1. INTRODUCTION:

Biocrystals are the valuable tools in medical field. Most of the living beings form crystals. The body fluids contain minerals at various levels of saturation. When the body fluids get supersaturated with minerals, crystallization takes place. These crystals have both beneficial as well as pathological effects on humans. The major beneficial roles of mineralization are the formation of bones and teeth. The pathological effects result in the crystal deposition diseases which may be defined as the pathological condition associated with the presence of crystals which contributes to the tissue damage and cause pain and sufferings. Uric acid crystals are the gout culprits. A saturated solution of sodium urate in the presence of urate seeds at the pH greater than 6 will form needle shape sodium urate crystals. Lowering of pH to 5.59 will result in the nucleation and growth of uric acid crystals. These crystallized uric acids in the joints are the main causative agents of gouty arthritis.

The uric acid crystals are grown by means of gel growth technique. It has gained considerable importance due to its simplicity and effectiveness in growing single crystals of certain compounds. The growth of single crystals in a gel is a self purifying process, free from thermal strains which is common in crystal grown from the melt. The gel method finds way to study the crystal formation in human system. Colchicine (Cc) is a medication of acute gout. It is effective in suppressing the imflammatory response caused by uric acid crystals. It is used to treat acute flares of gouty arthritis and to prevent recurrent acute attacks. It can arrest cell division and it is avoided in pregnancy since it affects the fetal growth. Nyctanthes arbor-tristis (Nat) is commonly known as knight jasmine are coral jasmine. It is a native of India and it is widely distributed throughout India, Sri Lanka, Bangaladesh and other parts of south East Asia [1-4]. The plant has been studied pharmacognostically and pharmacologically [5, 6]. The phytochemicals screening of the extract of Nyctanthes arbor-tristis leaves revealed the presence of important phytochemicals. Flavanoids are known to posses several therapeutic activities like anti-inflammatory, anti allergic, hepatoprotective, antithrombotic, antiviral and anticarcinogenic activities [7]. In view of the diverse pharmacological activities present in Nyctanthes arbor-tristis with its central effects hyperuricemia, the present investigation was undertaken to compare the pharmacologic property of Nyctanthes arbor-tristis with Colchicine on the growth of uric acid crystals.

2 MATERIALS AND METHODS:

2.1 Collection of Plant materials and Preparation of leaf extracts:

Leaves of Nyctanthes arbor-tristis were collected from Erode District, Tamil Nadu (India) during the month of June 2016. Collected fresh leaves were washed thoroughly, shade dried, powdered coarsely. The powder obtained (250 g) were extracted with ethanol in a soxlet extractor for 18 to 20 hours. The extract was concentrated using rotary flash evaporator and preserved at 4’C in air tight container.

2.2 Chemicals:

Sodium meta silicate and Uric acid were purchased from LOBA Chemie, Mumbai and all the chemicals used were of analytical grade.

2.3 Phytochemical Analysis:

The prepared extract was subjected to phytochemical screening by the method of Kokate [8].

2.4 Formation of Uric acid crystals:

Crystallized uric acids were formed by single diffusion gel growth technique. In a series of 3 test tubes, the gel medium is prepared by using 1% sodium meta silicate and 3 ml of 1% uric acid solution. One test tube is maintained as control (drug free) and the other two test tubes are considered as T1(treated with Nyctanthes arbor-tristis) and T2(treated with Colchicine). After gelation, 1 ml of 0.5mg of Nyctanthes arbor-tristis is added to T2 and 1 ml of 0.5 mg of Colchicine is added to T3 over the set jel.

3 RESULTS AND DISCUSSION:

Table 1 showed the presence of phytochemical constituents in Nyctanthes arbor-tristis. This showed the presence of alkaloids, flavanoids, saponins, phenols, tannins and glycosides. The presence of phytochemicals in Nyctanthes arbor-tristis reflected the complex mixture of total phenolic and flavanoid compounds. The flavanoids occur in a combined form as glycosides. In the flavanoid structure, attachment of sugar moiety takes place in any hydroxyl groups. The common positions are 7 hydroxyl group in flavones, flavanones and isoflavanones, 3 and 7 hydroxyls in flavanols and 3 and 5 hydroxyls in anthocyanidins [9].

table 1: Phytochemical analysis of Ethanolic extract of Nyctanthes Arbor. Tristis

Phytochemicals	Observations
Alkaloids	+
Saponins	+
Phenols	+
Tannins	+
Steroids	_
Terpenoids	_
Flavanoids	+
Glycosides	+

Studies have reported that the ethanolic extract of the leaves of Nyctanthes arbor-tristis exhibited significant anti inflammatory activity against acute inflammatory oedema produced in rats in hindpaw of the rats and the extract significantly reduces the acute inflammatory swellings in the knee joint of rats induced by the turpentine oil [10]. The scavenging potential of Nyctanthes arbor-tristis was also due to high phenolic and flavanoid content[11].

Table 2: Effects of Nyctanthes arbor-tristis and Colchicine on the Uric acid crystals.

	Vol. of 1% Uric acid (ml)	Concentration of drug (mg)	Size of Crystals	Time of disappearing of crystals (days)	Inference
Test 1	3.0	Nil	0.8cm(l) 0.4mm(t)	Nil	Presence of more number of crystal
Test 2	3.0	0.5	0.3cm(l) 0.2mm(t)	3	Presence of minute granule and absence of crystals
Test 3	3.0	0.5	0.5cm(l) 0.4mm(t)	8	Less number of broken crystals with minute granules

Table 2 and figure 1 illustrated the effects of Nyctanthes arbor-tristis and colchicine on the growth of uric acid crystals. In the present study, microscopic examination of the empty crystals (drug free) revealed that the crystals were 0.8 cm in length and 0.4mm in thickness. Size of crystals in T2 is 0.3 cm in length and 0.2 mm in thickness which was low when compared to T3(0.5 cm in length and 0.4 mm in thickness). When compared to T3, there was complete absence of crystals in T2 and also the time of disappearance of crystals in 3 days. This showed the higher efficiency of Nyctanthes arbor-tristis than colchicines.

The uric acid crystals have been considered as the important risk factors for gout which causes the deposits in the synovial fluid and synovial lining. The control of uric acid production is a key factor in the prevention and treatment of gout. Xanthine oxidase is a important enzyme involved in the catabolism of purines and it catalyses the oxidation of hypoxanthine to xanthine and xanthine to uric acid. Hence the inhibition of xanthine oxidase activity may decrease uric acid. In the present study, the reduction of uricacid crystals by Nyctanthes arbor-tristis proved the hypouricemic effects of Nyctanthes arbor-tristis leaves. This may be due to its flavanoid compounds which is considered to be the inhibitor of Xanthine oxidase and with anti oxidative action. Several invitro observations also confirmed the xanthine oxidase inhibitory activity of some phytochemicals such as flavanoids [12 - 15].

Figure:1 Effects of NAT and CC on the uric acid crystals

4 CONCLUSION:

In conclusion, the data reported in the present study indicate the enhanced hypouricemic effects of Nyctanthes arbor-tristis leaves than colchicines. It is therefore suggested that Nyctanthes arbor-tristis leaves with its flavanoids may represent a new type of hypouricemic agents and may have potent hypouricemic effect in clinical use.

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Received on 01.02.2017 Accepted on 25.04.2017

Asian J. Pharm. Ana. 2017; 7(2): 84-86.

DOI: 10.5958/2231-5675.2017.00014.X