Berberine as a chemical and pharmacokinetic marker of the butanol-extracted Food Allergy Herbal Formula-2 Nan Yang a , Kamal Srivastava a , Ying Song a , Changda Liu a , Sool Cho b , Yujuan Chen a,c , Xiu-Min Li a, ⁎ a Icahn School of Medicine at Mount Sinai, New York, NY 10029, United States b Hematology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, United States c Life Science and Technology, Changchun University of Science and Technology, Changchun 130022, China abstract article info Article history: Received 2 November 2016 Received in revised form 2 January 2017 Accepted 8 January 2017 Available online xxxx Rationale: Food Allergy Herbal Formula 2 (FAHF-2) provided protection against peanut anaphylaxis in a murine model and induced beneficial immune-modulation in humans. Butanol-refined FAHF-2, B-FAHF-2, retained safe- ty and efficacy in the peanut allergic murine model at only 1/5 of FAHF-2 dosage. One compound, berberine, was isolated and identified in vitro as a bioactive component present in FAHF-2 and B-FAHF-2. The aim of this study was to investigate berberine as a chemical and pharmacokinetic marker of B-FAHF-2. Methods: The consistency of constituents between B-FAHF-2 and FAHF-2 was tested. Peanut allergic C3H/HeJ mice were orally administered with 1 mg of berberine or B-FAHF-2 containing anequivalent amount of berberine, and the ability to protect against peanut anaphylaxis and pharmacokinetic parameters were determined. Human intestinal epithelial cells (Caco-2) were cultured with berberine with or without the nine individual herbal con- stituents in B-FAHF-2, and the absorbed berberine levels were determined. Results: Berberine is one of the major components in B-FAHF-2 and FAHF-2 formula. In a peanut allergic mouse model, B-FAHF-2, but not berberine, protected mice from anaphylaxis reactions. Pharmacokinetic profiles showed that the C max of B-FAHF-2 fed mice was 289.30 ± 185.40 ng/mL; whereas berberine alone showed very low bioavailability with C max value of 35.13 ± 47.90 ng/mL. Caco-2 cells influx assay showed that 7 of 9 herbal constituents in B-FAHF-2 increased berberine absorption at rates ranging from 18 to 205%. Conclusions: B-FAHF-2 remarkably increased the bioavailability of berberine. Berberine can be used as chemical and pharmacokinetic marker of B-FAHF-2. Other herbal components in B-FAHF-2 may facilitate the absorption of berberine. © 2017 Published by Elsevier B.V. Keywords: Butanol-extracted Food Allergy Herbal Formula-2 Food allergy Berberine Pharmacokinetics Caco-2 cells 1. Introduction Food allergy has been a growing and serious public health concern in the US [1]. No cure for food allergies has been developed, and no effec- tive prophylaxis exists other than specific food avoidance. Parenteral administration of epinephrine is the only approved rescue therapy for an anaphylactic event [2]. The combination of avoidance and emergency treatment substantially impairs the quality of life in food allergic pa- tients and their families. Consequently, research into developing food allergy treatment is a priority. More researchers have begun to investi- gate traditional Chinese medicine and use it as a source of alternative or complementary therapy for food allergies because of its demonstrated safety, low cost, and reputed effectiveness [3,4]. An herbal formula consisting of nine individual herbal extracts, called Food Allergy Herbal Formula 2 (FAHF-2), completely blocked peanut-induced anaphylaxis in a peanut allergic mouse model [3]. Also, FAHF-2 treatment produced at least 6 months protection against peanut specific anaphylaxis after discontinuing the therapy [5]. In an acute phase I study, FAHF-2 showed an immunomodulatory effect fol- lowing 1 week treatment [6]. However, the large amount of FAHF-2 tab- lets required for daily usage (30 tablets per day) made the medication adherence very difficult. In a double-blind, 6 months, phase II clinical trial of FAHF-2, 44% of the subjects had b 80% adherence for 2 months during the treatment period, and 30% did not adhere in the last 2 months of treatment [7]. This poor adherence rate severely affected the evalua- tion of the clinical trial. In order to reduce the dosage burden for pa- tients, a butanol-refined FAHF-2 product, B-FAHF-2, was generated. It showed the same effective therapeutic protection against peanut ana- phylaxis as FAHF-2 treatment at only 1/5 of the FAHF-2 dosage [8]. In vitro experiments showed that B-FAHF-2 was 9 times more potent than FAHF-2 on the inhibition of IgE production [9]. Toxicity experi- ments in mice showed that refined B-FAHF-2 formula was also as safe International Immunopharmacology 45 (2017) 120–127 ⁎ Corresponding author at: Pediatric Allergy and Immunology, Icahn School of Medicine at Mount Sinai, 1425 Madison Avenue, Box 1089, New York, NY 10029-6574, United States. E-mail address: xiu-min.li@mssm.edu (X.-M. Li). http://dx.doi.org/10.1016/j.intimp.2017.01.009 1567-5769/© 2017 Published by Elsevier B.V. Contents lists available at ScienceDirect International Immunopharmacology journal homepage: www.elsevier.com/locate/intimp