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Study of Tea Digitized Chromatographic Fingerprint Spectra Using Micellar Electrokinetic Chromatography

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Study of Tea Digitized Chromatographic Fingerprint Spectra Using Micellar Electrokinetic Chromatography CHINESE JOURNAL OF CHEMISTRY 2003, 21, 1325-1329 1325 Study of Tea Digitized Chromatographic Fingerprint Spectra Using Micellar Electrokinetic Chromatography SONG, Guan-Quna*b($%%) LIN, Jin-Ming' *'($$&%I)QU, Fengo(E%) DONG, Wen-Jub( 3 ) Research Center for Eco-Environmental Sciences , Chinese Academy of Sciences , Beijing 100085, China College of Chemistry and Environmental Science , Henun Normal University, Xinxiang , Henan 453002 , China This paper described the principle of digitized chromatographic tea quality or establishing the tea criterion by one or two fingerprint spectrum and established digitized chromatographic components as the guideline such as ( - )-epicatechin, fingerprint spectra of ten brands of Chinese famous tea by the ( - )-epigallocatechin , ( - )-epigallocatechin gallate. micellar electrokinetic chromatography, work was done Thii Therefore, they can not totally reflect the quality of tea. It using a 25 mmol L - sodium dodecylsulfate in a 20 mmol*L - is indispensable to establish a simple and convenient borate (pH 7.0) solution as running buffer, 20 kV applied po- tential and detection at 280 nm. The chromatographic finger- method for assessing the quality of tea by measuring sever- print spectra were digitized by the relative retention value (a) al components of tea simultaneously. and the relative area ( S,), and were analyzed to identify the The composition of tealeaves depends on many fac- tea samples. In the absence of the standard samples, the pre- tors, e.g . , agronomic condition, leaf age, storage time, sent method was easy setup and inexpensive, and provided the the degree of fermentation, etc . Therefore, the compo- applicable information for the quality assessment of teas. nents of tealeaves are very complex. The studies indicate that in the same chromatographic conditions, different Keywords tealeaves, micellar electrokinetic chromatography, digitized chromatographic fingerprint spectrum, quality assessment brands of tea result in different chromatograms, even though from the same brand, there have great difference Introduction between chromatograms because of resources and period. Based on these reasons, we establish the digitized chro- Tea is the most widely consumed beverage in matographic fingerprint spectrum consisted of relative-re- people's life. The heightened popularity of this beverage tention time (a) and corresponding peak area (S,) using throughout the world in recent years may be due in part to the micellar electrokinetic chromatography. The present the evidence of a relationship between tea consumption and method can transfer chromatograms to datasheets , resulting prevention of certain forms of human disease. The main in reducing the effect of operational factors and the fluctua- components of tea are polyphenolic compounds, commonly tion of peak location and improving the comparability among samples. The most important of all, this method known as catechins, which represent a group of com- pounds belonging to the flavonoid family. These com- can still obtain reliable results without standard samples. pounds are widely distributed in the tea leaves and consti- tute up to 30% of the dry leaf weight. Much interest has Experimental been focused on catechins, not only for their antioxidant activity,' but also for their known antimutagenic and anti- Apparatus tumorigenic properties. 3*4 Because of the above character- istics, the intake of tea becomes more and more. Accord- All experiments were carried out with a lab-built ing to data published by the Tea Council in the United (capillary electrophorecis) system equipped with a high Kingdom, global production is about 3 million tones each voltage power supply (Beijing New Tech. Inc . , China), a year,' the majority of which come from China. Therefore, UV spectrometer (Jasco, Japan) and a plexiglass box in the tea assessment is highly desirable, and many methods which the buffer and sample reservoirs were. A fused-sili- have been developed for the determination of catechins, ca capillary (Yongnian, China) with a total length of 60 e . g . , TLC ,6 HPLC ,' HPLC-MS ,* CZE9 and MEKC. lo cm and an I. D . of 50 pm was used. A detection window However, these methods are always used for assessing the was created at 45 cm from the capillary inlet by removing * E-mail : jmlin (3 mail. rcees . ac .cn Received January 22, 2003 ; revised April 8, 2003 ; accepted June 9, 2003. Project supported by the National Natural Science Fundation for Distinguished Young Scholars of Chino (No. 20125514) ontl the Nation- al Natural Science Foundation of China ( Nos. 207,75043 and 50273046) . 