Contents of Furanocoumarins in Grapefruit Juice and Health Foods

326 Vol. 49, No. 4 20 1 28 1, 2 1 1 1 1 Contents of Furanocoumarins in Grapefruit Juice and Health Foods Narue S6@6B6@> 1,,Mitsuo N6@6O6ID 2,Hiroko M6IHJBDID 1, Kayo H6<>CD 1,Keiko H>G6I6 1 and Hirofumi UH=>N6B6 1 1 Tama Branch Institute, Tokyo Metropolitan Institute of Public Health: 3 16 25 Shibasaki-cho, Tachikawa, Tokyo 190 0023, Japan; 2 Tokyo Metropolitan Institute of Public Health: 3 24 1 Hyakunin-cho, Shinjuku-ku, Tokyo 169 0073, Japan Interactions between grapefruit juice and medications have long been recognized. In recent years, several furanocoumarins (FCs) that inhibit P450 activity in intestinal microsomes have been isolated from grapefruit juice. FCs, i.e., bergamottin (BG), bergapten (BP), bergaptol (BT) and 6,7 -dihydroxybergamottin (DHB), in samples was extracted with acetonitrile, and separated on a Phenyl column using 0.1 phosphoric acid-acetonitrile gradient as a mobile phase, with monitoring at 311 nm. The recoveries of BG, BP, BT and DHB from lemon juice spiked at the level of 5.0 mg/g were 103 0.7, 99.5 0.2, 96.5 0.2 and 90.1 0.2, respectively. The quantification limits were 1.0 mg/g in samples. The contents of BG, BP and DHB in grapefruit juice (n 13), citrus fruit of 20 species and health food (n 16) were measured. The contents of BG were 0 16 mg/g, 0 16 mg/g and 0 5.6 mg/g, BT were 0 39 mg/g, 0 13 mg/g and 0 28 mg/g, DHB were 0 10 mg/g, 0 35 mg/g and 0 6.2 mg/g, respectively. BP was not detected. These results suggest that patients prescribed calcium antagonists or antiallergic agents should be cautions about their intake of FCs from grapefruit juice, citrus and health foods. (Received January 28, 2008) Key words: furanocoumarins; bergamottin; 6,7-6,7 -dihydroxybergamottin; grapefruit juice; health food; citrus fruit; drug interaction; HPLC 1) 5) (BG) 6,7 - (DHB) (FC) 1) 3) P450 3A4 (CYP3A4) 1 190 0023 3 16 25 2 169 0073 3 24 1 3 4 1) 3) 1 3 DHB 4 1 2 3 http://www.bayer.co.jp/hv/tenpu/data/al t.html http://med.astrazeneca.co.jp/product/pi/spl PIpdf http://med2.astellas.jp/med/jp/basic/details/prg-5/attached/tenpu-prg-5.pdf 4 Vol. 59, p. 287, 2005

August 2008 327 DHB FC 6) 13) BG (BP) (BT) FC FC 1. 17 11 19 1 13 16 20 25 2. FC BG EXTRASYNTHESE S. A. BP EXTRASYNTHESE S. A. BT EXTRASYNTHESE S.A. 10.0 mg 50 ml DHB Ultrafine Chemicals 4.0 mg 20 ml 1mL BG, BP, BT DHB 200 mg FC FC 5mL 20 ml HPLC 3. Agilent Technologies 1100 series 4. HPLC CAPCELL PAK SG-Phenyl 5 mm, 4.6 mm i.d. 150 mm A B 0.1 0 5 (A : B 50 : 50) 20 (80 : 20) 1.0 ml/min 40 311 nm 20 ml 5. 1) 6) 10 g 30 ml 10 50 ml 14,000 10 HPLC 2) 10 g HPLC 6. FC 0.2, 0.5, 1.0, 2.0, 5.0, 10.0, 20.0 50.0 mg/ml FC 20 ml HPLC 1. HPLC FC HPLC UV 6) FC UV Fig. 1 220 260 nm 2 3 311 nm 240 nm HPLC 311 nm 311 nm 6) BG DHB BT 3 HPLC 0.1 (4 : 6) 5 30 100 BP BP BP DHB 1 Fig. 1. UV spectra of standard BG, DHB, BT and BP

