A Quality Assessment of Forsythiae Fructus in the Taiwanese Market

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1 J Chin Med 26(2): , 11 pages, 2015 DOI: / E-ISSN: Journal of Chinese Medicine A Quality Assessment of Forsythiae Fructus in the Taiwanese Market Lie-Chwen Lin 1, *, Chao-Lin Kuo 2, Pei-Yi Tseng 1, Yao-Haur Kuo 1 1 National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taiwan 2 Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung, Taiwan Forsythiae fructus, the dry fruits of Forsythia suspense (Thunb.) Vahl. (Oleaceae), is a common traditional Chinese medicine that is used as an antipyretic and an anti-inflammatory agent in the treatment of bacterial infection. A series of analyses, including differentiation of the appearance characteristics, histological examination, water extract, dilute alcohol extract, thin layer chromatography, HPLC fingerprint and quantitation, are conducted to assess the quality of forsythiae fructus. A total of 69 samples were collected from Chinese hospitals, Chinese medical clinics and Chinese herbal stores in northern, central and southern Taiwan. The appearance characteristics clearly differentiate 6 samples of green forsythiae fructus (GF) and 63 samples of ripe forsythiae fructus (RF) in this collection. GF and RF have a similar organ structure as shown by the photomicrography examination but the former s soft cell is thicker and its smell is stronger. GF contains more water extract, dilute alcohol extract and key ingredients (forsythoside A, phillyrin, pinoresinol and arctigenin) than RF, but GF and RF have similar TLC and HPLC profiles. The results showed that samples from central Taiwan had higher amount of water extract and dilute alcohol extract than those from northern and southern Taiwan. For three workplaces in which the samples were collected, the samples from Chinese medical clinics showed lower amount of water extract and dilute alcohol extract than the samples from the other two workplaces. In conclusion, 69 samples are all normative for each item content, but GF and RF exhibit large differences in their amount of water extract and dilute alcohol extract and in their chemical content. Therefore, the classification of forsythia fructus into two grades, GF and RF, allows their safer clinical use. Key words: Quality assessment, Forsythiae fructus, green forsythiae fructus, Ripe forsythiae fructus, water extract, dilute alcohol extract, forsythoside A, phillyrin Received 24 Febuary 2015 Accepted 8 April 2015 Available online 1 December 2015 *Correspondence: Lie-Chwen Lin, National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taiwan, No , Li-Nong St., Sec. 2, Taipei 112, Taiwan, Tel: ext. 7101, lclin@nricm.edu.tw 1

2 Lin et al. / J Chin Med 26(2): , 11 pages, Introduction Chinese Medicine Yin-Pian (CMYP) is a Chinese herbal medicine in a ready-to-use form, which is made by a special process. CMYP is used clinically to treat or prevent diseases. The quality of CMYP directly impacts the therapeutic effects. Traditionally, the quality of CMYP is graded according to herbal shape, color, size, surface, cross-section, texture and smell. However, this type of grading is evaluated by human experience and not by modern scientific technology, which could provide a more effective and objective way to assess the quality of herbs. The differentiation of appearance characteristics, histological examination, water extract assessment, dilute alcohol extract assessment, thin layer chromatography examination, HPLC fingerprint analysis and HPLC quantitative analysis are the common methods used to determine the herbal quality. In Taiwan, more than 85% of Chinese herbal medicines are imported from mainland China and their qualities are seldom assessed. This study undertakes a qualitative and quantitative analysis for CMPY to assess the