1326 Micellar electrokinetic chromatography SONG et al. the polyimide coating. HW chromatography software However, in the present work, it was found that this solu- (Qianpu Inc . , China) was used for both the operation of tion caused a higher value of current, unacceptable Joule the detector and data processing. THZ-82 constant-temper- heating and peak broading, being presumably due to the ature shaker (Changzhou Inc., China) was used to dis- fact that the alteration of electroosmotic flow atrected the pose the sample. separation with different capillary. In order to achieve the rapid and valid separation, the buffer system was opti- Reagents mized. The results showed that the viscosity of the solution was increased with the increasing of the concentration of All reagents were of analytical-reagent grade. Sodium SDS and borate, resulting in the decreased velocity of dodecylsulfate ( SDS) was obtained from Wako Pure electroosmotic flow and longer separation time. Therefore, Chemival Industries Ltd ( Tokyo, Japan ) . All solutions a 25 rnmol-L-' SDS in a solution of 20 mmol*L-' borate were prepared with water, which was deionized using a (pH = 7.0) was chosen as the optimum buffer system, re- Milli-Q system with 0.2-pm fiber filter ( 18.3 Ma, Bam- sulting in the moderate separation time and the good sepa- stead, USA). The tea samples and their resources were ration. listed in Table 1. Establishment of digitized chromatographic fingerprint spec- Analytical conditions trwn A 25 mmol*L-'SDS in a 20 mmo1.L-' borate (pH Under the optimum conditions, the chromatograms of 7.0) solution was used as running solution. A potential of the ten brands of Chinese tea were obtained. As shown in 20 kV was applied and the detection was performed by UV Fig. 1, the chromatograms were characteristic but the absorption at 280 nm . The samples were injected by 1.5 quality of tea could not be identified only from them. kPa pressure for 10 s. Thus, it was necessary to establish the digitized chromato- The capillary was conditioned prior to use by flushing graphic fingerprint spectra according to the ten chro- for 10 min with 1 .O mo1.L-l HCl, 2 min with water, 10 matograms. min with 1 .0 mol * L-' NaOH , 2 min with water and finally The chromatographic peaks shown in Fig. 1 were 10 min with buffer. In order to increase the reproducibility dense. One of the peaks was selected as the reference of the migration time, between runs the capillary was peak, which was characteristic of strong intensity, mid-re- rinsed consecutively with 1.0 mol-L-' HCl (2 min) , wa- tention time and common in the ten chromatograms. Thus, ter ( 1 min) and finally with buffer (2 min) . based on the reference peak, the relative-retention time (a) can be obtained through the formulation : Preparation of tea sample solution 1 .O00 g of tealeaves were weighted and placed into a 250 mL of beaker, adding 100 mL of deionized hot water t, is absolute retention time of reference peak, ti is abso- at 90 T,stirring for 10 min by a constant temperature lute retention time of determined peaks. shaker at 90 T . When cooling at about 70 T , 20 mL of The chromatogram of Molihua tea (sample No. I11 in tea infusion filtered with common filter paper was diluted Table 1) was selected as the standard sample because of 10 times with 0.1% metaphosphoric acid. After passing its more peaks, stronger peak intensity and representative through the 0.45-pm membrane filter, the filtrates were peak shape. The relative area ( S,) of determined peak used as samples. can be obtained according to the total peak area of the standard. The equation was : Results Optimization of the separation condition A, was the total peak area of the standard sample, A" was Watababe et a1 . lo reported that catechins were sepa- the area of the determined peak. rated in SDS-borate-phosphate buffer solution at pH = 7.0. Table 1 Ten brand Chinese teas and their resources for the present experiment No. Sample Source No. Sample Source I Huangshan maofeng Anhui VI Qijing greentea Fujian I1 Xinyang maojian Henan VII Mingqian luzhen Fujian III Molihua tea Shanghai VIII Biluochun Jiangsu IV Dahongpao Fujian Ix Guihua woolong Anhui V Xihu longjing Zhejiang X Tieyanyin Fujinn Vol. 21 No. 10 2003 Chinese Journal of Chemistry 1327 120 T 9 100- 7 5 80- 1 .-5 60- E 40: 9 4 20: 0-- .... 2 4 6 8 10 12 14 16 2 4 6 8 10 12 14 16 Time (min) Time (min) I1 2 100- 6 9 80- YE E 60: .0 40- 20: 13 14 I 1 I I I I I IJ I I, I I I I 1 2 4 6 8 10 12 14 16 18 2 4 ,6 8 10 12 14 16 Time (min) Time (min) 2 1601 111 2 wn 140- 6 140- >^ 120- 120- 9 E 100- I E 100- 1 Y Y 8 c 80- 1 8 80- .p 60- .- 60- 40- f 40- 1 5 20- 1 a 0-1 0- II1llll1 111,1111 2 4 6 8 10 12 14 16 2 4 6 8 10 12 14 16 Time (min) Time (min) 140- IV 2 >^ 120- 100P 1' vE 100- 6 .-8 80- E 60- 5 40- a 20- II 0-- I I I1I 11 I I I I 2 4 6 8 10 12 14 16 Time (min) X 1 170- v 3 160- 6 8 54- 2 !% I E 4s- ;100- 36- o 80- 1 .g 'g 60- 40- 1 ::: B 20- 4 9- a0 0-y -.
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