diff --git a/CHANGELOG.md b/CHANGELOG.md
index 37a9ad6410..d09540a188 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -11,6 +11,11 @@ and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0
 
 ## Unreleased
 
+### Fixed
+
+- `opentelemetry-instrumentation-asgi`: fix excluded_urls in instrumentation-asgi
+  ([#3567](https://github.com/open-telemetry/opentelemetry-python-contrib/pull/3567))
+
 ## Version 1.34.0/0.55b0 (2025-06-04)
 
 ### Added
diff --git a/instrumentation/opentelemetry-instrumentation-asgi/src/opentelemetry/instrumentation/asgi/__init__.py b/instrumentation/opentelemetry-instrumentation-asgi/src/opentelemetry/instrumentation/asgi/__init__.py
index 8bf9c71aed..f4f84d1c58 100644
--- a/instrumentation/opentelemetry-instrumentation-asgi/src/opentelemetry/instrumentation/asgi/__init__.py
+++ b/instrumentation/opentelemetry-instrumentation-asgi/src/opentelemetry/instrumentation/asgi/__init__.py
@@ -254,11 +254,13 @@ def client_response_hook(span: Span, scope: dict[str, Any], message: dict[str, A
     OTEL_INSTRUMENTATION_HTTP_CAPTURE_HEADERS_SANITIZE_FIELDS,
     OTEL_INSTRUMENTATION_HTTP_CAPTURE_HEADERS_SERVER_REQUEST,
     OTEL_INSTRUMENTATION_HTTP_CAPTURE_HEADERS_SERVER_RESPONSE,
+    ExcludeList,
     SanitizeValue,
     _parse_url_query,
     get_custom_headers,
     normalise_request_header_name,
     normalise_response_header_name,
+    parse_excluded_urls,
     remove_url_credentials,
     sanitize_method,
 )
@@ -537,7 +539,7 @@ class OpenTelemetryMiddleware:
     def __init__(
         self,
         app,
-        excluded_urls=None,
+        excluded_urls: ExcludeList | str | None = None,
         default_span_details=None,
         server_request_hook: ServerRequestHook = None,
         client_request_hook: ClientRequestHook = None,
@@ -619,6 +621,8 @@ def __init__(
         self.active_requests_counter = create_http_server_active_requests(
             self.meter
         )
+        if isinstance(excluded_urls, str):
+            excluded_urls = parse_excluded_urls(excluded_urls)
         self.excluded_urls = excluded_urls
         self.default_span_details = (
             default_span_details or get_default_span_details
diff --git a/instrumentation/opentelemetry-instrumentation-asgi/tests/test_asgi_middleware.py b/instrumentation/opentelemetry-instrumentation-asgi/tests/test_asgi_middleware.py
index 95a874bcdb..c243ca0d9f 100644
--- a/instrumentation/opentelemetry-instrumentation-asgi/tests/test_asgi_middleware.py
+++ b/instrumentation/opentelemetry-instrumentation-asgi/tests/test_asgi_middleware.py
@@ -1633,6 +1633,29 @@ async def test_no_metric_for_websockets(self):
         await self.get_all_output()
         self.assertIsNone(self.memory_metrics_reader.get_metrics_data())
 
+    async def test_excluded_urls(self):
+        self.scope["path"] = "/test_excluded_urls"
+        app = otel_asgi.OpenTelemetryMiddleware(
+            simple_asgi, excluded_urls="test_excluded_urls"
+        )
+        self.seed_app(app)
+        await self.send_default_request()
+        await self.get_all_output()
+        spans = self.memory_exporter.get_finished_spans()
+        self.assertEqual(len(spans), 0)
+
+    async def test_no_excluded_urls(self):
+        self.scope["path"] = "/test_excluded_urls"
+        app = otel_asgi.OpenTelemetryMiddleware(
+            simple_asgi, excluded_urls="test_excluded_urls"
+        )
+        self.seed_app(app)
+        self.scope["path"] = "/test_no_excluded_urls"
+        await self.send_default_request()
+        await self.get_all_output()
+        spans = self.memory_exporter.get_finished_spans()
+        self.assertGreater(len(spans), 0)
+
 
 class TestAsgiAttributes(unittest.TestCase):
     def setUp(self):