| [objective] To establish a simple and quick, accurate and reliable method to determine the blood drug concentration of calcium folinate and 5-methyltetrahydrofolate .To explore the correlative studies of the blood drug concentration of calcium folinate,and 5-methyltetrahydrofolate in Children’s plasmaand acute lymphoblastic Leukemia’ consolidation therapy, to make clinical rational drug use and rescue plan for doctors who were in department of Pediatric hematology,[methods]1 .To establish a method of ultra high performance liquid chromatography to determine the blood drug concentration of calcium folinate and 5-methyltetrahydrofolate.2.Collected adequate amounts of specimens from department of pediatric hematology(44’MTX) and department of children’s health preventon (healthy children) , then extracted some specimens to determine as well as comparative analysis.3. To analyze the results with SPSS 17.0 statistical software by consulting medical records, collecting the basic information and adverse reactions of children.[results]1 .The methodology of ultra high performance liquid chromatography :chromatographic column: C18 column (100 mm x 2.1 mm, 1.7μm;Waters company);Column temperature: 40℃;Mobile phase: 0.05 mol/L acetic acid +0.07 mol/L EDTA acetate buffer:Methanol (88:12,buffer with acetic acid PH =4.41);Flow rate: 0.25 ml/min;Sample quantity: 5μL;Detection wavelength for FIR excitation wavelength (ex) : 308 nm, emission wavelength (em) : 365 nm.The stability of the method is better, high accuracy, CF’s linear equation was y = 4 x 106 x +106;82 =0.9984 (0.25～16 ug/mL),5,MTHF’s linear equation was y =4 x 107 x +661341;R 2=0.9998 (0.025 ～1.6ug/mL).2.① The level of CF,5-MTHF were 2.23～6.89 ug/mL,0.004～0.038 ug/mL (P 2.5～P 97.5) with 210 cases healthy children (not supplement calcium folinate).②The level of CF,5-MTHF were 2.04～8.63 ug/mL,0.016～0.34 ug/mL (P 2.5～P 97.5) ) with 223 cases from department of pediatric hematology (supplement calcium folinate).3.The value of MTX, CF, 5-MTHF with 160 cases were non-normal distribution.The range of MTX,CF,5-MTHF of them was 0.10～0.71 umol/L,1.90～8.57 ug/mL,0.018～0.36 ug/mL (P 2.5～P 97.5) .The value of CF was positively correlated withthe value of 5-MTHF (r = 0.012), the value of MTX was negatively correlated with the value of CF, 5-MTHF (r = 0.093, r ＝ 0.003).4.The results of CF value of boys and girls with 160 cases～the mean level were 3.99±1.40 ug/mL、 4.3811,75 ug/mL,z =-1.296,P=0.195,CF levels there was no statistically significant difference in gender (P > 0.05);< 1 year old group, 1 or more years, < 10 years old goup, 10 or more years group , the mean level were 3.36±0.77 ug/mL.4.30±1.62 ug/mL、3.72±1.36 ug/mL. Two comparison between them, there were no statistically significant difference;the Han nationality and non-Han nationality,the mean level were 4.19±1.58ug/mL、3.83± 1.48 ug/mL,CF levels there was no significant difference in ethnic (P > 0.05);Low risk, moderate risk and high risk, the mean level were 4.97± 1.53 ug/mL、3.8±1.39 ug/mL、3.81±1.65ug/mL,there was no statistically significant difference between moderate risk and high risk(P > 0.05), low risk was statistically significant between moderate risk and high risk;Adverse reactions and no adverse reactions,the mean level were3.94±1.46ug/mL、4.82±1.71ug/mL,z =-2.945,P=0.003,CF levels there was significant difference in adverse effects (P < 0.05).5.The results of 5-MTHF value of boys and girls with 160 cases,the mean level were 0.11 ±0.064 ug/mL、0.14±0.10 ug/mL,5,MTHF levels there was no significant difference in gender (P >0.05);< 1 year old group, 1 or more years, <10 years old goup,10 or more years group, the mean level were 0.28±0.098 ug/mL、0.12±0.075ug/mL 、 0.098±0.058 ug/mL,variance between groups together P>0.05);the difference was statistically significant (P < 0.05), there was a difference(among all age groups, by LSD-t test, there was no statistically significant difference between 1 .or more years, 1 < 10 years old goup and 10 or more years group, < 1 year old group was statistically significant difference between 1 or more years, < 10 years old goup and 10 or more years group;the Han nationality and non-Han nationality,the mean level were 0.12±0.085 ug/mL、0.13±0.056 ug/mL,5-MTHF levels there was no significant difference in ethnic (P>0.05);Low risk, moderate risk and high risk wereuneven normally distributed,the mean level were 0.11 ±0.066 ug/mL、0.14±0.098 ug/mL、0.099±0.059ug/mL,two comparison between them,there was statistically significant difference between moderate risk and high risk(P< 0.05), low risk was no statistically significant between moderate risk and high risk(P > 0.05);Adversereactions and no adverse reactions, the mean level were 0.12±0.087ug/mL、0.12±0.071ug/mL,z =-0.649,P=0.516,5-MTHF levels there was no significant difference in adverse effects (P >0.05).[conclusions]1. The method of ultra high performance liquid chromatography was simple,accurate and reliable, high sensitivity, wide linear range, low cost, it can be used for detecting the blood drug concentration of CF, 5-MTHF in human plasma and related clinical research.2.By statistical analysis of the value of CF, 5-MTHF with 210 cases healthy children in Kunming area, we suggest CF, 5-MTHF’s reference range of children’s plasma were 3.16～5.18 ug/mL,0.004～0.038 ug/mL in Kunming area.3.The value of CF was positively correlated with the value of 5-MTHF. Namely,the value of 5-MTHF with the value of CF increased; the value of MTX was negatively correlated with the value of CF, 5-MTHF. Namely, the value of CF,5-MTHF decreased with increasing the value of MTX;4.Gender, ethnic, geographical location had no effect on the value of CF,5-MTHF with Children’s acute lymphoblastic Leukemia in Yunnan province;5.Age had effect on the value of 5-MTHF in Children’s acute lymphoblastic Leukemia in Yunnan province, < 1 year old group’s mean level of 5-MTHF was least;6.Adverse reactions of MTX’s mean level was much higher than there was no adverse reaction of MTX’s average level, and adverse reactions of CF’s mean level was far less than that without adverse reactions of CF’s mean level, to illustrate that the rescue of the CF was significant. |
PDF Full Text Request