MAKALAH FITOKIMIA

TEKNIK ISOLASI SENYAWA METABOLIT SEKUNDER MINYAK ATSIRI PADA

TUMBUHAN AMORPHA FRUCTICOSA

Disusun Oleh :

1. Ahmad Misriani (164820144650002)

2. Eprilia Cahya Ainun
3. Nadia Krismanda Y S
4. Nur Khasanah
5. Olivia Febri Lestari
6. Tri Wulandari
(164820144730010)
(164820144950032)
(164820144980035)
(164820144990036)
(164820145060043)

SEKOLAH TINGGI ILMU KESEHATAN HARAPAN BANGSA PURWOKERTO

PRODI S1 FARMASI
2018

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 KATA PENGANTAR

Segala puji bagi Allah SWT yang telah memberikan nikmat serta hidayah-Nya terutama
nikmat kesempatan dan kesehatan sehingga penulis dapat menyelesaikan makalah“ISOLASI
MINYAK ATSIRI . Kemudian shalawat beserta salam kita sampaikan kepada Nabi besar kita
Muhammad SAW yang telah memberikan pedoman hidup yakni al-qur’an dan sunnah untuk
keselamatan umat di dunia. Makalah ini merupakan salah satu tugas mata kuliah
FITOKIMIA.Penulis menyadari bahwa banyak terdapat kekurangan-kekurangan dalam penulisan
makalah ini, maka dari itu penulis mengharapkan kritik dan saran yang konstruktif dari para
pembaca demi kesempurnaan makalah ini.

Purwokerto, 7 Juli 2018

Penulis

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 DAFTAR ISI

KATA PENGANTAR ....................................................................................................1

DAFTAR ISI ...................................................................................................................2

BAB I PENDAHULUAN

A. Latar belakang .........................................................................................................3

B. Rumusan masalah ...................................................................................................4

C. Tujuan……………………………………………………………………….…….4

BAB II PEMBAHASAN

2.1 Pengertian Minyak Atsiri........................................................................................5

2.2 Isolasi minyak atsiri pada Amorpha Fructicosa......................................................5

A. Penyulingan minyak atsiri ....... ......................................................................6

B. Destilasi…………………………………………………………………...…7

C. Ekstraksi……………………………………………………………………..8

BAB III PENUTUP

A. Kesimpulan .............................................................................................................13

DAFTAR PUSTAKA .....................................................................................................14

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 BAB I

PENDAHULUAN

A. Latar belakang

Hampir seluruh tanaman enghasil minyak atsiri yang saat ini tumbuh di wilayah Indonesia
sudah dikenal oleh sebagian masyarakat. Bahan beberapa jenis tanaman minyak atsiri menjadi
bahan yang sangat penting dalam kehidupan sehari-hari. Minyak atsiri dihasilkan dari bagian
jaringan tanaman tertentu seperti akar, batang, kulit, daun, bunga, buah, atau biji. Sifat minyak
atsiri yang menonjol antara lain mudah menguap pada suhu kamar, mempunyai rasa getir, berbau
wangi sesuai dengan aroma tanaman yang menghasilkannya, dan umumnya larut dalam pelarut
organic.

Banyak istilah yang digunakan untuk menyebut minyak atsiri. Misalnya dalam bahasa
inggris disebut assential oils, ethereal oils dan volatile oils. Dalam bahasa Indonesia
ada menyebutnya minyak terbang, bahkan ada pula yang menyebut minyak kabur. Mengapa
dikatakan demikian ? tidak lain karena minyak asiri mudah mengup apabila dibiarkan begitu dalam
keadaan terbuka. Dari sekitar 70 jenis minyak atsiri yang selama ini di perdagangkan di pasar
dunia, ternyata diantaranya dapat di produksi oleh Indonesia. Namun kenyataanya, sampai dengan
tahun 1993 barulah tercatat sekitar 14 jenis minyak asiri produksi Indonesia yang cukup nyata
peranannya sebagai komoditas ekspor. Hal ini tentunya merupakan tantangan karena potensi
Indonesia untuk mngembangkan minyak asiri sebenarnya luar biasa. Peluang pemasaran minyak
asiri juga tidak hanya terbuka untuk pasar luar negeri, melainkan sangat dibutuhkan didalam
negeri.

Didalam perdagangan internasional, masing-masing minyak asiri memiliki nama dagang

tersendiri. Hal ini bergantung pada jenis tanaman yang menghasilkannya. Misalnya, minyak asiri
yang berasal dari tanaman alpukat bernama dagang avocado oil, minyak jahe (ginger oil), minyak
akar wangi (vetiver oil), minyak cengkeh (clove oil), minyak adas (fennel oil ), dan
sebagainya.Kegunaan minyak asiri sangat luas dan spesifik, khususnya dalam berbagai bidang
industry. Banyak contoh kegunaan minyak asiri, antara lain dalam industry kosmetik ( sabun, pasta
gigi, sampo, losion ). Dalam industry makanan digunakan sebagai bahan penyedap atau penambah

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cita rasa, dalam industry parfum sebagai pewangi dalam berbagai produk minyak wangi, dalam
industry farmasi atau obat-obatan ( anti nyeri, antiinfeksi,pembunuh bakteri ) dalam industry bahan
pengawet, bahkan digunakan pula sebagai insektisida. Oleh karena itu, tidak heran jika minyak
asiri banyak di buru berbagai Negara.

B. Rumusan masalah

1. Apa pengertian dari minyak atsiri ?

2. Bagaimana isolasi minyak atsiri pada tanaman Amorpha Fructicosa ?

C.Tujuan

1. Dapat mengetahui pengertian minyak atsiri

2. Dapat mengetahui isolasi minyak atsiri pada Amorpha Fructicosa

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 BAB II

PEMBAHASAN

2.1 Pengertian Minyak Atsiri

Minyak atsiri adalah sejenis komponen berminyak yang mudah menguap yang
merupakan metabolit sekunder yang dihasilkan oleh tanaman aromatik; itu adalah air
mampu dan dapat disuling dengan uap untuk pemisahan. Minyak atsiri juga dikenal sebagai
minyak halus dan dicirikan sebagai komponen yang mudah menguap, alami, cair,
kompleks dengan bau aromatik yang kuat . Dalam beberapa tahun terakhir, minyak esensial
telah banyak digunakan dalam industri kosmetik dan kehidupan sehari-hari karena aroma,
serta di industri farmasi karena berbagai efek farmakologis positif . Karena aplikasi minyak
esensial yang luas, permintaan pasar akan minyak esensial telah meningkat luar biasa.
Metode yang umum dan paling sering digunakan untuk pemisahan minyak esensial adalah
hydrodistillation (HD) tetapi metode ini selalu disertai dengan efisiensi ekstraksi rendah
karena konsumsi waktu yang lama, hilangnya beberapa oksi-komponen terlarut dalam air
kondensat selama proses distilasi, seperti keton dan aldehida, dan beberapa komponen yang
sensitif secara termal yang ditemukan melalui efek panas, seperti -pinene dan camphene
Oleh karena itu, hydrodistillation dengan bantuan gelombang mikro (MHD) telah
dikembangkan untuk meningkatkan efisiensi ekstraksi untuk minyak esensial yang dapat
mengurangi durasi ekstraksi dan meningkatkan hasil minyak esensial karena karakteristik
iradiasi gelombang mikro . Secara umum, terpen dalam minyak esensial yang diperoleh
dengan dua metode yang disebutkan di atas relatif terkonsentrasi, tetapi komponen oksi
relatif kurang. Oksigen-komponen dalam minyak esensial cenderung memiliki nilai
aplikasi lebih tinggi daripada komponen bebas oksigen
2.2 Isolasi minyak atsiri pada Amorpha Fructicosa
Dari jurnal yang kami dapat diperoleh isolasi minyak atsiri dengan menggunakan
buah Amorpha Fruticosa. Amorpha fruticosa adalah semak gugur abadi dari keluarga
Leguminosae yang berasal dari Amerika Utara dan diperkenalkan di Cina sekitar tahun
1920-an.Tumbuhan Ini banyak ditanam di Cina untuk tujuan pengendalian erosi dan
restorasi gurun. A. fruticosa telah digunakan sebagai obat tradisional untuk pengobatan

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 luka bakar, ambustion, carbuncle dan eczema , dan pemanfaatan terapeutiknya dapat
berasal dari awal peradaban Penelitian sebelumnya telah mengungkapkan bahwa ia kaya
dalam sejumlah senyawa, termasuk rotenoids , stilbenes , flavanon dan isoflavon sehingga
menyebabkan antimikroba , antioksidan , antitumor antidiabetes dan aktivitas kekebalan.
Isolasi adalah proses pengambilan atau pemisahan senyawa bahan alam
dengan menggunakan pelarut yang sesuai (Djamal, 2008). Sejak abad ke-17 orang
telah dapat memisahkan berbagai jenis senyawa dari sumber-sumber organik.
Senyawa-senyawa tersebut dapat berupa senyawa metabolit primer dan senyawa
metabolit sekunder (Lenny, 2006).
Metabolit sekunder merupakan senyawa kimia yang terdapat dalam suatu
organisme yang tidak terlibat secara langsung dalam proses pertumbuhan,
perkembangan atau reproduksi organisme seperti terpenoid, steroid, kumarin,
flavonoid dan alkaloid. Senyawa metabolit sekunder dapat berasal dari tumbuhan,
hewan maupun mikro organisme (Herbert, 1996).
Teknik isolasi minyak atsiri terdiri dari beberapa metode, antara lain :

A. Penyulingan Minyak Atsiri

Salah satu cara untuk meng-isolasi minyak atsiri dari bahan tanaman penghasil minyak
atsiri adalah dengan penyulingan, yaitu pemisahan komponen yang berupa cairan dua macam
campuran atau lebih berdasarkan perbedaan titik didih. Proses tersebut dilakukan terhadap
minyak atsiri yang tidak larut dalam air. Berdasarkan kontak antara uap air dan bahan yang
akan disuling, metode penyulingan minyak atsiri dibedakan atas tiga cara, yaitu:
1. Penyulingan dengan air,
Bahan yang akan disuling kontak langsung dengan air mendidih. Mengapung atau
terendamnya bahan tersebut tergantung dari bobot jenis dan jumlah bahan yang
disuling. Metode ini dilakukan dengan panas langsung,mantel uap,pipa uap yang
berlingkar tertutup tatau dengan memakai pipa uap berlingkar terbuka atau berlubang

2. Penyulingan dengan uap dan air,

Bahan olah diletakkan di ata rak-rak atau saringan berlubang. Ketel suling diisi dengan
air sampai permukaan air berada tidak jauh di bawah saringan. Air juga dapat
dipanaskan dengan ap jenh yang basah dan bertekanan rendah. Ciri khas metode ini

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 yaitu uap selalu dalam keadaan basah, jenuh, dan tidak terlalu panas;bahan yang
disulng hanya berhubungan dengan uap dan tidak dengan air panas.
3. Penyulingan dengan uap.
Air tidak diisikan dalam ketel. Uap yang digunakan adalah uap jenuh atau uap kelewat
panas pada tekanan lebih dari 1 atmosfer. Uap dialirkan melalui pipa uap berlingkar
yang berpori dan terletak di bawah bahan dan uap bergeerak ke atas melalui bahan
yang terletak di atas saringan.
Proses utama yang terjadi pada peristiwa hidro destilasi yaitu :
a. difusi minyak atsiri dan air panas melalui membran tanaman (hidrodifusi)
b. Hidrolisa terhadap beberapa komponen minyak atsiri
c. Dekomposisi yang biasanya disebabkan oleh panas (Guenther,1987).

