| Objective With the development of the economy and the improvement of people’s awareness of health care and beauty,dietary supplements and cosmetics have been closely related to people’s lives.However,with the driving of economic benefits,many unscrupulous manufacturers have added various illegally added chemical medicines in order to highlight the functions of dietary supplements such as anti-impotence,anti-fatigue,immune enhancement,and the functions of cosmetics such as hair growth,wrinkle-removing,body building,and breast beautifying and so on in recent years.These illegal additions have brought great harm to public health,medical development and social stability.At present,only PDE-5 inhibitors are included in the official detection methods of relieving physical fatigue dietary supplements issued by the State Food and drug administration,while The Safety and Technical Standards for Cosmetics(2015 edition)only includes seven sex hormones.With the continuous emergence of new addition methods such as structural analogues and multiple additions,the existing detection methods can no longer meet the actual regulatory needs,and it is necessary to establish a wider,more accurate and sensitive screening method.Methods In this study,an ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)method was developed and validated for screening,confirmation and quantitation of 31 anti-impotence medicines potentially illegally added to dietary supplements.The Chromatographic separation was performed on an Agilent Eclipse Plus C18 column(100 mm×2.1 mm,1.8μm)and separated by the mobile phase consisted of 0.1%formic acid solution and acetonitrile with gradient elution at a flow rate of 0.3 mL/min.The electrospray ionization ionization source(ESI)was used to scan positive ions.The 31 anti-impotence medicines were analyzed under Scan MS and Targeted MS/MS mode.Basing on the information of retention time of liquid chromatography and characteristic fragments of mass spectrometry,the screening database of 31 anti-impotence medicines was established.In this study,a rapid screening method based on UPLC-Q-TOF-MS for illegal addition of 40 sex hormones in cosmetics was established.The gradient elution was carried out on Agilent SB-C18 column(2.1 mm×50 mm,1.8μm),0.1%formic acid water and acetonitrile were used as mobile phase in positive ion mode,water and acetonitrile were used as mobile phase in negative ion mode,the flow rate was 0.2 mL/min,the column temperature was 35℃.The electrospray ionization ionization source(ESI)was used to scan positive and negative ions.The 40 sex hormones were analyzed under Scan MS and Targeted MS/MS mode.Based on the information of retention time of liquid chromatography and characteristic fragments of mass spectrometry,a screening database of 40 sex hormones was established。Results The 31 anti-impotence medicines potentially illegally added to dietary supplements were separated within 17.5 minutes,and the mass error was less than 2.9 ppm.The screening database of 31 anti-impotence medicines in dietary supplements was established.The method validation showed good linearity with coefficients of determination(r2)higher than 0.9973 for all analytes.LODs and LOQs were in the ranges of 0.005-0.50 μg/g or μg/mL and 0.02-1.24μg/g or μg/mL,respectively.The accuracy was in the range of 86.6%-113.7%,while the intra-and inter-day precision were in the ranges of 0.9%-7.6%and 0.9%-11.4%,respectively.The absolute and relative matrix effect were in the range of 65.8%-115.6%and 0.6%-13.3%.The mean recoveries were in the range of 80.5%-116.9%.The RSD for stability was in the range of 0.4%-8.5%within 24 hours.Among 200 batches of dietary supplements,sildenafil and/or tadalafil were found to be added illegally in two samples,while not very high concentration of icariin was detected in one sample.The 40 sex hormones illegally added in cosmetics were separated in in positive and negative ion modes in 13.5 min and 7.5 min,respectively.The retention time of liquid phase and characteristic fragment ions of 40 sex hormones were obtained and screening database of 40 sex hormones was established.100 batches of cosmetics were screened and 1 batch of samples was found to have been added progesterone illegally.Conclusion Based on UPLC-Q-TOF-MS,the screening database and analysis method of 31 anti-impotence medicines and 40 sexual hormones were fast,sensitive,accurate and high-throughput,which could not only resist the illegal addition of fatigue health food and cosmetics for extensive screening,but also could effectively infer the suspected addition,providing strong technical support for administrative supervision. |