328 Vol. 49, No. 4 Table 1. Contents of furanocoumarins in grapefruit juice and health foods Sample Analytical result (mg/g) BG DHB BT Fig. 2. HPLC chromatograms of BG, DHB, BT and BP in sample extracts (A) Standard: 1.0 mg/ml, (B-1) lemon juice: control, (B-2) lemon juice: furanocoumarins added (5.0 mg/g) Operating conditions: CAPCELL PAK SG-Phenyl (5 mm, 4.6 mm i.d. 150 mm) column, mobile phase (a) acetonitrile (b) 0.1 phosphoric acid, gradient elution a/b 50 : 50 (0 5 min) a/b 80 :20 (20 min), column temperature 40, flow rate 1.0 ml/min, detection wavelength UV 311 nm, injection volume 20 ml 0.1 (5 : 5) BG, BP, BT DHB 4 Fig. 2 HPLC 0.2 50 mg/ml (r 0.9999) 1.0 mg/g 2. BT, DHB, BP BG 5.0 mg/g n 5 BT 96.5 0.2, DHB 90.1 0.2, BP 99.5 0.2, BG103 0.7 n 5 BT 98.7 8.1, DHB103 5.6, BP99.5 4.3, BG98.2 4.3 90 3. FC 13 16 FC Table 1 FC BG 7.7 mg/g DHB 3.7 mg/ g BT 8.8 mg/g BP 100 13 8 (a h) BG 0 7.2 mg/g DHB 0 11 mg/g BT 0 12 mg/g 200 (i) BG 16 mg/g BG Grapefruit Juice a reconstituted fruit juice 5.1 3.4 4.9 b reconstituted fruit juice ND 3.7 2.5 c reconstituted fruit juice 6.6 10 3.0 d reconstituted fruit juice 5.1 5.1 ND e reconstituted fruit juice 5.1 5.5 2.3 f reconstituted fruit juice 7.2 ND 6.8 g reconstituted fruit juice 6.2 11 6.8 h reconstituted fruit juice 4.5 ND 12 i reconstituted fruit juice (200 ) 16 ND 26 j 30 fruit juice beverage ND ND 3.1 k mix fruit juice 4.8 ND 5.5 l straight fruit juice 32 2.0 39 m straight fruit juice 7.4 7.9 2.9 Health Food 1 jelly drink ND ND ND 2 jelly drink ND ND ND 3 jelly drink ND ND 2.6 4 jelly drink ND ND ND 5 tablet 3.1 6.2 1.7 6 tablet ND 2.9 3.4 7 capsule ND ND ND 8 granule 5.6 ND 28 9 granule ND ND ND 10 granule ND ND ND 11 drink ND ND ND 12 drink ND ND ND 13 drink ND ND ND 14 candy ND 3.5 ND 15 jelly candy ND ND ND 16 biscuit ND ND ND ND: not detected : BPwas not detected. DHB 30 (j) BG DHB BT 3.1 mg/g 1 (k) BG 4.8 g/g, BT 5.5 mg/g DHB 2 (m) BG 7.4 mg/g, DHB 7.9 mg/g, BT 2.9mg/g (l) BG 32 g/g, DHB 2.0 mg/g, BT 39 mg/g BG 2 BT 1.5 10), 14) FC FC 6) 7 BG DHB 100