Penyulingan dengan air serta penyulingan dengan uap dan air lebih sesuai bagi industri
kecil karena lebih murah dan konstruksi alatnya sederhana. Namun penyulingan dengan uap
dan air memiliki kelemahan, yaitu membutuhkan uap air yang cukup besar. Hal ini karena
sejumlah besar uap akan mengembun dalam jaringan tanaman sehingga bahan bertambah
basah dan mengalami aglutinasi. Untuk mengatasi kelemahan ini, telah dikembangkan model
pe-nyulingan uap dan air yang dikombinasikan dengan sistem kohobasi. Pada sistem ini
pemanasan air dalam ketel penyulingan dilakukan secara langsung terhadap dasar ketel.
Dengan sistem ini, bahan bakar dapat dihemat sampai 25%, karena air yang digunakan hanya
40% dari yang normal.
B. Destilasi
Destilasi adalah suatu pemurnian senyawa organik cair yaitu suatu proses yang didahului
dengan penguapan senyawa cair, kemudian mengembunkan uap yang terbentuk sehingga
mencair kembali. Proses yang dilakukan yaitu larutan diuapkan pada alat uap yang kemudian
mengental kembali membentuk cairan. Itu jelas bahwa zat pengotor non-volatil mungkin dapat
dipisahkan dengan metode ini. Ketika dua atau lebih unsur volatil dari campuran bisa
dipisahkan dengan destilasi (Sugihara, 1961).
Macam-Macam Destilasi :
1) Destilasi Uap

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 Proses penyaringan suatu campuran air dan bahan yang tidak larut sempurna atau larut
sebagian dengan menurunkan tekanan sistem sehingga didapatkan hasil penyulingan jauh
dibawah titik didih awal.
(Cahyono, 1991)
2) Destilasi Vakum
Untuk memurnikan senyawa yang larut dalam air dengan titik didih tinggi sehingga
tekanan lingkungan harus diturunkan agar tekanan sistem turun.
(Cahyono, 1991)
3) Destilasi Biasa
Untuk memurnikan campuran senyawa dimana komponen-komponen yang akan
dipisahkan memiliki titik didih yang jauh berbeda.
(Cahyono, 1991)

Prinsip Destilasi Uap

Campuran substansi yang tidak larut menunjukkan reaksi yang sangat beda dalam
larutan homogen dan deskripsi sifatnya memerlukan hukum fisik yang berbeda. Dasar aturan
dapat dipakai dengan mempertimbangkan akibat naiknya deviasi pada hukum rault. Satu
gejala dari deviasi positif adalah dalam diagram hubungan antara tekanan dengan temperatur.
Pada batas deviasi positif besar dari hukum rault, dua komponen dapat larut dan komponen
tersebut menguap yang secara matematis memberikan tekanan total yang merupakan jumlah
total dari tekanan masing-masing (Wilcox, 1995).

C. Ekstraksi
Ekstraksi yang sering digunakan untuk memisahkan senyawa organik adalah
ekstraksi zat cair, yaitu pemisahan zat berdasarkan perbandingan distribusi zat tersebut yang
terlarut dalam dua pelarut yang tidak saling melarutkan. Yang paling baik adalah dimana
kelarutan tersebut dalam pelarut satu lebih besar daripada konsentrasi zat terlarut dalam
pelarut lainnya, harga K hendaknya lebih besar atau lebih kecil dari satu ekstraksi jangka
pendek disebut juga proses pengorokan, sedangkan pada proses jangka panjang menggunakan
soxhlot dan dengan pemanasan (Wasilah, 1978).

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 Kriteria pemilihan pelarut:
a. Pelarut mudah melarutkan bahan yang di ekstrak
b. Pelarut tidak bercampur dengan cairan yang di ekstrak
c. Pelarut mengekstrak sedikit atau tidak sama sekali pengotor yang ada
d. Pelarut mudah dipisahkan dari zat terlarut
e. Pelarut tidak bereaksi dengan zat terlarut melalui segala cara. (Cahyono, 1991)

Prinsip Ekstraksi pelarut

Ekstrasi adalah proses pemindahan suatu konstituen dalam suatu sample ke suatu pelarut
dengan cara mengocok atau melarutkannya. Ektraksi pelarut bisa disebut ekstraksi cair-cair yaitu
proses pemindahan solut dari pelarut satu ke pelarut lainnya dan tidak bercampur dengan cara
pengocokkan berulang. Prinsip dasar dari ekstraksi pelarut ini adalah distribusi zat terlarut dalam
dua pelarut yang tidak bercampur(Ibrahim,2009).
Di dalam penelitian ini dilakukan pengambilan minyak atsiri buah Amorpha Fruticosa dengan
metode microwave hydrodistillation. Metode microwave hydrodistillation menggunakan
gelombang mikro yang dihasilkan dari magnetron sebagai sumber pemanasan selama proses
ekstraksi. Gelombang mikro atau microwave adalah gelombang elektromagnetik dengan frekuensi
super tinggi (Super High Frequency, SHF), yaitu antara 300 Mhz – 300 Ghz. Microwave memiliki
rentang panjang gelombang dari 1 mm hingga 1 m.
Mekanisme dasar pemanasan microwave melibatkan pengadukan molekul polar atau ion yang
berosilasi karena pengaruh medan listrik dan magnet yang disebut polarisasi dipolar. Dengan
adanya medan yang berosilasi, partikel akan beradaptasi dimana gerakan partikel tersebut dibatasi
oleh gaya interaksi antar partikel dan tahanan listrik. Akibatnya partikel tersebut menghasilkan
gerakan acak yang menghasilkan panas. Keunggulan dalam pemilihan microwave sebagai media
pemanas karena microwave bisa bekerja cepat dan efisien. Hal ini dikarenakan adanya gelombang
elektromagnetik yang bisa menembus bahan dan mengeksitasi molekul-molekul bahan secara
merata. Gelombang pada frekuesnsi 2450MHz (2,45 GHz) ini diserap bahan. Saat diserap, atom-
atom akan tereksitasi dan menghasilkan panas. Proses ini tidak membutuhkan konduksi panas
seperti oven biasa. Maka dari itu, prosesnya bisa dilakukan sangat cepat. Disamping itu,
gelombang mikro pada frekuensi ini diserap oleh bahan gelas, keramik, dan sebagian jenis plastik.

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Selain itu, kelebihan lain metode microwave hydrodistillation yaitu lebih hemat biaya karena tidak
digunakan pelarut n-Hexane pada proses ekstraksi. Pelarut yang digunakan pada metode
microwave hydrodistillation adalah air karena memiliki konstanta dielektrik yang tinggi sehingga
penyerapan gelombang mikro lebih optimal.
Bahan yang diguunakan dalam jurnal kami adalah Etil eter, n-heksana, petroleum eter
(rentang didih 30–60 ◦C), sikloheksana dan n-pentana dibeli dari Tianjin Fuyu Fine Chemical Co.,
Ltd. (Tianjin, Cina). Natrium sulfat anhidrat dibeli dari Aladdin (Shanghai, Cina). Air osmosis
Reverse MilliQ (Millipore, Billerica, MA, USA) digunakan untuk persiapan dari semua solusi
sampel.uhu kamar, dan sampel bahan yang sama digunakan untuk semua tes.
Mempersiapkan peralatan Sebelum distilasi, MHD-LLE yang digunakan dalam penelitian
ini terdiri dari oven microwave dan peralatan ekstraksi cair cair hidrodistilasi yang digabungkan.
Microwave oven WP700 (Glanz, Zhongshan, China) digunakan untuk pemanasan, dengan
frekuensi penyinaran 2,45 GHz, dan ukuran rongga interior 215 mm x 350 mm x 330 mm.
Kekuatan iradiasi microwave dibagi menjadi lima tingkat, termasuk 100% (700W), 77% (540W),
55% (385W), 33% (230W), dan 17% (120W). Peralatan ekstraksi cair-cair penggabungan
hidrodistilasi yang dibentuk berdasarkan pada perangkat Clevenger. Diagram skematik dan
dimensinya yang rinci yang terdiri dari dua kolom pemisahan untuk pemisahan minyak esensial.
Kolom pemisahan pertama adalah proses pemisahan minyak-air konvensional, dan kolom
pemisahan kedua adalah proses ekstraksi cair-cair untuk isolasi komponen oleh pelarut organik.
1) Hydrodistillation dengan bantuan gelombang mikro

prosedur ekstraksi cair-cair

50,0 g buah A. fruticosa ditimbang secara akurat dan volume tertentu air deionisasi (untuk
mencapai rasio cairan-padat yang dibutuhkan) ditempatkan ke dalam blender Waring untuk waktu
homogenat yang berbeda, berdasarkan penelitian sebelumnya. Kemudian, suspensi benar-benar
dipindahkan ke labu alas bulat 1000 mL, yang ditempatkan ke dalam oven microwave dan
dilekatkan pada peralatan ekstraksi cair-cair terpadu yang dikembangkan dengan hidrodistilasi dan
kondensor melalui lubang di bagian atas oven microwave. Sebelum distilasi, volume dan jenis
pelarut organik yang berbeda ditambahkan ke kolom reparasi kedua untuk ekstraksi komponen
yang dilarutkan dalam air. Setelah campuran sampel mulai mendidih, suspensi kemudian disinari
oleh microwave untuk berbagai waktu di bawah kekuatan yang berbeda. Komponen volatil dapat

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dilepaskan dari sampel dan diuapkan dengan uap air, kemudian dikondensasikan dan dipisahkan
dari air, yang diselesaikan dalam kolom pemisahan pertama. Namun, beberapa komponen minyak
esensial sering dilarutkan dalam air untuk membentuk hidrosol. Ketika hidrosol mencapai banjir,
ia akan kembali ke kolom pemisahan kedua untuk ekstraksi beberapa komponen oleh pelarut
organik melalui kontak antara hidrosol dan ekstraksi pelarut. Air akan kembali ke labu sampel
secara konstan ketika mencapai tingkat banjir.

