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Kimia Makanan 339 (2001) 128046
Daftar isi tersedia di SainsLangsung
Kimia Makanan
beranda jurnal: www.elsevier.com/locate/foodchem
Pengembangan senyawa organik volatil dan prekursor glikosilasi mereka

di tamarillo (Solanum betaceum Cav.) selama pematangan buah: Prediksi
jalur biokimia
Xiao ChenSebuah, Bruno FedrizziSebuah, Paul A. KilmartinSebuah, Siew Young QuekSebuah,b,⁎
Sebuah Sekolah Ilmu Kimia, Universitas Auckland, Auckland 1010, Selandia Baru
b Riddet Institute, Pusat Penelitian Keunggulan dalam Penelitian Makanan, Palmerston North 4474, Selandia Baru
INFO ARTIKEL ABSTRAK
Kata kunci: Metabolit kunci dan jalur pengaturan rasa dalam terong belanda diselidiki untuk mengeksplorasi pengembangan
Tamarilo senyawa organik volatil (VOC) bebas dan terglikosilasi selama pematangan buah. Konsentrasi VOC bebas dan
Biosintesis rasa terikat ditentukan dengan analisis spektrometri massa kromatografi gas. Perubahan parameter fisik, konsentrasi
Glikosida
prekursor rasa, dan aktivitas enzim endogen kunci juga dipantau. Sebanyak 22 VOC bebas diidentifikasi dengan
Jalur senyawa organik volatil (VOC)
alkohol C6 dan ester sebagai senyawa utama. Dari 83 VOC terglikosilasi yang terdeteksi, fenol dan terpenoid
Lipoxygenase (LOX)
merupakan komponen yang dominan. Konsentrasi VOC terikat total meningkat hingga 4 kali lipat selama
pematangan buah. Jalur lipoxygenase dikonfirmasi sebagai mekanisme biosintetik penting untuk generasi VOC
bebas dan glikosilasi selama pematangan tamarillo.p < 0,05 atau p < 0,01).
1. Perkenalan jalur biosintesis penting, terkait dengan generasi alkohol, aldehida, dan ester
dalam banyak buah-buahan (misalnya pir, apel, dan stroberi) selama pematangan
Tamarilo (Solanum betaceum Cav.), juga disebut tomat pohon, adalah buah (Granell & Rambla, 2013). Secara khusus, aldehida dan alkohol C6 yang
subtropis yang memiliki kulit halus, bentuk lonjong, dan biji berair (Prohens & memberikan aroma "hijau dan seperti daun", sebagian besar berasal dari oksidasi
Nuez, 2001; Schotsmans, Timur, & Woolf, 2011). Sebagai kerabat botani tomat asam linoleat dan asam linolenat. Ester, mewakili volatil aromatik khas buah di
dan kentang, tamarillo berasal dari Amerika Selatan (Argentina, Chili, dan Peru). banyak buah matang, dihasilkan dari esterifikasi asil-KoA dan alkohol selama jalur
Buah ini tiba di Selandia Baru pada akhir abad ke-19 dan tumbuh dengan baik di lipoksigenase (LOX).Chen dkk., 2020; Defilippi dkk., 2009). Biosintesis senyawa
daerah utara termasuk Kerikeri dan Bay of Plenty (Schotsmans, Timur, & Woolf, aroma ini dipercepat selama pematangan buah-buahan seperti pada tomat (Klee,
2011). Kultivar tamarillo dapat diklasifikasikan menurut warna buahnya, yaitu 2010) dan pir (Chen dkk., 2020), karena akumulasi substrat dan peningkatan
kuning, merah, dan merah tua. Varietas berkulit merah adalah kultivar yang aktivitas lipase.
dominan di Selandia Baru. Kultivar ini memiliki bobot buah yang lebih besar
hingga 100 g, diameter 5-6 cm dan panjang 7-8 cm, serta memberikan rasa
sedikit astringen dan asam dibandingkan kultivar berkulit kuning (Prohens & Selain VOC bebas yang secara langsung berkontribusi pada aroma buah,
Nuez, 2001). volatil juga dapat melekat pada monosakarida atau disakarida melalui ikatan
-glikosidik, membentuk glikokonjugat yang tidak mudah menguap dan tanpa
Produksi senyawa organik volatil (VOC) diatur selama kematangan buah fruit rasa. Ini disebut VOC yang terikat secara glikosida, di mana aroma aglikon dapat
dalam hidup, dan ini telah berfungsi sebagai faktor yang mendominasi untuk dibebaskan selama pematangan buah, penyimpanan pascapanen dan dengan
membentuk sifat sensorik buah (Granell & Rambla, 2013). Selama pematangan proses hidrolisis asam atau enzimatik.Hjelmeland &
buah, VOC secara bertahap disintesismelalui berbagai jalur metabolisme dari Ebeler, 2015). Alkohol (misalnya cis-3-heksenol, heksanol), C13 norisoprenoid (
prekursornya, termasuk asam amino, karbohidrat, dan asam lemak.Defilippi, misalnya -damascenone, 3-hidroksi-β-ionon, 3-okso-α-ionol), terpenoid
Manríquez, Luengwilai, & González-Agüero, 2009). Jalur asam lemak adalah salah (misalnya linalool, -sitronellol, geraniol), dan fenol volatil (misalnya vanillin,
satu yang paling raspberry keton, eugenol) adalah aglikon yang paling banyak dilaporkan
⁎ Penulis yang sesuai.

