Simultaneous Determination Of7Sulfonylurea Type Oral Anti-diabetic Agents In Health Care Products

Glipizide, tolbutamide, tolazamide, gliclazide, glibenclamide,glimepiride and glurenor belong to sulfonylurea-type oral anti-diabeticagents which is one of major type of oral anti-diabetic agents, and they arewidely used for type Ⅱ diabetes. Common side effects from these drugsinclude hypoglycemic, obesity, gastrointestinal upset, skin allergic. Inrecent years, more and more people are interested in heath with the rapiddevelopment of society and economy. Most patients choose not only theoral anti-diabetic agents but also dietary supplements to control blood sugar.The consumption of dietary supplements has been growing in somecountries. However, to cater to consumer interest and get excess profits, theillegal adulterations of dietary supplements with synthetic anti-diabeticagents without labeling were frequently exposed，which bring a lot ofunexpected adverse effects and make public heath in danger. Thus rapid,reliable, sensitive and accurate methods are required for analyzing theillegal adulterants in dietary supplements to prevent the harmful effects ofsynthetic drugs.This study established a sensitive, rapid, accurate, reliabledetermination method to simultaneous determination of seven sulfonylureahypoglycemic agents in health care products, which detected by HPLC andUPLC-MS/MS respectively. The full text is composed of two parts. Part Ⅰ Determination of seven sulfonylurea-type oral anti-diabeticagents in Health Care Products by HPLCObjective: A method was established for the simultaneousdetermination of seven sulfonylurea hypoglycemic agents (Glipizide、Tolbutamide、Tolazamide、Gliclazide、Glibenclamide、Glimepiride andGlurenor)in health care products by solid phase extraction-highperformance liquid chromatography (SPE-HPLC). Methods: Samples wereextracted with methanol by ultrasound, cleaned-up and concentrated on anLC-C18SPE column (hypoglycemic capsule、hypoglycemic powder、hypoglycemic oral liquid、hypoglycemic milk powder、hypoglycemicbiscuits) and an NH2SPE column(hypoglycemic tea) separately, undernitrogen gas stream，followed by blowing to dryness，reconstitution withmethanol before the analysis by HPLC. The chromatographic separationwas performed on an Agilent Zorbax SB-C18column（250mm×4.6mm，5μm）and using acetonitrile (A)-0.02%phosphoric acid solution(B)withgradient elution(0～5min:40%A；5～10min:40%A→60%A；10～25min：60%A)as the mobile phase at a flow rate of1.0mL/min，columntemperature at30℃and ultraviolet(UV) detection at228nm. Results: Theseven sulfonylurea hypoglycemic agents had good linear relationships inthe range of0.2～20μg·mL-1with the correlation coefficients not less than0.9999; the recoveries of seven target compounds in five different healthcare products ranged from81.2%to114.5%and the relative standarddeviations were less than8.2%. Conclusion: High performance liquidchromatography and a solid phase extraction simultaneous determination ofseven sulfonylurea-type oral anti-diabetic agents in health care productshave been developed. The solid phase extraction column purification effectis better than ultrasonic extraction and dispersive solid phase extraction. The samples are analyzed by HPLC followed purification. The method issimple, accurate, rapid and repeatable. It can be used for the simultaneousdetermination of seven sulfonylurea hypoglycemic agents in health careproducts.Part Ⅱ Determination of seven sulfonylurea-type oral anti-diabeticagents in health care products by UPLC-MS/MSObjective: To develop an UPLC-MS/MS method for thedetermination of seven sulfonylurea hypoglycemic agents in health careproducts. Methods: The samples were extracted twice with methanol byultrasound and the extracts were combined, under nitrogen gas stream,followed by blowing to dryness, reconstitution with mobile phase of initialratio before the analysis by UPLC-MS/MS. The separation was achievedon a C18column with the mobile phase consisting of acetonitrile and water(0.2%formic acid (V/V) and5mM ammonium acetate), at a flow rate of0.3mL/min with gradient elution. The column temperature was35℃andthe injection volume was10μL. Conditions of MS/MS: electrospray wasoperated in positive ion mode and the ionization source parameters were:capillary voltage2.5kV, ion source temperature110℃and desolvationtemperature350℃. Nitrogen was used as the desolvation and cone gas witha flow rate of650and50L/h, respectively. Results: The standard curvehad good linear relationships in the range of5～200μg·L-1(r≥0.99). Therecoveries were between82.9%～109.2%, the RSD were less than6.0%for the high、 middle and low concentration. Conclusions: AnUPLC-MS/MS simultaneous determination of seven sulfonylureahypoglycemic agents in health care products has been developed. Thismethod is sensitive, accurate, and applied to determine the concentration ofseven sulfonylurea hypoglycemic agents in health care products.