Setelah setiap proses, minyak esensial yang diperoleh dalam kolom pemisahan pertama
dikumpulkan, dikeringkan dengan menggunakan natrium sulfat anhidrat, dan disimpan pada suhu
4 ◦C sampai analisis. Pelarut ekstraksi dalam kolom pemisahan kedua dikeringkan dengan natrium
sulfat anhidrat semalam dan kemudian dipekatkan untuk mendapatkan minyak esensial di bawah
tekanan yang dikurangi, yang disimpan pada 4 ◦ C sebelum analisis. Dalam penelitian ini, rasio
cair-padat, waktu homogenat, jenis dan volume pelarut ekstraksi di kolom pemisahan kedua, waktu
iradiasi microwave dan daya diselidiki secara sistematis. Hasil minyak esensial yang diperoleh
dalam dua kolom pemisahan dihitung sebagai berikut:

Y = M/ Mo

dimana Y (g / kg) adalah hasil dari minyak atsiri; M (g) adalah massa rata-rata minyak esensial
yang diperoleh dalam dua kolom pemisahan, dan Mo (kg) adalah massa rata-rata dari tiga sampel
buah sebelum distilasi.

2) Hidrodistilasi konvensional dan bantuan gelombang mikro

metode hidrodistilasi:

50,0 g serbuk buah A. fruticosa (40–80 mesh, digiling menggunakan disintegrator dan disaring
sebelumnya) ditimbang secara akurat dan 600 mL air deionisasi ditambahkan ke dalam labu
bundar 1000 mL, dan kemudian labu itu dilekatkan pada Clevenger.

Ketika labu mulai mendidih, untuk HD, labu dipanaskan dengan pemanasan jaket selama 4 jam di
bawah 540W, menurut Chinese Pharmacopoeia untuk MHD, labu itu diradiasi oleh microwave
selama 1 jam di bawah 540W. Setelah distilasi, minyak esensial yang diperoleh di atas air dalam
tabung koleksi dikumpulkan, didehidrasi, dikeringkan, ditutup dan disimpan pada suhu 4 ◦C
sampai analisis.

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A. faktor yang mempengaruhi
Dalam penelitian ini, beberapa faktor yang berpengaruh,yaitu :
a. Jenis dan volume pelarut ekstraksi di kolom pemisahan kedua.
b. Waktu homogenate
c. Rasio cair-padat
d. Daya iradiasi microwave
e. Waktu iradiasi microwave

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 BAB III
PENUTUP

A. KESIMPULAN
1. Minyak atsiri adalah sejenis komponen berminyak yang mudah menguap yang
merupakan metabolit sekunder yang dihasilkan oleh tanaman aromatic.
2. Isolasi adalah proses pengambilan atau pemisahan senyawa bahan alam dengan
menggunakan pelarut yang sesuai (Djamal, 2008). Sejak abad ke-17 orang telah dapat
memisahkan berbagai jenis senyawa dari sumber-sumber organik.
3. Metode microwave hydrodistillation menggunakan gelombang mikro yang dihasilkan
dari magnetron sebagai sumber pemanasan selama proses ekstraksi. Gelombang mikro
atau microwave adalah gelombang elektromagnetik dengan frekuensi super tinggi (Super
High Frequency, SHF), yaitu antara 300 Mhz – 300 Ghz. Microwave memiliki rentang
panjang gelombang dari 1 mm hingga 1 m.
4. Keunggulan dalam pemilihan microwave sebagai media pemanas karena microwave bisa
bekerja cepat dan efisien. Hal ini dikarenakan adanya gelombang elektromagnetik yang
bisa menembus bahan dan mengeksitasi molekul-molekul bahan secara merata. Selain
itu, kelebihan lain metode microwave hydrodistillation yaitu lebih hemat biaya karena
tidak digunakan pelarut n-Hexane pada proses ekstraksi. Pelarut yang digunakan pada
metode microwave hydrodistillation adalah air karena memiliki konstanta dielektrik yang
tinggi sehingga penyerapan gelombang mikro lebih optimal.

14
 DAFTAR PUSTAKA

Cahyono, Bambang, 1991, Segi praktisi dan Metode pemisahan senyawa organic, Kimia MIPA
UNDIP,Semarang
Guenther. 1987. Minyak Atsiri. Diterjemahkan oleh R.S. Ketaren dan R. Mulyono. Jakarta, UI
Press.
Herbert, R.B. 1996. Biosintesis Metabolit Sekunder. Alih Bahasa Bambang Srigandono. Penerbit
IKIP Semarang Press. Semarang. Hal. 103-123.
Lenny, S. 2006. Senyawa Flavanoida, Fenilpropanida dan Alkaloida, Karya Ilmiah Departemen
Kimia Fakultas MIPA Universitas Sumatera Utara
Sugihara, 1961. Distilasi Sederhana. Bandung : ITB Press. Wilcox, 1995, Experimental Organic
Chemistry, Prentice Hall, New jersey
Wasilah, Sudja. 1978. Penuntun Percobaan Pengantar Kimia Organik. Bandung: P.T Karya
Nusantara.

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Contents lists available at ScienceDirect

Journal of Chromatography A
journal homepage: www.elsevier.com/locate/chroma

A modified approach for isolation of essential oil from fruit of

Amorpha fruticosa Linn using microwave-assisted hydrodistillation
concatenated liquid-liquid extraction
Fengli Chen a , Jia Jia b , Qiang Zhang c , Huiyan Gu c,∗ , Lei Yang a,∗
a
Key Laboratory of Forest Plant Ecology, Ministry of Education, Northeast Forestry University, Harbin 150040, China
b
Pharmacy & Medical Laboratory Department, Daqing Medical College, Daqing 163312, China
c
School of Forestry, Northeast Forestry University, Harbin 150040, China

a r t i c l e i n f o a b s t r a c t

Article history: In this work, a modified technique was developed to separate essential oil from the fruit of Amorpha fru-
Received 29 September 2017 ticosa using microwave-assisted hydrodistillation concatenated liquid-liquid extraction (MHD-LLE). The
Accepted 3 October 2017 new apparatus consists of two series-wound separation columns for separating essential oil, one is the
Available online xxx
conventional oil-water separation column, and the other is the extraction column of components from
hydrosol using an organic solvent. Therefore, the apparatus can simultaneously collect the essential oil
Keywords:
separated on the top of hydrosol and the components extracted from hydrosol using an organic solvent.
Microwave-assisted hydrodistillation
Based on the yield of essential oil in the first and second separation columns, the effects of parameters
concatenated liquid-liquid extraction
(MHD-LLE)
were investigated by single factor experiments and Box–Behnken design. Under the optimum conditions
Essential oil (2 mL ethyl ether as the extraction solvent in the second separation column, 12 mL/g liquid-solid ratio,
Amorpha fruticosa 4.0 min homogenate time, 35 min microwave irradiation time and 540 W microwave irradiation power),
Fruit satisfactory yields for the essential oil in the first separation column (10.31 ± 0.33 g/kg) and second sep-
aration column (0.82 ± 0.03 g/kg) were obtained. Compared with traditional methods, the developed
method gave a higher yield of essential oil in a shorter time. In addition, GC–MS analysis of the essen-
tial oil indicated significant differences of the relative contents of individual volatile components in the
essential oils obtained in the two separation columns. Therefore, the MHD-LLE technique developed here
is a good alternative for the isolation of essential oil from A. fruticosa fruit as well as other herbs.
© 2017 Published by Elsevier B.V.

1. Introduction is always accompanied by low extraction efficiency due to long

Essential oil is a kind of volatile oily component which is a
secondary metabolite produced by aromatic plants; it is water-
immiscible and can be distilled with steam for separation. Essential
oil is also known as ethereal oil and is characterized as a volatile,
natural, liquid, complex component with strong aromatic odor [1].
In recent years, essential oil has been widely used in the cosmetic
industry and daily life due to its fragrance [2,3], as well as in the
pharmaceuticals industry because of its various positive pharma-
cological effects [4,5]. Due to the extensive applications of essential
oil, the market demand for essential oil has increased remarkably.
The common and most frequently used method for the separa-
tion of essential oil is hydrodistillation (HD) [6], but this method
∗ Corresponding authors.
E-mail addresses: hygnefu@126.com (H. Gu), ylchennefu@163.com (L. Yang).
time consumption, loss of some oxy-components dissolved in the
condensate water during the distillation process, such as ketones
and aldehydes, and some thermally sensitive components encoun-
tered through thermal effects, such as ␣-pinene and camphene
[7]. Hence, microwave-assisted hydrodistillation (MHD) has been
developed to improve the extraction efficiency for essential oil [8],
which can reduce the extraction duration and enhance the essential
oil yield because of the characteristics of microwave irradiation [9].
In general, terpenes in essential oils obtained by the two methods
mentioned above are relatively concentrated, but oxy-components
are relatively deficient. The oxy-components in essential oils tend
to have higher application values than the oxygen-free components
[10].
In the traditional Clevenger apparatus, the essential oil is sep-
arated on the top of condensed water, but some essential oil
components often dissolve in water to form hydrosol. Based on
the Clevenger apparatus, a new modified apparatus for the distilla-

https://doi.org/10.1016/j.chroma.2017.10.008
0021-9673/© 2017 Published by Elsevier B.V.