quality. Fructus forsythiae is selected for this study. Forsythiae fructus is the dry fruits of Forsythia suspense (Thunb.) Vahl. (Oleaceae) [1]. It is a common traditional Chinese medicine and is widely used as an antipyretic and an anti-inflammatory agent in the treatment of bacterial infections [2]. According to the Taiwanese herbal pharmacopoeia, there are two types of forsythiae fructus. One is the green forsythiae fructus (GF), which is collected when the fruits are still green and barely ripe, and the other is the ripe forsythiae fructus (RF), which is harvested when the fruits are fully ripe and yellow. This study collected forsythiae fructus samples from Chinese hospitals, Chinese medical clinics and herbal drug stores in northern, central and southern Taiwan for the evaluation of quality. The comprehensive examination to determine the quality of forsythiae fructus will support the use of Chinese medicine. 2. Materials and Methods 2.1. Sample collection Forsythiae fructus was purchased from Chinese hospitals, Chinese medical clinics and herbal drug stores in northern, central and southern Taiwan. Sixty-nine samples were collected and each weighed about 300 g Chemicals and reagents Forsythoside A standard, phillyrin standard and a reference herb, forsythiae fructus, were purchased from the National Institute for Food and Drug Control, China. Pinoresinol-4-β-Oglucopyranoside, pinoresinol and arctigenin standards were isolated in the laboratory. GR grade solvents were purchased from Duksan Pure Chemicals (Korea). Triply deionized water (Millipore, Bedford, MA, USA) was used for all preparations. Liquid chromatographic grade solvents were purchased from Merck (Darmstadt, Germany) Microscopic examination of forsythiae fructus In this study, transverse and longituinal sections of the pericarp of forsythiae fructus were studied using photomicrography. Staining reagents (phloroglucinol-hcl and Sudan III solution) and a fixed solution (glycerin water solution) were used with standard procedures [3,4]. The various identifying characters were studied, with or without staining, and recorded. A microscope and camera (Nikon photograph T-2, Japan) were used for the study of different characteristics Pretreatment of forsythiae fructus An aliquot of 50 g of forsythia samples was ground and passed through an 80-mesh sieve. The powder was collected for the following experiments Preparation of the sample solution for TLC analysis One gram of forsythia powder was extracted with 10 ml of MeOH under ultrasound for 30 minutes, followed by centrifugation (20 minutes). The dried supernatant was dissolved in 1 ml of MeOH as the test solution for TLC examination. The reference forsythia herbal solution was produced with the same process as described previously. Phillyrin standard was dissolved in MeOH to a concentration of 1.0 mg/ml. An aliquot 5 µl of test solution, a reference forsythia herbal solution, and a phillyrin standard solution were spotted on a TLC plate (Kieselgel 60 F254 silica gel, glass plate, 0.25 mm; Merck, Darmstadt, Germany) and developed with a solvent system of CH 2 Cl 2 : MeOH (6:1). The developed TLC plate was examined under UV at 254 nm and visualized by spraying with 10% H 2 SO 4 -EtOH solution, followed by heating Preparation of water and dilute alcohol 2

