半枝蓮中抗腫瘤活性成份之萃取研究

? 65 ? ??? ??? ??? ??? ???????????????? ??? ??? ??? ??? ??????????? ? ? ????????????????????????naringenin?luteolin? apigenin?wogonin?baicalein?baicalin???protocatechuic acid????????? ????????????????????????????????????? ????????????????????????????????????? ????????????????? (HPLC)????????????????? ????????????40-80%????50%????????????50??? ?1???naringenin????????????????????protocatechuic acid ??????????????????117-154 bar???40-50????????? ?????????????????? ???????????????????????wogonin?baicalein?baicalin ? ? ????????????????????????????????????????? ???????????????????????????(1)?????70-92??????? ?????????????????????????????????????????80- 92?????????????????????????????????????????? ???? ????????????????????????? ??????????????? ??????????????????????????????????????????? ??????????????????????????????????????????? ? (2)??????????????????(Scutellria barbata Don)??????????? ???? (3-4) ??????????????D- ???? (D-galactosamine)??????? (Acetaminophen)???????????????????????????????????? ? ? ? ? ? ? ? ? ? 65?7 4 ???????? C H I A - N A N A N N U A L B U L L E T I N V O L . 3 0 , P P . 6 5? 7 4 , 2 0 0 4 ? 66 ? ???????????????? ????????????(5)?????????????????????????????? ??Chou(6)???????(7-12)?????????????????????Baicalein?Baicalin ?Wogonin?? ???????????????????????Kimuya ??(13)?Abe??(14)????? ???? (15-19) ?????Baicalein ?Baicalin ?Wogonin ???????????????? (apigenin)????? (luteolin)???????????????(20)??????????? (protocatechuic acid)????????????????(21)????????????????? ??????????????????????????? ????????????????????????????????????????? ??????????????????????????????????????????? ??????????????????????????????????????????? ??????????????????????????????????????????? ???????????????1976?????(22)???????????????????? ????????3.5%????????????49.8%????46.7%??????????? ?????????????????????????????????? ????????????????(23-27)???????????(28)?????(29)???? ????(23-26,28,30-31)?????????????????????????????????? ??????????????????????????????????????????? ??????????????????????????????????????????? ?(304.2 K)????????(1070.7 psi)????????????????????????? ??????????????????????????????????????????? ??????????????????????????? ??????????????????????????????????naringenin? luteolin?protocatechuic acid?wogonin?baicalein?baicalin?apigenin????????????? ??????????????????????????????????????????? ?? ???? ?????? ????????????????????????????105??????????? ????????????????????12.1%???????????????????? ?(dry basis)????? ?????? ????????????????????????????????????????? ??????????????????? 10???????????(HPLC)?????? ? 67 ? ??? ??? ??? ??? naringenin?luteolin?protocatechuic acid?wogonin?baicalein?baicalin?apigenin???????? ??????????????????????????????????????????? ????????? ??????????? ?????????????????????????????ISCO????????? ???????????????????????40Mpa????????????????? ???????(cartridge)??????????????????????????????? ??????????????????????????????????????????? HPLC????????????? ?????? ????Naringenin???????????Aldrich????????HPLC??????? ?????????????HPLC?JASCO??????????UV/Vis?????(photodiode) ?????????Inertsil 5C18 ODS-2??????????4.6 mm???250 mm?????? ??45????????1 mL/min???protocatechuic acid?????0.1% H3PO4??????? ?270 nm???naringenin?luteolin?baicalin?apigenin?????0.1% H3PO4-CH3CN (80:20, v/v) ????????280 nm???baicalein?????0.1% H3PO4-CH3CN (75:25, v/v)??????? ?280 nm???wogonin?????0.1% H3PO4-CH3CN (70:30, v/v)????????280 nm? ????? ??HPLC?????????? ??0.5 mg/mL?Naringenin????????????????????