Three Australians featured in the first 10 out of 100 beauties, actress Cate Blanchett, who came in third, singer Natalie Imbruglia and model Elle Macpherson.
Princess Diana came in 12th.
Hepburn, the star of Roman Holiday and Breakfast at Tiffany's, topped the poll of beauty editors, make-up artists, fashion editors, model agencies and fashion photographers who were asked to choose their top 10 beauties from the list of 100 compiled by the drinks firm Evian.
The women were chosen for their "embodiment of natural beauty, healthy living, beautiful on the inside and out, with great skin and a natural glow to their personality, as well as their complexion", Evian said.
Hepburn was joined in the top five by current US film star Liv Tyler, Blanchett, Tomb Raider star Angelina Jolie and the '50sicon turned princess Grace Kelly.
Audrey Hepburn
"She has a rare charm and inner beauty that radiates when she smiles. Her skin looks fresh in all her films and her personality really shines through as someone warm and lively."
More than 76 per cent of the beauty experts who voted in the survey mentioned Hepburn in their top 10.
The survey also showed that while 32 per cent of the icons placed in the No. 1 spot by panel members were blondes, 22 per cent redheads and 46 per cent brunettes.
"People who are confident without a scrap of make-up and look healthy, happy and glowing, seem like natural beauties to me," said Eleonore Crompton, of Heat magazine.
Top 100 Beautiful Women:
2. Liv Tyler
3. Cate Blanchett
4. Angelina Jolie
5. Grace Kelly
6. Natalie Imbruglia
7. Juliette Binoche
8. Halle Berry
9. Helena Christensen
10. Elle Macpherson
11. Cameron Diaz
12. Princess Diana
13. Kate Moss
14. Charlize Theron
15. Scarlett Johansson
16. Isabella Rossellini
17. Nigella Lawson
18. Beyonce Knowles
19. Madonna
20. Jamelia
21. Nicole Kidman
22. Monica Bellucci
23. Audrey Tatou
24. Vanessa Paradis
25. Julianne Moore
26. Jennifer Lopez
27. Marilyn Monroe
28. Julia Roberts
29. Liberty Ross
30. Kylie Minogue
31. Estelle Warren
32. Gisele Bundchen
33. Gwyneth Paltrow
34. Kate Winslet
35. Katharine Hepburn
36. Ava Gardner
37. Keira Knightley
38. Iman
39. Jerry Hall
40. Heidi Klum
41. Ursula Andress
42. Virginie Ledoyen
43. Sophie Dahl
44. Michelle Pfeiffer
45. Uma Thurman
46. Kim Catrell
47. Jennifer Aniston
48. Eva Herzigova
49. Brigitte Bardot
50. Felicity Kendal
51. Claudia Schiffer
52. Jacqueline Kennedy
53. Marlene Dietrich
54. Milla Jovovitch
55. Lucy Liu
56. Penelope Cruz
57. Neve Campbell
58. Sharon Stone
59. Vivien Leigh
60. Sophie Marceau
61. Linda Evangelista
62. Dido
63. Catherine Zeta Jones
64. Jessica Lange
65. Ingrid Bergman
66. Greta Garbo
67. Jodie Kidd
68. Laetitia Casta
69. Princess Caroline of Monaco
70. Kathleen Turner
71. Rachel Weisz
72. Naomi Campbell
73. Grace Jones
74. Christie Turlington
75. Famke Jensen
76. Catherine Deneuve
77. Cindy Crawford
78. Heather Graham
79. Judy Garland
80. Ginger Rogers
81. Sophia Loren
82. Yasmin Le Bon
83. Kirsten Dunst
84. Sandra Bullock
85. Melanie Sykes
86. Cleopatra
87. Lisa Snowdon
88. Rita Hayworth
89. Katie Holmes
90. Honor Blackman
91. Joely Richardson
92. Joanna Lumley
93. Andie MacDowell
94. Alicia Silverstone
95. Cat Deeley
96. Rene Russo
97. Sienna Miller
98. Rachel Hunter
99. Jade Jagger
100. Kelly Brook
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