Alamat email: sy.quek@auckland.ac.nz (SY Quek).
https://doi.org/10.1016/j.foodchem.2020.128046
Diterima 6 Mei 2020; Diterima dalam bentuk revisi 1 September 2020; Diterima 4 September 2020
Tersedia online 16 September 2020
0308-8146/ © 2020 Elsevier Ltd. Hak cipta dilindungi undang-undang.
X.Chen, dkk. Kimia Makanan 339 (2001) 128046
(Winterhalter & Skoroumounis, 1997), yang seringkali lebih berlimpah daripada dicampur dengan air Milli-Q (1:1, b/b), diikuti dengan pencampuran dengan
rekan-rekan bebas mereka dalam buah. Konjugat ini berperan sebagai reservoir blender Vitamix (Total Nutrition Center, Cleveland, Ohio, USA) untuk memfasilitasi
yang potensial untuk menghasilkan aroma, sehingga kadar volatil yang pelepasan komponen metabolik intraseluler (Aurore dkk., 2011; Wang dkk., 2015).
terakumulasi dalam buah dapat dimodulasi selama pemasakan atau pemrosesan. Bubur yang diperoleh segera disentrifugasi dua kali pada suhu 4 °C selama 20
Akibatnya, ada minat yang berkembang untuk mempelajari prekursor terikat menit pada 16.993 × g. Supernatan dikumpulkan, dipulihkan, dan kemudian
glikosida dalam buah-buahan, dan untuk mengatur hidrolisis untuk melepaskan diklarifikasi dengan penyaringan vakum menggunakan corong Büchner dengan
aglikon yang melekat dan meningkatkan aroma buah. Banyak contoh VOC kertas saring (ukuran pori 11 m, Whatman, Maidstone, UK). Akhirnya, supernatan
terglikosilasi yang melimpah pada tanaman dapat ditemukan, seperti pada tomat ( bening disimpan sebagai 50 mL-alikuot dan dibiarkan pada suhu -18 °C untuk
zkaya, Sen, Aubert, Dundar, & Gunata, 2018), pisang (Aurore, Ginies, Ganou- analisis volatil. Sisa irisan tamarillo disimpan pada suhu -18 °C selama 12 jam,
parfait, Renard, & Fahrasmane, 2011), dan buah kiwi (Garcia, Stevenson, Atkinson, dipindahkan ke dalam freezer -80 °C selama 5 jam, diikuti dengan pengeringan
Winz, & Quek, 2013) juga dalam anggur (Hjelmeland & Ebeler, 2015), teh (Gui dkk., beku (pengering beku FreeZone 12 Plus, Labconco, MO, USA). Tamarillo yang
2015) dan tembakau (Dia dkk., 2015). Studi kami sebelumnya meneliti pengaruh diliofilisasi digiling menjadi bubuk halus dan disimpan pada suhu -80 ° C untuk
strategi hidrolisis dan waktu inkubasi pada pelepasan volatil terikat (Chen, Quek, analisis asam lemak dan uji aktivitas enzim.
Fedrizzi, & Kilmartin, 2020), dan merupakan satu-satunya penelitian yang
menyelidiki prekursor volatil terglikosilasi dalam buah tamarillo, sejauh yang
penulis ketahui. Namun, profil volatil terikat dalam kultivar terong yang berbeda
2.2. Bahan kimia dan reagen
dan bagaimana senyawa ini berubah selama periode pematangan belum
dieksplorasi.
Bahan kimia berikut diperoleh dari Sigma-Aldrich (St. Louis, MO, USA): standar
kimia untuk identifikasi dan kuantifikasi volatil (cis-3-heksenol (97%), 1-heksanol
Saat ini, perbedaan varietas senyawa organik non-volatil dalam terong
(99,9%), 2-etilheksanol (99,6%), pentanal (97,5%), heksanal (98%), nonanal (95%),
belanda telah diteliti dengan baik, termasuk vitamin C, karotenoid, fenolik, dan
metil butanoat (99,5%), -terpineol (90%), benzaldehida (99,5%), etil butanoat
antosianin.Acosta-Quezada dkk., 2015; Espin & Gonzalez-Manzano, 2016). Secara
(99,5%), 3-oktanon (98%), metil kaproat (99,8%), cis-3- heksenil asetat (98%), heksil
komparatif, hanya dua penelitian yang melaporkan komposisi volatil bebas dari
asetat (99,7%), metil benzoat
tamarillo tetapi tanpa data kuantitatif (Torrado dkk., 1995; Durant, Rodríguez,
Santana, Herrero, Rodríguez, & Gupta, 2013). Selain itu, ada kesenjangan
(99,5%), metil kaprilat (99,8%), metil salisilat (99%), D-limonene (97%), linalool (97%),
pengetahuan dalam biosintesis volatil bebas dan prekursor glikosilasinya selama
-terpinene (95%), eucalyptol (99%), 2-
pematangan buah tamarillo. Tujuan dari penelitian ini adalah untuk
metil-3-heptanon (99%), C7-C30 alkana jenuh); standar referensi kimia non-volatil
mengeksplorasi profil senyawa volatil bebas dan terikat glikosida dalam terong
(asam malat, arabitol, glukosa, asam quinic,
dan perubahannya selama pematangan buah dengan fokus lebih lanjut pada
asam sitrat, fruktosa, fenil -D-glukopiranosida, sukrosa, p-nitrofenil
mekanisme generasi volatil. Jalur LOX metabolisme asam lemak dipelajari dengan
(pNP), p-nitrofenil -D-glukopiranosida (pNPG), C4-C24 campuran metil ester asam
mengukur konsentrasi substrat dan aktivitas enzim terkait, untuk menyelidiki
lemak (FAMEs), asam linoleat (99%), metil heptade-
biosintesis alkohol C6, aldehida, dan ester turunan sepanjang pematangan buah.
kanoat (99%)); dan campuran sililasi (hexamethyldisilazane/ chlorotrimethylsilane/
Dengan demikian, penelitian saat ini dapat memberikan wawasan berharga
pyridine, 2:1:10, (v/v/v)) untuk derivatisasi GC.
tentang mekanisme pembentukan volatil selama pematangan buah untuk
Metanol (kelas HPLC) dan diklorometana (kelas HPLC) dibeli dari JT Baker
mengisi kesenjangan pengetahuan dan untuk memfasilitasi pemahaman tentang
(Phillipsburg, NJ, USA). Natrium klorida, natrium hidroksida, monosodium fosfat,
kualitas sensorik buah tamarillo.
dan dinatrium fosfat dengan grade analitis diperoleh dari ECP Ltd. (Auckland,
Selandia Baru).
Strata C18-E (500 mg/6 mL, 55 m, 70 A,) dibeli dari Phenomenex, Inc.
2. Bahan & metode
(Torrance, CA, USA). Enzim Rapidase AR2000 (glikosidase) dibeli dari DSM Food
Specialties (Alexander Fleminglaan, Delft, Belanda).
2.1. Pengumpulan buah dan persiapan sampel
buah tamarillo dari “Laird's Large”, “Mulligan” dan “Amber”, yang merupakan
kultivar yang umum dikenal di Selandia Baru (Prohens & Nuez, 2001; Schotsmans, 2.3. Analisis fisik
Timur, & Woolf, 2011), dipanen dari kebun buah-buahan di Whangarei, Selandia
Baru selama Mei 2019. Buah-buahan pada tiga tahap pematangan yang dapat Warna buah (L*, a*, b*, chroma, dan nilai sudut hue) diukur pada tiga bagian
dibedakan dipetik secara acak dari pohon menurut warna, kekencangan dan yang berbeda dari sampel buah: di bagian atas (1 cm dari kelopak buah), tengah
ukuran: hijau (hijau cerah, tekstur keras dan ukuran penuh), pembubutan (sekitar garis ekuator) dan bagian bawah (1 cm di sekitar bagian paling bawah) (
( anggur matang sebagian, tekstur keras dan warna ungu atau kuning muda), dan Gambar Tambahan 1B). Sebuah chroma meter genggam (CR-400, Konica Minolta,
matang (sulur matang sepenuhnya, tekstur lembut dan warna merah atau Tokyo, Jepang) dikalibrasi terhadap pelat putih standar digunakan untuk tujuan
oranye) (Gambar Tambahan 1SEBUAH). Hijau, balik dan matang ini. Indeks warna buah segar
Larid's Large buah disingkat LLG, LLT dan LLR, masing-masing, (CI) dihitung berdasarkan rumus berikut: CI = 2000 × a*/(L × (a*2 + b*2)1/2, seperti
Mulligan buah dengan tahap pematangan yang berbeda disingkat menjadi MG yang dilaporkan oleh Gomez, Varon, Amo, Tardaguila, dan Pardo (1998).
(buah hijau), MT (buah balik) dan MR (buah matang), sedangkan AG, AT dan AR Kekerasan buah dievaluasi di area tengah setiap buah menggunakan
mewakili buah mentah, balik dan matang Amber buah, masing-masing. Penganalisis Tekstur (Stable Micro Systems Ltd, Godalming, UK) yang dilengkapi
dengan probe 5 mm dengan kecepatan 1 mm/s. Ketegasan diwakili oleh gaya
Semua buah tamarillo yang dikumpulkan ditempatkan dalam karton dan maksimum (N) yang dibutuhkan oleh probe untuk menembus buah hingga
diangkut dengan mobil ke laboratorium dalam waktu 4 jam setelah pemetikan kedalaman 25 mm melalui kulitnya. Total padatan terlarut (TSS, %) dan nilai pH
buah. Buah segera dicuci dengan air keran dan dikeringkan di udara. Sebanyak 10 jus tamarillo ditentukan menggunakan refraktometer genggam digital (Atago Co.,
buah pada setiap tahap pematangan dan kultivar dipilih secara acak, dan segera Ltd., Tokyo, Jepang) dan PerpHecT LogR pH Meter (Orion Research, Inc. , Boston,
dilakukan pengukuran warna dan tekstur. Sisa buah disimpan pada suhu 4 °C MA, USA) pada 23 °C, masing-masing. Keasaman titrasi (TA) diukur dengan
(suhu penyimpanan optimal) tidak lebih dari tiga hari sebelum persiapan sampel ( mentitrasi 10 mL jus tamarillo dengan 0,1 mol/L NaOH sampai titik akhir pH 8,2.
Schotsmans, Timur, & Woolf, 2011). TA dinyatakan sebagai g asam sitrat anhidrat/L jus (zkaya dkk., 2018).
Buah tamarillo (1 kg) dipotong-potong dan sebagian 500 g
2
2.4. Analisis gula dan asam organik Clara, CA, AS). Suhu oven dinaikkan dengan kecepatan 10 °C/menit, dari 150 °C
menjadi 210 °C (ditahan selama 7 menit), dan akhirnya naik ke suhu 230 °C
Kandungan gula dan asam organik dalam sampel jus tamarillo ditentukan dengan laju 8 °C/menit, tetap konstan selama 20 menit. Kondisi berjalan
sebagai turunan yang mudah menguap menggunakan reagen derivatisasi kromatografi lainnya adalah sebagai berikut: volume injeksi, 1 L; rasio split, 30:1;
(campuran sililasi), seperti yang dilaporkan sebelumnya (Zheng, Kallio, suhu injeksi, 260 °C; suhu detektor, 300 °C. Komponen asam lemak adalah
Linderborg, & Yang, 2011) dengan sedikit modifikasi. Secara singkat, alikuot 0,25
mL jus terong belanda dicampur dengan volume standar internal yang sama, diidentifikasi berdasarkan C4-C24 Campuran FAME sebagai standar dan
yaitu, 0,49 g/100 mL asam tartarat (untuk kuantifikasi asam organik) dan 0,49 g/ dikuantifikasi menurut area puncak relatif.
100 mL sorbitol (untuk kuantifikasi gula). Campuran ini diencerkan menggunakan
air Milli-Q hingga volume akhir 5 mL, kemudian disaring dengan spuit filter 0,45 2.6. Ekstraksi dan uji aktivitas enzim
m. Sebuah alikuot dari 300 L dibawa ke dalam botol GC dan diuapkan sampai
kering di bawah aliran nitrogen lembut pada 50 °C. Sampel yang diperoleh Ekstraksi dan uji aktivitas enzim LOX dan alkohol dehidrogenase (ADH) dalam
disimpan dalam desikator semalaman. Derivatisasi silan gula dan asam organik berbagai tahap pematangan buah tamarillo dilakukan sesuai dengan metode
dibuat dengan menambahkan 600 L reagen campuran sililasi, dikocok kuat-kuat sebelumnya (Lagu & Bangerth, 2003). Secara singkat, larutan ekstraksi LOX
menggunakan Vortex (Vortexer, Heathrow Scientific, IL, US) selama 5 menit, dan adalah buffer fosfat (0,1 M, pH 7,5)
diinkubasi dalam blok pemanas (Labnet AccuBlock Digital Dry Bath, NJ, US) selama terdiri dari 1 mM EDTA-Na2, 2 mM dithiothreitol (DTT), 1% (b/v) polivinil
30 menit pada 60 °C. Sampel kemudian disentrifugasi selama 5 menit pada polipirolidon dan 0,1% Triton X-100. Sebanyak 1 g lyo-
jaringan tamarillo bubuk philized dihomogenisasi di atas es dengan 20 mL larutan
17.217 × g dan supernatan dikumpulkan dengan hati-hati dalam botol 1,5 mL GC ekstraksi LOX. Homogenat disentrifugasi pada suhu 4 °C pada 17.217 × g selama
untuk analisis lebih lanjut. 10 menit. Kemudian supernatan dikumpulkan dan didinginkan di atas es sebagai
Turunan volatil dianalisis dengan kromatografi gas (Agilent Technologies ekstrak enzim LOX mentah. Sistem uji reaksi mengandung 10 L enzim LOX
7890A GC System, Santa Clara, CA, USA) yang dilengkapi dengan Sistem MSD Seri mentah, 140 L buffer fosfat (pH 7,5,
5975C dan kapiler HP-5MS 0,1 M), dan 50 L larutan substrat (0,25% Tween-20 dan 8,6 mM asam linoleat
kolom (30 m× 0,25 mm id × 0,25 m df) dari Agilent Technologies (Santa Clara, CA, dalam buffer fosfat). Sedangkan untuk enzim ADH kasar, metode ekstraksinya
USA). Program suhu diatur sebagai berikut: mirip dengan enzim LOX, sedangkan sistem pengujiannya mengandung 10 L
150 °C selama 2 menit, 4 °C/menit meningkat menjadi 210 °C, dan akhirnya naik asetaldehida (80 Mm), 170 L larutan NADH (0,15 mM), dan 20 L enzim ADH kasar.
ke 275 °C dengan kecepatan 40 °C/menit, bertahan selama 5 menit. Helium
dengan kemurnian tinggi diaplikasikan sebagai gas pembawa pada laju aliran Aktivitas enzim diukur secara spektrofotometri menggunakan pembaca pelat
konstan (1,4 mL/menit). Sebuah alikuot dari 1 withL sampel dengan rasio split 15 (EnSpire Multilabel Reader, PerkinElmer, Inc., Waltham, MA, US), dan pemindaian
disuntikkan ke dalam instrumen setiap analisis. Suhu injektor diatur sebagai 250 dimulai 15 detik setelah menambahkan enzim kasar ke dalam sistem pengujian.
°C. Kuantifikasi gula dan asam organik dicapai dengan menghitung rasio luas Absorbansi dicatat selama 4 menit pada interval waktu 20 detik (Lagu & Bangerth,
puncak yang diberikan oleh analit di setiap sampel terhadap luas puncak standar 2003). Absorbansi untuk uji ADH yang diukur pada 340 nm menurun sebagai
internal, dengan mempertimbangkan faktor respon. Asam tartarat dan sorbitol akibat dari oksidasi NADH, sedangkan uji enzim LOX pada 234 nm meningkat
masing-masing digunakan sebagai standar internal untuk asam organik dan gula. dalam absorbansi karena asam linoleat dapat menghasilkan hidroperoksida
Faktor respons ditentukan dengan menganalisis larutan yang mengandung selama reaksi katalitik. Satu unit aktivitas (U) mencerminkan perubahan
konsentrasi analit tunggal yang diketahui dengan standar internal di atas. absorbansi per unit dalam satu menit, yang diwakili oleh U/mg protein dalam
sampel. Uji Bradford (Bradford, 1976) diterapkan untuk menentukan kandungan
protein dari enzim kasar, menggunakan bovine serum albumin (BSA) sebagai
2.5. Penentuan kandungan asam lemak standar.
Uji -glukosidase dilakukan seperti yang dijelaskan oleh Garcia dkk.
