill the blank of toxic animal and plant detection and evaluation in vivo, and make the component analysis technology of common toxic animal and plant in complex biological system of our country leap to the international advanced level. It will provide a technical platform for the scientific research of toxic animal and plant, poisoning identification, clinical diagnosis and treatment, food safety monitoring and so on.
【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：D919.1
【參考文獻】
相關期刊論文 前10條
1 王朝虹;文靜;陳義華;何毅;;液相色譜質(zhì)譜聯(lián)用測定烏頭堿在大鼠體內(nèi)代謝產(chǎn)物[J];中國法醫(yī)學雜志;2006年02期
2 張宏桂;滕坤;劉永剛;張群書;;LC-MS~n分析新烏頭堿在兔尿液中的代謝產(chǎn)物[J];中國法醫(yī)學雜志;2007年06期
3 越皓;皮子鳳;趙宇峰;宋鳳瑞;劉志強;劉淑瑩;;電噴霧串聯(lián)質(zhì)譜分析附子炮制中的化學成分變化[J];分析化學;2007年07期
4 向平;沈敏;卓先義;;生物樣品分析方法的有效性驗證[J];法醫(yī)學雜志;2008年01期
5 洪專,易瑞灶,許晨,楊志文,張揚揚;河豚毒素中毒機理與臨床救治探討[J];中國海洋藥物;2004年03期
6 樊楚明,張?zhí)毂?老年人烏頭堿類藥物中毒80例分析[J];急診醫(yī)學;1999年04期
7 張宏桂;史向國;孫瑩;鐘大放;張寒琦;;兔血液中烏頭堿代謝產(chǎn)物的研究[J];吉林大學學報(理學版);2006年02期
8 艾路;孫瑩;張宏桂;;復方中藥中烏頭生物堿在人體內(nèi)的代謝產(chǎn)物[J];北京中醫(yī)藥大學學報;2007年06期
9 項杰;王陽雪;侯大兵;陶宗婭;張宏;;反相高效液相色譜法測定附子有效成分含量的方法研究[J];四川大學學報(自然科學版);2006年01期
10 沈敏;向平;沈保華;馬棟;嚴慧;王萌燁;;LC-MS/MS多反應監(jiān)測篩選分析血液中132種毒藥物[J];中國司法鑒定;2006年01期
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