| OBJECTIVES:1. To establish and evaluate the electrochemical measurement method to determination the concentration of vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B9, vitamin B12, vitamin C, vitamin D and vitamin E in human serum.2. For regular collectin and testing the concentrations of the nine vitamins in newborns and infants and young children, pregnant women, older patients and postoperative, trauma and other debilitating diseases patients who can’t eat and other critical illness patients from March 2013 to November 2014.3. Statistic analysis the determination data of the 9 vitamins, to determine the difference between the measured value and the normal range of the nine vitamins of the different patient populations, and to provide expermental evidence for the diagnosis and treatment of the diseases which related to vitamin (vitamin deficiency or excess syndrome), and further promote rational use of vitamins.METHODS:1.3 mL blood of the newborns and infants and young children, pregnant women, elderly patients, and postoperative, trauma and other debilitating diseases patients who can’t eat of patients and other severe disease were drawed, placed in the normal serum vacuum blood vessels (red hat) which does not contain additives. After 4000 rpm centrifugal 4 min, take the corresponding serum into the vitamin sample processing solution, mixed, then determinated using LK3000V vitamin detector in the same day and statistical analysis the determination results.2. Using LK3000V vitamin detector to determinate the serum concentration of nine kinds of vitamins including vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B9, vitamin B12, vitamin C, vitamin D, vitamin E. Take the appropriate volume of serum (in which vitamin A, vitamin D, vitamin E sample processing solution were added 80μL serum; vitamin B9, vitamin B12 sample processing solution and vitamin B6 sample processing solution were added 40μL serum; vitamin B2 sample processing solution were added 20μL serum; and the vitamin B1 sample processing solution were dded 10 μL serum.) added into the sample processing solutionin, and determined after the electrode processing. The determination of vitamin B9 and vitamin B12 needs electrode activation before electrode processing.3. Statistical analysis the test data of nine kinds of vitamins in different patient populations, draw the scatter plot of each vitamin according to the test data to determine the difference between the measured value and the normal range of the nine vitamins of the different patient populations, analysis the cause of excess or lack of vitamin in different patient populations.4. Statistical analysis the blood concentration data of nine kinds of vitamins of 672 cases of newborn and infants patients, draw the scatter plot of each vitamin test data to determine the difference between the measured value and the normal range of the nine vitamins of the different patient populations, analysis the cause of excess or lack of vitamin in different patient populations and to diagnose vitamin deficiency or excess syndrome inside the body of the newborn and infant.RESULTS:1. Due to the different intakes, and the amounts that used to maintain the normal function of the body are different, so as to make the different vitamins have different linear range, and their determination should also needed to establish different standard curve. The linear range of vitamin A was 0.67-3.38μmol/L, and the standard curve was y=6.56131+0.57857 x (r=0.9999). The linear range of vitamin B1 was 33-265 nmol/L, and the standard curve was y= 48.8445 x+25.6258 (r=0.9999). The linear range of vitamin B2 was 194-968μg/L, and the standard curve was y=473.184 x+178.182 (r=0.9999). The linear range of vitamin B6 was 14.6-146 μmol/L, and the standard curve was y= 361.344 x-14.3965 (r=0.9985). The linear range of vitamin B9 was 11.8-118 nmol/L, and the standard curve was y=237.524 x-3.12678 (r=0.9999). The linear range of vitamin B12 was 203-1015 pg/mL, and the standard curve was y= 3575.17 x-52.1452 (r=0.9998). The linear range of vitamin C was 34-138 nmol/L, and the standard curve was y= 441.621 x+15.8955 (r=0.9999). The linear range of vitamin D was 20-200 mmol/L, and the standard curve was y= 530.894 x-4.29012 (r= 0.9999). The linear range of vitamin E was 10-30 ug/mL, and the standard curve was y= 27.5103 x+9.45439 (r= 0.9999). All the linear range of nine kinds of vitamins have good linear relationship, and the linear range could cover the clinical reference value range of vitamins, and that can be used for the determination of serum vitamin concentration in clinical patients.2. In March 2013 to November 2014, Shandong University Qilu Hospital Drug Monitoring Center received clinical routine inspection samples for testing. The concentration data collected including 2050 cases of vitamin A, vitamin D, vitamin E; 1894 cases of vitamin B1 and vitamin C; 1857 cases of vitamin B2 and vitamin B6; 2028 cases of vitamin B9, B12. Patients include newborns and infants and young children, pregnant women, older patients, and postoperative, trauma and other debilitating disease and patients with severe disease who cannot eat. The youngest patient was a newborn who was born three minutes, and the oldest one was a 99-year-old man.3. The mean value and standard deviation of vitamin A determinated concentration were 0.63±0.30 μmol/L (n=2050), in which the minimum value was 0.30μmol/L, the maximum value was 9μmol/L, and the maximum was 30 