Purification Methods For Serum Anticancer Polypeptide Drugs Triptorelin Using Solid Phase Extraction

Background:Cancers having various causes.However, all of them are controlledbythe expression and regulation of cancer gene and tumor-suppressor gene. With theimplementation of human genome project, the research of protein and peptide is moremature than before.In recent years, biological peptide is widely developed as newmedicines.Findingnew targets that have strong binding ability to biological peptides is ahot research area in the industry of anti-cancer drug.Solid phase extraction (SPE)was developed for the pretreatment of complexsamples from1970s.In recent years, SPE is widely used in biological research due to itshigh efficiency, reliable and less solvent cost.To guide the use of medicine,studyingdrug metabolism and pharmacokinetics is necessary.The concentration ofmedicines in blood andurine is too low to detect without effective pretreatment methods.Moreover, both blood and urine is complex systems, medicines in these matrixes aredifficult to be detected, even with mass spectroscopy used. SPE, an effectivepretreatment method, is widely used to simplify samples and concentrate analytes.Triptorelin is a kind of synthetic gonadotropins (gonadotropin-releasing hormone,GnRH) decapeptide analogue and its molecular mass is1311.5. In clinic, it is used forthe treatment of uterine fibroids, breast, endometrial cancer and prostate cancer. amedicine peptide, the establishment of the detective methods for triptorelinissignificanceand useful for other follow-up peptide drugs test.Propose: The hyphenated technique pretein precipitation with SPE was utilized asthe pretreatment method for serum. Reverse mode, hydrophilic mode and ion exchangemode were used in the development of SPE method. The hyphenated technique highperformance liquid chromatrography with Q-TOF was used to analyzesamples.Different original samples were used in this experiment and the results indicated that current method was useful for peptides detection from various kinds ofsamples.Methods:Serum from60patients were collected and used in thisexperiment.Precipitationwith acetonitrile andseveral of SPE method were developed forthese samples. Then,samples were separated and detected by nano-UPLC-MS. Thesoftware Markerlynxwas used for the analysis of data.Results:Three kinds of SPE mode were used forthe concentration oftriptorelinfrom various of samples. The result indicated that hydrophilic SPE method isthe most effective pretreatment method for triptorelin.Conclusions:Ansuccessful method was established forthe detection of triptorelin fromserum.1. An hyphenated technique method was establishedto detect peptides fromcomplex samples including serum, urine, saliva.2. Compared to previous study, moreeffective pretreatment method was developed for triptorelin.3. This method ismeaningful for the detection of other anti-cancer drugs and can be used for the study ofdrug metabolism and pharmacokinetics.