[논문]Determination of Sesamin and Sesamolin in Sesame (Sesamum indicum L.) Seeds Using UV Spectrophotometer and HPLC

Kim, Kwan-Su; Lee, Jung-Ro; Lee, Joon-Seol

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Sesamin and sesamolin, antioxidant lipidsoluble lignan compounds, are abundant in sesame (Sesamum indicum L.) seed oil and provide oxidative stability of oil related to sesame quality. The sesamin and sesamolin contents of 403 sesame land races of Korea were determined by HPLC analysis of methanol e...

Sesamin and sesamolin, antioxidant lipidsoluble lignan compounds, are abundant in sesame (Sesamum indicum L.) seed oil and provide oxidative stability of oil related to sesame quality. The sesamin and sesamolin contents of 403 sesame land races of Korea were determined by HPLC analysis of methanol extract (HPLC value), and their total lignan content was compared with those by using UV-Vis spectrophotometric analysis (UV method) of methanol (UV-MeOH value) and hexane (UV-Hexane value) extracts. HPLC values of total lignan content were strongly associated with UV-Hexane (r=0.705**) and UV-MeOH (r=0.811**) values. The UV values from both the extracts were 3.8-4.7 times higher than those of HPLC values. Lignan content was overestimated by UV method because total compounds in the mixture solution were quantified by absorbing at the same ultraviolet wavelength as in HPLC method. UV method could more rapidly analyze small amount of sample with higher sensitivity of detection than HPLC method. Average contents of lignans in sesame germplasm evaluated in this study were $2.09{\pm}1.02mg/g$ of sesamin, and $1.65{\pm}0.61mg/g$ of sesamolin, respectively, showing significant variation for lignan components. The results showed that UV method for the determination of sesamin and sesamolin could be practically used as a faster and easier method than HPLC by using the regression equations developed in this study.

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문제 정의

The objectives of this study were to investigate the variation of lignans such as sesamin and sesamolin contents among Korean sesame landraces using HPLC and UV methods, and to evaluate their efficiencies for a massive screening ofsesame seeds.

제안 방법

Running time was 20 minute for each sample. Each peak oflignan compounds was identified for further analysis with LC-MS (HCTultra, Bruker Daltonics Inc., USA), and comparing HPLC retention time and UV spectra with standard compounds ofsesamin and sesamolin. Chemical structures of sesamin and sesamolin, antioxidant lignans are shown in Fig.
In order to compare the linearity of calibration curves by using both UV and HPLC methods, triplicate calibration curves for sesamin and sesamolin in HPLC method and total lignans in UV method were obtained. Five levels of standard concentrations were measured in the ranges between 3.
UV method needed small amount (below 100 mg) of sample as compared to that (1-2 g) of HPLC method. Total lignans for calibration in HPLC metliod were calculated by summing the peak areas of sesamin and sesam_이in, and then linearity of calibration by HPLC was compared with that of the UV method. Linearity of calibration curves was obtained as shown in Fig.
However, small amount of sample is required along with short duration of analysis and high sensitivity of detection for UV method. Therefore, it is suggested that UV method can be used as a rapid and inexpensive technique with a reliable accuracy for massive screening of sesame breeding lines, provided that the regression equations developed in this study is applied for the determination of lignans content. Sesame breeding lines selected by UV method would need further confirmation by HPLC and then should be continuously evaluated through progeny test in breeding program of sesame.

대상 데이터

The 10 μL ofMeOH extract was injected without any dilution. The HPLC instrument (Agilent 1100 Series, Agilent Technologies Co., USA) equipped with an ultraviolet-visible detector(Diode-Array Detector) at 290 nm, which also provided the UV spectra of the peaks, and a reversed-phase column, Capcell Pak C18 UG 120, 54.6 x 150 mm (Shiseido Co., Japan) was used. The mobile phase was a mixture ofmethanol: water (75 : 25, v/v) as an isocratic condition and flow rate was set at 1.

이론/모형

The HPLC analysis was done for evaluation of the linearity of calibration curves to determine total lignans, and then its results were compared with those obtained by using the UV spectrophotometric method (UV method). The UV absorption spectra and HPLC chromatogram are shown in Fig.

성능/효과

01 level of probability in both methods. The results showed that UV and HPLC analysis could be sufficiently performed to determine the total lignan contents of sesame seeds with good accuracy.
In conclusion, although UV method has a shortcoming of imprecision and overestimation for measured values, it will be a beneficial and practical technique when the regression model has good agreement with HPLC values to adj_니st UV values. This method would be practically useful, especially in breeding program of sesame because the relative evaluation of chemical compo_니nds in large population was very important for rapid selection.

후속연구

The accessions displayed significant variation for lignan content and were selected as superior lines for sesame breeding. Those germplasms with high content of lignans, sesamin and sesamolin, should be further studied to confirm whether they are genetically useful for the quality breeding of sesame, in addition to testing variations of lignan contents according to different environment factors.

참고문헌 (13)

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상세보기
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Fukuda, Y., M. Nagata, T. Osawa, and M. Namiki. 1986. Chemical aspects of the antioxidative activity of roasted sesame seed oil, and the effect of using the oil for frying. Agric. Biol. Chem. 50 : 857-862

상세보기
Kamal-Eldin, A., L. A. Appelqvist, and G. Yousif. 1994. Lignan analysis in seed oils from four Sesamum species: comparison of different chromatographic methods. J. Am. Oil Chem. Soc. 71 : 141-147

상세보기
Katsuzaki, H., S. Kawakishi, and T. Osawa. 1994. Sesaminol glucosides in sesame seeds. Phytochemistry 35 : 773-776

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상세보기
Moazzami, A. A., R. E. Andersson, and A. Kamal-Edlin. 2006. HPLC analysis of sesaminol glucosides in sesame seeds. J. Agric. Food Chem. 54 : 633-638

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원문보기 상세보기
Shyu, Y. S. and L. S. Hwang. 2002. Antioxidative activity of the crude extract of lignan glycosides from unroasted Burma black sesame meal. Food Res. Int. 35 : 357-365

상세보기
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[국내논문] Determination of Sesamin and Sesamolin in Sesame (Sesamum indicum L.) Seeds Using UV Spectrophotometer and HPLC 원문보기

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[국내논문] Determination of Sesamin and Sesamolin in Sesame (Sesamum indicum L.) Seeds Using UV Spectrophotometer and HPLC 원문보기

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