ABSTRACT
Rosae Radix et Rhizoma is a herbal medicine in a variety of famous Chinese patent medicines, while the quality standard for this medicine remains to be developed due to the insufficient research on the quality of Rosae Radix et Rhizoma from different sources. Therefore, this study comprehensively analyzed the components in Rosae Radix et Rhizoma of different sources from the aspects of extract, component category content, identification based on thin-lay chromatography, active component content determination, and fingerprint, so as to improve the quality control. The results showed that the content of chemical components varied in the samples of different sources, while there was little difference in the chemical composition among the samples. The content of components in the roots of Rosa laevigata was higher than that in the other two species, and the content of components in the roots was higher than that in the stems. The fingerprints of triterpenoids and non-triterpenoids were established, and the content of five main triterpenoids including multiflorin, rosamultin, myrianthic acid, rosolic acid, and tormentic acid in Rosae Radix et Rhizoma was determined. The results were consistent with those of major component categories. In conclusion, the quality of Rosae Radix et Rhizoma is associated with the plant species, producing area, and medicinal parts. The method established in this study lays a foundation for improving the quality standard of Rosae Radix et Rhizoma and provides data support for the rational use of the stem.
Subject(s)
Drugs, Chinese Herbal/chemistry , Rhizome/chemistry , Plant Roots/chemistry , Plants, Medicinal , Quality ControlABSTRACT
In this paper, the newly isolated tannins were sorted after a review of the literature concerning tannins in recent 10 years, and their research progress was summarized in terms of extraction, isolation, pharmacological activity and metabolism. Hydrolysable tannins and condensed tannins are the main structural types. Modern research shows that tannins have many pharmacological effects, such as bacteriostasis, antioxidation, antitumor, antivirus and blood glucose reduction, and have broad development prospects. They are usually extracted by water, ethanol and acetone and isolated and purified by macroporous resin and gel column chromatography. The packings commonly adopted for the column chromatography mainly included Sephadex LH-20, Diaion HP-20, MCI-gel CHP-20 and Toyopearl HW-40. Modern analytical techniques such as nuclear magnetic resonance spectroscopy(NMR), fast atom bombardment mass spectrometry(FAB-MS) and circular dichroism(CD) are generally used for the structural identification of tannins. Howe-ver, their isolation, purification and structural identification are still challenging. It is necessary to use a variety of high-throughput screening methods to explore their pharmacological activities and to explore the material basis responsible for their functions through experiments in vivo.
Subject(s)
China , Hydrolyzable Tannins , Medicine, Chinese Traditional , Proanthocyanidins , TanninsABSTRACT
Prunella vulgaris(PV) is an edible and traditional medicinal herb which has a wide range application in fighting inflammation and oxidative stress, and protecting liver. Now it has been used to treat various types of liver diseases and has significant clinical efficacy. This study aims to investigate the effects of PV on ethanol-induced oxidative stress injury in rats and its metabolic mechanism. The rats were divided into control group, model group, PV group, and VC group. The liver protection of PV was identified by measuring pharmacological indexes such as antioxidant and anti-inflammatory activity. The metabolic mechanism of long-term ethanol exposure and the metabolic regulation mechanism of PV treatment were studied by LS-MS metabonomics. The pharmacological investigation indicated that ethanol could significantly decrease the contents of SOD, GSH-Px, CAT and other antioxidant enzymes in liver and increase the content of MDA. At the same time, PV could significantly reduce the contents of inflammatory factors(TNF-α, IL-6 and IL-1β) and liver function markers(ALT, AST, ALP) in serum. What's more, long-term ethanol exposure could significantly cause liver injury, while PV could protect liver. Metabolomics based on multiple statistical analyses showed that long-term ethanol exposure could cause significant metabolic disorder, and fatty acids, phospholipids, carnitines and sterols were the main biomarkers. Meanwhile, pathway analysis and enrichment analysis showed that the β oxidation of branched fatty acids was the main influencing pathway. Also, PV could improve metabolic disorder of liver injury induced by ethanol, and amino acids, fatty acids, and phospholi-pids were the main biomarkers in PV treatment. Metabolic pathway analysis showed that PV mainly regulated metabolic disorder of ethanol-induced liver injury through phenylalanine, tyrosine and tryptophan biosynthetic pathways. This study could provide a new perspective on the hepatoprotective effect of natural medicines, such as PV.
