Several methods have been described

Several methods have been described for the determination of
TOCS, CHLS and CARS in plant material, either separately [18–20]
or simultaneously [21–23], typically by reverse phase HPLC-DAD.
Fraser et al. [22] described a method using a C30 column which
enabled the simultaneous separation of CARS, TOCS, ubiquinones
and plastoquinones in 60 min. Burns et al. [21] described an extraction and a HPLC method for the determination of CARS, TOCS and
CHLS in fruits and vegetables in 40 min. Gleize et al. [23] described
a method to analyze eleven CARS, retinol, - and -tocopherol in
foods, human plasma and human adipose tissue in 35 min using a
C30 column.
The “traditional methods” mentioned above are appropriate for
the detailed profiling of such compounds. More specifically the
C30 column is very suitable for the separation of both geometrical
(reviewed in [24], and even some optical CARS isomers [25]). However, due to its retention characteristics the HPLC methods that use
it involve long analysis times and a higher use of solvents, with
the associated negative consequences for the environment. Consequently, they are not the most appropriate for screening purposes.
Recently, a clear improvement in chromatographic performance
has been achieved by the introduction of rapid-resolution liquid
chromatography (RRLC) or ultra-performance liquid chromatography (UPLC) that allows for a higher throughput of samples, often
without compromising resolution, and a considerable saving of solvents. Although they are widely used in the pharmaceutical area,
they are not as widespread for the analysis of food isoprenoids.
Granado et al. [26] compared HPLC vs. UPLC methods for the
simultaneous determination of biomarkers of the nutritional status.
Furthermore, they proposed a method that allowed the separation of vitamin A, E, and D, and major plasma carotenoids (lutein,
zeaxanthin, -carotene, -carotene, -cryptoxanthin, lycopene) in
4.5 min. A recent study describes the analysis of a mixture of up
to 16 CARS standards by UPLC–MS/MS in 12 min [27]. Similarly,
Bohoyo-Gil et al. [28] proposed a methodology by UPLC for the
determination of the seven carotenoids in honeybee pollen samples, pumpkin and nectarine flesh with satisfactory recoveries in
10 min.
Within this context, the main aims of the present work were
two:
1) To develop and validate a rapid and effective RRLC method for
the simultaneous determination of dietary plant isoprenoids of
interest in Food Science and Nutrition.
2) To apply it to their analysis in plant foods with different colors
as a result of their CARS and/or CHLS contents.
2. Materials and methods
2.1. Chemicals and standards
The extraction solvents were of analytical grade. Hexane, acetone and dichloromethane were purchased from Carlo-Erba (Milan,
Italy). The chromatographic solvents were methanol, acetonitrile, ethyl acetate (HPLC grade, procured from Merck, Darmstadt,
Germany). Water was purified in a NANOpure®
DIamondTM
system (Barnsted Inc., Dubuque, IO). -carotene, -cryptoxantin
and zeaxanthin were purchased from Sigma-Aldrich (Steinheim,
Germany). Violaxanthin, -carotene, lutein lycopene, phytoene
and phytofluene were isolated from appropriate sources in
accordance to standard procedures [29]. Chlorophylls a and b,
and trans--apo-8

-carotenal were purchased from Sigma-Aldrich
(Steinheim, Germany

Hasil (Bahasa Indonesia) 1:

Beberapa metode telah dijelaskan untuk penentuanTOCS, CHLS dan mobil di bahan tanaman, baik secara terpisah [18 – 20]atau secara bersamaan [21 – 23], biasanya oleh terbalik fase HPLC-ayah.Fraser et al. [22] menggambarkan metode yang menggunakan kolom C30 yangdiaktifkan pemisahan simultan Mobil, TOCS, ubiquinonesdan plastoquinones di 60menit Burns et al. [21] digambarkan ekstraksi dan metode KCKT untuk penentuan Mobil, TOCS danCHLS buah-buahan dan sayuran di 40 min. dijelaskan Gleize et al. [23]sebuah metode untuk menganalisis sebelas Mobil, retinol,- dan - tokoferol dimakanan, plasma manusia dan jaringan adiposa manusia dalam 35 menit menggunakanC30 kolom."Metode tradisional" disebutkan di atas sesuai untukProfil rinci senyawa tersebut. Lebih khusus lagiC30 kolom ini sangat cocok untuk pemisahan kedua geometris(dibahas dalam [24], dan bahkan beberapa optik mobil isomer [25]). Namun, karena karakteristiknya retensi metode KCKT yang menggunakanini melibatkan panjang analisis kali dan lebih tinggi penggunaan pelarut, dengankonsekuensi negatif yang terkait untuk lingkungan. Akibatnya, mereka yang tidak sesuai untuk tujuan skrining.Baru-baru ini, jelas perbaikan dalam kinerja kromatografitelah dicapai oleh pengenalan cepat-resolusi cairKromatografi (RRLC) atau ultra kromatografi cair (UPLC) yang memungkinkan untuk throughput yang lebih tinggi dari sampel, seringtanpa mengorbankan resolusi, dan penghematan cukup pelarut. Meskipun mereka secara luas digunakan di bidang farmasi,mereka tidak seperti luas untuk analisis makanan isoprenoids.Granado et al. [26] dibandingkan metode KCKT vs UPLCpenentuan simultan biomarker status gizi.Selain itu, mereka mengusulkan sebuah metode yang memungkinkan terjadinya pemisahan dari vitamin A, E, dan D, dan karotenoid utama plasma (lutein,zeaxanthin,-karoten, - karoten, - cryptoxanthin, lycopene) di4.5 min. Sebuah studi terbaru menggambarkan analisis campuran up16 mobil standar oleh UPLC-MS/MS di 12 min [27]. Demikian pula,Bohoyo-Gil et al. [28] diusulkan metodologi oleh UPLC untukpenentuan tujuh karotenoid dalam sampel serbuk sari lebah madu, labu dan Nektarin daging dengan memuaskan pemulihan di10 min.Dalam konteks ini, tujuan utama dari pekerjaan sekarang adalahdua:1) untuk mengembangkan dan memvalidasi metode RRLC yang cepat dan efektif untukpenentuan simultan makanan tanaman isoprenoids dariminat dalam ilmu makanan dan gizi.2) untuk menerapkannya ke analisis mereka dalam makanan nabati dengan warna yang berbedakarena isinya mobil dan/atau CHLS.2. bahan dan metode2.1. bahan kimia dan standarEkstraksi pelarut yang analitis kelas. Heksana, aseton dan diklorometana itu dibeli dari Carlo-Erba (Milan,Italia). Kromatografi pelarut yang metanol, asetonitril, etil asetat (kelas HPLC, Diperoleh dari Merck, Darmstadt,Jerman). Air dimurnikan dalam NANOpure ®DIamondTMsistem (Barnsted Inc, Dubuque, IO). -karoten, - cryptoxantindan zeaxanthin dibeli dari Sigma Aldrich (Steinheim,Jerman). Violaxanthin,-karoten, lutein lycopene, phytoenedan phytofluene diisolasi dari sumber-sumber yang sesuai disesuai dengan prosedur standar [29]. Chlorophylls a dan b,dan trans-- apo-8-carotenal dibeli dari Sigma Aldrich(Steinheim, Jerman