Ekstraksi dan penentuan asam lemak dilakukan sesuai dengan Chen dkk. (2013). Bubuk tamarillo beku-kering (0,5 g) diekstraksi dengan buffer fosfat 0,1 M
(2020)dengan sedikit modifikasi. Secara singkat, 2 g buah terong terliofilisasi sitrat-0,2 M (10 mL, pH 4,0). Ekstrak ini kemudian disentrifugasi selama 20 menit
diekstraksi dengan 20 mL campuran kloroform dan metanol (1:2,v:v), dan dikocok pada 8.609 × g dan 4 ° C. Supernatan dikumpulkan dan disimpan sebagai
kuat dengan Vortex (Vortexer, Heathrow Scientific, Vernon Hills, IL, US) selama 20 -glukosidase mentah. Untuk uji aktivitas enzim, 50 L ekstrak kasar dicampur
menit. Ekstrak disentrifugasi selama 10 menit pada 17.217 × g, diikuti dengan dengan 250 LpNPG (40 mM) dan 200 L buffer, inkubasi selama 30 menit pada 40
pemisahan supernatan. Pelet diekstraksi lagi dengan 5 mL larutan ekstraksi dan °C. Selanjutnya, 1 mL
disentrifugasi pada kondisi yang sama. Selanjutnya, supernatan diambil kembali NaCO3 larutan (3 M) ditambahkan ke dalam sistem untuk menghentikan seluruh
dan ditambahkan 10 mL larutan natrium klorida (0,76% (b/v)). Campuran ini reaksi. Sebuah alikuot dari 200 L campuran dimasukkan ke dalam 96-sumur
dikocok selama 15 menit, dan kemudian fase air dipindahkan. Fase organik lempeng mikro dan yang dibebaskan pNP diukur menggunakan pembaca pelat
bawah yang tersisa diuapkan sampai kering, menghasilkan fraksi lipid. Untuk mikro pada 405 nm dengan buffer sebagai blanko. Konsentrasi rilispNP dihitung
metilasi, 1 mL larutan kalium hidroksida dalam metanol (0,4 M), 1 mL benzena/ dari kurva standar eksternal. Aktivitas -glukosidase dinyatakan sebagai
petroleum eter (rentang didih 35–60 °C), 1/1, v/v, dan 50 L metil heptadekanoat pembebasanpNP (mM)/mg kandungan protein di bawah kondisi pengujian dan
(20,74 mg/mL, standar internal) ditambahkan ke dalam asam lemak yang kandungan protein ditentukan menurut uji Bradford (Bradford, 1976).
diperoleh, setelah itu campuran dikocok selama 15 menit. Mili-Q air (5 mL)
ditambahkan ke dalam larutan ini dan kemudian campuran dipindahkan ke dalam
corong pisah. Metil ester asam lemak yang terbentuk, disingkat FAMEs, 2.7. Analisis VOC gratis
diekstraksi menggunakan 2 mL heksana. Sebuah alikuot dari fase organik atas
dimasukkan ke dalam botol GC untuk analisis instrumental lebih lanjut. Jus tamarillo (5 mL) dengan 1,5 g natrium klorida, yang memfasilitasi
pelepasan volatil, dimasukkan ke dalam botol 20 mL tutup ulir. Lima L larutan 2-
metil-3-heptanon (64,28 g/mL) ditambahkan sebagai standar internal (hasil
pemulihan 84%-95%). Botol yang disegel diinkubasi di bawah 40 °C selama 15
menit dalam kubus yang dipanaskan dengan pengocokan (pengocok interval
Analisis FAME dilakukan pada kromatografi gas Agilent 6890N (Agilent orbital, 400 rpm), setelah itu volatil yang dilepaskan dikumpulkan oleh rakitan
Technologies, Santa Clara, CA, USA) yang dilengkapi dengan deteksi ionisasi serat SPME DVB/CAR/PDMS (2 cm, 24-Gauge , 50/30 m) dari Sigma Supelco
nyala, injektor otomatis Seri 7683, dan Agilent (Bellefonte, PA, USA) selama 45 menit. Serat ini dipilih karena memiliki kinerja
Kolom kapiler FFAP (30 m × 0,25 mm id × 0,25 m df, Sinterklas yang lebih baik dalam senyawa jejak
3
adsorpsi (Li, Zheng, Liang, & Sun, 2010), dan telah diterapkan secara luas untuk diekstraksi dengan 3 mL diklorometana/pentana sebanyak tiga kali. Itu
menganalisis volatil bebas buah (Wen dkk., 2014; Yu, Yang, Yang, Zheng, & Sun, lapisan organik dipulihkan, dikeringkan di atas NaSO anhidrat4, dan dipekatkan
2019; Fu et al., 2017). hingga volume akhir 200 L dalam konsentrator bertingkat 30 mL
Analisis volatil bebas dilakukan dengan menggunakan sistem spektrometri tabung di bawah aliran nitrogen yang lembut. Aglikon yang dibebaskan disimpan
massa kromatografi gas (Shimadzu GCMS-QP2010 Plus, Kyoto, Jepang) yang pada suhu -80 ° C untuk analisis instrumental dalam 3 hari isolasi.
dilengkapi dengan kolom polar DB-WAX kolom kapiler dan kolom non-polar
DB-5MS kolom kapiler. 2.8.2. Analisis VOC terglikosilasi yang dilepaskan
(30 m× 0,25 mm id × 0,25 m df, Agilent Technologies, Santa Clara, CA, USA), secara Sebuah alikuot dari 1 L sampel disuntikkan di bawah mode splitless ke dalam
terpisah. Desorpsi senyawa dilakukan pada sistem 7890A GC dari Agilent Technologies (Santa Clara, CA, USA) ditambah
port injeksi GC di bawah 250 °C. Penjelasan rinci tentang program GC/ MS dapat dengan sistem spektrometri massa (Agilent Technologies 5975 C, Santa Clara, CA,
ditemukan dalam penelitian kami sebelumnya (Chen dkk., 2020). Mode ionisasi USA), sebuah Rtx- Kolom kapiler WAX (30 m × 0,25 mm
elektron (EI) dilakukan untuk spektrometer massa pada 70 eV dengan mode id × 0,25 m df, Restek, Bellefonte, PA, USA) atau kapiler DB-5MS
pemindaian penuh darim/z 35 sampai 350. kolom (30 m × 0,25 mm id × 0,25 m df, Teknologi Agilent, Santa Clara, CA, AS).
Volatil bebas diidentifikasi melalui perbandingan spektrum massa yang Untuk kolom Rtx-WAX, suhu oven
diperoleh dengan yang ada di perpustakaan NIST14 dan selanjutnya dikonfirmasi meningkat dari 35 °C (5 menit) menjadi 150 °C pada 5 °C/menit selama 5 menit,
dengan standar otentik. Indeks retensi Kovats (RI) dihitung menurutLucero, Estell, dan akhirnya meningkat menjadi 230 °C (10 menit) dengan laju 5 °C/menit. Untuk
Tellez, dan Fredrickson kolom DB-5MS, suhu oven dimulai pada 35 °C selama 5 menit, dinaikkan pada 5
(2009). RI yang dihitung dibandingkan dengan yang disajikan dalam Buku Web °C/menit hingga 150 °C (5 menit), dan dinaikkan ke suhu akhir 280 °C pada 8 °C/
Kimia NIST (https://webbook.nist.gov/chemistry/casser/). menit selama 2 menit. Spektrometri massa dilakukan dalam mode tumbukan
elektron pada 70 eV dengan rentang pemindaian 40 hingga 500 Da. Identifikasi
Sebelum membuat kurva kalibrasi, matriks model disiapkan yang terdiri dari dilakukan dengan pencarian database melalui perpustakaan NIST14 Mass
5,6 g/L fruktosa, 7,2 g/L asam sitrat, 4,5 g/L glukosa, 0,3 g/L asam malat, dan 3 g/L Spectral, perbandingan standar referensi, dan perhitungan indeks retensi.
sukrosa dalam Milli-Q air. Rekonstitusi, yang mengandung semua volatil bebas Kuantifikasi relatif aglikon dalam tamarillo dilakukan dengan perbandingan
yang terdeteksi, disiapkan menggunakan standar referensi dan diencerkan langsung 2-metil-3-heptanon sebagai standar internal menurut penelitian kami
menjadi delapan konsentrasi berbeda dengan matriks model. Larutan yang sebelumnya (Chen dkk., 2020). Kandungan aglikon selanjutnya dikoreksi sesuai
diencerkan ditambahkan dengan jumlah IS yang sama dan dianalisis dalam dengan kandungan yang dihitung dari re-
kondisi yang sama seperti sampel terong. Kuantifikasi senyawa volatil dilakukan sewa fenol dan jumlah fenil -D-glucopyranoside (standar internal terglikosilasi)
dengan kurva kalibrasi (Tabel Tambahan 1), dimana x mewakili rasio ditambahkan.
konsentrasi (Cx/Caku s, konsentrasi volatil/konsentrasi IS), dan kamu 2.9. Analisis statistik
mewakili rasio luas puncak (SEBUAHx/SEBUAHaku s, area puncak volatil/area
puncak IS). Persamaan untuk mengukur konsentrasi volatil bebas adalah Eksperimen yang disebutkan di atas dilakukan dalam rangkap tiga. Data
ditentukan sebagai: kapak = Sebuah( cx masukTabel 1 dinyatakan sebagai nilai rata-rata ± standar deviasi, dan dalam
ais Ci) + b, dimana Sebuah mengacu pada faktor kemiringan untuk
kurva kalibrasi, b mengacu pada intersepsi kurva, SEBUAHx dan SEBUAH Tabel 2 dan 3 disajikan sebagai nilai rata-rata dan standar deviasi relatif (standar
menunjukkan daerah puncak masing-masing standar volatil dan IS, masing-masing, Caku s deviasi/konsentrasi rata-rata). Analisis statistik dilakukan dengan menggunakan
menunjukkan jumlah IS, dan Cx mewakili konsentrasisayafs SPSS Statistics 25 (IBM Corp, Armonk, NY, USA). Data dalam tabel dibandingkan
setiap senyawa volatil. Batas deteksi (LOD), rentang validasi, dengan analisis varians satu arah (ANOVA). Uji jarak berganda Duncan diterapkan
batas kuantisasi (LOQ), dan koefisien korelasi (r) untuk setiap kurva kalibrasi juga sebagaipasca hoc uji untuk setiap data yang menunjukkan signifikansi statistik (p
ditentukan. < 0,05). Efek kultivar dan tahap pematangan pada prekursor volatil dan profil VOC
diperiksa dengan ANOVA dua arah. Analisis komponen utama (PCA) dilakukan
dengan menggunakan perangkat lunak Un-scrambler X (Versi 10.4, Camo Inc.,
2.8. Analisis prekursor volatil terglikosilasi Oslo, Norwegia) untuk memvisualisasikan hubungan antara fitokimia dan sampel.
Angka dibangun oleh GraphPad Prism V5.01 (San Diego, CA, USA).
2.8.1. Isolasi dan hidrolisis prekursor volatil terglikosilasi
Isolasi prekursor volatil terikat glikosida diadopsi sesuai dengan metode
sebelumnya dengan sedikit modifikasi (Versini, Dellacassa, Carlin, Fedrizzi, &
Magno, 2008; Chen dkk., 2020). Pertama, 3. Hasil dan diskusi
20 L fenil -D-glucopyranoside (2,072 mg/mL) ditambahkan sebagai standar internal
terglikosilasi ke dalam 50 mL jus tamarillo bening. iso- 3.1. Perubahan parameter fisik dan fitokimia selama pematangan buah
lasi prekursor glikosilasi kemudian dilakukan sebagai berikut: 12 mL metanol dan
12 mL air Milli-Q diaplikasikan untuk prakondisi kartrid Strata C18-E 500 mg/6 mL
(55 m, 70 A). Campuran yang mengandung jus tamarillo dan standar internal Warna buah merupakan indikator yang paling signifikan untuk mengevaluasi
terglikosilasi perlahan-lahan dituangkan ke dalam kartrid yang diaktifkan. kematangan dan umur simpan pascapanen, juga memainkan peran penting
Selanjutnya, 6 mL air dan 6 mL diklorometana/pentana (1/2, v/v) masing-masing dalam menentukan keputusan pembelian konsumen. Nilai CI banyak digunakan
digunakan untuk mencuci komponen yang larut dalam air dan volatil bebas. untuk menentukan tanggal panen buah segar karena dapat mencerminkan
Solusi ini dibuang. Prekursor volatil terglikosilasi yang teradsorpsi pada kartrid proses derajat selama pematangan (Gomez et al., 1998). Saat buah tamarillo
dielusi dengan 6 mL metanol, yang kemudian dipindahkan ke dalam labu alas matang, nilai CI meningkat secara dramatis, misalnya, dari 18,7 untuk LLG
bulat, dan diuapkan hingga kering di bawah vakum pada suhu 37 °C dan 122 bar menjadi 40,8 untuk LLR (Tabel 1), sesuai dengan perubahan warna dari hijau
(Rotavapor R-210 dilengkapi dengan Heating Bath B- 491, Pompa Vakum V-700, menjadi merah (Gambar Tambahan 1SEBUAH). Perubahan warna ini dikaitkan
dan Recirculating Chiller F-105, BÜCHI, Flawil, Swiss). dengan degradasi klorofil dan biosintesis pigmen karakteristik, termasuk
karotenoid dan antosianin.Granell & Rambla, 2013), selama pertumbuhan buah.
TSS menunjukkan tren yang meningkat selama pematangan buah, terutama
Fraksi kering dilarutkan dalam 5 mL buffer sitrat-fosfat (pH 5,0) dan untuk
dihidrolisis secara enzimatik pada suhu 37 °C selama 48 jam setelah Laird Besar (1,7% untuk LLG hingga 3,6% untuk LLR) dan Mulligan (1,8% untuk MG
menambahkan 500 L larutan AR2000 25 mg/mL dan 20 L standar internal, hingga 3,4% untuk MR) (Tabel 1). Peningkatan TSS dikaitkan dengan akumulasi
yaitu, 2-metil-3-heptanon (0,6428 g/µL). Volatil yang dilepaskan adalah gula karena paralel dengan perubahan total gula
4
X.Chen, dkk.
Table 1
Physical and phytochemical characteristics of three tamarillo cultivars at different ripening stages.
BF-Values
Laird’s Large Mulligan Amber
AG
T R G T R G T R Cultivar (C) Ripening Stages C×R
(R)
Color
C 47.2 ± 1.5f
L* value 60.5 ± 1.4b 51.9 ± 0.9e 58.4 ± 0.2c 51.1 ± 1.4e 48.9 ± 0.7f 57.5 ± 0.6c 62.8 ± 0.4a 55.5 ± 1.2d 89.1*** 146*** 46.8***
a* value −8.1 ± 0.36f 10.2 ± 0.8c 20.9 ± 1.0a −8.4 ± 2.4f 4.8 ± 1.0e 16.9 ± 0.8b −7.8 ± 0.3f 6.7 ± 0.3d 16.1 ± 0.4b 26.0*** 1486*** 7.5***
b* value 11.8 ± 0.3c 9.4 ± 1.1de 6.0 ± 0.1f 8.8 ± 0.1e 2.9 ± 0.3 h 4.8 ± 0.5 g 10.3 ± 0.2d 22.5 ± 0.6a 14.5 ± 1.5b 505*** 46.8*** 151***
a*/b* ratio −0.68 ± 0.04e 1.1 ± 0.1c 3.5 ± 0.2a −0.96 ± 0.3e 1.7 ± 0.3b 3.5 ± 0.2a −0.76 ± 0.04e 0.30 ± 0.00d 1.1 ± 0.1c 140*** 992*** 59.1***
chroma 14.3 ± 0.3d 13.9 ± 1.4de 21.8 ± 0.9b 12.3 ± 1.6e 5.6 ± 0.9f 17.6 ± 0.9c 12.9 ± 0.1de 23.5 ± 0.7a 21.7 ± 1.1b 136*** 138*** 67***
hue angle 124 ± 2b 42.7 ± 1.4d 16.0 ± 0.8f 133 ± 9a 31.3 ± 4.0e 15.8 ± 0.9f 127 ± 1ab 73.4 ± 0.4c 42.0 ± 2.8d 109*** 2301*** 39.9***
Colour Index (CI) −18.7 ± 0.5e 28.3 ± 1.1c 40.8 ± 1.5a – 23.7 ± 3.7f 33.4 ± 1.8b 39.4 ± 0.5a −21.1 ± 0.8ef 9.1 ± 0.3d 26.7 ± 0.6c 166*** 3199*** 54.6***
Firmness (N) 103 ± 2a 76.2 ± 1.4c 34.9 ± 3.8d 98.7 ± 6.9ab 77.3 ± 1.1c 40.8 ± 4.0d 93.6 ± 4.5b 38.4 ± 3.3d 35.5 ± 1.7d 58.5*** 649*** 30.7***
pH 4.1 ± 0.0b 3.7 ± 0.0 g 3.8 ± 0.0e 4.1 ± 0.1a 3.7 ± 0.0 g 3.7 ± 0.0 g 3.8 ± 0.0f 3.9 ± 0.0d 3.9 ± 0.0c 6.5** 537*** 341***
SSC (%) 1.7 ± 0.1 h 2.8 ± 0.1d 3.6 ± 0.0b 1.8 ± 0.1 g 2.4 ± 0.1f 3.4 ± 0.0c 2.7 ± 0.0e 3.4 ± 0.1c 3.9 ± 0.1a 462*** 1636*** 46***
D Titratable acidity (TA)
5.2 ± 0.1f 8.2 ± 0.1b 7.1 ± 0.0c 4.6 ± 0.1 h 7.1 ± 0.1c 6.3 ± 0.1e 8.4 ± 0.1a 6.6 ± 0.1d 5.0 ± 0.1 g 287*** 696*** 1363***
Sugars (mg/100 mL juice)
Fructose 10.2 ± 0.5 g 282 ± 2.3e 1201 ± 8b 11.6 ± 0.1 g 163 ± 19f 979 ± 56c 142 ± 5f 693 ± 45d 1552 ± 67a 242*** 2030*** 28.0***
Glucose 15.3 ± 0.6 h 251 ± 2e 971 ± 20b 12.6 ± 0.7 h 164 ± 5f 847 ± 1.3c 121 ± 12 g 517 ± 21d 1078 ± 32a 375*** 5977*** 40.5***
Arabitol 0.36 ± 0.03a 0.48 ± 0.07a 0.37 ± 0.04a 0.44 ± 0.01a 0.36 ± 0.08a 0.39 ± 0.03a 0.42 ± 0.05a 0.37 ± 0.07a 0.45 ± 0.09a 0.11 0.01 2.2