Subject(s)
Animals , Rats , Antioxidants/metabolism , Ethanol/toxicity , Liver/metabolism , Metabolomics , Oxidative Stress , PrunellaABSTRACT
High performance liquid chromatography-ultraviolet(HPLC-UV) fingerprint is one of the most important methods for the quality control of Chinese medicines in Chinese Pharmacopoeia. However, certain subjectivity is present in selection of specific band of UV, and the inherent quality differences of Chinese medicine can't be well characterized by this method. Therefore, with different grades of Scrophulariae Radix were taken as the research object in this study, a new quality control model of HPLC-UV was established in this study based on the ultraviolet full-wavelength scanning spectrum. Firstly, different grades of Scrophulariae Radix samples were collected, and the full-wavelength ultraviolet absorption spectra of all the samples were established at the bands of 200-400 nm. In order to analyze the differences among samples, the analysis model was built following multivariate statistical analysis methods such as principal component analysis(PCA) and partial least squares discrimination analysis(PLS-DA) after the pretreatment of spectral data. The result showed that the ultraviolet band at 251 nm may contribute most to distinguish the quality differences among different grades of samples. Then, the HPLC fingerprints of samples were established with the band at 251 nm. The multivariate statistical analysis showed that there was a more significant classification trend in HPLC fingerprints than that in the original UV fingerprints, which could be used to distinguish different grades of samples, and could better reflect the differences among different grades. The method reported in this study can be of a great guidance and reference for the establishment of specific fingerprints of Chinese medicines as well as for the quality control of Chinese medicine.
Subject(s)
Drugs, Chinese Herbal , Chromatography, High Pressure Liquid , Plant Roots , Principal Component Analysis , Quality ControlABSTRACT
To explore the effect of Fuke Qianjin Capsules on anti-endometrial fibrosis in intrauterine adhesion(IUA) rats through TGF-ß1-PI3 K/Akt signaling pathway. With female SD rats as the object, IUA rat models were established through mechanical injury and infection, and they were randomly divided into normal group, sham operation group, Bujiale group(0.63 mg·kg~(-1)·d~(-1)), and high-dose Fuke Qianjin Capsules group(1.008 g·kg~(-1)·d~(-1)), medium-dose Fuke Qianjin Capsules group(0.504 g·kg~(-1)·d~(-1)), low-dose Fuke Qianjin Capsules group(0.252 g·kg~(-1)·d~(-1)). The rats were sacrificed 21 days after drug administration, and the uterus and liver were removed after blood collection from the abdominal aorta. The morphology of the uterus was observed with the naked eyes; the pathological and morphological changes of the uterine tissue and liver were observed by HE staining; the degree of fibrosis of the uterine tissue was observed by Masson staining; the expressions of TGF-ß1, TNF-α and IL-6 in serum were detected; the expressions of TGF-ß1, PI3 K, Akt, p-Akt protein in uterine tissue were detected by Western blot. The results showed that Fuke Qianjin Capsules could improve the pathological changes of uterine tissues in IUA rats, without damage to liver tissues, and reduce the expressions of TGF-ß1, TNF-α and IL-6 in serum(P<0.01); significantly reduce TGF-ß1, PI3 K, p-Akt protein expression in uterine tissues(P<0.05, P<0.01). It is indicated that Fuke Qianjin Capsules could exert the anti-endometrial fibrosis effect by regulating the TGF-ß1-PI3 K/Akt signal pathway, so as to achieve the effect in treating IUA rats, especially with the best effect in medium-dose Fuke Qianjin Capsules group.