Please cite this article in press as: F. Chen, et al., A modified approach for isolation of essential oil from fruit of Amor-
pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
https://doi.org/10.1016/j.chroma.2017.10.008
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tion and separation of essential oil from plant materials is designed
and developed, which consists of two series-wound separation pro-
cesses of essential oil: one is the conventional oil-water separation
process, and the other is the liquid-liquid extraction process of
components from hydrosol using an organic solvent. Therefore, the
apparatus can simultaneously collect the essential oil separated on
the top of hydrosol and the components extracted from hydrosol
using an organic solvent. Hopefully, the modified apparatus can be
useful and efficient for the separation of essential oil from plant
materials.
Amorpha fruticosa is a perennial deciduous shrub of Legumi-
nosae family [11], originating in North America and introduced in
China around the 1920s. It is widely planted in China for the pur-
pose of erosion control and wasteland restoration. A. fruticosa has
been used as a folk medicine for the treatment of burn, ambus-
tion, carbuncle and eczema [12], and its therapeutic utilization
can be dated back to the beginning of civilization [13]. Previous
studies have revealed that it is rich in a number of compounds,
including rotenoids [14], stilbenes [15], flavanones [16,17] and
isoflavones [11,14], thus leading to antimicrobial [15,18], antiox-
idant [19], antitumor [17], antidiabetic [20] and immune activities
[11]. In addition, essential oil with various pharmacological effects
and industrial utilizations has also been separated from the fruit of
A. fruticosa [21], but it has not been reported much. Thus, develop-
ing an effective method for the isolation of essential oil with high
yield is of great significance and can alleviate the demand tension
of essential oil.
The objective of this work was to separate essential oil from
the fruit of A. fruticosa by a developed apparatus combining with
microwave irradiation, namely, microwave-assisted hydrodistil-
lation concatenated liquid-liquid extraction (MHD-LLE). Many
influential parameters, including liquid-solid ratio, microwave
irradiation time, microwave irradiation power, type of extrac-
tion solvent and homogenate time (particle size), were optimized
systematically by single factor experiment and response surface
analysis (RSM) with Box–Behnken design (BBD). In addition, the
MHD-LLE procedure was compared with traditional HD and MHD
techniques. Hopefully, the developed technique is helpful and ben-
eficial for improving the yield of essential oil from A. fruticosa fruit
and other plants.
2. Experimental
2.1. Materials and chemicals
Fresh A. fruticosa fruits were hand-harvested in October 2016
from Maoershan town (Shangzhi, China). The fruits were dried nat-
urally at room temperature, and the same batch of materials sample
was used for all tests.
Ethyl ether, n-hexane, petroleum ether (boiling range 30–60 ◦ C),
cyclohexane and n-pentane were purchased from Tianjin Fuyu Fine
Chemical Co., Ltd. (Tianjin, China). Anhydrous sodium sulfate was
purchased from Aladdin (Shanghai, China). Reverse osmosis Milli-
Q water (Millipore, Billerica, MA, USA) was used for the preparation
of all sample solutions.
2.2. Apparatus
Prior to the distillation, the grinding process of raw materials
was performed in a Waring blender (Joyoung, Jinan, China). The
bath was a cylinder container with a volume of 1 L, and the reamer
rating was 10,000 r/min.
The MHD-LLE apparatus applied in this study consists of
a microwave oven and a hydrodistillation concatenated liquid-
liquid extraction apparatus. A WP700 microwave oven (Glanz,
Please cite this article in press as: F. Chen, et al., A modified approach for isolation of essential oil from fruit of Amor-
pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
https://doi.org/10.1016/j.chroma.2017.10.008
Zhongshan, China) was used for the heating, with an irra-
diation frequency of 2.45 GHz, and an interior cavity size of
215 mm × 350 mm × 330 mm. The microwave irradiation power
was divided into five levels, including 100% (700 W), 77% (540 W),
55% (385 W), 33% (230 W), and 17% (120 W). The hydrodistillation
concatenated liquid-liquid extraction apparatus was established
based on the Clevenger apparatus. The schematic diagram and its
detailed dimensions are shown in Fig. 1, which consists of two
separation columns for the separation of essential oil. The first sep-
aration column is the conventional oil-water separation process,
and the second separation column is the liquid-liquid extraction
process for the isolation of components by an organic solvent.
2.3. Microwave-assisted hydrodistillation concatenated
liquid-liquid extraction procedure
The MHD-LLE process was as follows: 50.0 g of A. fruticosa fruits
weighed accurately and a certain volume of deionized water (to
reach a required liquid-solid ratio) were placed into the War-
ing blender for different homogenate times, based on previous
studies [22–24]. Then, the suspension was completely transferred
to a 1000-mL round-bottom flask, which was placed into the
microwave oven and attached to the developed hydrodistillation
concatenated liquid-liquid extraction apparatus and a condenser
through a hole at the top of the microwave oven. Before distilla-
tion, different volumes and types of organic solvent were added
into the second separation column for the extraction of compo-
nents dissolved in water. After the sample mixture started to boil,
the suspension was then irradiated by microwave for various times
under different powers. The volatile components could be released
from the samples and evaporated with water steam, and then were
condensed and separated from the water, which was completed in
the first separation column. However, some essential oil compo-
nents often dissolved in water to form hydrosol. When the hydrosol
reached the flood, it would return to the second separation column
for extraction of some components by an organic solvent through
the contact between hydrosol and extraction solvent. The water
would return to the sample flask constantly when it reached the
flood level.
After each process, the essential oil obtained in the first separa-
tion column was collected, dried using anhydrous sodium sulphate,
and stored at 4 ◦ C until analysis. The extraction solvent in the
second separation column was dried by anhydrous sodium sul-
fate overnight and then concentrated to obtain essential oil under
reduced pressure, which was stored at 4 ◦ C before analysis. In this
study, the liquid–solid ratio, homogenate time, type and volume
of extraction solvent in the second separation column, microwave
irradiation time and power were investigated systematically. The
yield of essential oil obtained in the two separation columns was
calculated as follows:
M
Y= (1)
Mo
where Y (g/kg) is the yield of essential oil; M (g) is the mean mass of
essential oil obtained in the two separation columns, and Mo (kg)
is the mean mass of three fruit samples before distillation.
2.4. Conventional hydrodistillation and microwave-assisted
hydrodistillation methods
50.0 g of A. fruticosa fruit powders (40–80 mesh, milled using
a disintegrator and sieved in advance) weighed accurately and
600 mL of deionized water were added into a 1000-mL round-
bottom flask, and then the flask was attached to a Clevenger
apparatus. When the flask started to boil, for HD, the flask was
heated by heating jacket for 4 h under 540 W, according to the
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Fig. 1. Schematic diagram of the microwave-assisted hydrodistillation concatenated liquid-liquid extraction apparatus.

Chinese Pharmacopoeia [25]; for MHD, the flask was irradiated by oil obtained above the water in the collection tube was collected,
microwave for 1 h under 540 W. After the distillation, the essential dehydrated, dried, capped and kept at 4 ◦ C until analysis.

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pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
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2.5. Response surface methodology
Based on the single factor experiments, homogenate time (X1 ),
microwave irradiation time (X2 ) and microwave irradiation power
(X3 ) were chosen as the operational parameters for further opti-
mization. A three-factor-three-level BBD followed by response
surface analysis was performed here to evaluate the optimum oper-
ational parameters of the MHD-LLE process, in order to obtain the
maximum essential oil yield and to estimate the main effects, inter-
action effects and quadratic effects of the three variables. Seventeen
randomized combinations containing five replicates at the cen-
ter point were used to fit a full second-order polynomial equation
model. The fluctuation ranges for X1 , X2 and X3 were set as 3–5 min,
20–40 min and 385–700 W and were standardized to the interval.
The general full second-order polynomial equation was:

3

3

2

3
Y = ˇ0 + ˇi Xi + ˇii Xi2 + ˇij Xi Xj
i=1 i=1 i=1 j=i+1
where Y is the predicted response; Xi and Xj are the coded indepen-
dent variables; ˇ0 is the constant coefficient; ˇi , ˇii and ˇij are the
coefficients of the linear, quadratic and interaction terms, respec-
tively. Design-Expert (Version 8.0, Stat-Ease Inc., Minneapolis, USA)
software was adopted for statistical analyses of the experimental
design and data. One-way analysis of variance and Duncan’s multi-
ple range tests were applied to calculate the significant difference of
essential oil yield. All experiments were repeated three times, and
the result of which was expressed as the mean value ± standard
deviation.
2.6. Kinetic model
To describe the release kinetics of essential oil from A. fruti-
cosa fruits by different methods, second-order kinetic model was
applied. The general second-order model is as follows [26]:
dC t
= k · (Ce − Ct )2
dt
where Ct is the yield of essential oil (g/kg) obtained at any time t
(min), Ce is the yield of essential oil at saturation (g/kg) and k is the
second-order extraction rate constant (kg/g/min).
When considering the initial and boundary conditions, namely,
Ct = 0 and Ct = Ct at t = 0 and t = t, respectively, the integrated rate
law for a second-order process can be expressed as Eq. (4) or Eq.
(5) [27].
k · t · Ce 2
Ct =
1 + k · t · Ce
t 1 t 1 t
= + = +
Ct k · Ce 2 Ce h Ce
where h is the initial extraction rate (g/kg/min) when t approaches
0, and can be defined as:
h = k·C e 2
2.7. Gas chromatography–mass spectrometry analysis
The GC–MS system (6890N/5973, Agilent Technologies, Santa
Clara, CA) used in this study to determine the composition of essen-
tial oil consists of a gas chromatograph equipped with an HP-5MS
(Agilent Technologies) capillary column (30 mm × 0.25 mm, film
thickness of 0.25 ␮m) and a mass selective detector operating in
electron ionization mode (70 eV).
Before GC–MS analysis, the essential oil obtained was diluted
with n-hexane (1:100, v/v). The detailed detecting condition for
Please cite this article in press as: F. Chen, et al., A modified approach for isolation of essential oil from fruit of Amor-
pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
https://doi.org/10.1016/j.chroma.2017.10.008
the GC–MS analysis of essential oil was according to our prelimi-
nary study [28]. The identification of essential oil components was
based on the comparison between their GC retention indices on
the HP-5MS capillary column and literature data, combined with
the comparison of their mass spectra fragmentation patterns with
those of similar compounds from a database (Wiley, Mass Spectral
Library, NIST02) and MS literature data [4].
3. Results and discussion
3.1. Optimization of the MHD-LLE process
The parameters of the MHD-LLE procedure have a dominat-
ing effect on the distillation and extraction process for essential
oil. Thus, in this study, some influential factors, including type
and volume of extraction solvent in the second separation col-
(2) umn, homogenate time, liquid–solid ratio, microwave irradiation
power, microwave irradiation time, were evaluated systematically
in terms of the yields of EOf and EOs .
3.1.1. Effect of type of extraction solvent in the second separation
column
The extraction solvent in the second separation column plays
a key role for the extraction of essential oil components from
hydrosol; thus, the type of extraction solvent has to be compared
and chosen to facilitate the extraction yield.
In this study, five kinds of common extraction solvents were
selected to investigate the different extractive selectivities, includ-
ing ethyl ether, cyclohexane, petroleum ether (30–60 ◦ C), n-hexane
and n-pentane. As seen from Fig. 2a, the yield of essential oil in the
first separation column (EOf ) was similar for all five solvents, which
was only related to the separation process in the first separation
column and had nothing to do with the second liquid-liquid extrac-
tion column. However, obvious differences existed in the yield of
essential oil in the second separation column (EOs ) obtained by
(3) different extraction solvents. The yields of essential oil applying
the five extraction solvents in the second separation column were
0.63, 0.53, 0.41, 0.46 and 0.39 g/kg, respectively, which indicated
that the best result was obtained by ethyl ether. The differences
between ethyl ether and the other solvents were large, especially
for n-pentane. The result implied that the solubility of essential oil
components in ethyl ether was better than in others. Therefore,
ethyl ether was selected as the extraction solvent for the studies
performed subsequently.
(4)
3.1.2. Effect of extraction solvent volume in the second separation
column
(5) The extraction solvent volume in the second separation col-
umn is a crucial parameter for ensuring higher extraction result.
Too small solvent volumes will result in the incomplete extraction
of essential oil components; while too large solvent volumes are
always associated with high consumption of organic solvent and
(6) pollution.
The effect of various extraction solvent volumes (1, 2, 3, 4 and
5 mL) in the second separation column on the yields of EOf and EOs
was investigated, the result of which is shown in Fig. 2b. The EOf
presented a similar yield in terms of different extraction solvent
volumes, because which was separated in the first separation col-
umn. As for the yield of EOs , an initial remarkable enhancement
was followed by a non-obvious improvement with the increase in
extraction solvent volume from 1 to 2 mL, and 2 to 5 mL, respec-
tively. This phenomenon was speculated by that the extraction of
essential oil components was inadequate when using small vol-
ume. In terms of the yield of EOs and solvent consumption, 2 mL
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Fig. 2. Effect of extraction solvent type (a) and volume (b) on yield of essential oil in the first and second separation columns.

