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MATERIALS AND METHODS was refluxed 5 times with 110 l of an 8:2 mixture
of water and alcohol for 6 hr each time. It was then
Drugs Diakyur contains crude powder of Cas- concentrated under vacuum at 60 C. The herb to
sia javanica and dried extracts of Cassia auricu- extract ratio was 23:1. Total saponin content was
lata, Salacia reticulata, Gymnema sylvestre, Mu- determined by a gravimetric method.
cuna pruriens, Syzygium jambolanum and Termina- Cassia javanica bark: 15 kg crude powder was
lia arjuna, which were standardized using their re- used for the formulation and it was standardized by
spective bioactive marker compounds. HPTLC method of analysis. The raw material was
Extraction and Standardization of Plant Materi- compared with an in-house reference standard for
als: its total anthraquinone content.
Cassia auriculata seeds: 165 kg of plant ma- Standardization of the Diakyur Formulation:
terial was refluxed 5 times with 825 l of water for The extracts obtained from the authenticated
6 hr each time. It was then concentrated under vac- plant specimens were mixed in the right propor-
uum at 7075 C. The herb to extract ratio was tion and maintained as reference standard. The
15:1. It was standardized for its total emodin con- batches were compared with the reference standard
tent by High performance thin layer chromatogra- by HPTLC analysis. One g of the material was re-
phy (HPTLC). The stationary phase used was Sil- fluxed with 20 ml of alcohol in a water bath for 1 hr.
icagel 60F254 (Merck), (Mumbai, India). The mo- It was then filtered and the filtrate was concentrated
bile phase used was ethyl acetate: methanol: water to 5 ml. The concentrate was spotted and developed
(10:1.35:1.0) and it was detected by spraying 10% in a mobile phase consisting of toluene, ethyl ac-
ethanolic potassium hydroxide reagent. etate, and formic acid (6:4:0.4). The dried plate was
Syzygium jambolanum seeds: 162.5 kg of plant then scanned at 254 nm. The fingerprinting of the
material was refluxed 5 times with 815 l of water batches was compared with the reference standard.
for 6 hr each time. It was then concentrated un- The dried powder was dissolved in water by
der vacuum at 7075 C. The herb to extract ratio constant stirring in a water bath to reflux, filtered
was 8:1. It was standardized for its total tannin and administered to the animals by oral intubation.
content, which was determined by redox titration Animals Colony bred Wistar albino rats and
against 0.1 N potassium permanganate. Swiss albino mice were used. The animals were
Terminalia arjuna bark: 33.5 kg of plant mate- fed cereals, pulses, and green vegetables and had
rial was refluxed 5 times with 170 l of water for 6 hr free access to water. They were housed in Polyvinyl
each time. It was then concentrated under vacuum at Chloride (PVC) cages. Rats of either sex weighing
7075 C. The herb to extract ratio was 15:1. It was 125150 g and mice of either sex weighing 1822 g
standardized for its total tannin content, which was were used and approved by the Institutional Animal
determined by redox titration against 0.1 N potas- Ethical Committee (IAEC) No. 07/017/03, Institute
sium permanganate. of Basic Medical Sciences (IBMS), University of
Mucuna pruriens seeds: 50 kg of plant material Madras, India.
was refluxed 5 times with 250 l of water for 6 hr Acute Toxicity Study Swiss albino mice of
each time. It was then concentrated under vacuum either sex weighing 1822 g were randomly dis-
at 7075 C. The herb to extract ratio was 10:1. It tributed to 8 different groups with 6 animals in each
was standardized for its levodopa (L-DOPA) con- group. The animals were fasted overnight and the
tent, which was carried out by HPLC. The column drug was administered orally at dose levels of 100,
used was Phenomenex C18 and the mobile phase 200, 400, 800, 1600, 3200, 6400 and 12800 mg/kg
was 0.1% v/v phosphoric acid in water: acetonitrile of body weight. The animals were closely observed
(95:05). Peak detection was performed at 281 nm. for the first 12 hr for any toxic symptoms and for
Gymnema sylvestre leaves: 50 kg of plant mate- 72 hr for mortality rate.9)
rial was refluxed 5 times with 250 l of an 8:2 mix- Subacute Toxicity Study Wistar albino rats
ture of water and alcohol for 6 hr each time. It was of either sex weighing 125150 g were assigned to
then concentrated under vacuum at 7075 C. The each group (6 per group). Group I received distilled
herb to extract ratio was 10:1. Total gymnemic acid water for 28 days and Group II received the test drug
content was standardized by a gravimetric method Diakyur at the dose of 1600 mg/kg p.o., once daily
of determination. for 28 days. Body weight, food intake and water in-
Salacia reticulata bark: 22 kg of plant material take were monitored. The animals were sacrificed
No. 2 247
Table 4. Effects of 28 Day Administration of Diakyur on Kid- fate structure, from antidiabetic ayurvedic medicine
ney Function in Rats Salacia reticulata. Chem. Pharm. Bull., 46, 1339
Group Treatment Blood urea Serum creatinine 1340.
(mg%) (mg%) 3) Shanmugasundaram, E. R. B., Goliaths, K. L.
I Control 17.1 0.5 1.04 0.8 and Radhashanmugasundaram. K., (1990) Pos-
II Diakyur 16.9 0.4 0.98 0.7 sible regeneration of the islets of Langerhans
Data are the mean S.E. of 6 animals (One-way ANOVA). in streptozotocin-diabetic rats given Gymnema
sylvestre leaf extracts. J. Ethnopharmacol., 30, 265
279.
indicated hepatocyte damage.19) The normal levels 4) Prince, A. S. M., Menon, V. P. and Pari, L. (1998)
Hypoglycaemic activity of Syzigium cumini seeds:
of blood urea and serum creatinine (Table 3) indi-
effect on lipid peroxidation in alloxan diabetic rats.
cate that the test drug did not interfere with renal
J. Ethnopharmacol., 61, 17.
function and that renal integrity was preserved.20)
5) Karthikeyan, K., Bai, B. R., Gauthaman, K.,
Also, there were no significant changes in various
Sathish, K. S. and Devaraj, S. N. (2003) Cardiopro-
hematological parameters such as Hb, RBC, WBC,
tective effect of the alcoholic extract of Terminalia
ESR and differential count compared to the con-
arjuna bark in an in vivo model of myocardial is-
trol group, which indicates that Diakyur may not chemic reperfusion injury. Life Sci., 73, 27272739.
be toxic and does not affect circulating red cells, 6) Chander, R., Kavita, S., Khanna, A. K., Kaul, S.
hematopoiesis, or leukopoiesis. M., Anju, P., Rashmi, S., Gitika, B., Farha, R. and
The present findings suggest that Diakyur is Rastogi, A. K. (2004) Antidyslipidemic and antiox-
nontoxic since no marked changes in hematologi- idant activities of different fractions of Terminalia
cal, biochemical, and histopathological parameters arjuna stem bark. Indian J. Clin. Biochem., 19, 141
were observed. Thus, at normal therapeutic doses, 148.
Diakyur is considered to be safe for long-term treat- 7) Pari, L. and Latha, M. (2002) Effect of Cassia au-
ment in diabetic conditions. riculata flowers on blood sugar levels, serum and
tissue lipids in streptozotocin diabetic rats. Singa-
pore Med. J., 43, 617621.
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