Subject(s)
Drugs, Chinese Herbal , Transforming Growth Factor beta1 , Animals , Capsules , Female , Phosphatidylinositol 3-Kinases/genetics , Proto-Oncogene Proteins c-akt/genetics , Rats , Rats, Sprague-Dawley , Signal Transduction , Transforming Growth Factor beta1/geneticsABSTRACT
Idiosyncratic hepatotoxicity of Polygonum multiflorum has attracted a great attention in the world. The most toxic part of idiosyncratic hepatotoxicity was screened by MTT assay and flow cytometry, which was the 50% ethanol elute by macroporous adsorptive resins from alcohol-extraction of P. multiflorum. The fingerprints were collected by HPLC from 50% ethanol elute of crude and processed P. multiflorum from different habitats, then 14 common peaks were determined. Spectrum-toxicity relationship was analyzed by rough set theory(RST). Two main chemical components were predicted for idiosyncratic hepatotoxicity, in which TSG was the greater contributor. Idiosyncratic hepatotoxicity of TSG was tested in vitro, and the results indicated that TSG was the most important constituent contributed to idiosyncratic hepatotoxicity of P. multiflorum. The study showed the discovery of the main chemical components for idiosyncratic hepatotoxicity, and RST was effective for analyzing the spectrum-toxicity relationship, which could be a new method used in the effective/toxic constituents field of traditional Chinese medicine.
Subject(s)
Humans , Chemical and Drug Induced Liver Injury , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Fallopia multiflora , Chemistry , Medicine, Chinese Traditional , PhytochemicalsABSTRACT
Prunellae Spica is a perennial edible and medicinal plant, rich in antioxidant substances. Total flavonoids (TFC), Phenolics (TPC), triterpenoids (TSC), polysaccharides (PC) and their antioxidant capacities (by the FRAP, DPPH and ABTS⁺ methods) of ethyl acetate fraction, n-butanol fraction and other fractions of aqueous extract from Prunellae Spica were investigated in this study. Then the multivariate statistical method was adopted to analyze the relationship between the multiple pharmaceutical ingredients and antioxidant capacities of Prunellae Spica. The results showed that ethyl acetate fraction had relatively high concentration of TFC (0.61±0.10) g·g⁻¹DW, TPC (0.52±0.09) g·g⁻¹DW, and TSC (0.21±0.03) g·g⁻¹DW, with high scavenging capacity of DPPH (3.1±0.38) mmol·L⁻¹·g⁻¹DW and FRAP (2.56±0.35) mmol·L⁻¹·g⁻¹DW. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) results indicated the information from chemical compositions and antioxidant capacity can represent the "differences" of different fractions. Canonical correlation analysis (CCorA) revealed a high positive correlation between the amounts of multiple chemical compositions and the antioxidant capacities (r=0.970 0), and the first canonical variate had been reached. Moreover, ABTS⁺ method showed a low response to the compositions of different fractions, so this method may not be suitable for evaluation of Prunellae Spica antioxidant capacities, while DPPH evaluation method was more suitable for TSC and TPC. The results of this study have important reference significance for the evaluation method on antioxidant activity of Prunellae Spica in the field of food or medicine as well as for the development of related extracts.
Subject(s)
Antioxidants , Flavonoids , Phenols , Plant ExtractsABSTRACT
To establish a random forest algorithm for identifying and classifying different brands of Xiasangju granules, and provide effective reference for identifying multi-index complex fingerprint. HPLC method was used to collect the fingerprint of 83 batches of Xiasangju granules from different manufacturers. The classification of Xiasangju granules samples based on chromatographic fingerprints was identified by chemometric methods including principal component analysis (PCA), partial least squares discriminate analysis (PLS-DA) and random forest analysis (RF). The superiority of the above three chemometric methods was compared. The results showed that the fingerprints of 83 batches of Xiasangju granules were established in this study. PCA could only explicate 56.52% variance contribution rate and could not completely classify the samples; PLS-DA analysis was superior to PCA, explicating 63.43% variance contribution rate and could obtain certain separation; RF could well classify the samples into 3 types, and the predication accuracy of the proposed method was 96.5%. Therefore, The results indicate that RF combined with HPLC fingerprint could effectively construct traditional Chinese medicine quality control and analysis system.
