Gul et al / Chemistry International 1(1) (2015) 103-106

New validated method for analysis of salymarin in polyherbal formulation (aqueous extract, oral liquid and solid dosage form)

Somia Gul, Kashifa Khanum and Nusrat Mujtaba

 

Faculty of Pharmacy, Jinnah University for women, Karachi, Pakistan

*Corresponding author's e-mails: drsomi1983@yahoo.com (S.G), Kashifa_khanum@yahoo.com (K.K), Nusratafzal976@yahoo.com (N.M)

 

ABSTRACT

A UV/visible Spectrophotometric method for the determination of Silymarin in poly herbal formulation have been developed. Developed method is based on the simple concept of solubility of chemical i.e. Silymarin in methanol. This method is simple, rapid, accurate, sensitive and precise for the determination of  Silymarin in polyherbal formulation like aqueous extract, Oral liquid (i.e. Syrup) and solid (i.e. Capsules) dosage form containing crude herb Silybum marianum. The λmax. of Silymarin measured at 287nm.  The molecule (Silymarin) obeys Beer’s law in between range of 0.3 – 1.6 mg/ml. The method is reproducible, accurate and linear for routine quality control testing of phyto-pharmaceutical dosage forms.

Keywords: Poly herbal formulations, Aqueous extract, Oral liquid, Oral solids, Silymarin, UV spectrophotometric

 

Capsule Summary: Proposed method is potential method for the utilization in analysis of commercial batches of poly herbal formulation (Aqueous extract, oral liquid and solid dosage form).

Cite This Article As: Somia Gul, Kashifa Khanum and Nusrat Mujtaba. New validated method for analysis of salymarin in polyherbal formulation (aqueous extract, oral liquid and solid dosage form). Chemistry International 1(3) (2015) 103-106.


 

INTRODUCTION

 

Silymarin is hepatoprotective flavonoid drug available as bio marker in Silybun marianum (common name Milk thistle). In British pharmacopoeia individual monograph for Milk Thistle Fruit is available with claim of silymarin, expressed as silibinin (C25H22O10; MW 482.4) (British Pharmacopoeia, 2013). Silymarin is being used in treatment of various liver disease of different etiology because of its hepatoprotective action (Cavalieri, 1974; Salmi et al., 1982; Bosisio, 1992; Ferenci, 1992).

There are different analytical test method are available in pharmacopoeia or other source of information. HPLC method is available in British Pharmacopoeia 2013 on gradient system and other HPLC methods also available (Radijabian et al., 2008; Kvasnicka et al., 2003; Lee et al., 2007; Hadad et al., 2009).

Method on other analytical techniques are also available like thin layer chromatography and UV spectrophotometer (Abdel-Salam et al., 1982; Vanhaelen and Vanhaelen-Fastre, 1983; Dube and Vyas, 2009). Still little work has been done specially on simple analytical technique like UV spectrophotometry and no method available for poly herbal extract and phyto-pharmaceuticals.

 

EXPERIMENTAL

 

Reagent

 

Silymarin standard

Methanol

Chloroform

Polyherbal Aqueous Extract B # E14 001

Oral Liquid: Bonjigar Syrup B # 1314 001

Oral Solid: Bonjigar Capsule B # 1414 001

Instrumentation

 

UV Spectrophotometer: Perkin Elmer Lamba 25

 

Experiment conditions

 

Wavelength: 287 nm

 

Preparation of standard and sample solution

 

A 100 mg of standard were accurately weight and transfer into 100 mL volumetric flask, dissolve in methanol and make up the volume with methanol and mix well.

Total 6 volumetric flasks of 100 mL were taken and marked with 1, 2, …..,6, transfer 0.3, 0.4, 0.5, 0.6 and 0.7 mL of standard to volumetric flasks through pipette and make up the volume with methanol and mix well.

