J. Cosmet. Sci., 66, 175–187 (May/June 2015) 175 Clinical evaluation of Gac extract (Momordica cochinchinensis) in an antiwrinkle cream formulation PAKAPUN LEEVUTINUN, PANVIPA KRISADAPHONG, and AMORN PETSOM, Technopreneurship and Innovation Management, Graduate School, Chulalongkorn University, Bangkok 10330 (P.L.), School of Cosmetic Science, Mae Fah Luang University, Chiang Rai 57100 (P.K.), Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330 (A.P.), Thailand. Accepted for publication March 5, 2015. Synopsis The objective of this work was to evaluate the antioxidant and antityrosinase activities of Gac (Momordica cochinchinensis) extract and to clinically evaluate a Gac-containing antiwrinkle cream formulation. Gac extract exhibited higher antioxidant activity than vitamin C or E, as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH, 41.25 ± 0.34 mg TEAC/ml extract), 2, 2’-azinobis 3-ethylbenzothialine-6-sulfonic acid (ABTS, 47.70 ± 0.18 mg TEAC/ml extract), and ferric reducing antioxidant power (FRAP, 105.03 ± 2.326 mg TEAC/ml extract) scavenging. The antioxidant activity of Gac extract was 5.85- and 11.75-fold higher than that of vitamin E in the DPPH and ABTS assays, respectively. The FRAP assay indicated that the antioxidant activity of Gac extract was 2.91-fold higher than that of vitamin C. Gac extract also exhibited high tyrosinase inhibition (62.83% ± 1.99%). The tyrosinase inhibition activity of Gac extract was 1.51- and 2.06-fold greater than that of vitamins C and E, respectively. The safety and effi cacy of the formulated Gac extract cream as an antiwrinkle agent and its promotion of skin moisturization and smoothness were investigated. Evaluation of acute skin tolerance indicated nonirritating properties. A clinical study revealed increases in cutaneous hydration. Average roughness was decreased, while smoothness was increased. Skin overlook anal- ysis indicated skin roughness relief. These results indicate that the formulated Gac extract product is an ef- fective antiwrinkle cream. INTRODUCTION Wrinkles in the skin accumulate with aging. The skin develops wrinkles via two mecha- nisms: (a) intrinsic aging involving continuously deteriorating tissue function and (b) ex- trinsic aging caused by environmental factors such as light or UV radiation exposure (1). Momordica cochinchinensis (Lour.) Spreng or Gac is known as Phak Kao in Thailand. M. cochinchinensis is a tropical plant that is grown in countries such as Thailand, India, China, Laos, and, in particular, Vietnam, where Gac is used medicinally to treat dry eyes and Address all correspondence to Pakapun Leevutinun at vickypakapun@gmail.com.

JOURNAL OF COSMETIC SCIENCE 176 promote healthy vision. M. cochinchinensis is a good source of carotenoid, particularly beta- carotene and lycopene, as well as vitamins A and E (2,3). Vitamins A, C, and E are acceptable cosmetic components because of their antioxi- dant effi cacies. The oil in Gac aril is a valuable source of carotenoids (4,5) as well as signifi cant amounts of vitamin E and omega-3 fatty acids (6). The antioxidant con- tent of Gac aril is 5- to 10-fold higher than that of other sources such as grapefruit, guava, and tomato (7). The carotenoid compounds in Gac exhibit substantial free radical scavenging ability. The antioxidant activity of alpha tocopherol in Gac may play an important role in ameliorating antioxidative DNA damage (8) induced by H2O2 and Ultraviolet light in the C spectrum (UVC) in TK6 and L929 cells, as de- termined by cytotoxic activity measurements via 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT assay) (9). Moreover, Gac extract maintains antioxidant activity after transformation processes such as powdering, drying, and freezing (10). Gac extract is safe for human and animal con- sumption as applied in the food, beverage, pharmaceutical, and cosmetics industries (11). This study aimed to determine the antioxidant activity and tyrosinase inhibition activity of Gac extract and the effi cacy of Gac cream in 22 subjects after 28 and 56 days of use. Whitening, moisturizing, and antiwrinkle effects were evaluated. The skin tolerance and safety of this cream were also evaluated. MATERIALS AND METHODS MATERIALS Gac extract was a supplied by Specialty Natural Products Co. Ltd. (Bangkok, Thailand). The Gac extract, which is extracted by ethanol and water, contained 825.30 mg/100 g lycopene, 385.0 mg/kg beta-carotene, and 61.1 mg/kg alpha-tocopherol. Ethyl acetate, n-hexane, ethanol, methanol, and trichloroacetic acid were supplied by Merck (Bangkok, Thailand) ascorbic acid was supplied by Finechem (Bangkok, Thailand) linoleic acid was supplied by Sigma-Aldrich (Nonthaburi, Thailand) and 2,2’-diphenyl-1-picrylhydrazyl (DPPH) was supplied by Sigma. Ferric chloride (FeCl3), potassium ferricyanide (K3[Fe(CN)6]), ammonium thiocyanate (NH4SCN), and ferrous chloride tetrahydrate (FeCl2·4H2O) were supplied by Fisher Scientifi c (Bangkok, Thailand). Sodium dihydrogen phosphate (NaH2PO4·2H2O) and disodium hydrogen phosphate (Na2HPO4·12H2O) were supplied by Bdh-Prolabo (Bangkok, Thailand). The measurement was conducted by supplier provided Gac extract. EXTRACT PREPARATION Using 80% ethanol and 20% water to absorb plant materials, and a drying process using a rotary evaporator in a vacuum can enhance greater effi cacy of plants (12). The red mem- brane surrounding the seeds (seed membrane) of the Gac fruit were also extracted in ethanol and water measured by supplier properly mixing the solvent with the Gac fruit using a blender. This extract was evaporated to a cream paste consistency using the rotary evaporator in a vacuum at 37–40oC. The extract was diluted in propylene glycol to obtain Gac extract.