ABSTRACT
This paper aims to investigate the correlation between the antioxidant activity of Prunella vulgaris and its total phenolic acids content by measuring the antioxidant activity of different sources and different organs of P. vulgaris and the total contents of protocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid in these samples. Using the 50% methanol extract of P. vulgaris samples as the research object, DPPH method and HPLC method were used respectively to determine the antioxidant activities and the total contents of the above-mentioned five analytes in P. vulgaris samples. 0.5 mL of 50% methanol extract of P. vulgaris reacts with 0.1 mmol•L⁻¹ DPPH ethanol solution for 60 min, then the absorbance of the reaction solution was measured at 517 nm, scavenging rate and IC₅₀ values were calculated by the absorbance and the sample concentration for evaluating the antioxidant activity. HPLC analysis was made on a C₁₈ Epic column, with acetonitrile-0.1% formic acid aqueous solution as mobile phase (gradient elution), and the detection wavelength was set at 280 nm. The correlation between the antioxidant capacity of different habitats and different organs of P. vulgaris and the total contents of five kinds of phenolic acids was analyzed by partial least squares method. The reaction dose-response range of 50% methanol extract of P. vulgaris with 0.1 mmol•L⁻¹ DPPH ethanol solution was 0.300-1.65 g•L⁻¹. When the quantities of potocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid were respectively in 0.007 84-0.980, 0.011 5-1.44, 0.008 64-1.08, 0.080 0-1.00 and 0.079 8-0.998 μg range, their quantities were in good linear relationship with the corresponding peak areas. The average recovery of 5 components were 97.76%, 96.88%, 100.3%, 102.1%, 104.5%, with RSD of 1.8%, 1.6%, 1.7%, 1.6% and 1.7%, respectively. In a certain range of crude drug quantity, the antioxidant activity of each organ of P. vulgaris and total phenolic acids content inside has a good linear correlation. Therefore, in certain quality range of crude drug, DPPH bioassay combined with HPLC content determination can be used for the quality control of P. vulgaris, as is a new method for the quality control of P. vulgaris.
ABSTRACT
It was estimated that about 60 species and 15 varieties of genus Aconitum are distributed in China.These plants contain various kinds of chemical compounds, and the main compounds are diterpenoid alkaloids. In addition, there are flavonoids, phenolic acid and others.So far, phytochemical studies showed 339 compounds.This paper summarized the chemical compounds to provide the theoretical basis for the use of Tibetan medicinal plants of Aconitum genus.
ABSTRACT
This study is to develop a HPLC method for quality evaluation of Euodiae Fructus and related species by simultaneous determination limonin, indole alkaloids (14-fomyldihydroxyrutaecarpine, evodiamine, rutaecarpine), and quinolone alkaloids [1-methyl-2-undecyl-4 (1H)-quinolone, evocarpine, dihydroevocarpine] in the fruits of five Evodia species. Samples were analyzed on a YMC C18 column (4.6 mm x 250 mm, 5 microm) eluted with mobile phases of acetonitrile (A), tetrahydrofuran (B), and a buffer solution of 5 mmol x L(-1) ammonium acetate (pH 3.8) (C) in a linear gradient mode. The column temperature was 30 degrees C and the flow rate was 1.0 mL x min(-1). The PDA detector wavelengths were set at 220 and 250 nm. The seven compounds were well separated and showed good linearity (r = 0.999 9) within the concentration ranges tested. The mean recoveries were between 96.7%-102.4% (RSD 1.4%-3.1%). Through the validation, the method was proved to be accurate and repeatable. All the seven constituents were detected in the fruits of five species, but the contents of them varied widely in different samples. The total contents of seven constituents in 16 batches of Euodiae Fructus were 9.46-69.9 mg x g(-1), and the mean content was 28.2 mg x g(-1). The total content of seven constituents in E. compacta and E. fargesii was 25.8, 7.69 mg x g(-1), respectively.
Subject(s)
Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Evodia , Chemistry , Fruit , Chemistry , Time FactorsABSTRACT
Siraitia grosvenorii is a perennial herb endemic to Guangxi province of China. Its fruit, commonly known as Luo hanguo, and has been used for hundreds of years as a natural sweetener and as a traditional medicine for the treatment of pharyngitis, pharyngeal pain, as well as an anti-tussive remedy in China. Based on ninety-three literary sources, this review summarized the advances in chemistry, biological effects, and toxicity research of S. grosvenorii during the past 30 years. Several different classes of compounds have been isolated or detected from various parts of S. grosvenorii, mainly triterpenoids, flavonoids, polysaccharides, amino acids, and essential oils. Various types of extracts or individual compounds derived from this species exhibited a wide array of biological effects e.g. anti-tussive, phlegm-relieving, anti-oxidant, immunomodulatory, liver-protecting, glucose-lowering, and anti-microbial. The existing research has shown that extracts and individual compounds from S. grosvenorii are basically non-toxic. Finally, some suggestions for further research on specific chemical and pharmacological properties of S. grosvenorii are proposed in this review.