was enough and was selected as the extraction solvent volume for
the further optimization.
3.1.3. Effect of homogenate time
Homogenate time is strongly associated with the particle size
of plant materials, and a longer homogenate time will lead to a
decrease in particle size. Therefore, the influence of homogenate
time on the yields of EOf and EOs was evaluated with differ-
ent homogenate times, including 1–5 min. The other experimental
conditions applied were microwave irradiation power of 540 W
(specific absorbed power of 0.64 W/g), liquid-solid ratio of 10 mL/g
and microwave irradiation time of 30 min, respectively. As seen in
Fig. 3a, with the increase of homogenate time from 1 min to 4 min,
the yield of EOf increased gradually, but then the yield decreased
with further increase in homogenate time. A similar increase in
the yield of EOs was observed when the homogenate time was
increased from 1 min to 3 min, but the yield almost remained con-
stant and then decreased obviously when the homogenate time
further increased from 3 min to 4 min and 4 min to 5 min. The
enhancement of essential oil yield with a decrease in particle size
was because a small particle size can increase the interfacial area
between the matrix and water, and facilitate the release of essen-
tial oil from the broken cells of the plant matrix [29]. In contrast,
increasing the particle size was accompanied by a reduction in the
essential oil yield, which was speculated to be due to the essential
oil not being able to be transported completely and only the surface
oil could be removed due to the existence of unbroken cell walls
[30]. The reduction of essential oil yield with the homogenate time
beyond 4 min may be due to the very small particle size, resulting in
a mixture with high viscosity and low fluidity; another reason may
be the destruction of components. Thus, the range of homogenate
time from 3 to 5 min was selected for further optimization.
3.1.4. Effect of liquid-solid ratio
To avoid incomplete extraction caused by small solution vol-
umes and reduce the difficulty of the procedure and unnecessary
waste caused by large solution volumes, an appropriate liquid-solid
ratio was chosen after performing a series of experiments with

Please cite this article in press as: F. Chen, et al., A modified approach for isolation of essential oil from fruit of Amor-
pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
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Fig. 3. Effect of homogenate time (a) and liquid-solid ratio (b) on yield of essential oil in the first and second separation columns.

various liquid-solid ratios (6, 8, 10, 12, and 14 mL/g). The other
operational conditions were 30 min microwave irradiation time,
3 min homogenate time and 540 W microwave irradiation power
(0.64 W/g specific absorbed power). It can be seen from Fig. 3b
that the effect of liquid–solid ratio on the yield of EOf was smaller
than that on the yield of EOs . The yield of EOf gradually increased
and then almost remained constant when the liquid-solid ratio
increased from 6 mL/g to 10 mL/g and 10 mL/g to 12 mL/g. However,
a higher liquid-solid ratio exceeding 12 mL/g gave a slightly lower
yield. For EOs , an obvious improvement and then a slight reduc-
tion of yield were observed with the increase in the liquid-solid
ratio from 6 to 12 mL/g and 12 to 14 mL/g. An appropriate increase
in the liquid-solid ratio can increase the solute concentration dif-
ference between inside and outside of cell, thus contributing to the
penetration of water into the matrix and dissolution of essential oil.
In contrast, the slight reduction of oil yield may be attributed to the
dissolution of some essential oil components in water [8]. There-
fore, a liquid-solid ratio of 12 mL/g was considered as the optimal
value for further experiments.
3.1.5. Effect of microwave irradiation power
For investigating the effect of microwave irradiation power on
the yields of EOf and EOs , the MHD-LLE process was carried out
under various microwave irradiation powers (120, 230, 385, 560
and 700 W), with a liquid-solid ratio of 10 mL/g, microwave irradi-
ation time of 30 min and homogenate time of 3 min. To express
directly, the microwave irradiation powers were converted into
specific absorbed powers of 0.05, 0.26, 0.43, 0.64 and 1.02 W/g,
respectively. The calculation formula for absorbed microwave
power and specific absorbed power of the mixture (A. fruticosa fruit
and water) was based on Pongmalai et al. [31]:
T
Q = mCp (7)
t
Qs = Q/m (8)
where Q is the absorbed microwave power (W), Qs is the specific
absorbed power (W/g), m is the mass of mixture (g), Cp is the specific
heat of mixture (kJ/kg/◦ C), T is the temperature increased for the
mixture (◦ C), and t is the time period needed for the increase of
temperature (s), respectively.

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pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
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Fig. 4. Effect of microwave irradiation power (a) and microwave irradiation time (b) on yield of essential oil in the first and second separation columns.

The comparison between various microwave irradiation powers
on the yields of EOf and EOs is shown in Fig. 4a, which indicated an
obvious enhancement and then a decrease in both the yields of EOf
and EOs when the specific absorbed power ranged from 0.05 W/g to
0.64 W/g and 0.64 W/g to 1.02 W/g. However, the declining degree
of the yield of EOs was larger than that of EOf . Therefore, a specific
absorbed power above or below the optimum value would neg-
atively influence the essential oil yield. This phenomenon can be
attributed to the fact that increasing microwave irradiation power
can result in high specific absorbed power for rapid energy trans-
fer, thus promoting the destruction of cell wall and enhancing the
penetration of water into the matrix through molecular interac-
tion with the electromagnetic field, resulting in a higher yield of
essential oil. However, too high of a microwave irradiation power
with a high specific absorbed power would lead to the degradation,
condensation or isomerization of essential oil components. There-
fore, microwave irradiation power of 385–700 W (specific absorbed
power of 0.43–1.02 W/g) was selected as the optimum power range
for subsequent optimization.
3.1.6. Effect of microwave irradiation time
The whole process for obtaining essential oil in this study
included five steps: (1) diffusion of water into plant materials and
dissolution of essential oil under heating conditions; (2) diffusion
of water containing essential oil outside of the materials surface;
(3) distillation of water containing essential oil; (4) condensation
of water steam containing essential oil and separation of essential
oil on the water surface in the first separation column; (5) extrac-
tion of essential oil components from water-essential oil emulsion
into extraction solvent in the second separation column. Therefore,
an enough microwave irradiation time is crucial, which should be
investigated.
To evaluate the influence of microwave irradiation time on
essential oil yield in different separation columns, the MHD-LLE
procedure was carried out by distilling a slurry composed of 50.0 g
of fruit samples and 500 mL of water for different durations after
homogenizing for 3 min, with an applied microwave irradiation
power of 540 W (specific absorbed power of 0.64 W/g). The result
presented in Fig. 4b illustrated that the essential oil yield obtained
in both separation columns increased first and then remained