Sucrose 38.7 ± 0.9 g 453 ± 21e 538 ± 16b 26.6 ± 0.2 g 253 ± 55e 620 ± 26a 187 ± 16f 391 ± 21d 204 ± 14ef 17.5*** 382*** 106***
5
Total sugars 64.5 ± 0.9 h 986 ± 25e 2710 ± 12b 51.2 ± 0.4 h 580 ± 31f 2446 ± 29c 450 ± 1.0 g 1601 ± 86d 2834 ± 113a 211*** 3769*** 30.7***
Organic Acids (mg/100 mL
juice)
Malic acid 30.0 ± 0.43 cd 20.1 ± 0.0 fg 31.5 ± 1.4c 39.5 ± 0.6b 28.1 ± 2.0d 45.4 ± 1.1a 18.3 ± 0.4 g 21.6 ± 1.1e 27.5 ± 2.2d 228*** 125*** 23.9***
Citric acid 598 ± 9.8d 918 ± 20a 856 ± 6b 469 ± 21 791 ± 27c 595 ± 14d 925 ± 41a 797 ± 39c 549 ± 0.9d 88.1*** 102*** 110***
Quinic acid 0.66 ± 0.05d 1.3 ± 0.3c 2.8 ± 0.1b 0.47 ± 0.02d 1.2 ± 0.0c 2.7 ± 0.4b 0.71 ± 0.01d 1.3 ± 0.1c 4.9 ± 0.0a 46.4*** 501*** 35.7***
Total acids 629 ± 10 cd 940 ± 20a 890 ± 7a 509 ± 19e 820 ± 25b 643 ± 15c 944 ± 42a 820 ± 40b 581 ± 3d 74.9*** 90.2*** 105***
Sugars/Acids ratio 0.1 ± 0.0 h 1.0 ± 0.0e 3.0 ± 0.0c 0.1 ± 0.0 h 0.7 ± 0.0f 3.8 ± 0.1b 0.5 ± 0.0 g 2.0 ± 0.0d 4.9 ± 0.2a 351*** 4014*** 66.1***
Fatty Acids (mg/g lyophilized
fruit)
Palmitic acid (C16:0) 2.6 ± 0.1d 3.8 ± 0.3a 3.1 ± 0.2c 3.3 ± 0.1b 3.7 ± 0.2a 2.8 ± 0.0d 3.4 ± 0.1b 2.6 ± 0.1d 3.1 ± 0.1bc 5.7* 14.4*** 42.1***
Palmitoleic acid (C16:1n-7) 0.11 ± 0.00e 0.19 ± 0.02a 0.20 ± 0.01a 0.15 ± 0.01c 0.17 ± 0.01b 0.16 ± 0.01c 0.17 ± 0.00bc 0.13 ± 0.00d 0.16 ± 0.00bc 5.4* 32.6*** 49.0***
E N.D.
Heptadecenoic acid (C17:1n-7) 0.04 ± 0.00 N.D. 0.03 ± 0.00 0.04 ± 0.00 0.03 ± 0.00 0.04 ± 0.00 0.03 ± 0.00 N.D. 391*** 674*** 350***
Stearic acid (C18:0) 0.40 ± 0.03 g 0.84 ± 0.06bc 0.73 ± 0.03d 0.56 ± 0.03f 0.93 ± 0.05a 0.78 ± 0.01 cd 0.75 ± 0.03d 0.64 ± 0.01e 0.85 ± 0.02b 24.1*** 131*** 60.7***
Oleic acid (C18:1n-9) 4.6 ± 0.3d 8.3 ± 0.7a 5.9 ± 0.3c 4.7 ± 0.2d 6.6 ± 0.36b 4.9 ± 0.1d 6.9 ± 0.2b 4.9 ± 0.1d 5.8 ± 0.2c 16.5*** 38.5*** 66.2***
Linoleic acid (C18:2n-6) 14.2 ± 0.9d 24.1 ± 1.9a 15.5 ± 0.7 cd 23.4 ± 1.2a 24.8 ± 1.5a 16.8 ± 0.5c 21.4 ± 0.7b 15.5 ± 0.4 cd 17.4 ± 0.6c 37.1*** 50.9*** 52.1***
Linolenic acid (C18:3n-3) 1.8 ± 0.1b 1.8 ± 0.2bc 1.4 ± 0.1 cd 2.2 ± 0.1a 1.8 ± 0.1b 1.3 ± 0.1d 1.8 ± 0.1b 1.4 ± 0.0 cd 1.5 ± 0.0c 12.0*** 72.2*** 15.4***
Arachidic acid (C20:0) 0.10 ± 0.00c 0.15 ± 0.01a 0.12 ± 0.01b 0.11 ± 0.01c 0.15 ± 0.01a 0.13 ± 0.00b 0.12 ± 0.00b 0.10 ± 0.00c 0.13 ± 0.0b 9.3** 43.8*** 38.0***
Gondoic acid (C20:1n-9) 0.05 ± 0.00e 0.09 ± 0.01a 0.06 ± 0.01e 0.07 ± 0.01c 0.09 ± 0.01a 0.06 ± 0.01de 0.08 ± 0.00b 0.07 ± 0.00 cd 0.06 ± 0.00de 2.8* 50.5*** 18.8***
Behenic acid (C22:0) 0.09 ± 0.01d 0.19 ± 0.01a 0.15 ± 0.00bc 0.10 ± 0.01d 0.16 ± 0.01b 0.15 ± 0.01bc 0.14 ± 0.01c 0.13 ± 0.00d 0.16 ± 0.01b 1.4 101*** 41.6***
Lignoceric acid (C24:0) 0.13 ± 0.02e 0.29 ± 0.02a 0.28 ± 0.01a 0.15 ± 0.01e 0.21 ± 0.02 cd 0.23 ± 0.01bc 0.21 ± 0.01d 0.20 ± 0.00d 0.25 ± 0.01b 12.2*** 108*** 27.9***
Total fatty acids 21.4 ± 0.7e 35.7 ± 1.3a 24.1 ± 1.1 cd 31.3 ± 1.5b 34.7 ± 1.8a 24.3 ± 0.6 cd 31.4 ± 0.9b 22.8 ± 0.6de 26.1 ± 0.8c 25.0*** 69.9*** 90.7***
A R, ripe stage; T, turning stage; G, green stage. B *, **, *** indicates significant effects as p < 0.05, p < 0.01, and p < 0.001, respectively, using two-way ANOVA analysis; C Different letters in the same row indicate significant
statistical differences using Duncan’s multiple range test by one-way ANOVA analysis (p < 0.05). D Titratable acidity was expressed as g anhydrous citric acid/L juice; E N.D., not detected in samples.
Kimia Makanan 339 (2001) 128046
Meja 2
Konsentrasi VOC bebas (µg/L) dalam tiga kultivar terong belanda pada tahap pematangan yang berbeda.
SEBUAH LLG B Nilai-F
senyawa LLT LLR MG MT BAPAK AG DI AR
C jalan
jalan jalan jalan jalan jalan jalan jalan jalan Pematangan Kultivar C × R
(SD%) (SD%) (SD%) (SD%) (SD%) (SD%) (SD%) (SD%) (SD%) (C) (R)
Alkohol
cis-3-Heksenol 1537Sebuah 584e 338f 1429b 883c 342f 767d 508e 511e 77.0*** 628.0*** 85.5***
(3.8) (11) (4.9) (7.3) (7.4) (6.8) (1.5) (2.2) (3.0)
1-Heksanol 354b 20.6f 20.4f 467Sebuah 112c 33.4ef 60.9d 48.4de 106c 370*** 1574*** 491***
(4.6) (3.1) (6.7) (5.6) (2.1) (3.3) (1.67) (1.6) (0.4)
2-Etilheksanol 14.3c 11.6ef 13.4CD 27.3Sebuah 16.0b 12.3de 10.4g 10.6fg 11.2efg 382*** 181*** 157***
(7.3) (5.2) (3.3) (2.1) (2.6) (7.9) (2.0) (1.6) (4.3)
Total Alkohol 1904Sebuah 616d 372e (4.9) 1924Sebuah 1011b 387e (5.5) 838c (1.5) 567d (2.1) 628d (2.5) 155*** 1049*** 174***
(4.0) (10.8) (4.9) (6.6)
Aldehida
D ND
Pentanal 7.2b 3.5 CD 1.2ef 5.3SM 4.9c NQ 26,5Sebuah 2.4de 83.4*** 301*** 121***
(12.0) (15.6) (11.2) (19.8) (8.8) (-) (11.0) (37.1) (-)
E NQ
Heksanal 20.6b 1.1e 33.2Sebuah 3.7d 1.0e 5.9c 2.8d 6.1c 224*** 1439*** 360***
(1.9) (-) (11.1) (6.7) (7.9) (3.3) (9.6) (1.3) (5.9)
Nonanal ND 0,42c 0,61b 0,67b 0,65b 0.93Sebuah 0,66b 0,62b 0,97Sebuah 160.0*** 115*** 17.6***
(-) (2.1) (1.7) (14.3) (5.5) (7.4) (9.3) (9.3) (7.9)
Total Aldehida 27.8c 3.9 fg 2.9 g (8.2) 39.2Sebuah (8.0) 9.3d (3.5) 2.0 g (3.1) 33.1b 5.8ef 7.1de (4,5) 24,7*** 878.9*** 13.7***
(4.3) (13.8) (10.4) (15.9)
Ester
metil butanoat 4,5f 32.0d 32.1d 1.5f 102b 381Sebuah 7.3f 22.9e 51.7c 4306*** 4113*** 2515***
(9.5) (3.0) (7.4) (6.0) (7.2) (1.9) (3.4) (4.6) (2.9)
Etil butanoat 0.8f 1.9f 16.4c 0.8f 3.4e 142Sebuah 1.3f 6.4d 55.3b 2680*** 9172*** 2653***
(4.4) (2.0) (1.0) (2.3) (6.0) (2.4) (8.4) (7.3) (2.7)
metil kaproat NQ 24.1e 132c NQ 17.5ef 224Sebuah 8.5fg 71.3d 176b 96.3*** 2567*** 102,5***
(-) (7.5) (8.3) (-) (2.4) (3.8) (4.6) (3.6) (4.7)
Etil kaproat 1.5e 1.8e 3.9c 1.6e 1.6e 4.8b 1.7e 2.9d 8.9Sebuah 576*** 2342*** 268***
(0.28) (1.7) (3.8) (9.9) (7.1) (2.8) (1.7) (0.63) (3.7)
cis-3-Heksenil Asetat 4.1h 4.8ef 7.7b 4,5g 4.6fg 9.9Sebuah 5.0e 5.9d 6.6c 75.9*** 1840*** 265***
(6.1) (0.36) (0.78) (0.74) (2.3) (2.5) (1.0) (0.41) (1.7)
Heksil asetat 1.3e 1.3e 1.4c 1.3e 1.2e 1.6Sebuah 1.4c 1.4d 1.5b 26.4*** 185*** 20.7***
(2.0) (0.35) (2.7) (1.0) (0.7) (2.7) (1.1) (1.6) (1.8)
Metil benzoat 16.0 CD 13.4f 17.6b 16.7c 19.0Sebuah 19.1Sebuah 15.5d 15.6d 14.5e 92.3*** 12.2*** 32.4***
(3.2) (2.3) (5.9) (3.1) (2.4) (1.8) (2.1) (2.1) (2.9)
metil kaprilat 4.0d 4.0d 4.1c 4.0d 4.0d 4.7Sebuah 4.0d 4.1c 4.6b 63.4*** 292*** 49.2***
(1.4) (1.5) (2.9) (2.2) (2.1) (1.8) (1.1) (1.6) (3.4)
Jumlah Ester 32.2h 83.1f (1.8) 215c (4.0) 30.3h (1.9) 153d (5.2) 787Sebuah (1.6) 44.6g (1.3) 130e 319b (3.4) 2362*** 8748*** 1841***
(3.7) (0.63)
Keton
3-oktanon 3.5ab 3.3d 3.5abcd 3.4bcd 3.3 CD 3.4abcd 3.6Sebuah 3.5abc 3.5abcd 5.5* 2.5 1.3
(1.8) (0,75) (5.3) (0.81) (1.4) (1.5) (2.1) (3.6) (3.3)
Benzenoida
Benzaldehida 11.7b 6.3e 6.6e 16.8Sebuah 9.2c 6.8e 7.5de 6.4e 6.3e 26.0*** 48.5*** 10.3***
(3.2) (0.36) (1.7) (20.5) (16.0) (2.0) (5.1) (6.6) (2.8)
Metil salisilat 13.6Sebuah 8.5e 8.5e 12.3b 10.6c 9.3d 8.6e 8.5e 8.6e 362*** 604*** 231***
(2.8) (4.2) (5.5) (2.7) (3.6) (1.5) (9.2) (9.3) (8.1)
Jumlah Benzenoid 25.3b 14.7d 15.1d 29.1Sebuah 19.9c 16.1d (1.6) 16d (2.3) 14.9d 15.0d 51.0*** 96.4*** 19.7***
(2.5) (0,17) (0,38) (12.6) (7.2) (0,15) (0.36)
Terpenoid
D-limonene 1.6b 1.7b 1.9b 1.6b 1.6b 1.8b 3.1Sebuah 1.7b 1.6b 13.3*** 11.2*** 21.4***
(7.6) (2.9) (2.9) (1.1) (2.4) (9.8) (17.2) (14.9) (4.2)
Linalool 15.7Sebuah 8.2d NQ 11.9b 11.7b NQ 9.5c 12.0b NQ 5.9** 1275*** 65.2***
(8.1) (1.5) (-) (3.7) (8.5) (-) (1.3) (1.6) (-)
-Terpinena 0,98b 0,98b 1.0b 1.0b 1.0b 1.26Sebuah 1.1b 1.0b 1.3Sebuah 13.0*** 26.2*** 5.1**
(2.2) (1.5) (1.3) (8.5) (1.5) (14.1) (3.0) (1.1) (5.3)
kayu putih 0,86 g 19.4e 78.7d NQ 10.4f 108Sebuah 19.1e 82.9c 91.1b 1963*** 13447*** 1259***
(8.1) (2.1) (2.3) (-) (4.4) (1.1) (3.5) (1.4) (2.4)
-Terpineol 63.8 g 81.8f 146d 33.2 h 222c 499Sebuah 158d 130e 281b 735*** 1640*** 467***
(5.0) (1.1) (3.3) (3.4) (3.0) (4.7) (3.7) (1.2) (7.7)
Total Terpenoid 83g (5.2) 112f 228d (2.9) 47.6 h 247c (2.4) 610Sebuah (3.9) 190e (3.2) 227d 375b 820*** 2707*** 519***
(0.70) (1.4) (0,79) (0,46)
Total volatil gratis 2076Sebuah 833 g (7.8) 837 g (1.8) 2074Sebuah 1444c 1805b 1125e 948f (1.2) 1348d 374*** 388*** 160***
(3.9) (4.4) (4.7) (0.78) (1.4) (1.8)
SEBUAH LLG, LLT, LLR merupakan buah dari “Laird's Large” kultivar saat hijau, balik dan matang, masing-masing; MG, MT, MR ekspres”Mulligan” buah pada tahap hijau,
balik dan matang, masing-masing; AG, AT, AR artinya hijau, berputar dan matang”Amber” buah tamarillo. B*, **, *** menunjukkan pengaruh yang signifikan pada tingkat p <
0,05, p < 0,01, dan p < 0,001, masing-masing, menggunakan analisis ANOVA dua arah; C Konsentrasi disajikan sebagai sarana (atas) dan SD% (bawah, standar deviasi/
konsentrasi rata-rata); Nilai dengan huruf (ah) yang berbeda pada baris yang sama berbeda nyata pada taraf 0,05;D ND, tidak terdeteksi dalam sampel. E NQ, di luar LOQ,
tidak dihitung dalam sampel.
6
Tabel 3a
Alkohol, aldehida, asam, dan ester yang terikat secara glikosida dalam tiga kultivar tamarillo pada tiga tahap pematangan. Jumlah yang dinyatakan dalam g/L.
SEBUAH Indeks Retensi B Indo C Nilai-F
senyawa LLG LLT LLR MG MT BAPAK AG DI AR
Djalan
5-MS DBWAX jalan jalan jalan jalan jalan jalan jalan jalan Pematangan Kultivar C × R
Alkohol
708 1211 isoprenol RI, 22.4e 110d 1143b 23.9e 47.9de 1090b 75.5de 578c 1538Sebuah 258*** 3088*** 46.8***
NONA (7.7) (6.9) (3.1) (4.1) (1.8) (2.5) (1.3) (1.5) (6.6)
718 1159 2-Metilbutanol RI, 1.3f 2.3f 3.1f 15.2e 50.4d 122c 120c 202Sebuah 181b 4838*** 578*** 259***
NONA (9.1) (9.5) (6.9) (14,5) (1.2) (1.5) (0,55) (0.67) (5.7)
754 1178 cis-2-Penten-1-ol RI, 74.7c 160.8b 217Sebuah 12,7e 19.8de 29.3d 22.4de 28.5e 27.2d 794*** 110*** 71.6***
NONA (2.7) (0.45) (10.7) (15.3) (5.0) (6.1) (9.5) (4.8) (4.5)
850 1305 trans-3-Heksenol Std, 46.4e 43.1e 64.5 CD 23.1f 50.1de 66.3 CD 86.9b 68.8c 111Sebuah 49.9*** 23.5*** 4.6**
RI, (6.5) (6.3) (10.9) (8.9) (4.1) (19.4) (27.6) (8.5) (4.8)
NONA,
866 1299 1-Heksanol Std, 141b 51.5e 69.0d 128b 87.2c 79.2 cd 170a 141b 130b 119*** 109*** 10.1***
RI, (8.1) (7.6) (8.8) (5.1) (2.5) (3.2) (12.6) (1.3) (1.7)
NONA,
E N.D.
1012 1513 Norborneol RI, 2.5e 83.1b N.D. N.D. 60.8c N.D. 21.7d 102.7a 166*** 2973*** 57.7***
NONA (–) (10.4) (7.8) (-) (-) (5.6) (-) (0.84) (1.6)
1022 1442 2-Ethylhexanol Std, 12.3bc 27.0a 11.5bc 10.8 cd 14.3b 11.9bc 8.3de 9.5 cd 8.3d 58.4*** 47.8*** 22.5***
RI, (20.5) (7.8) (8.1) (11.2) (15) (11.7) (18.5) (6.9) (11.6)
NONA,
1157 1703 3,6-Nonadienol RI, 0.77d 1.4c 0.57d N.D. 0.80d N.D. 1.3c 16.4a 10.4b 1128*** 331.6*** 257***
NONA (18) (17.2) (18.3) (-) (12.8) (-) (22.6) (7.1) (5.3)
– 1064 3-Pentanol RI, 28.9 g 30.4 g 24.3 g 79.0f 145.3e 157.7d 232a 210b 190c 1917*** 14.5*** 77.9***
NONA (11.9) (22.3) (0.94) (10.3) (8.0) (0.86) (0.56) (2.2) (4.5)
– 1113 cis-3-Pentenol RI, N.D. N.D. N.D. N.D. N.D. N.D. 23.1b N.D. 32.7a 456.2*** 124*** 124***
NONA (-) (-) (-) (-) (-) (-) (0.93) (-) (13.8)
– 1198 Pentanol RI, N.D. N.D. N.D. 12.1b 26.6a N.D. N.D. N.D. N.D. 163*** 11.4*** 15.1***
NONA (-) (-) (-) (1.9) (22.7) (-) (-) (-) (-)
– 1324 cis-3-Hexenol Std, 856ef 988e 1689b 751f 1380c 1215d 1980a 1793b 1417c 156*** 22.5*** 77.2***
RI, (5.2) (9.2) (0.38) (5.5) (1.9) (4.9) (9.9) (1.5) (4.7)
NONA,
Total Alcohols 1,183 g 1,416f 3,304b 1,055 g 1,822e 2,831d 2,719d 3,069c 3,748a 441*** 615*** 37.0***
(3.5) (7.0) (1.2) (2.9) (0.91) (1.8) (7.4) (1.2) (5.1)
Aldehydes
793 1041 Hexanal Std, 66.2ab 68.5a 62.3ab 59.4b N.D. N.D. N.D. N.D. N.D. 531*** 63.2*** 60.7***
RI, (3.6) (2.3) (10.5) (18.6) (-) (-) (-) (-) (-)
MS,
1103 1346 Nonanal Std, 9.7e 18.8 cd 13.7de 0.75f 0.98f 48.7a 32.2b 23.2c 24.7bc 18.7*** 30.7*** 41.6***
RI, (11.4) (4.2) (4.7) (17.3) (5.2) (28.1) (1.6) (1.5) (4.3)
MS,
1163 trans-2-Hexenal Std, N.D. N.D. N.D. 18.3 N.D. N.D. N.D. N.D. N.D. – – –
RI, (-) (-) (-) 20.1 (-) (-) (-) (-) (-)
MS,
1535 trans-2, cis-6- RI, N.D. N.D. 17.7a N.D. N.D. N.D. N.D. 7.8b 7.8b 137*** 250*** 150***
Nonadienal MS (-) (-) (12.2) (-) (-) (-) (-) (3.8) (15.2)
Total Aldehydes 76.9b 87.3ab 93.7a 78.4b 1.0e 48.7c 32.5d 31.1d 32.2d 147*** 26.1*** 35.9***
(1.8) (1.8) (9.5) (16.8) (5.0) (28.1) (1.3) (2.1) (1.6)
Acids
1173 2072 Octanoic acid Std, 43.9a 17.5d 2.9e 29.3b 5.8e 16.7d 20.4 cd 18.1 cd 23.4c 5.0* 98.8*** 47.8***
RI, (8.8) (14) (4.9) (17.9) (13.8) (28.7) (9.1) (0.89) (9.1)
MS,
1268 2171 Nonanoic acid Std, 29.8d 31.4 cd 36.1bcd 54.4a 33.1 cd 37.2bc 33.6bcd 51.1a 40.3b 18.3*** 0.31 23.3***
RI, (8.5) (1.3) (8.0) (11.3) (2.6) (13.6) (8.5) (11.3) (5.4)
MS,
944 – 3-Methylvaleric RI, 3.4dc 4.1b 6.2a N.D. 2.1d 4.0b N.D. 3.89b 2.7 cd 115*** 157*** 20.7***
acid MS (10.7) (9.2) (12.2) (-) (14.2) (14.5) (-) (8.1) (4.4)
– 1605 2-Methylbutanoic RI, N.D. N.D. N.D. N.D. N.D. 23.8 N.D. N.D. N.D. – – –
acid MS (-) (-) (-) (-) (-) (7.7) (-) (-) (-)
– 1374 Acetic acid Std, 15.1c 29.2b 60.2a 50.2a 14.6c 14.6c 36.1b 15.6c 32.2b 4.8* 17.9*** 33.5***
RI, (24.5) (21.8) (14.5) (15.6) (29.5) (26.2) (27.5) (26.6) (3.2)
MS,
– 1563 Butanoic acid Std, 8.5e 15.6c 13.5 cd 14.9 cd 16.0c 15.5c 9.8de 21.7b 29.4a 18.0*** 23.6*** 10.3***
RI, (10.5) (2.7) (15.1) (16.1) (7.3) (14.6) (5.2) (22.8) (17.4)
MS,
– 1796 Hexanoic acid Std, 76.3c N.D. 95.4b 137a N.D. 31.4d N.D. 18.5e 102b 20.4*** 125*** 210***
RI, (11.3) (-) (6.8) (10.0) (-) (36.5) (-) (19.8) (1.5)
MS,
– 1881 2-Ethylhexanoic RI, N.D. N.D. N.D. 56.5 N.D. N.D. N.D. N.D. N.D. – – –
acid MS (-) (-) (-) (8.9) (-) (-) (-) (-) (-)
Total Acids 177c 97.7e 214b 341a 71.7f 143d 100e 129d 230b 28.2*** 347*** 291***
(1.6) (9.1) (7.9) (2.3) (6.3) (7.9) (6.8) (9.5) (2.3)
(continued on next page)
7
X. Chen, et al. Food Chemistry 339 (2021) 128046
Table 3a (continued)
A Retention Index B ID C F-Values