3 A Quality Assessment of Forsythiae Fructus in the Taiwanese Market extracts An aliquot of forsythia powder (2 g) was placed in a 125 ml flask and 70 ml of water or 50% ethanol was added. The mixture was shaken for 8 hours and allowed to stand for another 16 hours. The residues were filtered and washed with an additional 30 ml of water or 50% alcohol. The filtrate was then transferred to a volumetric flask, which was then filled with water or 50% alcohol to 100 ml. Fifty milliliter of the solution was placed in an evaporating dish and heated on a water bath to dryness. The extraction process was repeated three times for each sample. The percentage (%) of each extract was calculated by using the equation: percentage of extract (%) = [(weight of extract) 2 / 2 g] HPLC The analyses were performed on an Agilent 1100 HPLC system (Agilent, Waldbronn, Germany), which consisted of a chromatographic pump, an autosampler and a photodiode array detector. An Agilent HC-C18 column ( mm; 5 µm, Agilent, USA) was used to analyze the forsythia samples. The mobile phase consisted of acetonitrile (ACN) and water containing 0.4 % acetic acid. A gradient program was used as follows: a linear gradient from 10% to 25% ACN in the first 22 min; the next gradient from 25% to 65% ACN for an additional 20 min. The flow rate was 0.8 ml/min. The DAD detector recorded UV spectra in the range from 200 to 400 nm and the HPLC chromatogram was monitored at 277 nm. The output data from the detector were integrated using the Agilent ChemStations Calibration standards and method validation A stock solution of 0.2 and 2.0 mg/ml of forsythoside A and 0.2 mg/ml of phillyrin were prepared in 50% MeOH and stored at -4 C. Working solutions of 800, 500, 250, 100 and 50 µg/ml of forsythoside A and 100, 80, 50, 10 and 5 µg/ml of phillyrin were prepared from the stock solutions of forsythoside A and phillyrin. An aliquot of working solution (900 µl) was spiked with 100 µl of internal standard solution (0.1 mg/ml of luteolin). Each of the five working solutions (10 µl) was injected into the HPLC for analysis. The peak area ratios for each standard to the internal standard, monitored at 277 nm, were plotted against their individual concentrations to determine the calibration curves. The intra-assay and inter-assay variability were determined by quantitating five replicates on the same day and five successive days, respectively, in order to verify the precision and accuracy of the analytical method. The precision (% R.S.D.) was calculated using the standard deviation and the observed concentration (C obs ) as follows: (% R.S.D.) = [standard deviation (S.D.)/C obs ] 100. The accuracy (% bias) was calculated using the nominal concentrations (C nom ) and the mean value of the observed concentrations (C obs ) as follows: (% bias) = [(C obs C nom )/C nom ] Preparation of the sample solution for HPLC analysis The sample powder (0.25 g) was extracted with 50% MeOH under ultrasound for 30 minutes, followed by centrifugation of the extract for 20 minutes. This process was repeated one more time. Two extracts were transferred to a volumetric flask, which was then filled with 50% MeOH to 25 ml to produce the sample solution. An aliquot of the sample solution (900 µl) was spiked with 100 µl of the internal standard solution (0.1 mg/ml of luteolin). After filtration (Nylon syringe filter, 0.45 µm, Sterlitech, WA, USA), 10 µl of the sample solution was injected for HPLC analysis. 3. Results and Discussion A total of 69 forsythiae fructus samples were collected from the Taiwanese market. Depending on their appearance characteristics (Fig. 1), these samples were easily separated into two subgroups: 6 green forsythiae fructus (GF, including samples NA3, NA4, MB2, MC18, MC22 and SA2) and 63 samples of ripe forsythiae fructus (RF). GF has a long ellipsoid capsule that is green-brown in color and smells strongly. The capsule of RF has a yellow-brown or reddishbrown surface and a light brown inner and is less aromatic than GF. The transverse section of a pericarp of forsythiae fructus is shown in Fig. 2. The exocarp consists of one row of epidermis cells, which have thickened outer and lateral walls that are covered with cuticles. Mesocarp is composed of vascular bundles that are scattered in parenchyma on the outside and many layers of stone cells on the inner side. The stone cells are elongated, sub-rounded or oblong and have walls that vary in thickness. They are mostly tangentially arranged in parquet form. The endocarp consists of a layer of parenchymatous 3

