Study On The Detection Of Chemicals Illegally Added To Proprietary Chinese Medicines And Healthy Foods

OBJECTIVEThis issue aimed at the study on the detection of chemicals illegally added to proprietary Chinese medicines and healthy foods included the types of anti-rheumatic analgesic,antitussive antiasthmatic appeared on the market currently.For 14 kinds of common hormones,10 kinds of antipyretic analgesic category and 10 kinds of antitussive and antiasthmatic components,the fast and simple ways of TLC and HPLC-DAD are established,which could be used for screening rapidly in grass-roots units, and the establishment of HPLC-MS-MS detection methods can be applied to the confirmation of stRucture for the positive samples,which would provide technical basis for combating this act of illegally adding chemical substances to proprietary Chinese medicines and healthy foods.METHODS1 method of TLCHormonesTLC plate:Quickly seized dedication board;Developing Agent:【H-003】(dedicated developing agent for drug testing);Reagent:Blue TetRazolium alkaline test solution. Antipyretic analgesic categoryTLC plate:Quickly seized dedication board;Developing Agent:【H-042】(dedicated developing agent for drug testing);Observation conditions:UV lamp 254nm,365nm,sunlight,sprayed with Ninhydrin test solution,UV lamp 365nm after 120℃heating 20min,iodine,such as fumigation.Antitussive and antiasthmatic categoryTLC plate:Quickly seized dedication board;Developing Agent:【H-012】,【H-022】(dedicated developing agent for drug testing);Observation conditions:UV lamp 254nm,365nm,sunlight,iodine fumigation,spray with 1%ethanol solution of ninhydrin,105℃heating for coloring,spray to dilute bismuth potassium iodide test solution and etc.2 method of HPLC-DAD2.1 HormonesColumn:SHIMADZU Shim-pack VP-ODS(250mm×4.6mm,5μm) C₁₈(hereinafter the same),group A and B Mobile phase:Water: acetonitRile gradient elution,Column temperature:30℃,flow rate: 1.0ml·min^-1,DAD detector(200nm～400nm),detection wavelength:240nm, injection volume:20μl.2.2 Antipyretic analgesic categorygroup A:Mobile phase:A:10mmol·L^-1 Ammonium acetate solution, B:acetonitRile,gradient elution,Column temperature:30℃,flow rate: 1.0ml·min^-1,DAD detector(200nm～400nm),detection wavelength:280nm, injection volume:20μl.group B:Mobile phase:20mmol·L^-1 Sodium acetate solution(glacial acetic acid adjusted pH 5.0) - acetonitRile(60:40);Column temperature: 30℃,flow rate:1.0ml·min^-1,DAD detector(200nm～400nm),detection wavelength:230nm,injection volume:20μl.2.3 Antitussive and antiasthmatic categorygroup A:Mobile phase:A:10mmol·L^-1 Ammonium acetate solution,B: acetonitRile,gradient elution,Column temperature:30℃,flow rate: 1.0ml·min^-1,DAD detector(200nm～400nm),detection wavelength: 230nm,injection volume:20μl.group B:Mobile phase:0.1mol·L^-1 Phosphoric acid-acetonitRile (96:4);Column temperature:30℃,flow rate:1.0ml·min^-1,DAD detector(200nm～400nm),detection wavelength:207nm,injection volume: 20μl.3 method of HPLC-MS-MS3.1 Hormones3.1.1 Chromatographic conditions:Column:SHIMADZU Shim-pack VP-ODS(250mm×4.6mm,5μm) C₁₈(hereinafter the same);Mobile phase:A:20mmol·L^-1 Ammonium acetate solution(containing 0.1% formic acid,V/V) B:AcetonitRile(Methods with the same 2.1 Hormones group A and B of Mobile phase gradient elution procedure);Column temperature:30℃,flow rate:1.0ml·min^-1(Split ratio 4:1),injection volume:20μl.3.1.2 MS conditions:ESI positive ion detection mode(ESI⁺),Mass scan, daughter scan,source temperature:120℃,desolvation temperature:400℃,capillary voltage:3.00 kv,cone:15v,extRacor:3v,gas flow: desolvation:750L·h^-1,cone:50L·h^-1.3.2 Antipyretic analgesic category3.2.1 Chromatographic conditions:Mobile phase A:20mmol·L^-1 Ammonium acetate solution B:AcetonitRile(Methods with the same 3.2 Antipyretic analgesic category group A);Column temperature:30℃,flow rate:1.0ml·min^-1(Split ratio 4:1),injection volume:20μl.3.2.2 MS conditions:ESI-positive and negative ion detection mode(ESI⁺, ESI^-),Mass scan,daughter scan,source temperature:120℃,desolvation temperature:400℃,capillary voltage:3.00 kv,cone:15v,extRacor:3v, gas flow:desolvation:750L·h^-1 cone:50L·h^-1.3.3 Antitussive and antiasthmatic category3.3.1 Chromatographic conditions:Mobile phase A:20mmol·L^-1 Ammonium acetate solution(Containing 0.1%formic acid,V / V) B: AcetonitRile(Methods with the same 3.3 Antipyretic analgesic category group A);Column temperature:30℃,flow rate:1.0ml·min^-1(Split ratio 4:1),injection volume:20μl.3.3.2 MS conditions:electRospray ionization positive ion detection mode (ESI⁺),Mass scan,daughter scan,source temperature:120℃,desolvation temperature:400℃,capillary voltage:3.00 kv,cone:15v,extRacor:3v, gas flow:desolvation:750L·h^-1,cone:50L·h^-1.RESULTS50 kinds of market testing samples were tested with application of this technology,Seven kinds of samples were found to contain antipyretic analgesic and hormonal ingredients,and the above-mentioned 34 kinds of standard spectRa library reference substance information were established by the HPLC-MS-MS methods,And the stRucture of the positive samples were confirmed,Which found that a hormonal components was a miscarriage of justice by TLC and HPLC-DAD,and the other sample contained the composition of paracetamol were not detected,So that the test results is more accurate and reliable.CONCLUSIONThe detection technique is rapid,simple,reproducible and reliable results,can be applied to test screening review fastly for the drug testing vehicles and the grass-roots units,and the HPLC-MS-MS method can be used to confirmation of stRucture for the components illegally added.It has great practical significance for protecting the people's diet and drug safety.