| At present it has been proved by way of forensic medicine practice that forensic medicine technical appraisement has given full scope to maintain the people's legitimate rights and interests and medical error compensation when they were concerned medical negligence and it would provide a scientifically, the fair legal evidence.When it touches upon medical dispute of death,the evaluation quality of a patient's postmortem examination is a key link to cope with medical dispute。A high evaluation quality is a fair,objective,scientific prerequisite to the handle medical disputeand it will provide scientific basis。Medicolegal pharmaceutical analysis is one of forensic medicine evaluation .it uses principles of chemistry and pharmacy, adoptes methodology of science and the high tech examination method to separate,abstract and analyses the intoxicants, doubtable material, drugs, narcotics or metabolic product of them when they were absorbed in the body of people or animals in order to verify the toxicant reason and the usage the kind of medicine, to clarify the case nature.When we Carry on Medicolegal pharmaceutical analysis, First of all we should collect reasonable sample. besides the blood, the sample includes the urine, the saliva, the bile, the lymph, thecerebrospinal fluid, the sweat, the milk, the germ cells secretion and the excreta and so on.the organ, tissue as well as breath gas, is the analysis object,too.Because the material kind and the quantity to use Medicolegal pharmaceutical analysis is limited, , if they were consumed,it was difficult to get a new sample. In specific case, when the case needs to carry on in vivo analysis, there were no the material for to be examined actually, while only part organization organ which was fixed by formaldehyde(used in pathology examination )was left.it was the profound significance in certain case trying that separation and purification , quantitative and qualitative analysis to medicine (poison) from the organ,tissue fixed by formaldehyde . ObjectIn this research of animal experimentation,we could detect the content of Cefotaxime Sodium from the organ,tissue fixed by formaldehyde . through we carried on quantitative and qualitative analysis to it ,we observed the relation between the content of drugs and the formaldehyde so it could provide a convincing proof for the medicolegal practice .Methods1.The method was determined according to the physicochemical property of the drugs or intoxicants and their metabolic process in organism.For the tissure fixed by formaldehyde, the refining was applied in order to remove the protein, the organicsolvent was applied in order to abstract and extract, solid phase column extractor was applied to purify the ingredients so that it can be detected by high efficiency liquid phase chromatographic analysis systemGuinea pig fixed formaldehyde in experimental group was killed after they were gived the standard quantity Cefotaxime one hour, the fresh blood, the organ and tissure were obtained. All samples were dripped by the phosphate buffer before the perchloric acid was applied in order to remove the protein . then gentrifugalism and the concentration were used to the upper liquid, solid phase column extractor was applied to purify the ingredients so that it can be detected by HPLC . The equipment of HPLC is Agilent1100 Pickering5200 , ZORBAX Eclipse XDB C-18 chromatographic column was selected. For mobile phase, potassium dihydrogen phosphate 60mg and sodium hydrogen phosphate 1.g, watering dissolves and dilutes 1000ml) - the methyl alcohol (75:25) was used, the PH value was5.5, the detecting wave is 254nm, the temperature - room temperature 25 degrees. The chromatographic column was this: Agilent TC-C18 4.6×250mm,5μm;Agilent Technologies. its flow rate is 1.0ml/min and sample size is 20μl . Analysis method: The HPLC- external standard (1100HPLC series) examines.The organ and tissure fixed the formaldehyde after 1 month, 2 months, were separately detected in the scheduled time depends onthe method and the condition which introuduced in upper phase.Results1,standard curve: A series ofCefotaxime Sodium liquor of 5,10,20,50,100,200,500μg·ml–lwere detected,the regression equation corresponding the chromatograph peak area value and the density was: Y=-1.7397+1.0162X, r=0.997. the result showed that linear relationship is good when the density scope ofCefotaxime Sodium was 0.6--10μg·ml–l,the examination limit was 2.2ng( S/N=2 ) when the method to dilute Cefotaxime Sodium gradually adopted. In entire experiment we choose the quantitative measurement scope were 0.16-18.0μg·g-1;,the recurrence equation was:Y=-1.8463+1.0159X,γ=0.997。2,The content of Cefotaxime Sodiumca was detected in the tissue treated with formaldehyde after one month,two month and three month., The results are list as follows ( X±SD):7.20±2.70 ,5.79±2.37 , 4.78±2.47μg·ml-1 in liver, 6.17±2.44 ,4.89±2.14,4.67±2.43μg·g-1in kidney; 5.53±2.21 , 4.99±1.99 ,3.80±2.17μg·g-1in lung;4.85±0.71,3.44±1.97,4.44±2.18μg·g-1 in heart.The LOD of biosample is 0.023mg.kg-1, RSD% in one day and between days are both less than 4.0%. The absolute rate of Cefotaxime Sodium recovery for biosample fixed formaldehyde is(97.82±0.86)% , the relative rate of recovery is(96.91±3.34)%and abstracted rate of recovery is(94.75±2.19)%3,The result of SD and RSD% in one day and between days showed that the detection value was good identity , RSD% in one day and between days are both less than 4.0%.4,The absolute rate of Cefotaxime Sodium recovery is(97.82±0.86)% , the relative rate of recovery is(96.91±3.34)%and abstracted rate of recovery is(94.75±2.19)%ConclusionThe content of Cefotaxime Sodium in organ tissure treated with formaldehyde is in the affirmative. The detecting results of formalin fixed tissue will provide important proves for the correlation medical service dispute. Loss of Cefotaxime Sodium in organ tissure were more when it was fixed with formaldehyde one month,while the content of Cefotaxime Sodium in sample treated with formaldehyde two or three month had not been affected by formaldehyde.Gives extended application, other some antibiotics which has the similar the physicochemical property and their metabolic process in organism with Cefotaxime Sodium may be detected from organ tissure fixed with formaldehyde, but it still need further to test .This experimental significance lies in that The content of Cefotaxime Sodium in organ tissure treated with formaldehyde could be detected in short periods,this has filled blank which the antibiotic content determination in biological tissue organ after fixed the formaldehyde, simultaneously will also give a way for the forensicmedicine evaluation . |
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