Compounds LLG LLT LLR MG MT MR AG AT AR
DAve.
5-MS DBWAX Ave. Ave. Ave. Ave. Ave. Ave. Ave. Ave. Cultivar Ripening C × R
Esters
1516 2165 Ethyl 4- RI, 52.2e 79.9d 129bc N.D. 106c 186a 74.3de 107c 144b 5.9* 161*** 19.5***
ethoxybenzoate MS (7.6) (10.7) (23.7) (-) (1.0) (6.5) (19.5) (9.7) (5.2)
– 1406 Methyl 3- RI, 5.8 g 97.3e 750a N.D. 53.6f 517c 32.8f 203d 555b 106*** 4360*** 104***
hydroxybutyrate MS (19.4) (8.9) (3.0) (-) (3.2) (4.7) (1.6) (4.9) (4.4)
– 1456 Grape butyrate RI, N.D. 32.6e 590b N.D. 16.1 fg 200c 9.0 g 94.2d 723a 763*** 5470*** 517***
MS (-) (6.9) (0.15) (-) (14.5) (3.7) (21.4) (1.7) (4.5)
– 2067 Methyl 2- RI, N.D. N.D. N.D. 28.4 N.D. N.D. N.D. N.D. N.D. – – –
methoxybenzoate MS (-) (-) (-) (5.3) (-) (-) (-) (-) (-)
Total Esters 57.9 g 210e 1,468a 28.4 g 177e 903c 116f 404d 1,422b 377*** 7398*** 158***
(4.9) (9.2) (0.80) (5.3) (2.1) (2.1) (11.6) (4.2) (4.0)
content during fruit maturation for all cultivars. In contrast, TA of the “Amber” Sugars, organic acids and fatty acids play important roles not only for their
tamarillo decreased from 8.4 to 5.0 g anhydrous citric acid/L juice. Fruit firmness direct contribution to fruit flavour, but also as precursors for the biosynthesis of
showed a decrease in all cultivars from about 100 N at the early period to around many flavour-related aroma compounds during fruit ripening (Granell & Rambla,
35 N at the end stage, indicating the softening of the flesh throughout the fruit 2013). In this study, three sugars (fructose, glucose, and sucrose), one sugar
ripening. Besides, the pH showed a small variation over the lifetime of the fruit, alcohol (arabitol), three fruit acids (quinic acid, malic acid, and citric acid) and
which was related to the total organic acids’ concentration. eleven fatty acids were identified and quantified (Table 1). Sugars accumulated
continually
Table 3b
Glycosidically bound benzenoids, furans and C13-norisoprenoids in three cultivars of tamarillo at three ripening stages. Amount expressed in µg/L.
D Ave.
Benzenoids
956 1451 Benzaldehyde Std, 10.9 cd 13.7b 12.6bc 0.6e 16.3a 16.5a 11.1 cd 10.2d 10.9 cd 6.4** 95.7*** 72.2***
RI, (3.3) (5.3) (7.9) (3.6) (8.3) (13.7) (3.5) (5.4) (6.7)
MS,
1024 1821 Benzyl alcohol Std, 1277d 1962a 1707c 1179e 1765b 1972a 1304d 1595c 1713b 13.5*** 335*** 29.2***
RI, (5.3) (4.9) (2.8) (4.0) (0.7) (2.7) (2.2) (0.4) (1.7)
MS,
1032 1573 Benzeneacetaldehyde RI, 9.0e 10.3 cd 5.8f 6.6f 9.5de 10.7c 22.4 14.1b 10.8c 418*** 78.7*** 147.3***
MS (6.3) (3.1) (9.1) (3.7) (6.4) (2.6) (5.6) (4.2) (3.1)
1107 1839 Phenylethyl Alcohol Std, 359e 525d 710a 323f 521d 618c 542 619c 660b 79.2*** 352*** 28.7***
RI, (0.9) (5.9) (6.1) (3.2) (2.8) (3.0) (1.8) (0.9) (1.9)
MS,
1209 1743 2,4- RI, 28.7b 33.8b 55.1a 30.3b 39.6ab 53.6a 25.0b 27.6b 34.4b 5.1* 12.5*** 0.99
Dimethylbenzaldehyde MS (2.5) (23.0) (5.8) (2.5) (13.6) (6.2) (5.0) (4.8) (7.7)
1252 2147 2-Methoxybenzyl RI, 1.5e 1.7e 2.6d 11.3b 12.2a 1.7e 1.5e 2.9d 5.3c 455*** 56.8*** 214***
alcohol MS (1.9) (4.4) (6.9) (5.1) (7.3) (8.0) (22.6) (2.3) (16.7)
1288 2102 Cuminol RI, N.D. N.D. 20.9b N.D. N.D. 27.4a N.D. 19.2b 24.4a 42.6*** 443*** 41.0***
MS (-) (-) (0.8) (-) (-) (16.9) (-) (5.5) (10.1)
1650 – Springaldehyde RI, 5.9f 8.1ef 15.7 cd 4.60f 9.3def 13.8 cd 12.7de 28.5b 57.8a 189*** 121*** 37.7***
MS (9.8) (8.8) (13.0) (13.7) (10.8) (15.8) (11.1) (4.2) (13.9)
Total Benzenoids 1,692e 2,555b 2,529b 1,556f 2,372c 2,713a 1,918d 2,316c 2,516b 0.99 361*** 17.1***
(4.2) (5.9) (3.7) (3.7) (1.3) (2.4) (2.1) (0.32) (1.2)
Furans
1045 2030 Furaneol Std, N.D. N.D. 56.8a N.D. N.D. 25.7c N.D. 1.1d 50.7b 105*** 2207*** 101***
RI, (-) (-) (7.2) (-) (-) (8.7) (-) (14.9) (2.9)
MS
– 1595 2-Acetyl-5-methylfuran RI, N.D. N.D. 21.7 cd N.D. N.D. 32.2a N.D. 9.3d 27.3b 26.8*** 902*** 25.2***
MS (-) (-) (16.4) (-) (-) (7.8) (-) (2.1) (3.2)
Total Furans – – 78.5a – – 57.9b – 10.4c 78.0a 44.7*** 2697*** 25.9***
(8.1) (3.4) (3.2) (2.6)
Norisoprenoids
1604 2594 3-Hydroxy-β- RI, 26.9f 74.3c 119ab 52.3d 76.3c 129a 38.1e 43.8e 110b 30.6*** 425*** 9.5***
damascone MS (1.6) (10.4) (7.7) (9.7) (7.6) (0.6) (5.8) (17.3) (6.5)
1623 2700 3-Hydroxy-7,8- RI, 1.4d 36.9c 112a 0.87d 52.6b 103a 3.6d 36.2c 58.8b 26.3*** 471*** 22.1***
dihydro-β-ionol MS (5.8) (2.3) (2.2) (16.3) (2.1) (17.1) (13.4) 2.8 (8.9)
1693 2732 3-oxo-7,8-Dihydro-α- RI, 587f 1217c 1502a 824e 1288bc 1338b 1072d 987d 1210c 2.1 138*** 33.5***
ionol MS (8.6) (2.2) (8.0) (1.3) (9.9) (1.5) (1.7) (6.4) (3.8)
1681 – 3-Hydroxy-5,6-epoxy- RI, 3.5e 7.1d 12.6b 4.4e 6.9d 13.6b 10.3c 13.6b 25.9a 131*** 182*** 8.1***
β-ionone MS (14.6) (5.2) (12.9) (33.6) (1.6) (16.6) (10.7) (11.2) (4.5)
Total Norisoprenoids 619f 1,335c 1,745a 882e 1,423c 1,583b 1,124d 1,080d 1,404c 4.1*** 226*** 35.6***
(8.1) (2.6) (7.3) (0.66) (9.2) (2.1) (1.6) (6.5) (2.5)
8
Table 3c
Glycosidically bound terpenoids, volatile phenols and others in three cultivars of tamarillo at three ripening stages. Amount expressed in µg/L.
X. Chen, et al.
D Ave.
Terpenoids
759 1263 Prenol Std, RI, 241 fg 379e 1448 cd 231 g 345ef 1637b 327 fg 955d 1762a 73.2*** 1197*** 20.9***
MS, (2.4) (6.5) (1.2) (8.9) (20.1) (7.6) (4.1) (2.0) (6.2)
1080 1424 trans-Linalool oxide (furanoids) RI, MS 71.1b 45.1e 54.3d 98.4a 64.8c 46.1e 25.4 g 24.3f 24.3f 484*** 144*** 558.7***
(5.0) 6.9 (4.2) (6.7) (4.4) (2.2) (8.5) (2.4) (9.5)
1097 1509 Linalool Std, RI, 38.2d 52.3c 35.3d 37.4d 107a 94.3b 11.5e 7.9e N.D. 1195*** 160*** 143***
MS, (2.8) (10.9) (0.5) (1.5) 5.5 (4.3) (18.6) (12.4) (-)
1171 1667 α-Phellandren-8-ol RI, MS 2.5f 15.2e 135.2c N.D. 2.6f 159b 8.9ef 47.8d 177a 83.2*** 2628*** 21.7***
(15.5) (9.5) (9.9) (-) (13.3) (2.9) (6.0) (1.5) (12.0)
1184 1804 8-Hydroxy-p-cymene RI, MS 1.9de 1.7e 2.9d N.D. N.D. 16.0a N.D. 10.7b 4.6c 70.1*** 296*** 286***
(13.0) (5.5) (1.7) (-) (-) (7.0) (-) (13.5) (8.5)
1192 1643 α-Terpineol Std, RI, 10.0e 39.9d 407b 3.6e 34.1d 406b 45.1d 122c 545a 352*** 7822*** 38.7***
MS, (5.4) (1.1) (3.3) (16.0) (0.3) (3.7) (7.1) (4.0) (2.2)
1208 1659 2,3-Pinanediol RI, MS N.D. 24.7e 103b N.D. 5.0 g 132a 12.2f 54.8d 84.7c 45.1*** 7653*** 532***
(-) (1.9) (3.0) (-) (14.7) (2.7) (5.8) (2.3) (2.7)
1224 1818 2-Hydroxy-1,8-cineole RI, MS N.D. 5.5d 211b N.D. 2.2d 237a 18.9d 101c 234a 35.6*** 846*** 16.1***
(-) (16.2) (15.5) (-) (5.7) (2.3) (10.1) (7.2) (6.4)
1241 1846 3-exo-Hydroxy-1,8-cineol RI, MS 1.1e 0.79e 65.6c N.D. N.D. 104b N.D. 34.3d 131a 74.3*** 818*** 29.6***
(18.7) (13.5) (13.9) (-) (-) (10.9) (-) (9.4) (6.6)
1326 2064 p-Mentha-1,4-dien-7-ol 8- RI, MS N.D. N.D. 16.3b N.D. N.D. 16.5b N.D. 0.6c 29.7a 35.6*** 710*** 31***
(-) (-) (9.6) (-) (-) (18.2) (-) (2.6) (7.5)
1339 2260 Hydroxylinalool RI, MS 76.2d 39.3f 78.9d 13.1 g 61.3e 101c 92.7c 127b 167a 322*** 173*** 50***
9
(13.7) (10.7) (4.6) (0.3) (15.3) (5.5) (4.8) (5.8) (4.1)
1377 2292 Sobrerol RI, MS 0.21f 69.9e 174c N.D. 1.7f 220a 70.3e 139d 199b 104*** 738*** 52***
(27.9) (8.1) (4.4) (-) (14.5) (9.9) (17.4) (8.2) (3.7)
987 – 2,4(10)-Thujadiene RI, MS N.D. N.D. N.D. N.D. N.D. 6.1a N.D. N.D. 5.9a 26.1*** 104*** 26.1***
(-) (-) (-) (-) (-) (25.4) (-) (-) (22.4)
1255 – 2,5-Bornanediol MS N.D. N.D. N.D. N.D. 3.1 cd 31.5b 1.2d 6.2c 48.0a 212*** 504*** 140***
(-) (-) (-) (-) (7.1) (0.3) (4.1) (14.5) (11.8)
1330 – 2-Acetoxy-1,8-cineole RI, MS N.D. N.D. 9.6c 3.9d 2.9e 14.0a N.D. N.D. 11.6b 165*** 1537*** 5.7**
(-) (-) (3.4) (21.2) (16.5) (2.8) (-) (-) (7.6)
1285 – 6-(1-Hydroxy-1- methylethyl)-3-methyl-2- MS N.D. 13.0e 112c N.D. N.D. 161b 9.2e 55.0d 233a 71*** 633*** 23***
cyclohexen-1-yl acetate (-) (5.4) (8.0) (-) (-) (13.2) (5.6) (2.5) (9.7)
1311 – Isogeraniol RI, MS 1.9c 1.7d 2.0c N.D. 1.8d 21.8b 1.5d 1.3d 57.8a 810*** 2060*** 835***
(14.9) (6.8) (18.5) (-) (12.9) (2.1) (18.1) (14.9) (4.9)
1342 – Limonen-1,2-diol RI, MS N.D. N.D. 3.2c N.D. N.D. 4.3b N.D. N.D. 5.0a 10.5*** 627*** 10.5***
(-) (-) (25.0) (-) (-) (7.1) (-) (-) (3.4)
1346 – Geranic acid Std, RI, N.D. N.D. 8.2b N.D. 4.2d 9.7a N.D. 1.9e 5.2c 72.5*** 779*** 42.3***
MS, (-) (-) (9.1) (-) (15.2) (4.8) (-) (11.3) (12.8)
1352 – 6-(1-Hydroxy-1-methylethyl)-3-methyl-2- RI, MS N.D. N.D. 14.8a N.D. N.D. N.D. N.D. 8.0c 38.1b 267*** 391*** 149***
cyclohexen-1-ol (-) (-) (22.8) (-) (-) (-) (-) (15.4) (6.3)
– 1149 2,3-Dehydro-1,8-cineole RI, MS N.D. N.D. N.D. N.D. N.D. N.D. N.D. N.D. 43.5 – – –
(-) (-) (-) (-) (-) (-) (-) (-) (23.1)
– 1618 Myrcenol RI, MS N.D. N.D. 39.5c N.D. N.D. 55.2b N.D. 21.5d 67.5a 342*** 2671*** 179***
(-) (-) (2.5) (-) (-) (3.4) (-) (6.1) (3.8)
– 1878 2,6-Dimethyl-3,7-octadiene-2,6-diol RI, MS N.D. 23.2e 129b N.D. N.D. 160a 15.2e 36.4d 107c 48.4*** 568*** 17.5***
(-) (5.9) (11.8) (-) (-) (1.4) (10.9) (9.3) (0.7)
Total Terpenoids 444f(2.7) 711e(3.4) 3,048c 387f(4.1) 637e(12.2) 3,630b 639e(2.1) 1,754d 3,981a 362*** 6633*** 79.7***
(2.8) (2.8) (1.6) (2.1)
(continued on next page)