4 Lin et al. / J Chin Med 26(2): , 11 pages, 2015 cells [1,5]. A comparison of the internal structures of GF and RF shows that they are similar, but the former soft cell is thicker. Fig. 1. Herbal picture of forsythiae fructus. (A) green forsythiae fructus (GF) and (B) ripe forsythiae fructus (RF). In the TLC examination, the reference herb, forsythiae fructus, and standard phillyrin were used as references. The TLC developing condition was CH 2 Cl 2 :MeOH (6:1). A total of 69 samples show similar TLC profiles to that of the reference herb, forsythiae fructus, with the corresponding spot of phillyrin at Rf 0.46 (Fig. 3). They all passed the qualitative assessment in the TLC examination. Forsythiae fructus samples were extracted with water and diluted alcohol to evaluate the amounts of both crude extracts. As shown in Tables 1-3, the amount of the water extracts ranges from 12 % to 32% for the GF group and from 10 % to 21% for the RF group. The amount of dilute alcohol extracts ranges from 21 % to 37% for the GF group and from 16 % to 25% for the RF group. The amount of dilute alcohol extract is larger than that of the water extract for each sample. The HPLC separation of the forsythiae fructus extract was performed with a C18 column, using a mixture of acetonitrile and water with 0.4% acetic acid as the mobile phase. Good separation was achieved by a gradient elution program at a flow rate of 0.8 ml/min. Multiple components of the herbal extract have different chromophoric characteristics, which respond with different sensitivity under a single monitoring wavelength. Using a DAD detector to record the UV spectra in the range from 200 to 400 nm gives sufficient information for multiple components. The HPLC chromatogram was monitored at 277 nm, which was sufficient for most components in the forsythiae fructus extract, in terms of chromatographic sensitivity. The HPLC fingerprint of forsythiae fructus was constructed as shown in Fig. 4, which allows identification for herbal regulation. A total of 69 samples show similar HPLC fingerprints, but the peak intensity for individual components in each sample is different. Forsythoside A and phillyrin are the bioactive or characteristic components of forsythiae fructus [6,7] that are used as quantification targets to monitor the quality of forsythiae fructus. In order to quantify the content of forsythoside A and phillyrin, a calibration curve for each standard was constructed and tested for linearity. There is good linearity for forsythoside A and phillyrin over the ranges, µg/ ml and µg/ml respectively, and the coefficients of determination (r 2 ) are all greater than The intra- and inter-assay accuracy and precision are acceptable for the quantitative analysis of forsythiae fructus samples. The precision (R.S.D., %) of forsythoside A ranges from 7.90 to 0.82 %, and the precision of phillyrin ranges from to 1.42 % (Table 4). The analytical accuracy (bias, %) varies from 8.84 to % for forsythoside A and from 2.16 to % for phillyrin (Table 4). The regression equations for forsythoside A and phillyrin are respectively derived as: Y= X (r 2 = ) and Y= X (r 2 = ). An optimized chromatographic method was used to determine the concentrations of forsythoside A and phillyrin in the fructus forsythia samples. The results of northern, central, and southern Taiwanese samples are respectively shown in Tables 1-3. The concentration of forsythoside A ranges from 0.25 to 8.22 % and that of phillyrin ranges from 0.09 to 2.54 %. All 69 samples show the same tendency, with a higher concentration of forsythoside A than phillyrin. Table 5 shows the summary of the concentrations of water extract, diluted alcohol