苦參堿的毒理及HPLC法測定綿羊血漿苦參堿的研究
發(fā)布時間：2019-02-13 07:38
【摘要】：本文目的為研究苦豆子的主要有效成分苦參堿的毒理以及苦參堿經(jīng)綿羊皮膚給藥的吸收作用。試驗所用到的苦豆子均采自于鄂爾多斯市鄂托克前旗,并經(jīng)由內(nèi)蒙古農(nóng)業(yè)科學院獸醫(yī)研究所化學提純后得到苦參堿樣品。實驗一為苦參堿毒性試驗；該實驗用一定濃度的苦參堿溶液對小白鼠進行灌胃,記錄小白鼠的急性中毒癥狀、探尋苦參堿作用的主要靶器官、測定苦參堿對小白鼠的半數(shù)致死量以及觀察停止灌胃苦參堿后主要靶器官的毒性恢復程度。借鑒Bliss的公式來計算苦參堿的半數(shù)致死量。結(jié)果表明：通過眼觀與病理組織學對比發(fā)現(xiàn)苦參堿主要作用靶器官有肺臟、肝臟、腦、腎臟和心臟。在恢復程度上肺臟的情況較差,腦組織、肝臟和腎臟造成的損害恢復情況較為理想。實驗二為刺激性試驗,試驗從研究苦參堿對皮膚的刺激性和眼的刺激性兩方面來進行。皮膚刺激性試驗分為單次皮膚刺激性試驗和長期皮膚刺激性試驗兩部分,所用藥物分別是苦參堿制劑和苦參堿透皮制劑兩種規(guī)格。其中苦參堿透皮制劑組中添加有透皮促進劑(氮酮和丙二醇),所用到的試驗動物為大白兔。實驗結(jié)果顯示：未添加透皮劑的苦參堿組在單次與多次刺激試驗中對皮膚與眼結(jié)膜均無刺激性；而添加了透皮促進劑的苦參堿透皮制劑組在單次與多次刺激試驗中對皮膚存在中等強度刺激性,刺激作用的眼觀變化表現(xiàn)為眼皮紅腫、皮增厚、皮結(jié)痂、痂皮脫落、新皮生成等；病理切片上則表現(xiàn)為皮膚角質(zhì)層的增生脫落,真皮層的纖維水腫斷裂,刺激小血管充血、出血,炎性細胞侵潤等。實驗三苦參堿液相學研究,采用島津LC-2010C液相色譜儀,建立血漿中苦參堿的HPLC檢測方法,其中用槐定堿作為內(nèi)標。血漿前處理以甲醇為蛋白沉淀劑,通過紫外檢測器進行檢測,檢測波長220nm,流動相為A-0.05moL/L的磷酸二氫鉀(磷酸氫二鉀調(diào)pH到6.46)和B-甲醇(A/B,65：35),等濃度洗脫,流速0.8mL·min-1,建立在羊血漿檢測苦參堿的高效液相色譜方法,目標生物堿色譜峰出峰良好�？鄥A的線性關(guān)系良好R20.999。其中MT使用非經(jīng)脈管模型計算其藥效動力學各參數(shù)。實驗證明苦參堿外用時,藥物的主要作用部位為體表,添加透皮促進劑也僅有極小量的藥物會進入全身血液循環(huán),因此透皮吸收進入血液的有效成分少,本實驗建立了在綿羊血漿內(nèi)檢測苦參生物堿的方法。
[Abstract]:Objective to study the toxicology of matrine, the main active component of Sophora flavescens, and the absorption of matrine through sheep skin. The samples of Sophora flavescens collected from Qianqi, Ordos city, were purified by chemical purification from Institute of Veterinary Medicine, Inner Mongolia Academy of Agricultural Sciences, and the samples of matrine were obtained. The first experiment is matrine toxicity test. In this experiment, a certain concentration of matrine solution was used to gavage the mice, to record the acute toxic symptoms of the mice, and to explore the main target organs of matrine action. The median lethal dose of matrine on mice and the recovery of toxicity of the main target organs after the cessation of oral administration of matrine were determined. The Bliss formula is used to calculate the 50% lethal dose of matrine. The results showed that the main target organs of matrine were lung, liver, brain, kidney and heart. The recovery degree of lung was poor, and the damage caused by brain tissue, liver and kidney was better. The second experiment was an irritation test, which studied the skin irritation of matrine and the irritation of the eye. The skin irritation test is divided into two parts: single skin irritation test and long-term skin irritation test. The drugs used are matrine preparation and matrine transdermal preparation respectively. Transdermal enhancers (azone and propylene glycol) were added in matrine transdermal preparation group. The results showed that there was no irritation to the skin and conjunctiva in the matrine group without the addition of transdermal agents. In the single and multiple irritation tests, the skin was irritated with moderate intensity in the matrine transdermal preparation group. The ocular changes of the irritation were as follows: swelling of the eyelid, thickening of the skin, scabbing of the skin, exfoliation of the scab, and exfoliation of the skin. New skin formation; The histopathological sections showed the proliferation and exfoliation of the cuticular layer of the skin, the fibrous edema and rupture of the dermis, the stimulation of small blood vessel congestion, hemorrhage, inflammatory cell infiltration and so on. The HPLC method for the determination of matrine in plasma was established by using LC-2010C liquid chromatograph of Shimadzu, in which Sophorine was used as the internal standard. In plasma pretreatment, methanol was used as protein precipitator and detected by UV detector. The detection wavelength was 220 nm. The mobile phase consisted of A-0.05moL/L potassium dihydrogen phosphate (pH to 6.46) and B- methanol (A / B). 65:35), iso-concentration elution, flow rate 0.8mL min-1, was established in sheep plasma for the detection of matrine by high performance liquid chromatography, the target alkaloid chromatographic peak produced a good peak. The linear relationship of matrine is good R20.999. The pharmacokinetic parameters of MT were calculated by non-vessel model. The experiment proved that when matrine was used for external use, the main action site of the drug was the body surface, and the addition of transdermal accelerator only a minimal amount of drugs would enter the whole body blood circulation, so transdermal absorption into the blood of the effective components less, A method for the determination of alkaloids of Sophora flavescens in sheep plasma was established.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S853.7
