Ultra high performance liquid chromatography

To detect the pharmacokinetic parameters of MG and CA in vivo, quantification analysis was performed by an ultra-high-performance liquid chromatography (Shimadzu Corp., Japan) tandem ion trap quadrupole QTRAP 6500 plus mass spectrometry (AB Sciex, USA). The quantitative analyses for MG and CA were approved by the Guidance for Bioanalytical Method Validation issued by the Chinese Pharmacopoeia Commission in 2020. The pharmacokinetic parameters were calculated by Drug and Statistic software (Shanghai, China). Detailed information on the collection of blood samples and UHPLC-QTRAP-MS/MS conditions is provided in Supplementary Materials Section 13.

PAHs standards of benzo[b]fluoranthene (99.4%), ben-zo[a]anthracene (99.0%), fluorene, anthracene (99.9%), and pyrene (98.5%), as shown in Table 1, were acquired from Manhage Bio-technology (Beijing, China). Analytical grade sodium hydroxide (NaOH), Iron (Ⅱ) chloride tetrahydrate, and Iron (III) chloride hexahydrate were obtained from Macklin. Tetraethyl orthosilicate (TEOS), isopropyl alcohol, ammonia, anhydrous toluene, triethylamine, chloro(diethyl)octadecylsilane (ODS, 95% v/v), hydrochloric acid (HCl, 37% v/v), and potassium bromide (KBr) came from Sigma Macau. Methanol (CH₄O, polarity:6.6), 95% ethanol (C₂H₆O, polarity:4.3), acetonitrile (CH₃CN, polarity:6.2), and acetic acid (CH₃COOH, polarity:6.2) came from Macklin (Shanghai, China). Acidified acetonitrile (3% acetic acid, v/v) was prepared personally. Ultra-high performance liquid chromatography was from Shimadzu. HPLC-grade acetonitrile and methanol were provided by Anaqua (Cleveland, OH, USA).