(intraday)????? ??????????(interday)????????HPLC???????????????wogonin ?baicalein?baicalin naringenin?luteolin?apigenin ?protocatechuic acid?????intraday??? ????(RSD)???3.41?2.92? 3.71? 2.18?3.06? 2.87?3.64??wogonin?baicalein?baicalin ?naringenin?luteolin?apigenin ?protocatechuic acid?????interday???????(RSD)?? ?3.83? 3.04?4.19? 2.73?3.53?3.05?3.97??????????(RSD)????4%????? ?????????????????? ?????? 1.??????????????????? ???50????1??????????????????????????????1?? 2?????protocatechuic acid??????????????40-80%???????????? ???????protocatechuic acid????????(benzoic acid)?????????????? ??????????????????????????????????????????? ??????????????????????????????????????????? ?????????????luteolin?apigenin?wogonin?????????????????? ? 68 ? ???????????????? ?baicalin?baicalein?naringenin???????????????????????40?80%?? ??????????????????????????????????????????? ???????????baicalein????????????3??? 0 20 40 60 80 100 Percentage of alcohol (%) 0 0.02 0.04 0.06 0.08 C on te nt s of c om po ne nt s ( m g/ g) naringenin protocatechuic acid baicalein wogonin 0 20 40 60 80 100 Percentage of alcohol (%) 0 0.3 0.6 0.9 1.2 1.5 1.8 C on te nt s of c om po ne nt s (m g/ g) apigenin luteolin baicalin ?1 ???????????????? naringenin??????????(50?, 1h) ?2 ???????????????? luteolin??????????(50?, 1h) ?3 baicalein?????????? ? 69 ? ??? ??? ??? ??? 2.??????????? ???30-80????60%???????1???????4??5?????????50-80? ???????naringenin?protocatechuic acid??????????apigenin?baicalein???? ????50?????????????????????????????????????? ?????????????????50????????? 20 30 40 50 60 70 80 90 Temperature (oC) 0 0.02 0.04 0.06 0.08 C on en ts o f c om po ne nt s ( m g/ g) naringenin protocatechuic acid baicalein wogonin 20 30 40 50 60 70 80 90 Temperature (oC) 0 0.4 0.8 1.2 1.6 C on en ts o f c om po ne nt s ( m g/ g) apigenin luteolin baicalin ?4 ?????????naringenin???? ??????(60%???1h) ?5 ?????????luteolin????? ?????(60%???1h) 3.??????????? ?50????60%????????????10?120????????????????6? ?7?????protocatechuic acid?????????70-90???baicalein?naringenin?wogonin? ???????60?????apigenin?????????????????????????? ????????10??????????? 0 30 60 90 120 150 Time (min) 0 0.02 0.04 0.06 0.08 C on en ts o f c om po ne nt s (m g/ g) naringenin protocatuic acid baicalein wogonin 0 30 60 90 120 150 Time (min) 0 0.4 0.8 1.2 1.6 C on en ts o f c om po ne nt s ( m g/ g) apigenin luteolin baicalin ?6 ?????????naringenin???? ??????(60%???50?) ?7 ?????????luteolin????? ????? (60%???50?) ? 70 ? ???????????????? 4.??????????? ?50??????????????????????8??11????????????? ????????????baicalein??????????8??9?????????????? ???baicalein?baicalin???????????????????????????????? ??????????????????????????????????????????? ???? 0 30 60 90 120 150 Time (min) 0 0.01 0.02 0.03 C on en ts o f c om po ne nt s ( m g/ g) narigenin protocatuic acid baicalein wognin 0 30 60 90 120 150 Time (min) 0 0.2 0.4 0.6 0.8 C on en ts o f c om po ne nt s ( m g/ g) apigenin luteolin baicalin ?8 ???????naringenin?????? ????(50?) ?9 ???????luteolin??????? ??? (50?) 0 30 60 90 120 150 Time (min) 0 0.02 0.04 0.06 0.08 C on en ts o f c om po ne nt s (m g/ g) naringenin protocatuic acid baicalein wogonin 0 30 60 90 120 150 Time (min) 0 0.3 0.6 0.9 1.2 1.5 1.8 C on en ts o f c om po ne nt s ( m g/ g) apigenin luteolin baicalin ?10 50%???????naringenin??? ???????(50?) ?11 50%???????luteolin????? ?????