| Diabetes is a kind of chronic lifelong disease which is related to thepeople health. Metformin hydrochloride has become first choice oftreatment medicine by the diabetic, especially for the obese patients.Therefore, development a method with high selectivity, accuracy and lowdetection limit will have profound significance to the humanity medicalresearch. However, when determined directly the selectivity is not sosatisfactory because of low or non-absorption of metforminhydrochloride.This paper use p-chlorobenzoyl chloride as a derivatization reagentto react with metformin hydrochloride for the first time. By this way, thepre-column derivatization can be received. According to the experiment,the optimum experimental conditions are as follows: the wavelength is at249.0nm; methanol: buffer solution: (KH2PO4+H3PO4, pH=4)=80:20;the flow rate is 0.8mL/min; the column temperature is at 25℃; thesamples size is 20μL; the optimum solvent is dichloridethane; theoptimum concentration of NaOH is 2.5mol/L; the optimum reaction time is 60min; the optimum throw material of than is 4:1; the optimumtemperature is room temperature. Under the optimum experimentalconditions, the linear range of calibration curve is 0.0050mg/L～50.0mg/L, the minimum detecting concentration is 5.0μg/L, the within-dayand inter-day precision RSD of 0.20mg/L,5.0 mg/L,20.0 mg/L are2.81％,0.92％,0.47％and 3.54％,3.32％,1.41％respectively. Therecovery are 98.5％～103.3％,96.13％～102.9％,93.93％～96.38％respectively.According to the experiment, the metformin hydrochloride has aclear ultraviolet absorption after derivation. A method named columnchromatography chromatography is used to purify the derivatizationproduct. Then UV,IR,NMR and MS are used to identified the structure.The mechanism of the reaction is also be studied. After reaction,the main structure of metformin hydrochloride is changed. Themechanism of the reaction is also be studied.Finally, the method is used to determine the simulation blood sample.Under the optimum preprocessing, the linear range of calibration curve is1.0mg/L～20.0mg/L, the minimum detecting concentration is 5.0μg/L,the within-day and inter-day precision RSD of 2.0 mg/L,5.0 mg/L,20.0mg/L are 5.40％,1.98％,3.22％and 5.59％,2.52％,4.17％respectively.The recovery are 94.92％～101.68％,98.23％～102.78％,96.88％～100.08％respectively.
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