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pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
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constant with the increase in microwave irradiation time. The equi-
librium point for EOs was almost 20 min, which was slightly earlier
than that for EOf (approximately 30 min). The phenomenon could
be explained by the fact that the whole distillation process of essen-
tial oil required enough time for the isolation of water, dissolution
of essential oil and evaporation. The rapid phase for the increase
in yield was mainly from the surface of the matrix, and essential
oil existed in the interior of deep cells needed longer time for dis-
tilling out, thus leading to the slight improvement of yield with
further increasing the microwave irradiation time. The faster time
for reaching equilibrium of EOs yield may be attributed to that the
essential oil itself separated on the water in the first separation
column can form an extraction process and appropriately reduce
the content of essential oil dissolved in water, with the amount
of EOf increasing gradually. Hence, microwave irradiation time of
20–40 min was selected as the optimum range for further tests.
3.2. Parameter optimization by RSM
3.2.1. Model building and statistical analysis
To evaluate the interaction effects between independent vari-
ables (homogenate time X1 , microwave irradiation time X2 and
microwave irradiation power X3 ) on the yields of EOf and EOs , sev-
enteen experiments were carried out based on BBD design. The
experimental result is listed in Table 1, and the variance analysis
of parameters applied for the response surface model is given in
Table 2. Coefficient of determination (R2 ) can represent the rela-
tionship between predicted and actual values [32], and a higher
R2 demonstrates that the model is workable. R2 of 0.9899 and
0.9823 for EOf and EOs noted that high degrees of correlation were
existed between actual and predicted values. F-test was used to
determine the statistical significances and regression coefficients.
Generally, the significance of terms is estimated based on the P-
value, the model term is significant when P < 0.05 [33]; larger F
value and smaller P-value is desirable [34]. The models with F-
values of 76.37 and 43.15 for EOf and EOs and low P-values (less
than 0.0001 for both EOf and EOs ) noted that the models were sig-
nificant and appeared to reasonably represent the data. The chance
for model F-value of this size to occur due to statistical noise was
only <0.01%. The “lack of fit” can measure whether the model fitting
of data is good, and a lack of fit with P > 0.1 is desirable [35]. In this
model, the “lack of fit” gave F-values of 0.86 and 0.29, and P-values
of 0.5325 and 0.8295 for EOf and EOs , respectively. These values
suggested a non-significant “lack of fit,” and the probability for a
“lack of fit” this large to occur due to statistical noise was 53.25%
and 82.95%, indicating better agreement of the experimental data
with predicted values.
For both the yields of EOf and EOs , the linear terms of
homogenate time (X1 ), microwave irradiation time (X2 ) and
microwave irradiation power (X3 ); and the quadratic terms of
homogenate time (X1 2 ), microwave irradiation time (X2 2 ) and
microwave irradiation power (X3 2 ) could reach significance. In con-
trast, all of the interaction terms among the different variables were
non-significant in terms of the yields of EOf and EOs .
The predicted R2 values of 0.9274 and 0.9263 for EOf and EOs ,
respectively, were in reasonable agreement with the adjusted R2
values of 0.9770 and 0.9595. The coefficient of variation (CV) is
used to determine the reproducibility of a model [36], and a model
with high CV can give high variation in the mean value and is not an
adequate response model [37]. The CV obtained in the model for EOf
and EOs was 1.34% and 1.58%, which was low relatively and ensured
a better precision for the experiments conducted. “Adeq precision”
(24.29 for EOf and 17.96 for EOs ) is an indicator to measure the
signal-to-noise ratio, where a value greater than 4 represented that
an adequate signal was obtained.
Please cite this article in press as: F. Chen, et al., A modified approach for isolation of essential oil from fruit of Amor-
pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
https://doi.org/10.1016/j.chroma.2017.10.008
Fig. 5. Dynamic curves for the total essential oil obtained by various methods (a)
MHD-LLE; (b) MHD; (c) HD. The points represent actual values, and lines represent
fitting behavior predicted by second order models.
The mathematical model was expressed by the mathematical
regression model as follows:
EOf (%) = −18.85 + 6.43X 1 + 0.26X 2 + 0.04X 3 − 0.85X 1 2 (9)
2
EOs (%) = −0.73 + 0.19X 1 + 0.02X 2 − 0.03X 1 (10)
3.2.2. The optimum conditions derived
After the fitting and point prediction by software, the optimum
conditions were homogenate time of 4.0 min, microwave irradi-
ation time of 35 min and microwave irradiation power of 540 W
(specific absorbed power of 0.64 W/g). The theoretical yields of EOf
and EOs were 10.19 g/kg and 0.80 g/kg, respectively.
3.3. Verification tests
To validate the reliability of BBD, verification experiments
were conducted five times under the optimum conditions pre-
dicted (homogenate time of 4.0 min, microwave irradiation time
of 35 min and microwave irradiation power of 540 W (specific
absorbed power of 0.64 W/g)). The actual yields of EOf and EOs were
10.31 ± 0.33 g/kg and 0.82 ± 0.03 g/kg, respectively. The very simi-
lar results between the actual yields and predicted yields suggested
that the BBD was reliable.
3.4. Comparison with different traditional procedures
To highlight the superiority of the developed MHD-LLE process,
traditional HD and MHD methods were selected for comparison.
The extraction kinetics of the three techniques was plotted based
on the yield of essential oil versus time, and second-order kinetic
model was applied for the fitting. For obvious comparison, the total
essential oil yield (the sum yield of EOf and EOs ) was used as the
response in the developed MHD-LLE. The MHD-LLE was carried out
based on the optimum conditions, and the operational conditions
for MHD and HD were presented in Section 2.4.
As shown by the result in Fig. 5, the essential oil yield for the tra-
ditional HD method increased gradually and reached equilibrium
at approximately 180 min. For MHD and the developed MHD-
LLE method, the enhancement of essential oil yield was basically
ended after 30 min. The essential oil yields achieved by MHD-LLE,
MHD and HD were 10.92, 10.25 and 10.18 g/kg, respectively, which
needed 30 min, 30 min and 180 min to reach equilibrium. Consider-
ing the higher yield of essential oil and shorter duration, MHD-LLE
performed better than conventional HD and could be a promising
alternative to HD. The yield of essential oil achieved by the pro-
posed MHD-LLE in this study was higher than that of MHD, and
the time required was similar. The higher yield of MHD-LLE was
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Table 1
Box–Behnken design (BBD) for the experimental values and predicted values for essential oil yield in the first and second separation column (g/kg).a

Run No. Factors Response 1 Response 2

Homogenate Microwave Microwave Predicted EOf b Actual EOf yield Predicted EOs Actual EOs yield
time (min) irradiation time irradiation power yield (g/kg) (g/kg) yield (g/kg) (g/kg)
(min) (W)

1 5 30 385 7.84 7.81 0.68 0.68

2 4 40 700 9.30 9.37 0.70 0.70
3 3 30 385 8.31 8.37 0.73 0.74
4 3 30 700 8.56 8.59 0.70 0.70
5 4 30 543 10.10 10.13 0.79 0.81
6 4 20 385 8.21 8.15 0.67 0.67
7 4 20 700 8.38 8.48 0.63 0.64
8 3 20 543 8.69 8.70 0.72 0.72
9 5 30 700 8.31 8.26 0.64 0.64
10 4 30 543 10.10 10.27 0.79 0.80
11 5 40 543 9.05 9.04 0.74 0.75
12 4 40 385 8.88 8.93 0.74 0.73
13 3 40 543 9.47 9.37 0.79 0.79
14 4 30 543 10.10 10.04 0.79 0.78
15 4 30 543 10.10 9.93 0.79 0.77
16 4 30 543 10.10 10.15 0.79 0.79
17 5 20 543 8.52 8.48 0.66 0.66
a
The results were obtained with Design Expert 8.0 software.
b
EOf : Essential oil in first separation column; EOs : Essential oil in second separation column.

Table 2
Estimated regression coefficients and analysis of variance (ANOVA) for the quadratic polynomial model of essential oil in first and second separation column determined
from BBD.a

Source Sum of squares Degree of Mean square F-value P-valued

freedom

EOf c EOs EOf EOs EOf EOs EOf EOs

−2 −3
Model 10.17 5.10 × 10 9 1.13 5.66 × 10 76.37 43.15 <0.0001 *
<0.0001*
X1 b 0.26 6.05 × 10−3 1 0.26 6.05 × 10−3 17.78 46.07 0.0040* 0.0003*
X2 1.05 1.01 × 10−2 1 1.05 1.01 × 10−2 71.12 76.66 <0.0001* <0.0001*
X3 0.26 2.39 × 10−3 1 0.26 2.39 × 10−3 17.78 18.21 0.0040* 0.0037*
X1 X2 0.00 5.16 × 10−5 1 0.00 5.16 × 10−5 0.21 0.39 0.6603 0.5504
X1 X3 0.01 3.31 × 10−6 1 0.01 3.31 × 10−6 0.84 0.03 0.3894 0.8784
X2 X3 0.00 1.32 × 10−6 1 0.00 1.32 × 10−6 0.21 0.01 0.6603 0.9229
X1 2 2.97 3.29 × 10−3 1 2.97 3.29 × 10−3 200.88 25.08 <0.0001* 0.0016*
X2 2 0.56 4.93 × 10−3 1 0.56 4.93 × 10−3 38.10 37.61 0.0005* 0.0005*
X3 2 4.27 2.15 × 10−2 1 4.27 2.15 × 10−2 288.90 163.60 <0.0001* <0.0001*
Residual 0.10 9.19 × 10−4 7 0.01 1.31 × 10−4
Lack of fit 0.04 1.66 × 10−4 3 0.01 5.52 × 10−5 0.86 0.29 0.5325 0.8295
Pure error 0.06 7.53 × 10−4 4 0.02 1.88 × 10−4
Cor total 10.27 5.19 × 10−2 16

Credibility analysis of Index mark Standard deviation Mean CV%e Press R2 Adjust R2 Predicted R2 Adequacy precision
the regression EOf 0.12 9.06 1.34 0.75 0.9899 0.9770 0.9274 24.29
equations EOs 0.01 0.73 1.58 0.00 0.9823 0.9595 0.9263 17.96
a
The results were obtained with Design Expert 8.0 software.
b
X1 : Homogenate time (min); X2 : Microwave irradiation time (min); X3 : Microwave irradiation power (W).
c
EOf : Essential oil in first separation column; EOs : Essential oil in second separation column.
d
P-value: Probability value.
e
CV%: Coefficient of variation (%); R2 : Coefficient of determination.
*
Significant at P < 0.05.

attributed to the extraction of some essential oil components from
water-essential oil emulsions by the organic solvent in the sec-
ond separation column. The advantage of MHD in the essential oil
yield compared to HD was speculated to be due to the degradation,
condensation or isomerization of essential oil occurring during the
longer time required for HD.
The process for obtaining essential oil could be divided into three
different periods. The first period was accompanied by a constant
extraction rate and sharp enhancement of yield, because the extrac-
tion of essential oil occurred in the superficial layer of particles.
The second period was characterized by a falling extraction rate
and a slight improvement of yield, and the phenomenon was inter-
preted by the fact that a large amount of solute available on the
surface had been released and the solute inside the particles needed
to diffuse into the surface. The third period was represented by a
nearly horizontal line, in which almost no improvement of essen-
tial oil yield was observed. The tendency of essential oil yield over
time is typical for a second-order process [26]; thus, the experi-
mental data were fitted and interpreted by a second-order kinetic
model. To analyze the experimental data and the fitted result, the
second-order kinetic model was deformed to a linear curve plotted
by t/Ct versus time. The analysis result of the linear curve is also
presented in Fig. 5, including the essential oil yield at saturation,
Ce , the extraction rate constant, k, the initial extraction rate, h, and
the coefficient of determination, R2 . All of the R2 values for MHD-
LLE (0.9949), MHD (0.9961) and HD (0.9969) were higher than 0.99,
which illustrated that the values of extraction kinetics fitted by the
second order kinetic model were highly consistent with the actual

Please cite this article in press as: F. Chen, et al., A modified approach for isolation of essential oil from fruit of Amor-
pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
https://doi.org/10.1016/j.chroma.2017.10.008
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Table 3
Gas chromatography–mass spectrometry results for the chemical compositions of essential oil from fruit of Amorpha fruticosa.