(50?) ?10??11???50%??????????????60%??????????????? ????????????????????8??9??? ? ? ?? ??????????? LuquedeCastro?Tena(32)?????????????????????????????? ??????????????????????????????????????????? ? 71 ? ??? ??? ??? ??? ?????????????????????????????????????? (cosolvent or entrainer)?????????????????????????????????????? ??????????????????????????????????????????? ??????????????????????????????????????????? ?????????????????????????????????117-154 bar???40- 50?????4??????????????????????????????????? CO2????????????????????????????????????????? ??????? ? ? ????????????????????????????????????????? ????????????????????????50????40-80%????50%????? ???????1???????????????????????117-154 bar???40-50?? ???????????????????????????protocatechuic acid? ? ? ??????????(NSC 91-2214-E-006-008)??????????? ???? 1. ???(http://www.doh.gov.tw/)??????? 2. Chiu, H. F., Lin, C. C., Yang, C. C. and Yang, F., “The Pharmacological and Pathological Studies on Several Hepatic Protecttive Crude Drugs from Taiwan(I)”, American Journal of Chinese Medicine, 16(3-4), 127-137, 1988. 3. Chiu, F. H., Lin, C. C., Yen, M. H., Wu, P. S. and Yang, C. Y., “The Pharmacological and Pathological Studies on Several Hepatic Protective Crude Drugs from Taiwan (V): The Effects of Bombax Malabrica and Scutellaria Rivularis”, Am. J. Chin. Med., 4, 257-264, 1992. 4. Lin, S. C., Lin, C. C., Lin, Y. H. and Chen, C. H., “Protective and Therapeutic Effects of Ban-Zhi-lian on Hepatotoxin-induced Liver Injuries”, Am. J. Chin. 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ABSTRACT Extraction of Anti-Tumor Activities from Scutellaria Barbata Don Ming-Chi Wei, Chin-Haw Han, Jwu-Maw Yang and Li-Yin Chen Department of Pharmacy, Chia Nan University of Pharmacy and Science, Tainan, Taiwan 71710, R.O.C. ABSTRACT This study extracted the hepatoprotective and antitumor activities, such as naringenin, luteolin, protocatechuic acid, wogonin, baicalein, baicalin and apigenin from Scutellaria Barbata Don using solvent and supercritical carbon dioxide extraction. With pure water, different percentage of alcohol aqueous solution, acetone and n-hexane as extractants, the effects of extraction factors, temperature and extraction time on extraction yield were studied. On the other hand, the effects of temperature and pressure on extraction yield were also discussed using supercritical fluid extraction. Analyses of the extracts were performed by high-performance liquid chromatography to determine the contents of components. For all the analytes of the experimental conditions providing the highest extraction yield inside the experimental domain considered were found. Solvent extraction results showed that the optimum conditions for six flavonids constituents, such as naringenin etc. were achieved by using 40-80% ethanol and 50% acetone at 50? for extraction 1 h. The content of protocatechuic acid in ethanol and acetone extracts was low, but water extracts showed higher content. On the other hand, using carbon dioxide alone in supercritical fluid extraction at pressures from117 to 154 bar and temperatures from 40 to 50? , the extraction yield of Scutellaria Barbata Don was not satisfactory. Key words: Scutellaria barbata don, Supercritical carbon dioxide extraction, Solvent extraction, Wogonin, Baicalein, Baicalin C H I A - N A N A N N U A L B U L L E T I N V O L . 3 0 , P P . 6 5 ? 7 4 , 2 0 0 4