LC-APCI+-MS/MS method for the analysis of ten hormones and two endocannabinoids in plasma and hair from the mice with different gut microbiota

• Published in: Journal of pharmaceutical and biomedical analysis 2020-06, Vol.185, p.113223-113223, Article 113223
• Main Authors: Chu, Liuxi, Li, Na, Deng, Jia, Wu, Yan, Yang, Haoran, Wang, Wei, Zhou, Dongrui, Deng, Huihua
• Format: Article
• Language: English
• Subjects: Animals; Biomarkers - analysis; Biomarkers - metabolism; Chromatography, High Pressure Liquid - methods; Corticosterone - analysis; Corticosterone - metabolism; Endocannabinoids; Endocannabinoids - analysis; Endocannabinoids - metabolism; Endogenous hormones; Female; Gastrointestinal Microbiome - physiology; Gonadal Steroid Hormones - analysis; Gonadal Steroid Hormones - metabolism; Gut microbiota; Hair; Hair - chemistry; LC-APCI+-MS/MS; Male; Mice; Plasma; Reproducibility of Results; Sex Factors; Solid Phase Extraction; Specific Pathogen-Free Organisms; Tandem Mass Spectrometry - methods
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2020.113223

•LC-APCI-MS/MS was developed for quantitation of 12 compounds in rodent’s plasma and hair.•The mice having more-diversity gut microbiota had higher hair levels of CORT, 11-DHC and DHEA.•Gut microbiota possibly influence long-term activity of the HPA and HPG axes and ECS.•The mice with less-diversity gut microbiota show sex-specific pattern in plasma hormones. The effect of gut microbiota on the activity of the HPA and HPG axes and ECS is a short-term or long-lasting process remains unclear in rodents. However, the extant studies focused only on its short-term effect on the HPA activity because there is lack of reliable biomarkers characterizing short-term activity of the HPG axis and ECS and long-term activities of the three endocrine systems. The endogenous levels of aldosterone (ALD), 11-dehydrocorticosterone (11-DHC), estradiol (E2), estrone (E1), androstenedione (A4), dihydrotestosterone (DHT), corticosterone (CORT), dehydroepiandrosterone (DHEA), testosterone (T), progesterone (P), N-arachidonoyl ethanoamide (AEA) and 1-arachydonoyl glycerol (1-AG) in hair and plasma are the potential long-term and short-term biomarkers of the three systems. This study aimed to develop the sensitive and selective methods for simultaneous quantitation of the twelve compounds in rodent’s hair and plasma. Then the methods were used to explore the differences in the hair levels of the twelve compounds between the mice in XZ group possibly having gut microbiota with more diversity and SPF group possibly having gut microbiota with less diversity and the inter-group differences in the plasma levels in the response to 1-h restraint stress. The methods were based on high performance liquid chromatography tandem mass spectrometry with atmospheric pressure chemical ionization in positive mode. The methods were adopted for 20 mg hair and 100 μL plasma, respectively. Hair samples were incubated in methanol at 40 ℃ for 24 h, and were performed by solid phase extraction. Plasma samples were implemented by liquid-liquid extraction with ethyl acetate. The methods showed limit of quantification at 0.06–1.3 pg/mg and 0.03-0.6 ng/mL and recovery ranging between 87.7–115.1 % and 86.1–114.6 % for all compounds in rodent’s hair and plasma, and the intra-day and inter-day coefficients of variation less than 15 %, and good freeze/thaw and short-term stability. The present methods also had good reliability as demonstrated by the sex difference in the testosterone levels in hair and plasma. The