Quality Control And Screening Of Illegal Additives In Fur Seal Ginseng Pills By Multi-dimensional HPLC Fingerprints

As an irreplaceable consumer product in daily life,health care products gradually attracted people’s attention.Currently,there are many brands of dietary supplements on the market,the diversity of illegal additives and the complexity of the sample matrix pose challenges for monitoring.Fur seal ginseng pills(FSGP)was approved by CFDA in 2004(No.G20040972),which has a therapeutic effect on sexual dysfunction,and it is favored by people.In this paper,FSGP was selected as the object of this study.Based on the research of the properties of active ingredients in the FSGP,combined with high-performance chromatographic separation and multi-dimensional ultraviolet spectrometry detection techniques,and the optimized sample preparation methods,the fingerprint characteristics of FSGP were studied.A multidimensional fingerprinting screening method for illegal additives in the FSGP was established combination with chemometrics method and the spectral characteristics of illegal additives.The main research content was as follows:1.HPLC analysis methods for the effective components in FSGP were established with reversed-phase chromatography and gradient elution.The analysis of 11 batches of samples and the fitting of standard fingerprints were used to construct the multidimensional fingerprints of FSGP which was used for quality evaluation.The detection results of some commercial samples showed that three of them unqualified products.2.Principal component analysis and systematic cluster analysis were performed on the chromatographic data using Simca-P and SPSS software respectively.According to the classification method,11 batches of samples could be roughly divided into three groups,the first group S1,S2,S3,S4,S8 and S9;the second group S5,S6,S7,S10;the third group S11.It was found that the chromatographic peaks with retention times of 74.8 min,76.6 min,and 111.3 min were recognized as the quality control components of the FSGP.3.Based on the fingerprint characteristics of FSGP,chromatographic behaviors of 16 potential illegal additives including PDE-5 inhibitors,androgen,alpha-receptor antagonists,and yohimbine were also studied using multi-dimensional HPLC fingerprinting technology.Combined with multiple wavelength detection to reduce or remove sample matrix interference,A new screening method for screening illegal additives in FSGP was established and it has been successfully applied to the screening of illegal additives in FSGP.One batch of samples suspected to contain testosterone in 5 batches of sampleand the content was 18.8 μg/g.The recovery rate and relative standard deviation are 83.6% and 6.82%,respectively.The primary and secondary fingerprints combined with the spectral purity and LC-MS verification greatly improve the accuracy and reliability of the detection.4.The dimethyl carbonate/water/ammonium sulfate multiphase extraction system,combined with microwave-assisted extraction,was used for the extraction.The conditions for the formation of the system and the extraction conditions were investigated.The results show that when the ratio of dimethyl carbonate to water was 1:7.5(V/V),salting-out agent was ammonium sulfate and added with 8 g,the extraction temperature was 70° C,the extraction time was 10 min and the ratio of liquid to material was 25,the extraction ratereached the maximum.Compared with single solvent extraction,salting phase-separation extraction significantly reduces the interference of the sample matrix,shortens the analysis time.Combined with multi-stage fingerprinting and spectrum and LC-MS verification,the sensitivity,accuracy,and reliability of screening for illegal additives can be further improved.