No.a Compounds RIb ID Molecular formula RAe (%)

MHD-LLD MHD HD
f
EOf EOs

1 ␣-Pinene 924 RIc , MSd C10 H16 5.55 – 4.75 5.07

2 ␤- Pinene 980 RI, MS C10 H16 0.89 – 0.83 0.52
3 3-Carene 1007 RI, MS C10 H16 3.84 – 2.92 3.51
4 ␣-Terpineol 1188 RI, MS C10 H18 O – 1.91 – –
5 ␥-Elemene 1418 RI, MS C15 H24 0.61 – 0.61 0.56
6 ␣-Cubebene 1343 RI, MS C15 H24 1.26 – 1.11 1.21
7 Ylangene 1364 RI, MS C15 H24 0.92 – 0.99 0.84
8 ␣-Copaene 1376 RI, MS C15 H24 7.65 1.70 8.11 6.80
9 ␤-Cubebene 1388 RI, MS C15 H24 2.01 – 2.02 1.84
10 ␣-Gurjunene 1408 RI, MS C15 H24 0.71 – 0.62 0.65
11 Caryophyllene 1423 RI, MS C15 H24 9.62 – 8.74 10.67
12 ␣-Bergamotene 1433 RI, MS C15 H24 3.98 – 3.97 3.64
13 ␥-Gurjunene 1472 RI, MS C15 H24 1.29 1.44 1.56 1.71
14 Humulene 1488 RI, MS C15 H24 2.98 – 2.62 2.72
15 Bicyclosesquiphellandrene 1498 RI, MS C15 H24 8.98 – 7.44 8.09
16 ␥-Muurolene 1477 RI, MS C15 H24 10.61 9.61 13.88 11.52
17 ␤-Selinene 1493 RI, MS C15 H24 0.72 – 0.63 0.66
18 ␣-Muurolene 1504 RI, MS C15 H24 1.12 – 1.26 1.05
19 ␥-Cadinene 1524 RI, MS C15 H24 6.97 9.84 7.99 6.65
20 ␦-Cadinene 1528 RI, MS C15 H24 12.10 3.32 12.22 15.01
21 1,2,3,4,4a,7-hexahydro-1,6-dimethyl- MS C15 H24 0.60 – 0.49 0.49
4-(1-methylethyl)-Naphthalene
22 ␣-Calacorene 1536 RI, MS C15 H20 0.48 – 0.53 0.42
23 2-isopropyl-5-methyl-9-methylene- MS C15 H24 1.49 – 1.22 1.46
Bicyclo[4.4.0]dec-1-ene
24 7,8-dehydro-8a-hydroxy- MS C15 H24 O – 7.51 – –
Isolongifolene
25 Isolongifolol MS C15 H26 O 0.51 8.34 0.63 0.50
26 4-ethenyl-␣,␣,4-trimethyl-3-(1- 1547 RI, MS C15 H26 O 0.40 4.00 0.52 0.38
methylethenyl)-,
[1R-(1␣,3␣,4␤)]-
Cyclohexanemethanol
27 2,2,5,5-tetramethyl- MS C15 H26 O 3.28 8.85 3.35 3.09
Bicyclo[6.3.0]undec-1(8)-en-3-ol
28 2,3,4,4a,5,6,7,8-octahydro- 1613 RI, MS C15 H26 O 0.52 4.63 0.78 0.66
.alpha.,.alpha.,4a,8-tetramethl-,
[2R-(2.alpha.,4a.beta.,8.beta.)]-
2-Naphthalenemethanol
29 ␥-Eudesmol MS C15 H26 O 2.11 15.12 2.28 2.02
30 Necahydro-1, 4a-dimethyl-7-(1- 1682 RI, MS C15 H26 O 0.51 3.04 0.61 0.47
methylethylidene)-,
[1R-(1.alpha.,4a.beta.,
8a.alpha.)]-1-Naphthalenol
31 Methyl ether isolongifolol MS C16 H28 O 0.61 7.74 0.72 0.56
32 2-Hexyl-1-decanol MS C16 H34 O – 1.31 – –
Total identified compounds 92.34 88.37 93.42 92.76
Total terpene hydrocarbons 83.90 25.92 84.53 85.07
Total oxygenated terpenes 8.44 62.46 8.90 7.69
a
Compounds listed in order of elution from HP-5MS capillary column.
b
Retention indices for each component were achieved from the reference data of the same component using HP-5MS capillary column.
c
Tentative identification by comparison with RI on HP-5MS capillary column with literature data.
d
Confirmed by comparison with mass data obtained from NIST02 mass spectra library.
e
Relative area percentage (peak area relative to the total peak area, %).
f
EOf : Essential oil in first separation column; EOs : Essential oil in second separation column.

test results, and the extraction kinetics could broadly represent the
experimental data.
The dominance of MHD-LLE compared with MHD and HD was
mainly attributed to the excellent mechanism of microwave irra-
diation and hydrodistillation concatenated liquid-liquid extraction
apparatus. Microwave irradiation with localized and inhomoge-
neous heating can greatly promote the rupture of plant cells and
completely release the essential oil from those broken cells [38,39].
Moreover, in addition to the essential oil-water separation in the
first separation column, some water-soluble components can also
be extracted in the second separation column.
In recent years, MHD technique has been widely applied in the
separation of essential oil [40–42], but loss of essential oil was
caused due to some essential oil components dissolved in water.
Driven by this reason, a rapid extraction method for volatile com-
pounds was developed by Ferhat et al. [43]. They added a small
volume of an extraction solvent in the phase separation tube of the
Clevenger apparatus, thus the components dissolved in water could
also be separated. Following, microwave-assisted simultaneous
distillation and extraction of essential oil was developed [9,28,39],
in which fresh extraction solvent was constantly refluxed for ade-
quate extraction of essential oil. Compared with these methods, the
technique developed in this work includes two series-wound sep-
aration processes of essential oil, one is the conventional oil-water
separation process, and the other is the extraction process of essen-
tial oil components using organic solvent. Therefore, two types of

Please cite this article in press as: F. Chen, et al., A modified approach for isolation of essential oil from fruit of Amor-
pha fruticosa Linn using microwave-assisted hydrodistillation concatenated liquid-liquid extraction, J. Chromatogr. A (2017),
https://doi.org/10.1016/j.chroma.2017.10.008
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essential oils containing components with different proportions
were achieved.
3.5. Chemical composition of essential oil
Through GC–MS analysis, the components of essential oil
obtained by MHD-LLE, MHD and HD are shown in Table 3, including
the identified components and their retention indices, molecu-
lar formulas and relative percentages. A total of 32 components
were detected, among which EOf consisted of 29 components, EOs
owned 15 components, and the essential oil obtained by MHD and
HD was composed of 29 components. The detected components
represented 92.34%, 88.37%, 93.42% and 92.76% of the total essen-
tial oils obtained in the first and second separation columns by
MHD-LLE, and MHD and HD. The relative area percentage (RA%)
of individual volatile component is taken as an indicator, which
is expressed as the percentage of their peak areas in total peak
areas. As seen from Table 3, significant differences existed in the
RA% of individual volatile components in EOf and EOs . In general,
␥-muurolene and ␦-cadinene are the two important compounds in
the essential oil of A. fruticosa fruit, with high RA% in EOf (10.61% and
12.10%). Three components, including 2-Hexyl-1-decanol (1.31%),
␣-Terpineol (1.91%) and 7,8-dehydro-8a-hydroxy-Isolongifolene
(7.51%), were only detected in EOs . All of the oxy-components could
be obtained with higher RA% in EOs than EOf due to their high
polarity, for example, ␥-eudesmol with only 2.11% in EOf , which
accounted for 15.12% in EOs . Therefore, the EOf and EOs with dif-
ferent kinds and proportions of components could provide different
values for application. The essential oils obtained by MHD and HD
had similar compositions to the EOf obtained by MHD-LLE.
4. Conclusion
A modified approach was developed to isolate and quantify
essential oil from the fruit of A. fruticosa using MHD-LLE, includ-
ing conventional oil-water separation process and liquid-liquid
extraction process for some components by an organic solvent.
After optimization by single factor experiments and response sur-
face analysis, satisfactory yields for the EOf (10.31 ± 0.33 g/kg) and
EOs (0.82 ± 0.03 g/kg) were obtained under the optimum conditions
(2 mL ethyl ether as the extraction solvent in the second separation
column, 12 mL/g liquid-solid ratio, homogenate time of 4.0 min,
microwave irradiation time of 35 min and microwave irradiation
power of 540 W). Compared with other traditional methods, MHD-
LLE gave a higher yield of essential oil in a shorter time. Additionally,
GC–MS analysis implied that the relative contents of individual
volatile components in EOf and EOs were markedly different. The
oxy-components of essential oil could be obtained with a higher
relative content in the second separation column than in the first
one. In the future, MHD-LLE could be applied to effectively isolate
essential oil from plant materials.
Acknowledgement
The authors also thank the National Natural Science Foundation
of China (Grant no. NSFC31670580) for financial support.
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KELOMPOK 5
1. AHMAD MISRIANI
2. EPRILIA CAHYA .A
3. NADIA KRISMANDA Y.S
4. NUR KHASANAH
5. OLIVIA VEBRI .L
6. TRI WULANDARI
Apa itu minyak atsiri????

Minyak atsiri adalah sejenis

komponen berminyak yang mudah
menguap yang merupakan metabolit
sekunder yang dihasilkan oleh
tanaman aromatik.
 Metode yang umum dan paling sering digunakan untuk
pemisahan minyak esensial adalah hydrodistillation (HD) namun
metode ini selalu disertai dengan efisiensi ekstraksi rendah
karena konsumsi waktu yang lama, hilangnya beberapa oksi-
komponen terlarut dalam air kondensat selama proses distilasi,
seperti keton dan aldehida, dan beberapa komponen yang
sensitif secara termal yang ditemukan melalui efek panas,
seperti -pinene dan camphene.

