Journal of Pharmacy Research Vol.4.Issue 4. April 2011 Nilakshi V. Gambhir et al. / Journal of Pharmacy Research 2011,4(4),1197-1198 1197-1198 Research Article ISSN: 0974-6943 Available online through * Corresponding author. Nilakshi V. Gambhir Department of Botany, GDM Arts, KRN Commerce and MD Science College, Jamner, Dist. Jalgaon, Pin. 424 206, India I NTRODUCTI ON The family Fabaceae with 730 genera and over 19,400 species is an economically important family found throughout the world [1] . Pithecellobium dulce benth is one of the common road side fabaceae member found in Asia. The plant is extensively used in number of traditional medicine treatments and is reported to be a folk remedy for ear ache, leprosy, peptic ulcer and tooth ache. Infusions of different plant parts have been used traditionally to treat dysentry, intestinal disorders, ulcer, and indigestion etc. Pithecellobium dulce pods are customarily sold on roadside stands in tropical countries, contain a thick sweetish, but also acidic pulp, eaten raw or made into a drink by tribal peoples and also used in traditional medicines but still most of the chemical constituents of the pods are remain unexplored and underutilized [2,3,4] . Vitamine C is an effective anti-oxidant and is necessary for treatment of scurvy, in humans. It performs numerous physiological functions in human body. The richest natural sources of vitamin C are fruits and vegetabels [5,7] . The present investigation has therefore been made to find out Vitamin C con- tent of the fresh Pithecellobium dulce pods by simple and reproducible HPTLC method. MATERIALS AND METHODS : Plant Material: Fresh and healthy Pithecellobium dulce pods were collected from the forests of Thane district, Maharashtra, India. The plant was authenticated at Department of Botany, MD Science College, Jamner, Maharashtra. A herbarium specimen, number MDSC/PS/20 was also preserved in the same college. Fresh pod pulp was then separated from the seeds and husk. Extraction of Plant Material: Fresh pulp weighing 100gm were macerated with 150 ml of 50% methanol for 24 hrs with occasional shaking. The extract was separated and mark was again extracted twice with 50 ml of 50% methanol (fresh). The extract was pooled (mixed) and concentrated in a rotary vaccum evaporator to get 100 ml of sample [6] . HPTLC analysis of vitamin C from Pithecellobium dulce, Benth (Fabaceae) Nilakshi V. Gambhir* and V. V. Bhaskar. Department of Botany, GDM Arts, KRN Commerce and MD Science College, Jamner, Dist. Jalgaon, Pin. 424 206, India Received on: 04-01-2011; Revised on: 17-02-2011; Accepted on:16-03-2011 ABSTRACT This work describes the simplest HPTLC method developed for quantification of vitamin C from methanolic extract of Pithecellobium dulce pods, a traditional medicinal plant widely used in Asia. A good separation was achieved with mobile phase ethanol: water (2:1) on precoated silica gel 60 F 254 HPTLC plates. Quantitative analysis was carried out in the absorbance at 254nm. A good linear relationship, r 2 = 0.992 with respect to peak area was observed between the concentration ranges of 1.0 -6.0 μg. Pithecellobium dulce pods are a rich source of vitamin C and can be considered as a functional food. The method developed is reproducible and can be used for routine analysis of vitamin C in crude drugs and in herbal and traditional dosage containing Pithecellobium dulce as an ingredient. Key words: Pithecellobium dulce pod, HPTLC analysis, Vitamin C, traditional medicine. One ml of sample extract was further diluted to 10 ml with 50% Methanol which was used for quantitative estimation. Preparation of Standard Vitamin C and calibration curve: Ten mg of std. vitamin C sample dissolved in 10 ml of 50% of Methanol for the preparation of Standard Curve. The six standard levels (1, 2, 3, 4, 5 and 6 μg) of Std. vitamin C were used for the calibration curve for which 1, 2, 3, 4, 5 and 6 μl of standard solution was applied in triplicate on a TLC Plate. The chromatogram was developed for 15 mts, dried at room temperature and scanned at 254 nm; average peak areas of 3 standards were calculated. The calibration curve of the standard drug concentration (X-axis) over the average peak area (Y-axis) was prepared to get a regression equation by Win Cats Figure 1. HPTLC Chromatogram of vitamin C Figure 2. Standard curve of vitamin C with respect to area http://jprsolutions.info