extract, forsythoside A and phillyrin in 69 forsythiae fructus samples. The samples from central Taiwan had higher water extract and dilute alcohol extract than those of northern and southern Taiwan. For three collecting workplaces, Chinese herbal stores, Chinese medical clinics, and Chinese hospitals, the samples from Chineses medical clinics showed lower concentrations of water extract and dilute alcohol extract than the samples from the other two. When the 69 samples are divided into 6 GF and 63 RF, the forsythoside A concentration in the GF group ranges from 2.02 to 8.22 % and the phillyrin concentration ranges from 0.90 to 2.54 %. In the RF group, the 4

5 A Quality Assessment of Forsythiae Fructus in the Taiwanese Market Fig. 2. Microscopic observations of pericarp of forsythiae fructus. (A) green forsythiae fructus (GF); (B) ripe forsythiae fructus (RF). (a) a sketch of fructus forsythia, (b) transverse section of pericarp, (c) a detailed drawing of the transverse sections of pericarp, (d) longitudinal sections of pericarp [(d1) peel surface view of pericarp, (d2) longitudinal view of exocarp, spiral tracheis and spiral vessel, (d3) longitudinal view of mesocarp, parenchyma, vascular bundle, spiral tracheis and spiral vessel, (d4) longitudinal view of stone cell populations and embedded fiber]. cul: cuticular layer; ep: epidermis; enc: endocarp; epc: epicarp; fb: fiber; mec: mesocarp ; ph: phloem; vb: vascular bundle; sto: stone cell ; xy: xylem forsythoside A concentration ranges from 0.25 to 1.40 % and the concentration of phillyrin ranges from 0.09 to 1.02 %. It is obvious from these results that there is a great disparity in terms of the quantities of forsythoside A and phillyrin found in the RF and GF groups. In order to calculate the content of other constituents, the HPLC fingerprint was divided into six zones, A-F (with a retention time of 15, 19, 22, 30, 32 and 37 minutes as shown in figure 5A), and the area of each zone was recorded. The average of the zone areas, A-F, was calculated for the GF group and the RF group, and each was individually compared with that in the other group. The results are shown in Fig. 5B. The average areas of zones B, D, E and F in the GF group are higher than those in the RF group. The differences between the GF and RF groups are significant, with P <0.01 for zones B, D and F and P <0.05 for zone E. The components in zones B, D, E and F are respectively confirmed to be forsythoside A (2), phillyrin (5), pinoresinol (6) and arctigenin (7), by comparison with the HPLC retention time and the UV spectrum of Fig. 3. TLC profile of forsythiae fructus. Developing in solvent system of CH2Cl2: MeOH (6:1); P: phillyrin (Rf = 0.46); S: herbal reference of forsythiae fructus; MA1 ~MC2: collected forsythiae fructus. 5

6 Lin et al. / J Chin Med 26(2): , 11 pages, 2015 Fig. 4. HPLC profiles of forsythiae fructus (A) and forsythiae fructus spiked with internal standard (B). IS: internal standard, luteolin; F: forthysoside A; P: phillyrin Table 1. Contents of water extract, dilute alcohol extract, forsythoside A, and phillyrin in forsythiae fructus samples collected from northern Taiwan. Sample Water ext. (%) Dil. alc. ext. (%) Forsythoside A (%) Phillyrin (%) NA ± ± ± ± 0.01 NA ± ± ± ± 0.00 NA3* ± ± ± ± 0.01 NA4* ± ± ± ± 0.00 NA ± ± ± ± 0.01 NB ± ± ± ± 0.00 NB ± ± ± ± 0.00 NB ± ± ± ± 0.01 NB ± ± ± ± 0.00 NC ± ± ± ± 0.00 NC ± ± ± ± 0.00 NC ± ± ± ± 0.00 NC ± ± ± ± 0.02 NC ± ± ± ± 0.01 NC ± ± ± ± 0.01 NC ± ± ± ± 0.01 Data are expressed as mean + standard deviation (n=3). N: samples collected from northern Taiwan, A: from Chinese hospitals, B: from Chinese medical clinics, and C: from herbal drug stores; *: GF. Table 2. Contents of water extract, dilute alcohol extract, forsythoside A, and phillyrin in forsythiae fructus samples collected from central Taiwan. Sample Water ext. (%) Dil. alc. ext. (%) Forsythoside A (%) Phillyrin (%) MA ± ± ± ± 0.00 MB ± ± ± ± 0.01 MB2* ± ± ± ± 0.01 MB ± ± ± ± 0.00 MB ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.01 MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ±

