Determination Of Vitamin D 3 Level By LC-MS/MS Method And Analysis Of Serum Vitamin D Level In Patients With Pulmonary Tuberculosis

Background:Determination of25-OHD has no uniform method currently, mainly with radioimmunoassay (RIA), enzyme immunoassay (ELISA), liquid chromatography (HPLC), chemiluminescence method and liquid chromatography tandem mass spectrometry (LC-MS/MS) which developed very rapidly in recent years. Because LC-MS/MS has high sensitivity, a lot of research in it as the" golden standard". Vitamin D has certain relations on tuberculosis and diabetes mellitus.There is many tuberculosis and diabetes in China, but there is little research about vitamin D in tuberculosis and diabetes, and no reports about vitamin D level in pulmonary tuberculosis complicated with diabetes mellitus or pulmonary tuberculosis complicated with hyperglycemia crowd in China.Objective:To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of25hydroxy vitamin D3(25OHD3) in human serum. And to observe serum vitamin D concentration changes in pulmonary tuberculosis patients with or without elevated blood glucose, analysis of factors.Methods:The general data of578subjects was collected. The TB case group and control group were divided into two groups respectively based on fasting blood glucose of6.1mmol/L, healthy group:(controls whith glucose<6.1mmol/L); hyperglycemia group:(controls whith glucose≥6.1mmol/L); pulmonary tuberculosis group:(TB case group whith glucose <6.1mmol/L); pulmonary tuberculosis complicated with hyperglycemia group:(TB cases whith glucose≥6.1mmol/L). Using LC-MS/MS method for measuring25-OHD3in serum.Results:The LC-MS/MS assay had a limit of quantitation of1.0ng/ml for25-OHD3with a linear response between6ng/ml and120ng/ml (r=0.999).53serum samples were measured by LC-MS/MS and RIA respectively, correlations was:RIA=LC-MS/MS-3.57, r=0.883.25-OHD3levels in healthy group (16.75ng/ml) and hyperglycemia group (19.70ng/ml) were significantly higher than that of pulmonary tuberculosis group (11.10ng/ml)(P<0.001); the hyperglycemia group was significantly higher than that of pulmonary tuberculosis complicated with hyperglycemia group (16.70ng/ml)(P=0.008); The research subjects whith glucose≥6.1mmol/L (18.83ng/ml) has no difference whith the healthy group (P>0.05), and was significantly higher than that of pulmonary tuberculosis group (P <0.001);25-OHD3levels in pulmonary tuberculosis patients (13.70ng/ml) was significantly lower than that in normal group (P<0.001) and high glucose group (P<0.001). Vitamin D levels increased with sun exposure time, but the difference was not statistically significant. Vitamin D in different age, sex, body mass index did not differ significantly.Conclusion:The LC MS/MS assay and compares well with the RIA.25-OHD3levels in patients with pulmonary tuberculosis is lower.25-OHD3levels is higher in high blood glucose in cases and controls than normal blood sugar. Physical activity on25-OHD3deficiency rate has certain effect.