Food Chemistry 339 (2021) 128046
Table 3c (continued)

X. Chen, et al.
D Ave.
Volatile Phenols
1042 2077 p-Cresol RI, MS 2.1d 7.2c 12.1b N.D. N.D. 6.8c N.D. 0.23e 14.0a 81.5*** 413*** 33.4***
(3.0) (22.4) (7.2) (-) (-) (20.3) (-) (20.0) (5.9)
1075 1800 Guaiacol Std, RI, 38.4b 54.0a 29.1c 28.9c 24.8d 28.0c 13.1e 9.2f 7.8f 1248*** 82.6*** 111***
MS, (1.1) (2.6) (2.9) (6.2) (3.8) (4.7) (18.9) (1.8) 6.4
1188 1701 Methyl salicylate Std, RI, 67.3a 70.6a 59.0b 25.9c 26.0c 23.6c 7.1d 20.6c 25.8c 270*** 3.2 7.3***
MS, (2.7) (12.8) (5.9) (4.5) (7.3) (7.3) (21.0) (45.1) (9.9)
1213 2130 2-Phenoxyethanol Std, RI, 1.5d 0.5d N.D. 34.2a 3.9c 1.7d N.D. N.D. 20.2b 263*** 182*** 425***
MS, (7.2) (4.2) (-) (5.1) (1.7) (12.5) (-) (-) (15.0)
1304 2176 2-Methoxy-4-vinylphenol Std, RI, 23.5f 45.9e 231b 16.6f 51.2e 214c 45.7e 83.5d 421a 350*** 2827*** 148***
MS, (5.1) (3.7) (2.5) (18.9) (9.0) (4.9) (9.1) (2.4) (4.6)
1348 2156 Eugenol Std, RI, N.D. N.D. 39.9b N.D. 0.27e 24.0c 0.21e 5.7d 70.8a 181*** 1441*** 125***
MS, (-) (-) (13.0) (-) (6.5) (6.3) (28.1) (29.4) (2.8)
1388 2592 Vanillin Std, RI, 3.2d 65.9c 80.8bc 67.7c 90.4b 111a 88.7b 67.5c 110a 48.5*** 53.4*** 14.5***
MS, (6.0) (13.5) (3.2) (19.0) (14.5) (15.3) (8.7) (13.9) (0.2)
1515 – Methyl gentisate RI, MS 235b 266a 118d 161c 159c 152c 119d 92.5e 52.4f 182*** 71.4*** 25.9***
(14.1) (4.6) (4.8) (3.2) (8.1) (1.4) (4.7) (9.6) (1.1)
1641 – Dihydroconiferyl alcohol RI, MS 296f 381e 1124b 449d 598c 1243a 9.2 h 138 g 373e 1320*** 1768*** 111***
(11.5) (6.9) (1.8) (3.7) (6.7) (0.8) (9.8) (10.4) (9.4)
1729 2992 Coniferol RI, MS 614e 586e 3214b 740e 1246d 3712a 1357 cd 1529c 3870a 60.2*** 883*** 3.3***
(3.2) (5.7) (4.4) (8.8) (1.3) (6.2) (6.9) (10.6) (7.8)
1586 – Methoxyeugenol RI, MS N.D. N.D. 1.3b N.D. N.D. 4.0b N.D. N.D. 23.5a 5244*** 9880*** 5244***
(-) (-) (19.4) (-) (-) (2.3) (-) (-) (1.8)
10
1594 – Antiarol RI, MS 1.9e 2.9e 4.4d N.D. 6.2c 8.4b 5.1 cd 6.1c 12.6a 79.5*** 126*** 19.7***
(11.6) (5.5) (3.4) (-) (18.9) (13.5) (22.6) (13.1) (9.3)
– 1603 2-Hydroxybenzaldehyde RI, MS N.D. 9.1d 18.3b N.D. 12.3c 22.3a N.D. 4.8e 3.3e 77.0*** 77.5*** 23.5***
(-) (24.0) (16.1) (-) (17.5) (11.0) (-) (8.8) (4.8)
1968 – Sinapyl aldehyde RI, MS 90.5gh 115 fg 217d 61.0 h 145ef 296c 172de 551b 1151a 793*** 548*** 195***
(2.3) (4.4) (2.1) (18.0) (9.0) (13.7) (23.6) (0.9) (5.4)
– 1917 o-Cresol RI, MS N.D. N.D. 21.7b N.D. N.D. 36.4a N.D. N.D. N.D. 130*** 39.9*** 39.9***
(-) (-) (13.9) (-) (-) (15.6) (-) (-) (-)
Total Phenols 1,374e 1,604de 5,169b 1,584de 2,363c 5,882a 1,816d(5.7) 2,508c 6,156a 56.3*** 1766*** 3.5*
(0.41) (0.77) (2.2) (5.5) (2.5) (3.5) (8.1) (5.5)
Others
1467 2576 Unknown 1 (59, 79, 93, 109,152,169) – N.D. 75.7c 526.1b N.D. N.D. 614.3a N.D. N.D. 105.0c 138*** 797*** 117***
(-) (4.9) (9.0) (-) (-) (9.1) (-) (-) (11.3)
1484 – Unknown 2 (55, 70, 79, 91, 95, 105, 121, – N.D. 1.7d 45.9b N.D. N.D. 2.0c N.D. 19.5d 94.3a 169*** 315*** 96.0***
135) (-) (13.9) (7.7) (-) (-) (0.7) (-) (6.1) (13.2)
1936 – Scopoletin RI, MS 8.5e 3177b 4456a 2258c 2154c 4128a 390d 2254c 1920c 32.2*** 117*** 24.7***
(6.2) (9.3) (16.2) (5.8) (31.1) (6.6) (3.4) (4.7) (12.6)
Total Others 8.5d(6.2) 3,255b(9.1) 5,028a 2,258c 2,154c 4,744a 390d(3.4) 2,274c 2,119c 38.3*** 153*** 27.9***
(14.9) (5.8) (31.1) (5.7) (4.6) (11.5)
Total Bound Volatiles 5,632e 11,270c 22,680a 8,172d 11,019c 22,536a 8,855d(3.6) 13,576b 21,688a 16.5*** 1651*** 16.5***
(2.5) (3.64) (4.3) (2.3) (7.1) (2.3) (1.9) (3.6)
A Calculated retention index on DB-5MS capillary column and RTx-WAX capillary column. B Compounds identification methods: MS-mass spectra comparison with published data and MS library (NIST14); RI-retention index
agreed with the data reported in previous papers or a database on the web (http://www.odour.org.uk/lriindex.html); Std-confirmed by authentic standards. Compounds identified without authentic standards are considered as
tentatively identified. C *, **, *** indicates significant effects at the level of p < 0.05, p < 0.01, and p < 0.001, respectively, using two-way ANOVA analysis; D Values with different superscript letters (a-h) in the same row are significantly
different at the level of 0.05. E N.D., not detected in samples.
Food Chemistry 339 (2021) 128046
throughout fruit maturation and the total sugars content reached a maximum those for LLT and LLR (833 and 837 µg/L, respectively). Among many types of free
value ranging from 2446 to 2834 mg/100 mL in ripe tamarillo, depending on the volatiles, the alcohols decreased significantly, from 1904 µg/L in LLG to 372 µg/L
cultivars. Fructose was the predominant sugar for all varieties and the generation in LLR (Table 2); As the most abundant compounds in tamarillo, cis-3-hexenol
of fructose from unripened to fully ripened stage of Laird’s Large fruit was 2- and decreased approximately 4.5- fold during the “Laird’s Large” ripening process,
8-fold greater than that of glucose and sucrose, respectively. Among the from 1537 µg/L to 338 µg/L. Besides, a relatively small decrease was observed for
identified organic acids, citric acid was the major one for all ripening stages; the aldehydes and phenols. Among all of the classes, the content of ketones (3-
especially in the turning stage, its concentration (791–918 mg/100 mL) was more octanone) was not changed significantly.
than 28-fold higher than the second most abundant acid, namely malic acid (20.1
to 28.1 mg/100 mL). The sugars/acid ratio was measured by dividing the total Esters were plentiful as free volatile compounds in all the ripe tamarillo
sugars content by the total acids content, which has been reported as one of the cultivars. In total, eight esters were detected, with concentrations varying from
most important indicators reflecting the desirable taste and sweetness of the 215 μg/L to 787 μg/L, depending on the cultivars. Among them, ethyl butanoate,
fruits (Zheng et al., 2011). It is evident that eating quality of tamarillo harvested at methyl butanoate, and methyl caproate accounted for the most abundant esters,
the ripe stage is better than the turning and green fruit because of the significant providing powerful pineapple- like, apple-like, and fruity odours in tamarillo fruit.
higher sugars/acid ratio. The formation of esters is related to acyl esterification of alcohols derived from
fatty acids or amino acids via the action of alcohol acyltransferase (Granell &
Rambla, 2013). The substrate availability and endogenous enzyme activities are
Fatty acids have multiple biochemical and physical functions including important factors in this process, all of which are highly regulated by fruit
energy storage, metabolic promotion, and biological structure construction (Song maturation (Granell & Rambla, 2013; Defilippi et al., 2009). The total contents of
& Bangerth, 2003). Besides, fatty acids are involved in the biosynthesis of many fatty acids and free alcohols generally decreased during the ripening of “Amber”
types of free volatiles, especially the “green leaf volatiles”, a family of six carbon and “Mulligan” fruit (Tables 1
aldehydes, alcohols, and derived esters, which related to fresh “green” and typical
fruity notes during fruit maturation (Defilippi et al., 2009). In this study, eleven and 2), and this indicates the consumption of precursors during metabolic
fatty acids were determined after methyl esterification, and three of them were pathways to form the final product, i.e., esters, which was consistent with the
reported for the first time, namely heptadecenoic acid (C17:1n-7), gondoic acid increased content of free esters as the fruit developed (Table 2). This result was
(C20:1n-9) and behenic acid (C22:0). The majority of fatty acids in tamarillo were also supported by previous research (Yang, Zheng, Yu, & Sun, 2019). Notably,
linoleic acid (C18:2n-6), oleic acid (C18:1n-9), palmitic acid (C16:0), and linolenic since high concentrations of straight chain C6 compounds (cis-3-hexenol, 1-
acid (C18:3n-3), which is consistent with previous findings (Dorado Achicanoy, hexanol, and hexanal) were also observed in the present research, it provides
Hurtado Benavides, & Martínez-Correa, 2018). The changes of fatty acids content more support to the earlier speculation that the LOX pathway could be one of the
during fruit ripening were different among the cultivars. For the “Larid’s Large” main volatile formation mechanisms.
and “Mulligan” cultivars, the highest total concentration was observed at the
turning stage (35.7 mg/g and 34.7 mg/g, respectively), while for the “Amber” Volatile terpenoids are an important group of aroma compounds in fruits due
cultivar, the fatty acid content peaked at the green period (31.4 mg/g). Since to their appreciated floral notes, although they only comprise a small proportion
linoleic acid was present in the highest concentration in all samples regardless of of free volatiles in most fruits. In this study, five terpenoid compounds were
cultivars and ripening stages, it was hypothesised that fatty acid metabolism, detected, including limonene, linalool, γ- terpinene, eucalyptol, and α-terpineol.
especially the LOX pathway, could be the most critical mechanism for generating Generally, the concentration of the individual terpenoids increased during
the free volatile compounds in tamarillo. To confirm this, the free volatiles’ maturation and peaked at the ripe stage. The highest increase was recorded for
contents and the activities of enzymes involved in LOX metabolism need to be in- eucalyptol as ripening progressed, and this was particularly notable for the “
vestigated. Laird’s Large” cultivar, which showed a 91-fold increase of content from LLG (0.86
μg/L) to LLR (78.7 μg/L). The exception to this maturation-related increase was
linalool, observed for all “Laird’s Large”, and limonene, noticed for “Amber”, and
the contents of these two terpenoids decreased significantly with the growth of
the fruit. It is worthwhile to note that terpenoids are widely reported as glycosidic
3.2. Analysis of free and glycosylated VOCs during fruit ripening bound volatiles in fruit, and their concentrations are dramatically higher than
those of the related free forms. For example, Özkaya et al. (2018) discovered six
3.2.1. Development of free VOCs terpenoids in tomato cultivars grown in Turkey and among them, only one
The free volatile compounds in the “Laird’s Large”, “Mulligan” and “Amber” (geranial) was present as the free form. Another study showed geranyl acetone, β-
cultivars at three ripening stages were analysed by gas chromatography-mass damascenone and neral present at significantly higher concentrations as bound
spectrometry. A total of 22 free volatile metabolites were detected and quantified form in raisins prepared from different grape cultivars (Thompson, Flam and
using the corresponding calibration curves (Supplementary Table 1). These Crimson) (Wang et al., 2015). Additionally, Wen et al. (2014) discovered a 13-fold
compounds included 3 alcohols, 3 aldehydes, 8 esters, 1 ketone, 2 phenols, and 5 higher content of bound geraniol than its free counterpart in the “Rainier” cherry
terpenoids, with significant difference of concentrations among cultivars (Table 2 and this strengthened the floral note of the fruit after liberation. Our previous
). For the ripe fruit, the predominant volatile classes were alcohols, esters and research (Chen et al., 2020) has shown that bound volatiles in ripe “Laird’s Large”
terpenoids, accounting for 21% to 47%, 24% to 44%, and 27% to 34% of total free fruit are more abundant than their free forms (30 free volatiles vs. 49 bound
volatiles, respectively, for the three cultivars of “Laird’s Large”, “Mulligan” and “ volatiles); especially, the concentrations of linalool and α-terpineol were three
Amber”. times higher compared with their free counterparts after enzymatic hydrolysis.
The effect of ripening on the accumulation of glycosylated volatile metabolites is
The ripening process has been extensively studied in fruits like tomatoes ( reported in the following section.
Ortiz-Serrano & Gil, 2010), raspberry (Yu et al., 2019), and mandarins (Gao et al.,
2018), and the change of individual volatile compounds was observed to be
different during fruit maturation. Generally, the concentration of alcohols and
aldehydes was found to decrease, while esters accumulated significantly toward
the ripening of fruit, as also observed in current study (Table 2). During tamarillo
fruit ripening, the total content of free volatiles showed a decreasing trend in the 3.2.2. Development of glycosylated VOCs
cultivars of “Laird’s Large” and “Mulligan”; for example, the free volatile The glycosidically bound volatile compounds were much more abundant than
compounds in LLG was 2076 µg/L, about 2.5 times higher than the free volatiles in the tamarillo fruit (Table 3). A total of 83 compounds were
detected in three cultivars of tamarillo digested
11
with glycosidase AR2000. These volatiles can be classified into ten groups, transformation process is mainly dependent on endogenous β-glucosidase
including 12 alcohols, 4 aldehydes, 8 acids, 4 esters, 8 benzenoids, 2 furans, 4 activity and substrate availability.
norisoprenoids, 23 terpenoids, 15 volatile phenols, and 3 defined as others. The The profile of glycosylated esters was totally different from that of the free
number of bound volatiles identified in tamarillo was much higher than those ester metabolites: a total of four esters were identified in bound fraction with
reported in literature, for example: 51 aglycones detected in strawberries (Ubeda methyl-3-hydroxybutyrate as the most abundant compound (Table 3a). While for
et al., 2012), 56 in mandarins (Gao et al., 2018), 32 presence in tomatoes (Özkaya the free volatile metabolites shown in
et al., 2018), and 25 reported for banana cultivars grown in French West In- dies ( Table 2, eight esters were identified and mainly dominated by methyl butanoate
Aurore et al., 2011). and methyl caproate. However, the change of esters in both free and bound
forms showed a consistent increasing trend with ripening of the fruit.
Glycosylated volatile metabolites composition and its changes throughout
fruit maturation have been found to vary according to fruit tissue, cultivar, and Benzenoids, derived from the amino acid pathway, have played an important
species. In milk bubble (Rubus corchorifolius), the bound alcohols were found to role in defense systems of plant against pathogens (Granell & Rambla, 2013). In
be predominate, but their total concentration decreased significantly as the fruit this study, eight benzenoids were identified as re- leased in the bound fraction,
developed (Yang et al., 2019). Acids are an important glycosidic class in kiwifruit. with the highest concentrations found for benzyl alcohol and phenylethyl alcohol,
In “Hay- ward” kiwi cultivar, some of the detected acids increased in con- which showed an obvious increase as fruit ripened (Table 3b). Benzyl alcohol has
centration during fruit ripening from the under-ripe stage to the over- ripe stage, been described as having flowery and sweet notes, but its odour thresholds (2546
while in “Hort16A” variety, the content of most acids increased at the early ripe μg/L) is higher than the concentrations detected in all the cultivars. Compar- ably,
stage and later dropped during the postharvest period (Garcia et al., 2013). For “ phenylethyl alcohol is capable of contributing the honey- and rose-like odour to
Kendington Pride” mangoes, 85 glycosidic volatiles were present in the skin, tamarillo juice due to its low threshold (60 μg/L) (Burdock, 2016; Van Gemert, 2011
much more than those in the pulp (29 aglycones detected). The most plentiful ).
bound volatiles in the skin of mango were aromatic amino acid metabolites,
including ben- zenemethanol, 2-xylyl ethanol and phenylacetaldehyde, whilst in Norisoprenoids are important aroma compounds with extremely low sensory
the mango pulp, terpenoids were observed to be predominant, accounting for thresholds derived from the degradation of carotenoids (Liu, Zhu, Ullah, & Tao,
38% of total liberated aglycones (Lalel, Singh, & Tan, 2003). The majority of 2017), contributing to the varietal character- istic of many wines, especially
glycosylated volatile precursors in mango pulp increased in concentration during aromatic varieties such as Riesling (Skouroumounis & Winterhalter, 1994). There
fruit ripening, which were similar to the trend observed here with tamarillo. were four norisoprenoids identified in the bound fraction, including 3-
hydroxy-7,8- dihydro-β-ionol, 3-oxo-7,8-dihydro-α-ionol, 3-hydroxy-β-damascone,
and 3-hydroxy-5,6-epoxy-β-ionone (Table 3b), which are firstly reported for “
Mulligan” and “Amber” cultivars. These norisoprenoids, except, 3-hydroxy-β-
Considering the ripe tamarillo fruit, “Laird’s Large” and “Mulligan” were similar damascone, are discovered for the first time in “Laird’s Large” fruit. Glycosylated 3-
in terms of bound volatiles numerically represented (both 73 types, and 68 types oxo-7,8-dihydro-α-ionol was the most plentiful norisoprenoids in all cultivars, and
in common), as well as the total concentrations, it was continually accumulated during fruit maturation, for example, from 587 μg/
22,680 μg/L and 22,536 μg/L, respectively. The cultivar “Amber” was found to be L in LLG increased to 1502 μg/L in LLR. However, the odour thresholds for these
mainly composed of volatile phenols (28.4%), followed by terpenoids (18.4%), norisoprenoids are not available in the literature, thus their influence on the
alcohols (17.3%) and benzenoids (11.6%). For all cultivars, compounds with the aroma cannot be readily assessed.
highest contents (≥1000 μg/L) were glycosides of isoprenol, cis-3-hexenol, benzyl
alcohol, 3-oxo-7,8-dihydro-α-ionol, prenol, coniferol, and scopoletin. All
compound classes except acids increased in concentration during fruit ripening, Furaneol has been reported as a main odour-active compound for strawberry
which was especially significant from the ‘turning’ to ‘ripe’ stage. This variation and pineapple (Ubeda et al., 2012), and is characterised by sweet, brown and
was obvious for volatile classes of esters, terpenoids, and volatile phenols, whose caramel-like aroma with a extremely low odour threshold of 0.03 μg/L (Burdock,
accumulation between middle-stage (turning) to ripen were 2 to 3 folds greater 2016). Furaneol was more likely to be existed as glycosylated form in fruit, for
than that from early season (green) to the ‘turning’ stage, with the consideration example, 39 μg/ kg in “Ca- marosa” strawberry (Ubeda et al., 2012), 60.5 μg/ kg in
given to the “Laird’s Large” cultivar. Garcia et al. (2013) reported similar findings ripe“Hayward” kiwifruit (Garcia et al., 2013) and 3.2 μg/kg in “Vitis cinerea” grape (
when studying bound aroma profile in “Hayward” kiwifruit during ripening. They Ghaste et al., 2015), but it has not been previously reported in tamarillo. It is
discovered that the contents of all compound classes except acids showed surprising to discover such high concentrations of bound furaneol (25.7–56.8 μg/
increased trends during ripening stage, followed by subsequent decreases at the L) in the ripe tamarillo juice (Table 3b), as this fruit is recognised for acidic, fresh,
postharvest stage. and green flavours rather than su- gary and candy like. This might be related to
the fact that furaneol remained mainly as an odourless glycosidically bound form
throughout the fruit ripening process, as shown by the free volatile profile (Table 2
There were 12 alcohols detected in the glycosidically bound fraction but only ), and therefore it cannot contribute immediately to the aroma of the fruit. On the
three (cis-3-hexenol, 1-hexanol and 2-ethylhexanol) were present as free forms in other hand, furaneol was not detected in the green
the tamarillo juice. Among the bound alcohols, three of them (isoprenol, 1-
hexanol and cis-3-hexenol) had concentration that exceeded their thresholds
(550 µg/L, 5.6 µg/L and 3.9 µg/L, respectively) (Burdock, 2016; Van Gemert, 2011) “Amber” tamarillo fruit, but an obvious increase in concentration was detected
in all stages of fruits or only in ripe fruit. These three alcohols were significantly during the end of “Amber” fruit ripening, from 1.1 μg/L in AT to 50.7 μg/L in AR.
different in concentrations among the three cultivars, and the highest concentra- This might be related to the key genes that regulate furaneol generation, for
tions of 1-hexanol and cis-3-hexenol were found in AG (170 µg/L and 1980 µg/L example, the FaQR gene, which participated in the biosynthesis of furaneol in
respectively). 1-hexanol and cis-3-hexenol can provide strong “green”, strawberry fruits. The expression level of
“herbaceous” and “leaf” notes (Burdock, 2016) to tamarillo juice once liberated by FaQR gene increased significantly during strawberry fruit maturation, which led to
glycosidase. The free cis-3-hexenol decreased with the ripening in the “Laird’s accumulation of furaneol dramatically at a later stage (Fu et al., 2017).
Large” and “Amber” fruits (Table 2), while in the bound fraction the trends were
the opposite (Table 3a). This suggests that cis-3-hexenol is inclined to attach to the Volatile phenols and terpenoids were the most abundant bound volatiles
hydroxyl groups of sugar compounds via glycosidic bonds and accu- mulates as present, accounting for 22.8% to 28.4% and 13.4% to 18.4% of bound extracts,
an aroma reservoir with growth of the fruit. The respectively, for the ripe fruit of all cultivars. The three ripe tamarillo cultivars
could be distinguished by the concentrations of volatile phenols: 6,156 μg/L for “
Amber”, 5,882 μg/L for “Mulligan” and
12
linalool increased dramatically as the “Mulligan” fruit ripened, from