7 A Quality Assessment of Forsythiae Fructus in the Taiwanese Market Table 2. Contents of water extract, dilute alcohol extract, forsythoside A, and phillyrin in forsythiae fructus samples collected from central Taiwan. Sample Water ext. (%) Dil. alc. ext. (%) Forsythoside A (%) Phillyrin (%) MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.01 MC ± ± ± ± 0.01 MC ± ± ± ± 0.00 MC ± ± ± ± 0.01 MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC18* ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC22* ± ± ± ± 0.00 MC ± ± ± ± 0.00 MC ± ± ± ± 0.01 MC ± ± ± ± 0.01 MC ± ± ± ± 0.01 MC ± ± ± ± 0.00 Data are expressed as mean + standard deviation (n=3). M: samples collected from central Taiwan, A: from Chinese hospitals, B: from Chinese medical clinics, and C: from herbal drug stores; *: GF. Table 3. Contents of water extract, dilute alcohol extract, forsythoside A, and phillyrin in forsythiae fructus samples collected from southern Taiwan. Sample Water ext. (%) Dil. alc. ext. (%) Forsythoside A (%) Phillyrin (%) SA ± ± ± ± 0.00 SA2* ± ± ± ± 0.01 SB ± ± ± ± 0.00 SB ± ± ± ± 0.01 SB ± ± ± ± 0.02 SC ± ± ± ± 0.01 SC ± ± ± ± 0.00 SC ± ± ± ± 0.01 SC ± ± ± ± 0.00 SC ± ± ± ± 0.01 SC ± ± ± ± 0.00 SC ± ± ± ± 0.01 SC ± ± ± ± 0.00 SC ± ± ± ± 0.00 SC ± ± ± ± 0.01 SC ± ± ± ± 0.00 SC ± ± ± ± 0.00 SC ± ± ± ± 0.00 SC ± ± ± ± 0.00 SC ± ± ± ± 0.00 SC ± ± ± ± 0.00 Data are expressed as mean + standard deviation (n=3). N: samples collected from southern Taiwan, A: from Chinese hospitals, B: from Chinese medical clinics, and C: from herbal drug stores; *: GF. 7

8 Lin et al. / J Chin Med 26(2): , 11 pages, 2015 Table 4. Precision (R.S.D. %) and accuracy (bias %) for quantifications of forsythoside A and phillyrin in fructus forsythia samples. Nominal conc. Observed conc. R.S.D. (%) bias (%) Observed conc. R.S.D. (%) bias (%) (μg/ml) (μg/ml) (μg/ml) Forsythoside A Intra-day Inter-day ± ± ± ± ± ± ± ± ± ± Phillyrin Intra-day Inter-day ± ± ± ± ± ± ± ± Data are expressed as mean ± standard deviation (n=5) Table 5. Summary of the contents of water extract, diluted alcohol extract, forsythoside A and phillyrin in 69 forsythiae fructus samples. collected from (No. of Water ext. (%) Dil. alc. ext. (%) Forsythoside A (%) Phillyrin (%) samples) Northern Taiwan % % % % (16) Central Taiwan (32) % % % % Southern Taiwan (21) % % % % Chinese herbal stores % % % % (50) Chinese medical clinics (11) % % % % Chinese hospitals (8) % % % % GF ( 6 ) % % % % RF ( 63 ) % % % % authentic samples. These compounds are all important components of forsythia fructus [6 9]. Li s report [10] indicated that the concentration of forsythoside A was higher than phillyrin in the seeds of Forsythia suspensa. Further comparison of the concentration of forsythoside A between seeds and nutshells of Forsythia suspensa showed that seeds had higher concentration than that in nutshells. GF is collected when the fruits are barely ripe. GF has a complete capsule and full of seeds inside. Moreover, RF is harvested when the fruits are fully ripe, where the capsule of RF is always split into two halves and most seeds are lost. Therefore, our finding that GF has higher concentrations of forsythoside A and phillyrin than those in RF is reasonable and consistent with the previous study. 4. Conclusion Sixty-nine samples are all normative for the content of each item, as specified in the Taiwan Herbal Pharmacopeia 1 ; however, the variation in quality between forsythiae fructus is obvious. The 69 samples include 6 GF and 63 RF. Three of the six GF samples came from Chinese hospitals, which reflects the fact that GF is used in clinics. GF and RF are derived from the same plant species, but have different harvesting times. GF is collected when the fruits are still green and barely ripe and RF is harvested when the fruits are fully ripe and yellow. Our results showed that GF and RF had similar TLC and HPLC profiles, which indicates that 8