Subject(s)
Animals , Humans , Amino Acids , Cucurbitaceae , Chemistry , Flavonoids , Plant Extracts , Pharmacology , Polysaccharides , TriterpenesABSTRACT
Nineteen compounds were isolated from the whole plants of Aconitum tanguticum by means of various of chromatographic techniques such as silica gel, ODS, sephadex LH-20 and preparative HPLC, and their structures were elucidated as syringin (1), vanillic acid-4-O-beta-D-allopyranoside (2), (E) -ferulic acid 4-O-beta-D-allopyranoside (3), (E) -ferulic acid-4-O-beta-glucopysoside (4), (E) -sinapic acid-4-O-beta-glucopyranoside (5), (E) 4-hydroxycinnamyl alcohol 4-O-beta-D-glucopyranoside (6), quercetin 3-O-alpha-L-rhamnopyranosyl-(1 --> 2) -[alpha-L-rhamnopyranosyl-(1 --> 6)] -beta-D-galactopyranoside-7-O-alpha-L-rhamnopyranoside (7), kaempferol 3-O-alpha-L-rhamnopyranosyl-(1 --> 2) - [alpha-L-rhamnopyranosyl-(1 --> 6)] -beta-D-galactopyranside-7-O-alpha-L-rhamnopyranoside (8), quercetin 3-O-alpha-L-rhamnopyranosyl-(1 --> 6) -beta-D-glucopyranoside-7-O-alpha-L-rhamnopyranoside (9), kaempferol 3-O-[beta-D-glucopyranosyl-(1 --> 3)-(4-O-trans-p-coumaroyl) ] -alpha-L-rhamnopyranosyl-(1 --> 6) -beta-D-galactopyranside-7-O-alpha-L-rhamnopyranoside (10), quercetin 3-O- [beta-D-glucopyranosyl-(1 --> 3 ) -(4-O-trans-p-coumaroyl)] -alpha-L-rhamnopyranosyl-(1--> 6) -beta-D-galactopyranoside-7-O-alpha-L-rhamnopyranoside (11), salidroside (12), 2-(3,4-dihydroxyphenyl) ethanol 1-O-beta-D-glucopyranoside (13), (7S, 8R) -dehydrodiconiferyl alcohol-9'-O-beta-D-glucopyranoside (14), citrusin B (15), heteratisine (16), tanaconitine (17), shanzhiside methyl ester (18) and icariside B1 (19). Except compounds 4, 13, 16 and 17, the other compounds were separated from the species for the first time.
Subject(s)
Aconitum , Chemistry , Drugs, Chinese Herbal , Chemistry , Magnetic Resonance Spectroscopy , Molecular Structure , Spectrometry, Mass, Electrospray IonizationABSTRACT
<p><b>OBJECTIVE</b>To study chemical constituents of Ardisia punctata,in order to find pioneering compounds.</p><p><b>METHOD</b>95% ethanol extracts of A. punctata was separated and purified by using normal phase silica gel column chromatographies, Sephadex LH-20 gel column chromatography and high-pressure preparative HPLC,and their structures were identified by such spectroscopic techniques as NMR and MS.</p><p><b>RESULT</b>Eight compounds were separated from 95% ethanol extract of A. punctata and identified as 6-methoxy-8-hydroxy-benzoic acid butylester-5-O-beta-D-glucoside (1), aridisiacrispin B (2), ardisicrenoside A (3), dibutyl phthalate (4), bergenin (5), quercetin-3-O-alpha-L-rhamnoside (6),3-methoxy-4-acetoxy-6-tridecyl-phenol(7) and belamcandaquinone C(8).</p><p><b>CONCLUSION</b>Compound 1 was a new compound, and compounds 4 and 6 were separated from this plant for the first time.</p>
Subject(s)
Ardisia , Chemistry , Drugs, Chinese Herbal , Chemistry , Mass Spectrometry , Molecular StructureABSTRACT
The research patterns for the pharmacodynamic chemical substances in compound prescriptions of Chinese medicinal materials in recent years were summarized, and the deficiencies of the commonly used patterns were commented on. A research pattern for the pharmacodynamic chemical substances in compound prescriptions of Chinese medicinal materials was raised, which is suitable for the characteristics of the Chinese medicine. The trend of the research work was predicted, which would provide some thinking for the pharmacodynamic chemical substances in compound prescription of Chinese medicinal materials.