Table 1: Calibration data and linearity

Concentration

Absorbance ± SD

(mg/mL)

 

0.3

0.2075 ± 0.0001

0.4

0.2937 ± 0.0005

0.5

0.3642 ± 0.0003

0.6

0.4614 ± 0.0003

0.7

0.5277 ± 0.0003

1

0.7108 ± 0.0001

 

Table 2: LOD and LOQ results

Concentration

LOD

LOQ

Range

(mg/mL)

(mg/mL)

0.3 – 1 g

0.015

0.06

 

Table 3: Inter day precision results of day 1

Concentration

Absorbance ± SD

(mg/mL)

 

0.3

0.2075 ± 0.0001

0.4

0.2937 ± 0.0005

0.5

0.3642 ± 0.0003

0.6

0.4614 ± 0.0003

0.7

0.5277 ± 0.0003

1

0.7108 ± 0.0001

 

Table 4: Inter day precision results of day 2

Concentration

Absorbance ± SD

(mg/mL)

 

0.3

0.1975 ± 0.0001

0.4

0.2836 ± 0.0003

0.5

0.3541 ± 0.0002

0.6

0.4513 ± 0.0001

0.7

0.5175 ± 0.0001

1

0.7008 ± 0.0001

 

 

Table 5: Results of commercial products

Products

Results

Extract

99.80%

Bonjigar Capsule

101.00%

Bonjigar Syrup

100.70%

 

 

Preparation of sample solution

 

Extract

 

Accurately weight and transfer 200mg of extract in separating funnel, mix with 20ml water, extract the content with using 50ml chloroform. Collect the aliquot in round bottom flask through filter paper containing sodium phosphate. Repeat the same procedure 3 times. Evaporate the combine filtrate on water bath. Dissolve the residue in 10ml methanol and transfer to 50ml volumetric flask quantatively. Make –up the volume with methanol and mix well.

 

Oral liquid

 

Accurately transfer 10ml of syrup in separating funnel, mix with 20ml water, and extract the content with using 50ml chloroform. Collect the aliquot in round bottom flask through filter paper containing sodium phosphate. Repeat the same procedure 3 times. Evaporate the combine filtrate on water bath to dryness. Dissolve the residue in 10ml methanol and transfer to 50ml volumetric flask quantatively. Make –up the volume with methanol and mix well. 

 

Oral solid

 

Accurately transfer content of capsule in separating funnel, mix with 20ml water, and extract the content with using 50ml chloroform. Collect the aliquot in round bottom flask through filter paper containing sodium phosphate. Repeat the same procedure 3 times. Evaporate the combine filtrate on water bath to dryness. Dissolve the residue in 10ml methanol and transfer to 50ml volumetric flask quantatively. Make –up the volume with methanol and mix well. 

 

RESULT AND DISCUSSION

 

Method development and optimization

 

Standard and sample solutions were separately scan in the range of 200–400 nm to measure the absorbance maximum of standard and sample. Both samples showed absorbance maximum at 287nm. Placebo sample was also run in the same range and no peak observed at ʎmax 287 nm.

 

Calibration data and linearity

 

Readings of 6 dilutions were taken. Results are summarized in Table 1.

 

Specificity/selectivity

 

Method specificity or selectivity was performed by running placebo samples with standard. There is no peak at 287nm.

 

Limit of detection (LOD) and Limit of quantification (LOQ)

 

The limit of detection (LOD) and limit of quantification (LOQ) of this method were determined from the known concentrations of Silymarin. The LOD and LOQ for this analysis were calculated from using three and ten times the noise level of the response, respectively. Results are given in Table 2.

 

Precision (Inter-day)

 

Same samples were kept at ambient room temperature and again readings were taken and results are shown in Table 3 and 4.

 

Fig. 1: Linearity plot

 

Application of method on poly herbal formulation

 

Samples of commercial batches were tested using this method. Results are shown in Table 5.

The method was developed and optimized by scanning on UV spectrophotometer in range of 200 – 400nm. Data showed that method is reproducible and precise. 

