Study On Analysis Method And Application Of34Elements In Human Blood And Urine

As the further study on the relationship between elements and human health anddisease, accurate measurement of elements levels in human body has become more andmore important. Facing the heavy metal pollution in China, the treating for metal pollutionhas been on the government’s agendas. In2011.2, the “12thfive-year plan for theprevention and control of heavy metal pollution” became the first “12thfive-year” nationalplanning formally approved by the state council’. Our research is one component of the keytechniques of detection and evaluation of metal poisoning which belongs to national “12thfive-year” science and technology support plan.People have paid great attention to the levels of elements in human body in recentyears. Elements analyses have been included in general medical checkup of some medicalcare and health institutions. But now there is no an accurate and united criteria fordetermination of elements. The recent study is to establish an inductively coupledplasma-mass spectrometry （ICP-MS） method with wet digestion, direct diluted aspretreatment method for determination of34elements (⁷Li,⁹Be,¹¹B,²⁴Mg,²⁷Al,⁴³Ca,⁴⁷Ti,⁵¹V,⁵³Cr,⁵⁵Mn,⁵⁷Fe,⁵⁹Co,⁶⁰Ni,⁶³Cu,⁶⁶Zn,⁶⁹Ga,⁷⁵As,⁸²Se,⁸⁵Rb,⁸⁸Sr,⁹⁰Zr,⁹⁵Mo,¹⁰⁷Ag,¹¹¹Cd,¹¹⁸Sn,¹²¹Sb,¹³³Cs,¹³⁷Ba,¹⁹⁷Au,²⁰²Hg,²⁰⁵Tl,²⁰⁸Pb,²³²Th,²³⁸U) in human blood andurine, which will provide technical support for developing normal value of biologicalsamples inorganic element database in Chinese population. The study results will improvethe testing of metal elements of public safety, provide reference data for detection andresults evaluation of elements in human body. The main research contents include thefollowing3parts:1.Establishment of ICP-MS method for34elements determination in bloodWith Scandium (⁴⁵Sc), Germanium (⁷²Ge), Yttrium (⁸⁹Y), Indium (¹¹⁵In), Terbium (¹⁵⁹Tb) as internal standard, a wet digestion method for blood as well as the operatingparameters for ICP-MS was optimized to establish ICP-MS method for determination of34elements in human blood.65%HNO₃and29-32%H₂O₂was used as digestion reagents in our study. Thepretreatment method was optimized in digestion reagents, digestion time and temperature.The optimized pretreatment method was0.25mL blood was mixed with0.8mL65%HNO₃and0.2mL29-32%H₂O₂and digested in90℃for3h using a heating plate andthen diluted to10mL with ionized water. Our method was compared with microwavedigestion and found no difference. In consideration of the need of a lager sample size, thenew method was more practical.The limits of detection of34elements ranged for0.0002ng/mL （Cs）-21.8ng/mL （Ca）;the precision and accuracy were obtained by standard addition method, accuracy shown inadded recovery ranged from91%-112%, precision of intra-day and inter-day ranged from0.5%-8.9%and0.4%-10.9%, respectively. The results can meet detection requirements.2.Establishment of ICP-MS method for34elements determination in bloodWith Scandium (⁴⁵Sc), Germanium (⁷²Ge), Yttrium (⁸⁹Y), Indium (¹¹⁵In), Terbium(¹⁵⁹Tb) as internal standard, a direct dilution technique for urine as well as the operatingparameters for ICP-MS was optimized to establish ICP-MS method for determination of34elements in human urine. The diluted reagent concentration of5%HNO₃and diluted factorof40were selected.The limits of detection of34elements ranged for0.0001ng/mL （Th）-4.64ng/mL （Ca）;the added recovery ranged from90%-109%, precision of intra-day and inter-day rangedfrom0.4%-8.0%and1.4%-8.7%, respectively. The results can meet detectionrequirements.3.Establishment of normal reference range and analysis of influence factorsMeasure284blood and urine samples from Jinan of Shandong Province and SanmenCounty of Zhejiang Province. The34elements normal reference ranges in blood of Jinanwas established. The34elements normal ranges in blood and urine of Sanmen County were calculated, too.The statistical analysis shows there exists diversity in body elements contents fromdifferent regions. Some of the elements is different due to different diet, like the As, Hg.Sanmen County is rich in seafood, so local residents eat more seafood. The blood Ascontent was significantly higher than that in Jinan blood samples. Some elements are dueto the different geographical environment. Ba content in Sanmen County blood and urinesamples is much higher than that in Jinan blood and urine samples. This may be due to thedifferent geological or pollution of the environment.Gender and age are potential determinants of elements levels in blood and urine. Theresults show there were some effect of gender on levels of Cr, Mn, Co, Rb, Mo and Pb andage on levels of Se, Cd, Cs, Hg, Pb in blood. For urine, there were Zn, had statisticallyrelationship with gender and Cu and Co with age.The validated method was successfully used in the judicial authentication practice.100ng/mL of Ag was measured from a sufferer’s blood and1.35ng/mL in the urinesample. This level has well above that documented in the reference.Overall, the innovation points and the significance of our research summarized asfollows:1.Developed inductively coupled plasma-mass spectrometry （ICP-MS） with wetdigestion method and direct diluted method for simultaneous analysis of34elements inhuman blood and urine.2.The establish method was applied to measure healthy people’s blood and urinesamples in Jinan and Sanmen in order to obtain normal ranges.3.Explore the diversity of elements level between the two regions and effects ofgender and age on elements levels in blood and urine.