August 2008 329 i k l 9 b BG f h 2 DHB 6 (a, c, d, e, g, m) BG DHB 1 200 ml FC Table 2 FC BG 0 6.4 mg, DHB 0 2.2 mg, BT 0 7.8 mg 200 250 ml 70 225 4) 100 a h l m 10 200 ml FC mg 10 mg 16 11 2 BG 5.6 mg/g 3 DHB 6.2 mg/g 4 BT 28 mg/g BP FC 7 1 FC Table 2 200 ml 1/20 FC FC 4. FC 20 Table 3 DHB BG FC De Castro 10) Fukuda Table 2. Daily intakes of BG, DHB and BT in grapefruit juice and health foods Sample Furanocoumarins (mg/day) BG DHB BT Grapefruit juice a reconstituted fruit juice 1.0 0.68 0.98 b reconstituted fruit juice ND 0.74 0.50 c reconstituted fruit juice 1.3 2.0 0.60 d reconstituted fruit juice 1.0 1.0 ND e reconstituted fruit juice 1.0 1.1 0.46 f reconstituted fruit juice 1.4 ND 1.4 g reconstituted fruit juice 1.2 2.2 1.4 h reconstituted fruit juice 0.90 ND 2.4 i reconstituted fruit juice (200 ) 3.2 ND 5.2 j 30 fruit juice beverage ND ND 0.62 k mix fruit juice 0.96 ND 1.1 l straight fruit juice 6.4 0.40 7.8 m straight fruit juice 1.5 1.6 0.58 Health food (daily intake) 1 jelly drink (180 g) ND ND ND 2 jelly drink (180 g) ND ND ND 3 jelly drink (130 g) ND ND 0.34 4 jelly drink (150 g) ND ND ND 5 tablet (4 g) 0.012 0.025 0.0068 6 tablet (38 g) ND 0.11 0.13 7 capsule (2.1 g) ND ND ND 8 granule (4 g) 0.022 ND 0.11 9 granule (8 g) ND ND ND 10 granule (4 g) ND ND ND 11 drink (120 g) ND ND ND 12 drink, powder (12 g) ND ND ND 13 drink (150 g) ND ND ND 14 candy (30 g) ND 0.11 ND 15 jelly candy (31 g) ND ND ND 16 biscuit (80 g) ND ND ND ND: not detected : To estimate the daily intakes, it was assumed that a person takes 200 ml of grapefruit juice in a day. : Toestimate the daily intakes, it was assumed that a person takes one package or the recommended serving size per day as shown on a package. 14) FC FC Saita 15) 15 red pummelo 8 Saita 15) FC FC 200 ml 200 g FC

330 Vol. 49, No. 4 Table 3. Quantitative comparison of furanocoumarins in grapefruit and other citrus fruits No. of samples Analytical result (mg/g) BG DHB BT Grapefruit (white) 2 6.3, 7.7 3.8, 7.9 ND Grapefruit (ruby) 2 4.1, 4.9 ND ND Grapefruit (white), Canned in light syrup 1 11 ND 13 Oroblanco 2 3.7, 4.7 3.2, 4.9 ND Melogold 1 16 35 ND Banpeiyu pummelo 1 3.5 ND ND Akunebuntan pummelo 1 ND ND ND Suishobuntan pummelo 1 ND ND ND Tosabuntan pummelo 1 ND ND ND Iyokan 1 ND ND ND Satsuma mandarins 1 ND ND ND Kumquats 1 ND ND ND Setoka 1 ND ND ND Sweet spring 1 ND ND ND Dekopon tangor 2 ND ND ND Natsumikan 1 ND ND ND Hassaku 1 ND ND ND Pearl-kan 1 ND ND ND Hyuganatsu 1 ND ND ND Ponkan 1 ND ND ND Lemons 1 ND ND ND ND: not detected 200 g 1 1 200 g 400 g FC 13 16 FC 14,000 HPLC FC 311 nm FC BG 0 32 mg/g DHB 0 11 mg/g BT 0 39 g/g 100 200 ml mg 10 mg FC BG 0 5.6 mg/g, DHB 0 6.2 mg/g, BT 0 28 mg/g 11 0 0.34 mg FC FC 20 FC FC 92 2006 11 1) 2005. (ISBN 4-7532-2136-9) 2) 109, 781 788, (2006). 3) Yamazoe, Y. Properties of grapefruit juice-drug interaction. Rinsho-Yakuri No Shimpo (Recent Advances in Clinical Pharmacology), 21, 68 77 (2000). 4) 3 1999. (ISBN 4-8407-2482-2) 5) Boullata, J. I., Armenti, V. T. Handbook of drugnutrient interactions 2005. (ISBN 4-621-07548-9)

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