The method selectivity/specificity was performed by scanning standard, sample and placebo and no peak found in placebo at λmax i.e. 287nm.

Test for Linearity was performed in the range 0.3 – 1 mg mL -1 of Silymarin with a correlation coefficient ± 0.99.  Results reveal that method is linear for the range of assay i.e. 80 – 120% (Fig. 1).

Method precision (inter-day) was checked by performing test on same samples and system on next day and results RSD found less than 2%.

A commercial batch of poly herbal extract, product Bonjigar Syrup and Bonjigar capsules was randomly selected for analysis and method was applied to quantify the content of Silymarin in product.

 

CONCLUSION

 

A simple, rapid, sensitive and reliable UV Spectrophotometric method for analysis of the Silymarin has been developed and method validity verified by performing linearity, and inter-day precision. Proposed method is potential method for the utilization in analysis of commercial batches of poly herbal formulation (Aqueous extract, oral liquid & solid dosage form).

 

REFERENCES

 

British Pharmacopoeia, 2013, Volume I & II, Medicinal & Pharmaceutical Substance, Milk Thistle Fruit.

Cavalieri, S,A., 1974. controlled clinical trial of Legalon in 40 patients, Gazzette MedicaItaliana 1974; 133 628.

Salmi, H.A., Sarna, S., 1982. Scand. Effect of silymarin on chemical, functional, and morphological alterations of the liver. A double-blind controlled study. Journal of Gasteroenterology 17, 517.

Ferenci, P., Dragosics, B., Dittrich, H., Frank, H., Benda, L., Lochs, H., Meryn, S., Base, W., Schneider, B., 1989. Randomized control trial of silymarin in patients with cirrhosis. Journal of Hepatology 9, 105.

Bosisio, E., Benelli, C., Pirola, O., 1992. Effect of the flavanolignans of Silybum marianum L. on lipid, Pharmacological. Research 25, 147.

Radijabian, T., Rezadeh, Sh., Huseini, H.F., 20008 Trends in publication of evidence-based Traditional Iranian medicine in endocrinology and metabolic disorders. Iranian Journal of Science & Technology A 32, A2.

Kvasnicka, F., Bba, B., Sevck, R., Voldrich, M., Kratka, J., 2003. Analysis of the active component of Silymarin, Journal of Chromatography A, 2003, 990; 239–245.9.

Lee, J.I., Narayan, M., Barrett, J.S., 2007. Analysis and comparison of active constituent in commercial standardized Silymarin extract by liquid chromatography. Journal of Chromatography B 845, 95–103.

Hadad, G.M., Emara, S., Abdel-Salam, R.A., 2009. Validated Optimized Method for Simultaneous Analysis of Active Silymarin Components and Dimethyl-4,4′-dimethoxy-5,6,5′,6′-dimethylene Dioxybiphenyl-2,2′-dicarboxylate in a Pharmaceutical Preparation by Use of a Monolithic Silica C18 Column, Chromatographia 70, 217-221

Abdel-Salam, N.A., Abdel-Salam, M.A., Elsayed, M.A., 1982. Optimized and Validated Kinetic Spectrophotometric Method for the Determination of Silymarin in drug formulations. Pharmazie 37, 74.             

Vanhaelen, M., Vanhaelen-Fastre, R., 1983. Quantitative determination of biologically active constituents in medicinal plant crude extracts by thin-layer chromatography—densitometry I. Aesculus hippocastaneum L., Arctostaphyllos uva-ursi Spreng, Fraxinus excelsior L., Gentiana lutea L., Glycyrrhiza glabra L., Hamamelis virginiana L., Hypericum perforatum L., Olea europea L., Salix alba L. and Silybum marianum Gaertn, Journal of Chromatography 281, 263.

Dube, D., Vyas, S.P., 2009. Simultaneous spectrophotometric estimation of Lamivudine and silymarin in mixture, International Journal of Pharmacy and Pharmaceutical Sciences 1(2), 108-111.