Subject(s)
Biomedical Research , Drug Prescriptions , Drugs, Chinese Herbal , Chemistry , Medicine, Chinese TraditionalABSTRACT
<p><b>OBJECTIVE</b>To develop an HPLC method for determination of loniceroside A and loniceroside C in Lonicerae japonicae.</p><p><b>METHOD</b>The analysis was carried out on an Alltech C18 column (4.6 mm x 250 mm, 5 microm) evaluated with methanol-acetonitrile-0.1% glacial acetic acid as mobile phase. Flow rate was at 1.0 mL x min(-1) and the detection wavelength was at 210 nm for UV detection.</p><p><b>RESULT</b>The calibration curves were linear over the range from 0.15 to 2.25 microg (r = 0.999 9) for loniceroside A and 0.11 to 1.65 microg (r = 0.999 1) for loniceroside C. The average recoveries were 99.9% and 98.3%, respectively.</p><p><b>CONCLUSION</b>The contents of Loniceroside A&C are diverse in Flos Lonicerae japonicae in different regions.</p>
Subject(s)
Chromatography, High Pressure Liquid , Methods , Flowers , Chemistry , Lonicera , Chemistry , Plants, Medicinal , Chemistry , Reproducibility of Results , SaponinsABSTRACT
<p><b>OBJECTIVE</b>Utilizing RP-HPLC analysis of single, merging and simultaneously boiled Lonicera japonica and Forsythia suspense to find out their identity and difference.</p><p><b>METHOD</b>Samples were separated by Alltech-C18 column (4. 6 mm x 250 mm, 5 microm) with a mixture solution of acetonitrile-water (containing 0.2% ethanoic acid) as mobile phase, 1.0 mL x min(-1) as flow-rate, 280 nm as detected wave-length, room temperature as temperature of column, 10 microL as injected volume.</p><p><b>RESULT</b>The linear ranges of chlorogenic acid and forsythoside A were 0.980-9.800 microg (r = 0.999 9) and 0.972-9. 720 microg (r = 0.999 9), respectively. The average recoveries of chlorogenic acid and forsythoside A were 99.85% (n = 6, RSD 2.0%) and 100.87% (n = 6, RSD 1.6%), respectively. Major peaks were confirmed to the two component herbs, contents of chlorogenic acid and forsythoside A were determinated, ingredients were represented by following peaks were compared qualitatively: peaks k, m, n: L. japonica > merging sample > boiled sample; peaks g, h: Forsythia suspense > merging sample > boiled sample; peaks d, e, i: boiled sample > L. japonica, boiled sample > merging sample.</p><p><b>CONCLUSION</b>Difference and identities among 4 experimental samples of RP-HPLC separations show that Yinqiao, L. japonica and F. suspense' substances are difference and identities, which offered direction for investigating further the topic-pharmacodynamic substance of Yinqiao, provided increasing experimental evidence for clinic medicine and pharmaceutics and supplied reference for the complex prescription of traditional Chinese medicine.</p>
Subject(s)
Chlorogenic Acid , Chromatography, High Pressure Liquid , Methods , Drug Combinations , Forsythia , Chemistry , Glycosides , Hot Temperature , Lonicera , Chemistry , Plants, Medicinal , Chemistry , Reproducibility of ResultsABSTRACT
<p><b>OBJECTIVE</b>To compare the therapeutic effects of Radix Tripterygium hypoglaucum tablet (THT) and Tripterygium glycosides tablet (TGT) in treating erosive oral lichen planus (EOLP).</p><p><b>METHODS</b>The patients were randomized into two groups, and they were treated with THT (n = 47) or TGT (n = 47), respectively. The therapeutic effects were evaluated after 3 months treatment.</p><p><b>RESULTS</b>For the patients of grade 1, the total efficacy in TGT group was 85.71%, compared with 52.38% in THT group, the efficacy was statistically greater in the group receiving TGT (P = 0.043). However, for the patients of grade 2, the difference was not statistically significant (P = 0.173).</p><p><b>CONCLUSION</b>TGT is more effective in treating EOLP than THT for grade 1 patients. However, TGT is not suitable for patients of child bearing age.</p>