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2 protocols using carbamazepine

1

Composition and Characterization of TAZF

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Carbamazepine (CAS No.: 298–46-4), berberine (CAS No.: 2086-83-1), coptisine (CAS No.: 6020-18-4), epiberberine (CAS No.: 6873-09-2), palmatine (CAS No.: 3486-67-7), jatrorrhizine (CAS No.: 3621-38-3), columbamine (CAS No.: 483–34-1), evodiamine (CAS No.: 518–17-2) and rutaecarpine (CAS No.: 20575–76-2), all > 98% purity, were purchased from Shanghai Yuanye Bio-Technology Company. Acetonitrile and methanol (LC grade) were purchased from Fisher Chemical, USA. Formaldehyde (AR) was purchased from Beijing Tong Guang Fine Chemicals Company. Isoflurane was provided by HARVEYBIO Company.
TAZF was provided by the State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University. The concentrations of the compounds in TAZF were measured using high-performance liquid chromatography (HPLC), with berberine accounting for the majority of TAZF at a ratio of 14.73% (g/g), followed by palmatine (3.14%), coptisine (2.52%), evodiamine (1.41%), epiberberine (1.30%), rutaecarpine (1.27%), columbamine (0.90%) and jatrorrhizine (0.47%).
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2

Quantitative Analysis of Peimine and Peiminine

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Peimine and peiminine were purchased from the National Institutes for Food and Drug Control (Beijing, China). Carbamazepine (internal standard; IS) was obtained from Shanghai Yuan Ye Bio-Technology Co., Ltd. (Shanghai, China). These reference substances had a purity of >98%. The raw material peimine (purity ≥ 90%) was obtained from Xi'an Hui Lin Bio-Technology Co., Ltd. (Shaanxi, China). The raw material peiminine (purity ≥ 90%) was provided by Chengdu Master Biotechnology Co., Ltd. (Chengdu, China). Methanol and acetonitrile were provided by the Tedia Company (USA). Ammonium formate was obtained from Fisher (USA). All solutions were mass spectrum grade. Fetal bovine serum, trypsin, methyl thiazole tetrazolium (MTT), and Roswell Park Memorial Institute (RPMI) medium 1640 were purchased from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). TNF-α was purchased from Peprotech (USA). The ultrapure water was produced by the Millipore Mill-Q system (Bedford, MA, USA). Human IL-8, MMP-9, and TIMP-1 ELISA kits were purchased from Baxter Biological Co., Ltd. The A549 cell line was obtained from Shanghai Institutes for Biological Sciences (Shanghai, China).
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