37.4 μg/L at MG to 107 μg/L for MT (Table 3c), while its free counterpart
remained unchanged for this period and decreased to zero in ripe “Mulligan” fruit (
Table 2). Throughout the fruit maturation process, both the bound and free α-
terpineol accumulated (Tables 2 and 3c), but the content of bound formwas more
than two times higher than the free version in the ripe “Laird’s Large” and “Amber
” fruits. These results suggest that linalool and α-terpineol are more likely to be
bound as glycosylated volatile precursors as the fruit ripened, and these gener-
ated bound forms can give a floral and fruity aroma when liberated by hydrolysis
during fruit processing, due to the low thresholds of aglycones (2.0 μg/L for
linalool and 4.6 μg/L for α-terpineol) (Burdock, 2016; Van Gemert, 2011).
3.3. Distribution of phytochemicals in different samples
Principal component analysis (PCA) was conducted to visualize the

relationships between phytochemicals and tamarillo samples (Supplementary Fig.
2). Different classes of phytochemicals were coded in Supplementary Table 2. The
first three principal components (PCs) explained 73% of the total variance, with
PC-1 and PC-2 accounting for 47% and 14%, respectively.
A clear separation of the nine different samples was observed along PC-1.
Ripe fruits (LLR, MR and AR), located at the right part of PC-1, was characterized by
high abundance of most glycosylated VOCs, particularly higher bound
terpenoids, alcohols, and phenols. Unripe fruits including MG and LLG, were
distributed at the upper left corner which showed correlations with most of the
free VOCs, especially free aldehydes, and alcohols. This is in accordance with
high concentrations of free C6 alcohols (cis-3-hexenol and 1-hexanol) in green
fruits. It is worth noting that the turning fruits (MT and LLT) and green “Amber”
which occupied the lower left quadrant, were correlated with most of fatty acid
compounds, particularly linoleic acid (18:2n6). Also, the presence of linolenic acid
(C18:3n3) in the upper left quadrant indicated possible correlations with green
fruits.
The cis-3-hexenol, 1-hexenol and hexanal are all known as “green leaf
volatiles (GLVs)”, capable of imparting “fresh green” “leaf-like” and “herbaceous”
odours to most of fruits and vegetables. The fatty acid derivatives pathway,
particularly the LOX pathway, is the main source of GLVs and their derived esters
in plants (Granell & Rambla, 2013).
This pathway starts with C18 polyunsaturated free fatty acids (FFAs), linolenic
(C18:3n3) and linoleic acid (18:2n6), which are oxidized by
LOX into 9- or 13-hydroperoxides (HDPOs) according to specific cata- lytic position
of the substrates by the enzyme. This is followed by the subsequent catalysis of
HDPOs to produce aldehydes with six or nine carbons by hydroperoxide lyase
(HPL). The aldehydes produced can be further reduced to alcohols, catalysed by
ADH. In the last step of the LOX pathway, alcohol acyltransferase (AAT) catalyses
the biosynthesis of esters, which contribute to the generation of typical fruity
aroma in many fruits (Chen et al., 2020; Defilippi et al., 2009). During fruit ri-
pening, the LOX pathway is favourable because of the changes in endogenous
enzyme activity and/or substrate levels. For example, higher availability of free
Fig. 1. Changes in the activity of key enzymes during tamarillo fruit ripening. LOX,
fatty acids with the increase of membrane perme- ability have been reported
lipoxygenase; ADH, alcohol dehydrogenase. Statistically significant differences as
during the maturation of pear (Chen et al., 2020) and tomatoes (Klee, 2010).
determined by one-way ANOVA with Tukey’s multiple comparison test are
indicated (*, p < 0.05; **, p < 0.01; ***, p < 0.001).
5,169 μg/L for “Laird’s Large” (Table 3c). Some of the compounds, namely p-cresol,
3.4. Key enzyme activities and their correlations with other metabolites during
guaiacol, and eugenol had significantly higher concentrations exceeded their
the entire maturation period
thresholds (3.9 μg/L, 0.48 μg/L and 25 μg/L, respectively), and thus they provide
phenol-like, sweet, and strong clove-like odour to tamarillo fruit, respectively (
Combining the results obtained, the LOX pathway of fatty acid metabolism is
Burdock, 2016). The content of coniferol was higher than 3,000 μg/L in all ripe
suggested to be one of the most important biosynthesis mechanisms to produce
tamarillo varieties, but it is impossible to evaluate its contribution to the overall
free volatiles, as well as glycosylated volatile precursors. To confirm this, key
aroma profile because its odour threshold has not been reported. Li- nalool and α-
enzyme activities including LOX, ADH and β-glucosidase, were examined during
terpineol were the only two terpenoids detected in both free and bound fractions
tamarillo fruit ripening (Fig. 1). Generally, the activities of LOX and ADH showed a
among all cultivars. The content of glycosidic
consistent trend among all cultivars, increased as the fruit ripened and peaked in
13
the ripe fruit. However, the activity of β-glucosidase in tamarillo had the opposite
trend, where the highest activity was found in the unripen tamarillo, especially in
the “Mulligan” and “Amber” cultivars (Fig. 1). The result obtained for LOX activity
resembles that recorded for Korla
fragrant pears, but the activity of ADH in pear showed a later decrease instead of
a continuous increase as the fruit maturated (Chen et al., 2020). Unlike the results
1
obtained by Garcia et al. (2013), in which β- glucosidase activity kept almost
constant throughout the “Hayward” kiwifruit maturation, the activity of β-
glucosidase in tamarillo changed significantly for all cultivars during fruit
−0.783*
ripening, especially from ‘green stage’ to ‘turning stage’ (Fig. 1). This suggests
1
that, different from kiwifruit, the β-glucosidase activity and the content of
Bound Alcohols Bound Aldehydes Bound Esters Linoleic Acid Linolenic Acid Palmitic Acid Oleic Acid ADH Activity LOX Activity Glycosidase Activity
substrates, for example glycosylated precursors, could be crucial factors for