9 A Quality Assessment of Forsythiae Fructus in the Taiwanese Market Fig. 5. Chemical comparisons of GF and RF. (A) The HPLC chromatogram of forsythiae fructus is divided into six zones A-F, (B) the average peak area for each zone in the GF group and the RF group samples; *, #, : p <0.01; : p <0.05 their secondary metabolite profiles are close. However, GF and RF exhibit large differences in the contents of water extract, the dilute alcohol extract, and the key ingredients, forsythoside A, phillyrin, pinoresinol and arctigenin. For considering the content difference on GF and RF, the classification of fructus forsythiae into two grades, GF and RF, for clinical use should improve safety. Acknowledgments This work was supported by the grant from Ministry of Health and Welfare, ROC (CCMP102- RD-203). The authors would like to thank Dr. Chien-Chang Shen, National Research Institute of Chinese Medicine for his assistance on polishing the manuscript. References 1. Editorial Committee of the Taiwan herbal Pharmacopeia. Taiwan Herbal Pharmacopeia. 2nd ed. Taipei: Executive Yuan Press, Juangsu New Medical College. Zhong Yao Da Ci Dian (Dictionary of Chinese Materia Medica), Vol. 1. Shanghai: Shanghai Scientific and Technological Publishers, Zhao ZZ. Application of microscopic techniques for the authentication of herbal medicines. In: Mendez-Vilas A, Diaz J editors. Microscopy: Science, Technology, Applications and Education. No. 4, vol. 2, Badajoz: Formatex, pp , Zhao ZZ, Shimomura H, Sashida Y, Ishikawa R, Okamoto T, Kazami T. Identification of crude drugs in traditional Chinese patent medicines by means of microscope and polariscope (2): polariscopic characteristics of stone cells, vessels and fibres. Nat. Med., 51: , Japanese Pharmacopoeia Committee. The Japanese Pharmacopoeia. 15th ed. Tokyo: Society of Japanese Pharmacopoeia,

10 6. Endo K, Takahashi K, Abe T, et al. Structure of forsythoside A, an antibacterial principle of Forsythia suspensa leaves. Heterocycles, 16: , Li HB, Chen F. Preparative isolation and purification of phillyrin from the medicinal plant Forsythia suspensa by high-speed counter-current chromatography. J. Chromatogr. A, 1083: , Chen CC, Chen HY, Shiao MS, Lin YL, Kuo YH, Ou JC. Inhibition of low density lipoprotein oxidation by tetrahydrofurofuran lignans from Forsythia suspensa and Magnolia coco. Planta Med., 65: , Sedlák É, Borsodi L, Boldizsár I, Preininger É, László M, Szôke É, Gyurján I. Bioactive phenols in leaves of Forsythia species. Int. J. Hortic. Sci., 14:57 59, Li WJ, Li XE, Qi JJ. Studies on diversity of quality of Forsythia suspensa. Zhongguo Zhong Yao Za Zhi, 32: , Lin et al. / J Chin Med 26(2): , 11 pages,

11 J Chin Med 26(2): , 11 pages, 2015 DOI: / Journal of Chinese Medicine E-ISSN: 台灣市場中藥連翹之品質分析 林麗純 1,* 郭昭麟 2 曾佩儀 1 郭曜豪 衛生福利部國家中醫藥研究所 台北 台灣 中國醫藥大學 中國藥學暨中藥資源學系 台中 台灣 中藥連翹的基原為木犀科 Oleaceae 連翹 Forsythia suspense Thunb. Vahl 的乾燥 果實 為常用中藥 具有清熱作用與對抗細菌感染所引起的發炎作用 本實驗收集台灣北 中 南部中醫院 中醫診所及中藥房的連翹藥材飲片共 69 件樣品 利用性狀特徵 組織切片 顯微鏡檢 水抽提物 稀醇抽提物 薄層層析 指紋圖譜分析 及定量等方法 進行台灣市 場中藥連翹的品質分析 由樣品外觀特點可區分 69 個樣品中有 6 件青翹和 63 件老翹樣本 由切片顯微觀察 青翹 GF 和老翹 RF 有相似的組織結構 但青翹具有較肥厚的軟組織 切片的同時 青翹散發出較濃烈的氣味 青翹與老翹有相似的 TLC 和 HPLC 指紋圖譜 但比 較其水提取物 稀醇提取物和主要成分 連翹酯苷 A 連翹苷 松脂醇及牛蒡苷元 的含量 青翹含量遠高於老翹 比較台灣北 中 南部樣品 發現中部樣品具有較高含量的水提取物 與稀醇提取物 以 3 類工作場域做比較 來自中醫診所的樣品有較低的水提取物 及稀醇提 取物含量 69 樣品都符合藥典各規範項目標準 但青翹與老翹的水提取物 稀醇提取物 及 重要化學成分含量差異大 因此建議連翹飲片 分青翹與老翹二個分級使用 可增加臨床用 藥安全 關鍵字 品質分析 連翹 青翹 老翹 水提取物 稀醇提取物 連翹酯苷 A 連翹苷 104 年 2 月 24 日受理 104 年 4 月 8 日接受刊載 104 年 12 月 1 日線上出版 * 聯絡人 林麗純 衛生福利部國家中醫藥研究所 台北市北投區立農街二段 號 電話 分機 7101 電子郵件信箱 lclin@nricm.edu.tw 11