[Abstract]:Objective to study the toxicology of matrine, the main active component of Sophora flavescens, and the absorption of matrine through sheep skin. The samples of Sophora flavescens collected from Qianqi, Ordos city, were purified by chemical purification from Institute of Veterinary Medicine, Inner Mongolia Academy of Agricultural Sciences, and the samples of matrine were obtained. The first experiment is matrine toxicity test. In this experiment, a certain concentration of matrine solution was used to gavage the mice, to record the acute toxic symptoms of the mice, and to explore the main target organs of matrine action. The median lethal dose of matrine on mice and the recovery of toxicity of the main target organs after the cessation of oral administration of matrine were determined. The Bliss formula is used to calculate the 50% lethal dose of matrine. The results showed that the main target organs of matrine were lung, liver, brain, kidney and heart. The recovery degree of lung was poor, and the damage caused by brain tissue, liver and kidney was better. The second experiment was an irritation test, which studied the skin irritation of matrine and the irritation of the eye. The skin irritation test is divided into two parts: single skin irritation test and long-term skin irritation test. The drugs used are matrine preparation and matrine transdermal preparation respectively. Transdermal enhancers (azone and propylene glycol) were added in matrine transdermal preparation group. The results showed that there was no irritation to the skin and conjunctiva in the matrine group without the addition of transdermal agents. In the single and multiple irritation tests, the skin was irritated with moderate intensity in the matrine transdermal preparation group. The ocular changes of the irritation were as follows: swelling of the eyelid, thickening of the skin, scabbing of the skin, exfoliation of the scab, and exfoliation of the skin. New skin formation; The histopathological sections showed the proliferation and exfoliation of the cuticular layer of the skin, the fibrous edema and rupture of the dermis, the stimulation of small blood vessel congestion, hemorrhage, inflammatory cell infiltration and so on. The HPLC method for the determination of matrine in plasma was established by using LC-2010C liquid chromatograph of Shimadzu, in which Sophorine was used as the internal standard. In plasma pretreatment, methanol was used as protein precipitator and detected by UV detector. The detection wavelength was 220 nm. The mobile phase consisted of A-0.05moL/L potassium dihydrogen phosphate (pH to 6.46) and B- methanol (A / B). 65:35), iso-concentration elution, flow rate 0.8mL min-1, was established in sheep plasma for the detection of matrine by high performance liquid chromatography, the target alkaloid chromatographic peak produced a good peak. The linear relationship of matrine is good R20.999. The pharmacokinetic parameters of MT were calculated by non-vessel model. The experiment proved that when matrine was used for external use, the main action site of the drug was the body surface, and the addition of transdermal accelerator only a minimal amount of drugs would enter the whole body blood circulation, so transdermal absorption into the blood of the effective components less, A method for the determination of alkaloids of Sophora flavescens in sheep plasma was established.
【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S853.7
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相關(guān)期刊論文 前10條
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