Oleh karena itu, hydrodistillation dengan bantuan

gelombang mikro (MHD) telah dikembangkan untuk
meningkatkan efisiensi ekstraksi untuk minyak esensial yang
dapat mengurangi durasi ekstraksi dan meningkatkan hasil
minyak esensial karena karakteristik iradiasi gelombang mikro .
Secara umum, terpen dalam minyak esensial yang diperoleh
dengan dua metode yang disebutkan di atas relatif
terkonsentrasi, tetapi komponen oksi relatif kurang. Oksigen-
komponen dalam minyak esensial cenderung memiliki nilai
aplikasi lebih tinggi daripada komponen bebas oksigen
Isolasi minyak atsiri pada Amorpha Fructicosa

• Dari jurnal yang kami dapat diperoleh isolasi minyak atsiri dengan
menggunakan buah Amorpha Fruticosa.
• Amorpha fruticosa adalah semak gugur abadi dari keluarga
Leguminosae yang berasal dari Amerika Utara dan diperkenalkan di
Cina sekitar tahun 1920-an.
• Tumbuhan Ini banyak ditanam di Cina untuk tujuan pengendalian
erosi dan restorasi gurun.
• A. fruticosa telah digunakan sebagai obat tradisional untuk
pengobatan luka bakar, ambustion, carbuncle dan eczema , dan
pemanfaatan terapeutiknya dapat berasal dari awal peradaban
Penelitian sebelumnya telah mengungkapkan bahwa ia kaya dalam
sejumlah senyawa, termasuk rotenoids , stilbenes , flavanon dan
isoflavon sehingga menyebabkan antimikroba , antioksidan ,
antitumor antidiabetes dan aktivitas kekebalan.
Apa itu isolasi????
Isolasi adalah proses pengambilan atau pemisahan senyawa
bahan alam dengan menggunakan pelarut yang sesuai (Djamal,
2008).
Teknik isolasi minyak atsiri terdiri dari
beberapa metode antara lain :
• Penyulingan Minyak Atsiri
1. Penyulingan dengan air
2. Penyulingan dengan uap dan air
3. Penyulingan dengan uap.
• Destilasi
1. Destilasi Uap
2. Destilasi Vakum
3. Destilasi Biasa
• Ekstraksi
Penyulingan Minyak Atsiri
yaitu pemisahan komponen yang berupa cairan dua macam campuran atau
lebih berdasarkan perbedaan titik didih. Proses tersebut dilakukan terhadap
minyak atsiri yang tidak larut dalam air.

metode penyulingan minyak atsiri dibedakan atas tiga cara, yaitu:

1. Penyulingan dengan air
Bahan yang akan disuling kontak langsung dengan air mendidih. Metode ini
dilakukan dengan panas langsung,mantel uap,pipa uap yang berlingkar
tertutup tatau dengan memakai pipa uap berlingkar terbuka atau berlubang.

2. Penyulingan dengan uap dan air

Ciri khas metode ini yaitu uap selalu dalam keadaan basah, jenuh, dan tidak
terlalu panas;bahan yang disulng hanya berhubungan dengan uap dan tidak
dengan air panas.

3. Penyulingan dengan uap.

Air tidak diisikan dalam ketel. Uap yang digunakan adalah uap jenuh atau
uap kelewat panas pada tekanan lebih dari 1 atmosfer. Uap dialirkan melalui
pipa uap berlingkar yang berpori dan terletak di bawah bahan dan uap
bergeerak ke atas melalui bahan yang terletak di atas saringan.
Proses utama yang terjadi pada
peristiwa hidro destilasi yaitu :
a. difusi minyak atsiri dan air panas melalui membran tanaman
(hidrodifusi)
b. Hidrolisa terhadap beberapa komponen minyak atsiri
c. Dekomposisi yang biasanya disebabkan oleh panas
(Guenther,1987).
 Destilasi adalah suatu pemurnian senyawa organik cair yaitu suatu
proses yang didahului dengan penguapan senyawa cair, kemudian
mengembunkan uap yang terbentuk sehingga mencair kembali.

Macam-Macam Destilasi :
1. Destilasi Uap
Proses penyaringan suatu campuran air dan bahan yang tidak larut
sempurna atau larut sebagian dengan menurunkan tekanan sistem sehingga
didapatkan hasil penyulingan jauh dibawah titik didih awal. (Cahyono, 1991)

2. Destilasi Vakum
Untuk memurnikan senyawa yang larut dalam air dengan titik didih tinggi
sehingga tekanan lingkungan harus diturunkan agar tekanan sistem
turun.(Cahyono, 1991)

3. Destilasi Biasa
Untuk memurnikan campuran senyawa dimana komponen-komponen yang
akan dipisahkan memiliki titik didih yang jauh berbeda. (Cahyono, 1991)
Prinsip Destilasi Uap

Campuran substansi yang tidak larut

menunjukkan reaksi yang sangat beda dalam larutan homogen
dan deskripsi sifatnya memerlukan hukum fisik yang berbeda.
Dasar aturan dapat dipakai dengan mempertimbangkan akibat
naiknya deviasi pada hukum rault. Satu gejala dari deviasi positif
adalah dalam diagram hubungan antara tekanan dengan
temperatur. Pada batas deviasi positif besar dari hukum rault, dua
komponen dapat larut dan komponen tersebut menguap yang
secara matematis memberikan tekanan total yang merupakan
jumlah total dari tekanan masing-masing (Wilcox, 1995).
ekstraksi
• Ekstraksi yang sering digunakan untuk memisahkan senyawa
organik adalah ekstraksi zat cair; yaitu pemisahan zat
berdasarkan perbandingan distribusi zat tersebut yang
terlarut dalam dua pelarut yang tidak saling melarutkan.

• Kriteria pemilihan pelarut:

a. Pelarut mudah melarutkan bahan yang di ekstrak
b. Pelarut tidak bercampur dengan cairan yang di ekstrak
c. Pelarut mengekstrak sedikit atau tidak sama sekali pengotor
yang ada
d. Pelarut mudah dipisahkan dari zat terlarut
e. Pelarut tidak bereaksi dengan zat terlarut melalui segala cara.
(Cahyono, 1991)
• Di dalam penelitian ini dilakukan pengambilan minyak atsiri buah
Amorpha Fruticosa dengan metode microwave hydrodistillation.

• Metode microwave hydrodistillation menggunakan gelombang mikro

yang dihasilkan dari magnetron sebagai sumber pemanasan selama
proses ekstraksi.

• Gelombang mikro atau microwave adalah gelombang elektromagnetik

dengan frekuensi super tinggi (Super High Frequency, SHF), yaitu antara
300 Mhz – 300 Ghz. Microwave memiliki rentang panjang gelombang dari
1 mm hingga 1 m.

• Mekanisme dasar pemanasan microwave melibatkan pengadukan molekul

polar atau ion yang berosilasi karena pengaruh medan listrik dan magnet
yang disebut polarisasi dipolar.

• Pemilihan metode microwave hydrodistillation karena metode ini

memiliki Keunggulan dalam pemilihan microwave bisa bekerja cepat dan
efisien. Hal ini dikarenakan adanya gelombang elektromagnetik yang bisa
menembus bahan dan mengeksitasi molekul-molekul bahan secara
merata
Bahan yang diguunakan dalam jurnal kami adalah
1. Etil eter
2. n-heksana
3. petroleum eter (rentang didih 30–60 ◦C)
4. sikloheksana
5. n-pentana dibeli dari Tianjin Fuyu Fine Chemical Co., Ltd.
(Tianjin, Cina)

alat yang di gunakan yaitu:

1. oven microwave
2. peralatan ekstraksi cair cair hidrodistilasi yang digabungkan.
 Hydrodistillationdengan bantuan gelombang mikro prosedur ekstraksi
cair-cair

• 50,0 g buah A. fruticosa ditimbang secara akurat dan di campur air

deionisasi (untuk mencapai rasio cairan-padat yang dibutuhkan)
• ditempatkan ke dalam blender Waring untuk waktu homogenat
yang berbeda, berdasarkan penelitian sebelumnya.
• Kemudian, suspensi dipindahkan ke labu alas bulat 1000 mL, yang
ditempatkan ke dalam oven microwave dan dilekatkan pada
peralatan ekstraksi cair-cair terpadu yang dikembangkan dengan
hidrodistilasi dan kondensor melalui lubang di bagian atas oven
microwave.
• Sebelum distilasi, volume dan jenis pelarut organik yang berbeda
ditambahkan ke kolom reparasi kedua untuk ekstraksi komponen
yang dilarutkan dalam air.
• Setelah campuran sampel mulai mendidih, suspensi kemudian
disinari oleh microwave untuk berbagai waktu di bawah kekuatan
yang berbeda. ;
• Komponen volatil dapat dilepaskan dari sampel dan
diuapkan dengan uap air,
• kemudian dikondensasikan dan dipisahkan dari air, yang
diselesaikan dalam kolom pemisahan pertama. Air akan
kembali ke labu sampel secara konstan ketika mencapai
tingkat banjir.
• Setelah setiap proses, minyak esensial yang diperoleh dalam
(kolom pemisahan pertama) dikumpulkan, dikeringkan
dengan menggunakan natrium sulfat anhidrat, dan disimpan
pada suhu 4 ◦C sampai analisis.
• Pelarut ekstraksi dalam (kolom pemisahan kedua)
dikeringkan dengan natrium sulfat anhidrat semalam dan
kemudian dipekatkan untuk mendapatkan minyak esensial di
bawah tekanan yang dikurangi, yang disimpan pada 4 ◦ C
sebelum analisis.
Hasil minyak esensial yang diperoleh dalam dua kolom
pemisahan dihitung sebagai berikut

Y = M/ Mo
• dimana Y (g / kg) adalah hasil dari minyak atsiri;
• M (g) adalah massa rata-rata minyak esensial yang diperoleh
dalam dua kolom pemisahan,
• Mo (kg) adalah massa rata-rata dari tiga sampel buah sebelum
distilasi.
faktor yang mempengaruhi Dalam penelitian
ini, beberapa faktor yang berpengaruh,yaitu :

1. Jenis dan volume pelarut ekstraksi di kolom

pemisahan kedua.
2. Waktu homogenate
3. Rasio cair-padat
4. Daya iradiasi microwave
5. Waktu iradiasi microwave
Hasil
• Dari jurnal dapat diketahui bahwa hasil yang di peroleh yaitu
• Hasil EOF (kolom pemisahan 1): 10,31 ± 0,33g/kg
• Hasil EOS ( kolom pemisahan 2): 0,82 ± 0,03g/kg
• Di peroleh pada kondisi optimum (2 ml etil eter sebagai
pelarut ekstraksi dalam kolom pemisahan , rasio cair padat
12ml/g wktu homogent 4 menit, waktu penyinaran 35 menit,
dan daya iridasi gelombang mikro 540w. Di bandi ng metode
tradisional lainnya MHDLLE memberikan hasil yang lebih
tinggi.
ALAT YANG DI GUNAKAN
TERIMAKASIH