−0.800** aroma development in tamarillo fruit.
0.617
1
To further explore the relationship between enzyme activities and the

concentrations of metabolites (fatty acids, free volatiles, and glycosidic volatiles)
during the entire ripening period, pairwise correlation analysis was carried out
−0.410
0.092
0.293
using the Spearman rank (Table 4,

1
Spearman rank correlation test was applied, and correlations were statistically significant (*, p < 0.05; **, p < 0.01); LOX, lipoxygenase; ADH, alcohol dehydrogenase.
Supplementary Tables 3 and 4). Overall, the correlations of the free alcohols and
aldehydes with linolenic acid were noticeably greater than those with other fatty
acids (p < 0.01) in all three cultivars. Generally, the “Mulligan” cultivar (Table 4)
−0.544
−0.577
0.563
0.377
presented significantly higher correlation coefficients and noticeable trends

Correlation analysis for free volatiles, glycosidically bound volatiles, fatty acids and key enzymes involved in LOX pathway during “Mulligan” fruit ripening.
regarding enzyme activities and the changes of free and bound volatiles than the “
Larid’s Large” (Supplementary Table 3) and “Amber” (Supplementary Table 4) culti-
vars. Referring to the “Mulligan” cultivar, the ADH and LOX activities showed
−0.879**
−0.745*
0.879**
strong negative correlations with the free alcohols and aldehydes (p < 0.05 and p
−0.361
0.546
< 0.01, respectively) and strong positive correlations with free esters and bound
1
alcohols (p < 0.01), while the glycosidase activity was at the opposite trend (Table 4
). These results suggest that in “Mulligan” tamarillo, the free alcohols and
aldehydes tend to be derived from linolenic acid, but they could also be further
0.983**
−0.494
−0.661
0.597
0.479
0.427
consumed during the subsequent LOX pathway for the production of the final
1
product, i.e., the free esters. Interestingly, the absence of free alcohols and
aldehydes is more likely to be compensated from glycosylated volatile
precursors under the action of endogenous β-glucosidase. Based on these
−0.870**
−0.867**
0.967**
−0.586
−0.502
results, we have proposed a schematic biosynthesis of volatile metabolites in the “

0.351
0.650
Mulligan” tamarillo fruit involving fatty acids pathway, as shown in Fig. 2.

1
−0.908**
−0.477
−0.252
−0.370
−0.075
−0.444
0.513
0.536
1
4. Conclusion
Tamarillo fruit maturation involved an increment in colour index, softening of

−0.904**
−0.900**
0.883**
0.833**
0.850**
flesh, enhancement of taste, and generation of free VOCs and their glycosylated
−0.460
−0.469
0.452
0.368
precursors. Quantitative differences in free and glycosidically bound aroma

1
compounds were observed for the three tamarillo cultivars studied. The LOX
pathway is a crucial biosynthesis mechanism for the generation of free and
−0.929**
−0.917**
0.950**
0.883**
0.883**
0.800**
−0.460
−0.444
−0.427
glycosylated volatile metabolites during tamarillo ripening. The biosynthesis is

0.410
Free Alcohols Free Aldehydes Free Esters
highly correlated with the key enzymes’ activities, including LOX, ADH and β-
1
glucosidase, as well as the concentration of substrates, especially linolenic acid.

Future studies are required to illuminate further details, such as the expression of
genes associated with the LOX pathway and studies on the function of other
−0.883**
−0.883**
−0.917**
−0.917**
−0.767*
0.812**
0.817**
−0.343
precursors (e.g. amino acids and carotenoids) in the generation of odour-active

0.385
0.510
0.460
compounds in tamarillo.
1
−0.917**
−0.917**
−0.833**
−0.883**
−0.783*
0.883**
0.929**
−0.259
0.427
0.611
0.577
CRediT authorship contribution statement

Glycosidase Activity 0.833**
1
Xiao Chen: Investigation, Formal analysis, Writing - original draft.

Bound Aldehydes
Bruno Fedrizzi: Methodology, Supervision, Writing - review & editing.

Bound Alcohols
Free Aldehydes
Linolenic Acid
Free Alcohols
Bound Esters
Palmitic Acid
Paul A. Kilmartin: Supervision, Writing - review & editing. Siew Young Quek:
Linoleic Acid
ADH Activity
LOX Activity
Free Esters
Oleic Acid
Project admiration, Supervision, Writing - review & editing.

Table 4
14
Fig. 2. Biosynthesis of volatile compounds derived from the fatty acid pathway in tamarillo fruit. LOX, lipoxygenase; ADH, alcohol dehydrogenase; AAT, alcohol
acyltransferase; HPL, hydroperoxide lyase.
Declaration of Competing Interest quantities of protein utilizing the principle of protein-dye binding. Analytical
Biochemistry, 72, 248–254. https://doi.org/10.1016/0003-2697(76)90527-3.
Burdock, G. A. (2016). Fenaroli's handbook of flavor ingredients (15th ed.). New York: CRC
The authors declare that they have no known competing financial interests or Press.
personal relationships that could have appeared to influence the work reported Chen, J., Lu, J., He, Z., Zhang, F., Zhang, S., & Zhang, H. (2020). Investigations into the
production of volatile compounds in Korla fragrant pears (Pyrus sinkiangensis Yu). Food
in this paper.
Chemistry, 302, 125337. https://doi.org/10.1016/j.foodchem.2019.125337.
Chen, X., Quek, S. Y., Fedrizzi, B., & Kilmartin, P. A. (2020). Characterization of free and
Acknowledgements glycosidically bound volatile compounds from tamarillo (Solanum betaceum Cav.) with
considerations on hydrolysis strategies and incubation time. LWT-Food Science and
Technology, 124. https://doi.org/10.1016/j.lwt.2020.109178.
The authors wish to thank Mr. Craig Watson for providing the tamarillo fruits Defilippi, B. G., Manríquez, D., Luengwilai, K., & González-Agüero, M. (2009). Aroma
for the research and the technical support given by Mr. Tony Chen, Ms. Mansa volatiles: Biosynthesis and mechanisms of modulation during fruit ripening. Advances in
Botanical Research, 50, 1–37. https://doi.org/10.1016/s0065-2296(08)00801-x.
Nair and Ms. Sreeni Pathirana. The authors also wish to acknowledge the China
Dorado Achicanoy, D., Hurtado Benavides, A., & Martínez-Correa, H. A. (2018). Study of
Scholarship Council (P.R. China) for awarding a PhD scholarship to the first author supercritical CO2 extraction of tamarillo (Cyphomandra Betacea) seed oil containing high
(Xiao Chen) and the PBRF fund from the School of Chemical Sciences, Faculty of added value compounds. Electrophoresis, 39, 1917–1925. https://doi.org/10.
1002/elps.201700430.
Science, the University of Auckland, for supporting the purchase of chemical
Durant, A. A., Rodríguez, C., Santana, A. I., Herrero, C., Rodríguez, J. C., & Gupta, M. P.
standards for this work. (2013). Analysis of volatile compounds from Solanum betaceum Cav. fruits from pa- nama
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of yellow and purple-red Ecuadorian cultivars of tree tomato (Solanum betaceum
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07.131.
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responses of four biosynthetic pathways of aroma compounds in postharvest strawberry (
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beranda jurnal: www.elsevier.com/locate/foodchem

Pengembangan senyawa organik volatil dan prekursor glikosilasi mereka

INFO ARTIKEL ABSTRAK

⁎ Penulis yang sesuai.

Buah tamarillo (1 kg) dipotong-potong dan sebagian 500 g

(continued on next page)

A Retention Index B ID C F-Values

Compounds LLG LLT LLR MG MT MR AG AT AR

(continued on next page)

A Retention Index B ID C F-Values

linalool increased dramatically as the “Mulligan” fruit ripened, from

3.3. Distribution of phytochemicals in different samples

Principal component analysis (PCA) was conducted to visualize the

substrates, for example glycosylated precursors, could be crucial factors for

To further explore the relationship between enzyme activities and the

using the Spearman rank (Table 4,

presented significantly higher corre- lation coefficients and noticeable trends

results, we have proposed a schematic biosynthesis of volatile metabolites in the “

Mulligan” tamarillo fruit involving fatty acids pathway, as shown in Fig. 2.

Tamarillo fruit maturation involved an increment in colour index, softening of

precursors. Quantitative differences in free and glycosidically bound aroma

glycosylated volatile meta- bolites during tamarillo ripening. The biosynthesis is

glucosi- dase, as well as the concentration of substrates, especially linolenic acid.

precursors (e.g. amino acids and carotenoids) in the generation of odour-active

CRediT authorship contribution statement

Xiao Chen: Investigation, Formal analysis, Writing - original draft.

Bruno Fedrizzi: Methodology, Supervision, Writing - review & editing.

Project admiration, Supervision, Writing - review & editing.

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