times to the minimum. The mean value and standard deviation of vitamin B1 determinated concentration were 125.72±60.67 nmol/L (n=1894), in which the minimum value was 11.01 nmol/L, the maximum value was 640.42 nmol/L, and the maximum was 58.17 times to the minimum. The mean value and standard deviation of vitamin B2 determinated concentration were 227.58±73.39μg/L (n=1857), the minimum value was 8.01 μg/L, the maximum value was 1348.84μg/L, and the maximum was 168.39 times to the minimum. The mean value and standard deviation of vitamin B6 determinated concentration were 36.18±23.43μmol/L (n=1857), the minimum value was 0.03 nmol/L, the maximum value was 160.51 nmol/L, and the maximum was 5350.33 times to the minimum. The mean value and standard deviation of vitamin B9 determinated concentration were 9.43±6.58 nmol/L (n=2028), the minimum value was 0.03 nmol/L, the maximum value was 106.51 nmol/L, and the maximum was 5350.33 times to the minimum. The mean value and standard deviation of vitamin B12 determinated concentration were 279.65±142.44 pg/mL (n=2028), the minimum value was 3.63 pg/mL, the maximum value was 1014.7 pg/mL, and the maximum was 279.53 times to the minimum. The mean value and standard deviation of vitamin C determinate concentration was 40.48±12.66μmol/L (n=1894). The minimum value was 0.39μmol/L, the maximum value was 209.46μmol/L, and the maximum was 537.08 times to the minimum. The mean value and standard deviation of vitamin D determinate concentration were 38.71±26.56μmol/L (n=2050). The minimum value was 0.36 nmol/L, the maximum value was 485.65 nmol/L, and the maximum was 1349.03 times to the minimum. The mean value and standard deviation of vitamin E determinate concentration were 12.58±7.90 μg/mL (n=2050), the minimum value was 0.63 μg/mL, the maximum value was 300.18 μg/mL, and the maximum was 476.48 times to the minimum.4. Through the analyzed the vitamin testing results of 672 cases of newborn and infant patients, the vitamin A deficiency patients accounted for 36% of the total number of determination of vitamin A, and the vitamin A overdose patients accounted for 0.3%. Vitamin B1 deficiency patients accounted for 7% of the totle number of determination of vitamin B1, and the vitamin B1 overdose patients accounted for 28.1%. Vitamin B2 deficiency patients accounted for 20.5% of the totle number of determination of vitamin B2. Vitamin B6 deficiency patients accounted for 7.2% of the totle number of determination of vitamin B6, and the vitamin B6 overdose patients accounted for 8.1%. Vitamin B9 deficiency patients accounted for 49.8% of the totle number of determination of vitamin B9, and the vitamin B9 overdose patients accounted for 0.6%. Vitamin B12 deficiency patients accounted for 52.1% of the totle number of determination of vitamin B12. Vitamin C deficiency patients accounted for 42.5% of the totle number of determination of vitamin C, and the vitamin C overdose patients accounted for 0.9%. Vitamin D deficiency patients accounted for 28.4% of the totle number of determination of vitamin D, and the vitamin D overdose patients accounted for 0.3%. Vitamin E deficiency patients accounted for 21.8% of the totle number of determination of vitamin E, and the vitamin E overdose patients accounted for 8.8%.CONCLUSIONS:In this paper, the electrochemical method that used for simultaneous determination of serum concentration of vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B9, vitamin B12, vitamin C, vitamin D and vitamin E was investigated. The method is sensitive, accurate, rapid and reproducible, the linear range could cover the clinical reference value range of vitamins, and can be used for the routine monitoring the blood concentration of the relecance diseases (vitamin deficiency or excess syndrome) of the nine kinds vitamine. The testing data can help the diagnose and individualized treatment of certain vitamins related diseases.The difference between the maximum measured values and the minimum values of 9 kinds vitamin is comparatively large, in which the most was vitamin B6, the maximum was 5350.33 times to the minimum. The difference of measured values between each classification of the population was large, and clinically normal reference value cannot be fully applicable to all people, it is necessary to establish the normal range of each vitamin of each classification of the population respectively.The lack of vitamin A, vitamin B2, vitamin B9, vitamin B12, vitamin C, vitamin D and vitamin E accounted for the majority of the population of newborns and infants. While the vitamin Bi overdose accounted for a larger proportion. Therefore, during the growth and development process of newborns and infants should be noted the supplement of vitamin A, vitamin B2, vitamin B9, vitamin B12, vitamin C, vitamin D and vitamin E, and should prevent excessive vitamin B1.The electrochemical method that used for simultaneous determination of serum concentration of nine kinds of vitamin can be applied for the diagnosis of vitamin correlation disease under certain conditions. But there are certain limitations, such as the information about the correlation of the method itself and generally accepted methods, for example, the LC-MS method; and the correlation of some abnormal values and the clinical diagnosis of vitamin correlation disease is not yet clear. It remains needed the large sample of clinical patients data for correlation analysis. |
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