and quantity of the active principl

and quantity of the active principle in the
herbal extract and herbal formulations.
Standardization is a basic prerequisite of
quality. Irrespective of the question of
whether the active ingredient of an herbal
drug are known or not, every manufacturing
process should be subjected to
standardization. The extract can be
standardized to a specified percentage of
active principle by appropriate dilution with
adjuvant carrier materials. In practice most
botanicals owe their activity to a group of
structurally related active compounds rather
than to a single compound and the extract
may be standardized according to its total
content of active ingredients. The
presumption is that when the extract is
properly standardized to a key constituent,
the other constituents in the extract which
are responsible for the activity are also
present in sufficient quantity. In these cases
the constituent in the question is a marker
substance; while practical experiences have
shown that extracts containing the marker
compound at a specified percentage have the
desired activity. Standardization of botanical
extract necessary because strong
standardized extracts may cause stomach
upset and should be used for no more than
two weeks continuously[19,20]. So the present
work is carried out for preliminary
Phytochemical screening, isolation of
phytomarker and standardization of aqueous
extract of Berberis vulgaris.
2. Materials and Methods
2.1 Plant Material and Extract
Preparation
Fresh root of Berberis vulgaris L. were
collected from Jammu, in the month of
November. Roots were cleaned with running
tap water were chopped into pieces. They
were dried under shade at ambient
temperature for 5 days and the air-dried
roots were then ground to powder for
extraction. The aqueous extract was
prepared by cold maceration of 1.5Kg of
powdered root in 5lts of distilled water for
48 hr. Then the extract was filtered,
concentrated, dried in vacuum (yield
142gm).
2.2 Preliminary Phytochemical
Investigation
Preliminary Phytochemical analysis was
performed through standard official
procedure for the identification of different
classes of components present in extract[21].
2.3 Isolation of Berberine
The aqueous extract was dissolved in 1%
HCl. The solution was filtered, alkalinized
with a concentrated NH4OH to pH 8 and
extracted with chloroform from which
tertiary alkaloids were obtained after
evaporation of the solvent (21.22 gm). The
pure phytoconstituents from the chloroform
fraction isolated by column chromatography
on silica gel 100-200 mesh and eluted with
chloroform and gradient with methanol
(CHCl3:MeOH, 9:1; 8:2) to isolated a
yellow needles shape crystal compound
(1.021gm) which was identified by TLC, 1H
NMR, 13C NMR and compared with the
spectral data from literature values[22].
2.4 Standardization of Aqueous Extracts
by RP-HPLC
2.4.1 Preparation of Standard Solution
A stock solution of 10mg/ml is prepared by
taking 100mg accurately weight pure
standard Berberine and transferred to a 10
ml volumetric flask. 7ml of hplc grade
methanol was added and sonicated for 10
min. Final volume is made up to 10ml of
hplc grade methanol to a concentration of
10mg/ml. The working standard solution of
concentration 400, 800, 1200, 1600,
2000ppm are prepared by transferring 0.4,
0.8, 1.2, 1.8, 2ml solution from stock
solution and volume made up to 10ml with
hplc grade methanol.

Hasil (Bahasa Indonesia) 1:

dan jumlah prinsip aktif dalamekstrak herbal dan formulasi herbal.Standardisasi merupakan prasyarat dasar darikualitas. Terlepas dari pertanyaanApakah bahan aktif herbalobat yang dikenal atau tidak, setiap manufakturproses harus tunduk kepadaStandardisasi. Ekstrak dapatstandar untuk persentase tertentuprinsip aktif oleh diencerkan denganajuvan pembawa bahan. Dalam prakteknya kebanyakantumbuhan berutang aktivitas mereka ke grupstruktural terkait senyawa aktif agakdaripada senyawa tunggal dan ekstrakmungkin standar menurut totalnyakandungan bahan aktif. TheAnggapan ini adalah bahwa ketika ekstrakbenar standar untuk Konstituante kunci,konstituen lain dalam ekstrak yangbertanggung jawab untuk aktivitas jugahadir dalam jumlah yang memadai. Dalam kasus inikonstituen dalam pertanyaan adalah penandasubstansi; sementara memiliki pengalaman praktismenunjukkan bahwa ekstrak yang mengandung penandasenyawa persentase tertentu memilikikegiatan yang diinginkan. Standardisasi botaniekstrak diperlukan karena kuatstandar ekstrak dapat menyebabkan perutmarah dan harus digunakan untuk tidak lebih daridua minggu terus menerus [19,20]. Jadi sekarangpekerjaan dilakukan untuk awalFitokimia skrining, isolasiphytomarker dan standardisasi berairekstrak Berberis vulgaris.2. bahan dan metode2.1 tanaman bahan dan ekstrakPersiapanAkar segar Berberis vulgaris L. yangdikumpulkan dari Jammu, di bulanBulan November. Akar dibersihkan dengan menjalankanair keran yang dicincang potong. Merekayang kering di bawah naungan di ambientsuhu selama 5 hari dan kering udaraakar yang kemudian digiling menjadi bubuk untukekstraksi. Ekstrak berair adalahdisiapkan oleh dingin maserasi 1,5 kgbubuk akar dalam 5lts air suling untuk48 jam. Kemudian ekstrak disaring,terkonsentrasi, kering dalam kekosongan (hasil142 gm).2.2 awal fitokimiaPenyelidikanAnalisis fitokimia awal adalahdilakukan melalui standar resmiprosedur untuk identifikasi berbedakelas komponen hadir dalam ekstrak [21].2.3 isolasi BerberineEkstrak berair dilarutkan dalam 1%HCl. Larutan disaring, alkalinizeddengan NH4OH terkonsentrasi menjadi pH 8 dandiekstrak dengan kloroform yangalkaloid tersier diperoleh setelahpenguapan pelarut (21.22 gm). Thephytoconstituents murni dari kloroformsebagian kecil yang terisolasi oleh Kromatografi kolomsilika gel 100-200 mesh dan eluted dengankloroform dan gradien dengan metanol(CHCl3:MeOH, 9:1; 8:2) untuk terisolasikuning jarum bentuk kristal senyawa(1.021 gm) yang diidentifikasi oleh TLC, 1 HNMR, 13C NMR dan dibandingkan denganspektral data dari nilai-nilai sastra [22].2.4 Standardisasi ekstrak cairdengan RP-HPLC2.4.1 Persiapan larutan standarLarutan saham 10mg/ml disiapkan olehmengambil 100mg akurat berat murniBerberine standar dan ditransfer ke 10labu Volumetrik ml. 7ml hplc kelasmetanol ditambahkan dan sonicated untuk 10minimal volume akhir dibuat sampai dengan 10mlHPLC kelas metanol konsentrasi10mg/ml. Kerja standar solusikonsentrasi 400, 800, 1200, 1600,2000ppm disiapkan oleh mentransfer 0.4,0,8 1.2, 1.8, 2ml larutan dari sahamsolusi dan volume dibuat hingga 10ml denganHPLC kelas metanol.